detection progress of selected drugs in tlc (thin layer chromatography)

Review ArticleDetection Progress of Selected Drugs in TLC

Alina Pyka

Department of Analytical Chemistry Faculty of Pharmacy Medical University of Silesia 4 Jagiellonska Street41-200 Sosnowiec Poland

Correspondence should be addressed to Alina Pyka apykasumedupl

Received 28 April 2013 Revised 9 October 2013 Accepted 11 October 2013 Published 16 January 2014

Academic Editor Hidetoshi Arima

Copyright copy 2014 Alina Pyka This is an open access article distributed under the Creative Commons Attribution License whichpermits unrestricted use distribution and reproduction in any medium provided the original work is properly cited

This entry describes applications of known indicators and dyes as new visualizing reagents and various visualizing systems as wellas photocatalytic reactions and bioautography method for the detection of bioactive compounds including drugs and compoundsisolated from herbal extracts Broadening index detection index characteristics of densitometric band modified contrast indexlimit of detection densitometric visualizing index and linearity range of detected compounds were used for the evaluationof visualizing effects of applied visualizing reagents It was shown that visualizing effect depends on the chemical structure ofthe visualizing reagent the structure of the substance detected and the chromatographic adsorbent applied The usefulness ofdensitometry to direct detection of some drugs was also shown Quoted papers indicate the detection progress of selected drugsinvestigated by thin-layer chromatography (TLC)

1 Introduction

Among the different chromatographic techniques one of themost popular in biological and pharmaceutical analysis ofbioactive substances and recommended byUSP (United StatePharmacopia) is HPLC with respective detection systemfor example HPLC-UV It is well known that the choice ofthe analytical method (including chromatographic methods)depends on the chemical and physicochemical propertiesof the compound being separated and also on the availableapparatus and sensitivity needed for the analysis Moreoverthe total time of chromatographic analysis and its low costare also important In many papers concerning thin-layerchromatography (TLC) analysis of various drugs TLC is animportant method for qualitative and quantitative analysis ofdrugs because it indicates some advantages in comparison toHPLC and GC methods which are listed below

(i) TLC can be used in those situations when HPLC-UVand GC are not suitable for example absence of UVactivity of examined compound (important forHPLCanalysis) or when the absence of volatility (importantfor GC analysis) is observed

(ii) in comparison to HPLC in the case of TLC the highpurity and high concentration of examined samples

are not required Unlike HPLC there is no danger thatthe samples impurities influence column damage andits separation property

(iii) TLC needs no expensive equipment and is easy towork in comparison to HPLC and GC

(iv) TLC allows a parallel separation and quantitativedetermination of many samples at the same time

(v) it is possible to put on TLC plates a large volume ofsample because the solvent excess removing duringthe sample is spotted on chromatographic plates

For these purposes TLC greatly contributes to the analysisof different groups of drugs The last decade shows thatinterest in TLC application in pharmaceutical analysis (egcontrol purity of drugs) has increased with improvements inTLC instrumentation such as TLC combined with densito-metry or with MS and IR respectively If the standard com-pounds of drug are not available identification of unknowndrug components has to be done Moreover TLC combinedwith densitometer or MS and also detector enables thedetermination of drug components in a low range pmol orfmol For this reason TLC is used as one of the promisingmethods of separation and quantitative determination for

Hindawi Publishing CorporationBioMed Research InternationalVolume 2014 Article ID 732078 19 pageshttpdxdoiorg1011552014732078

2 BioMed Research International

these types of compounds for which GC or HPLC is not suit-able The above-mentioned arguments explain enough whythe TLC-densitometry is proposed in many drug analyses asan alternative method to the pharmacopoeias methods

Important step of thin-layer analysis of compounds is thedetection of investigated substances including drugsThe sep-arated substances by thin-layer chromatography (TLC) canbe detected by the following methods physical (individualcolour of substance or fluorescence of substance in UV light)chemical (coloured reactions of separated substances withvisualizing reagents) physicochemical (eg the applicationof isotopes as visualizing reagent) biological (the applicationof biodetectors) [1ndash10] Many drugs are active in the rangeof UV light and can be directly detected and determinedon chromatographic plate for example by densitometricanalysis in short or long wavelength UV light [11ndash39] Thevisualizing reagents have the special significance to detectseparated compounds on thin layers In view of the detectionmechanism of the compound the visualizing reagents canbe sorted as follows conservative reagents which do notdestroy separated substances destructive reagents whichdestroy or change the structure of separated substances Cur-rently the most important field of application of thin-layerchromatography is pharmacy The number of publications inthe field of pharmacy steadily increases It results from thefact that contemporary thin-layer chromatography is a fullyinstrumentalized and automated technique [1ndash3 29]

The applications of many reagents for the detection oforganic compounds including drugs after their separationon thin-layer were presented in books [1 2] on the basisof manuscripts published up to 1992 The objectives of thisreview are the presentation and discussion of the significanceof known indicators and dyes as new visualizing reagentsand various visualizing systems as well as photocatalytic reac-tions and bioautography method for detection of bioactivecompounds including drugs and compounds isolated fromherbal extracts in TLC investigations that were described inthe selected manuscripts from 1993

2 Parameters Describingthe Visualizing Effects

The estimation of visualizing effects of the detected com-pounds must be objective Therefore Gregorowicz and Sli-wiok [39 40] introduced the indexes for estimation ofrhodamine B new fuchsine basic fuchsine and crystal violetfor detection of cholesterol and higher fatty acids To comparethe developing effects of the above-mentioned dyes it wassuggested [39 40] to estimate the chromatographic spotsobtained by means of indexes given bellow

(i) detection index which indicates the ratio of the mini-mal number of micrograms of the substance detectedto the area of the chromatographic spot in squaremillimeters obtained by the planimetric method

119868det =1198981

1199011

[120583g

mm2] (1)

where 1198981is the smallest quantity of substances

detected [120583g] with the visualizing reagent (limit ofdetection) and 119901

1is the spot area of the substance

[mm2] at the limit of detection of the substance(ii) developing index which gives the numerical value of

the part of the chromatographic spot area relating to1 120583g

(iii) contrast index which represents two independentvalues namely the angle between tangents drawn tothe arms of the densitometric band formulated inthe degrees and the densitometric band height incentimeters

(iv) durability of the spot in order to formulate the dura-bility of chromatographic spot colors The followingrelative scale was proposed spot of minor durabilityvisible up to 2 hours spot of mean durability visiblefrom 2 to 6 hours and durable spot durability visiblefor over 6 hours

Today the area of chromatographic spot is determinedby densitometric analysis Therefore Pyka et al modifiedthe above-mentioned indexes and also proposed new indexeswhich characterized the visualizing effects [41ndash45] Thelimit of detection (detectability) detection index broadeningindex modified contrast index densitometric visualizingindex and linearity range were determined and tested forestradiol tocopherol tocopherol acetate ibuprofen stearicacid stearyl alcohol and salicylanilide detected by the useof visualizing reagents The broadening index [41 42] wasdefined as

119868broad =119886

1199012

times 1000 [120583gAU] (2)

where 119886 is the mass [120583g] of the detected substance and 1199012is

the spot area [AU] of 119886 120583g of this analyzed substanceThe detection index was defined as

119868det =1198981

1199011

[120583gAU] (3)

where 1198981is the smallest quantity of substance detected [120583g]

(limit of detection) and 1199011is the spot area of substance [AU]

at the limit of detection of substanceThe modified contrast index [43] was defined as

119868contr(mod if) =ℎ

120572[AU∘] (4)

where ℎ is the height of the densitometric band [AU] of119886 120583g of analyzed substance and 120572 is the angle [∘] betweenthe tangents at the inflection points to the curves of thedensitometric band of substance

The densitometric visualizing index (DVI) [44] wasproposed and defined as

DVI =1199012

1198981times 120572times 10minus4[AU120583g sdot∘] (5)

where 1198981is the limit of detection of the analyzed substance

[120583g] 1199012is the spot area [AU] of 119886 120583g of analyzed substance

BioMed Research International 3

and 120572 is the angle [∘] between the tangents at the inflectionpoints to the curves of the densitometric band of 119886 120583g ofanalyzed substance

21 Application of New Parameters Characterizing the Visu-alizing Effects New visualizing reagents namely gentianviolet methylene violet methylene blue methyl green mala-chite green and Janus blue have been used to detect estradioltocopherol tocopherol acetate ibuprofen stearic acid andstearyl alcohol [41ndash43 45] Brilliant green was applied todetect salicylanilide [44] Estradiol was chromatographed onneutral aluminum oxide 60F

254and neutral aluminum oxide

150F254

using a mixture of toluene and ethyl acetate (1 1vv) as mobile phase Bartonrsquos reagent rhodamine B andsulphuric acid were administered as the comparative visual-izing reagents [43] (plusmn)-120572-Tocopherol and (+)-120572-tocopherolacetate were chromatographed on silica gel 60 using tolueneas mobile phase [41] Rhodamine B and 221015840-bipyridine-iron (III) chloride reagent were applied as the comparativevisualizing reagents Ibuprofen was chromatographed onsilica gel 60F

254using a mixture of chloroform and methanol

(50 14 vv) as a mobile phase Rhodamine B was usedas the comparative visualizing reagent [42] Stearic acidand stearyl alcohol were chromatographed on silica gel 60using a mixture of n-hexane-ethyl acetate (46 4 vv) asmobile phase Rhodamine B was used as the comparativevisualizing reagent [45] Fundamental absorption band ofstudied drugs color of spot of detected drugs backgroundcolor of chromatogram limit of detection and linearity rangeare presented in Table 1 Characteristics of densitometricband of detected drugs modified contrast index densito-metric visualizing index modified broadening index anddetection index are presented in Table 2 It was stated that theproposed modified contrast index is the objective parameterdescribing the applied visualizing reagents Influence of solidsupport on the obtained visualizing effects was found Theangles (120572) between the tangents at the inflection points tothe curves of the densitometric peaks were more compacton neutral aluminum oxide 60F

254than on neutral alu-

minum oxide 150F254

This observation indicates that theutility of any particular visualizing reagent depends on thetype of chromatographic support applied For quantitativeresearch of estradiol investigated relatively good propertieshad sulphuric acid (VI) Bartonrsquos reagent gentian violetand methylene violet [43] 221015840-Bipyridine-iron (III) chloridereagent can be used only to detect (plusmn)-120572-tocopherol Amongall studied new visualizing reagents methylene violet andmethyl green were the best to detect (plusmn)-120572-tocopherol Thebest detection way of (+)-120572-tocopherol acetate was densito-metric method without using a visualizing reagent whereasamong all studied new visualizing reagents gentian violetmethyl green and Janus blue were the best to detect (+)-120572-tocopherol acetate These visualizing reagents have similardetection properties of (+)-120572-tocopherol acetate in relationto rhodamine B [41]The best detection way of ibuprofen wasdensitometric method without using a visualizing reagentAmong all studied new visualizing reagents methylene violetwas the best to detect ibuprofen [42] New visualizing

reagents for detection of stearic acid and stearyl alcohol werebetter visualizing reagents in comparisonwith the universallyapplied rodamine B to detect the lipophilic compounds Thebest visualizing reagents to quantitative determination ofstearic acid were methylene blue and Janus blue The bestvisualizing reagents to quantitative determination of stearylalcohol were malachite green and Janus blue [45]

3 Visualizing Effects for Different Substancesincluding Drugs

31 Detection by the Use of Visualizing Reagents andVarious Visualizing Systems

311 Detection of Steroids Compounds including Drugs Hue-tos et al [46] investigated the specificity of chromatographicconditions of 35 anabolic steroids and 10 other veterinarydrugs The best sensitivity of the different corticosteroidswas performed with silica gel plates eluted with a mixtureof chloroform and methanol in volume composition 92 8Four spray reagents were compared for selectivity sensitivityand specificity The best results were obtained using reagents1 and 2 Reagent 1 was a 1 1 mixture of a solution of 24-dihydroxybenzaldehyde (25mg) in glacial acetic acid (50mL)and a mixture of sulfuric acid (125mL) and glacial aceticacid (375mL) Reagent 2 was a 1 1 mixture of a solutionof tetrazolium blue (025mg) in methanol (50mL) and asolution of sodium hydroxide (10mg) in water (50mL) [46]The plates after spraying were heated at 85ndash90∘C for 10minTable 3 shows 119877

119865values color of spots and detection limits

for the selected seven most common corticosteroids afterreaction with reagents 1 and 2 Reagent 2 can be administeredfor sensitive screening for the presence of corticosteroids inillegal cocktails Reagent 1 is more suitable Reagent 1 givesmore information about the corticosteroids owing to thedevelopment of different colors with reasonable sensitivity[46]

Stigmasterol and 120573-sitosterol (10 120583g and 5 120583g) were visu-alized after separation on silica gel using a mixture of tolueneand ethyl acetate (7 3 vv) [47] The results obtained forthe stigmasterol and 120573-sitosterol by use of the individualvisualizing reagents (gentian violet methylene violet methylgreen malachite green and Janus blue) depend on themethod of applicationmdashspraying or dipping Rhodamine Bwas used as comparative visualizing reagent The best resultsof detection of stigmasterol and 120573-sitosterol were obtained byspraying the visualizing reagents (Tables 4 and 5) [47]

The alkacymetric indicators namely thymol blue bro-mophenol blue and bromothymol blue were also used inthe detection of different classes including the compoundsof biological significance namely sterols (cholesterol stig-masterol) on silica gel 60F

254after separation using the

n-hexane + acetone (8 2 vv) mobile phase The colors ofspots were durable and sufficient contrast between spotsand background was observed The detection limits withthese visualizing reagents were from 05 120583g to 20 120583g forsterols [48] Alkaline blue aniline blue brilliant greenbromothymol blue thymol blue phenol red helasol green


Table1Fu

ndam

entalabsorptionband

ofdrugscoloro

fspo

tofd

etecteddrugsbackgrou

ndcoloro

fchrom

atogramlim

itof

detection

andlin

earityrange

Detecteddrug

Stationary

phase

Detectio

nway

Fund

amental

absorptio

nband

ofdrug120582max(nm)

Color

ofspot

ofdetected

drug

Backgrou

ndcoloro

fchromatogram

Limitof

detection

(120583g)

Linearity

range

(120583gspo

tminus1 )

(rcorrelatio

ncoeffi

cient)

Reference

Estradiol

Neutralalum

inum

oxide6

0F254

toluene+

ethylacetate(11vv)

With

outu

sing

visualizingreagent

205

Lack

ofcoloredspot

invisib

lelight

White

032

110divide2500

(119903=09951)

[43]

Estradiol

Bartonrsquosreagent

233

Darkbluegreen

Willow

-green

032

053divide1024

(119903=09975)

[43]

Estradiol

Sulphu

ricacid

278

Lighto

range

Lightb

eige

010

088divide1280

(119903=09974)

[43]

Estradiol

Gentia

nviolet

226

Violetblue

Violet

032

053divide2500

(119903=09943)

[43]

Estradiol

Methylene

violet

227

Greyblue

Lightg

rey

032

088divide336

(119903=09975)

[43]

Estradiol

Neutralalum

inum

oxide150F 2

54toluene+

ethylacetate(11vv)

With

outu

sing

visualizingreagent

207

Lack

ofcoloredspot

invisib

lelight

White

032

137divide2000

(119903=09972)

[43]

Estradiol

Gentia

nviolet

598

Darkviolet

Violet

032

088divide2000

(119903=09879)

[43]

Estradiol

Methylene

violet

204

Greyblue

Lightg

rey

053

137divide2500

(119903=09983)

[43]

Salicylanilide

Silicag

el60F 2

54(10

5744

)chloroform

With

outu

sing

visualizingreagent

307

Lack

ofcoloredspot

invisib

lelight

White

007

500divide3000

(119903=09802)

[44]

Salicylanilide

Brilliant

green

597

Darkgreen

Green

070

300divide3000

(119903=09929)

[44]

(plusmn)-120572-Tocop

herol

Silicag

el60

toluene

With

outu

sing

visualizingreagent

270

Lack

ofcoloredspot

invisib

lelight

White

045

346divide2000

(119903=09894)

[41]


herol

221015840-Bipyridine-iro

n(III)chloride

524

Raspberryred

Lightb

eige

208

576divide3000

(119903=09927)

[41]


herol

Methylen

eviolet

270

Azure

with

white

border

Lightg

rey-blue

030

075divide576

(119903=09949)

[41]


herol

Methylgreen

270

Lightg

reen

Green

030

075divide576

(119903=09863)

[41]

(+)-120572-Tocop

herol

acetate

Silicag

el60

toluene

With

outu

sing

visualizingreagent

203

Lack

ofcoloredspot

invisib

lelight

White

030

075divide2000

(119903=09978)

[41]

(+)-120572-Tocop

herol

acetate

Rhod

amineB

203

Darkpink

Pink

045

125divide1600

(119903=09936)

[41]

(+)-120572-Tocop

herol

acetate

Gentia

nviolet

203

Darkviolet

Violet

075

208divide2500

(119903=09908)

[41]

(+)-120572-Tocop

herol

acetate

Methylgreen

203

Green

(smallcon

trast

with

backgrou

ndcolor)

Green

075

125divide2500

(119903=09943)

[41]

(+)-120572-Tocop

herol

acetate

Janu

sblue

203

Darkblue

Blue

075

125divide2500

(119903=09938)

[41]


Table1Con

tinued

Detecteddrug

Stationary

phase

Detectio

nway

Fund

amental

absorptio

nband

ofdrug120582max(nm)

Color

ofspot

ofdetected

drug

Backgrou

ndcoloro

fchromatogram

Limitof

detection

(120583g)

Linearity

range

(120583gspo

tminus1 )

(rcorrelatio

ncoeffi

cient)

Reference

Ibup

rofen

Silicag

el60F 2

54chloroform

+methano

l(50

14

vv)

With

outu

sing

visualizingreagent

200

Lack

ofcoloredspot

invisib

lelight

White

062

125divide1000

(119903=09958)

[42]

Ibup

rofen

Methylene

violet

200

White-bluew

ithdark

blue

border

Light

125

500divide2500

(119903=099549)

[42]

Stearic

acid

Silicag

el60

n-hexane

+aceton

e(4

1vv)

Rhod

amineB

585

Darkpink

Pink

125

156divide2840

(119903=09913)

[45]

Stearic

acid

Methylene

blue

687

Blue

with

light

border

Blue

007

041divide744

(119903=09975)

[45]

Stearic

acid

Janu

sblue

617

Navyblue

with

white

border

Blue

012

156divide744

(119903=09928)

[45]

Stearylalcoh

olSilicag

el60

n-hexane

+ethylacetate+

methano

l(46

3505vvv)

Rhod

amineB

586

Darkpink

Pink

148

290divide2700

(119903=09964)

[45]

Stearylalcoh

olSilicag

el60

n-hexane

+ethylacetate+

methano

l(46

3505vvv)

Malachitegreen

644

Green

with

light

border

Green

032

089divide885

(119903=09959)

[45]

Stearylalcoh

olSilicag

el60

n-hexane

+ethylacetate+

methano

l(46

3505vvv)

Janu

sblue

655

Navyblue

Blue

032

053divide708

(119903=09995)

[45]


Table2Ch

aracteris

ticso

fdensitom

etric

band

ofdetected

drugsmod

ified

contrastindexdensito

metric

visualizingindexmod

ified

broadening

indexanddetectionindex

Detecteddrug

Stationary

phasemobile

phase

Detectio

nway

Densitom

etric

band

characteris

ticof

detected

drug

Mod

ified

contrast

index(AU∘ )

Densitom

etric

visualizing

index(Au120583gsdot∘)

Mod

ified

broadening

index(120583gAU

)

Detectio

nindex

(120583gAU

)Re

ference

Area

(AU)

Height

(AU)120572(∘ )

Estradiol

Neutralalum

inum

oxide

60F 2

54toluene+

ethylacetate

(11vv)

With

outu

sing

visualizingreagent

1304

8246

145

16966

mdash19

16032495

[43]

Estradiol

Bartonrsquosreagent

23786

255

1319615

mdash10

5103210

70[43]

Estradiol

Sulphu

ricacid

44709

497

1338231

mdash0559

010850

[43]

Estradiol

Gentia

nviolet

11348

220

1514667

mdash2203

032760

[43]

Estradiol

Methylene

violet

13622

221

1713000

mdash18

35032705

[43]

Estradiol

Neutralalum

inum

oxide

150F

254toluene+

ethylacetate

(11vv)

With

outu

sing

visualizingreagent

11424

192

131476

9mdash

2188

032409

[43]

Estradiol

Gentia

nviolet

10906

127

2077

00mdash

2292

032796

[43]

Estradiol

Methylene

violet

8466

154

334667

mdash2953

053490

[43]

Salicylanilide

Silicag

el60F 2

54(10

5744

)chloroform

With

outu

sing

visualizingreagent

72889

707

9785

11570

0686

00715

90[44]

Salicylanilide

Brilliant

green

91842

704

8880

1640

0544

07011

112

[44]


herol

Silicag

el60

toluene

With

outu

sing

visualizingreagent

34519

459

162869

0479

0724

0453100

[41]


herol

221015840-Bipyridine-

iron(III)

chlorid

e60

474

698

97756

0323

0413

2088915

[41]


herol

Methylene

violet

40426

636

87950

1684

0618

0303456

[41]


herol

Methylgreen

37196

576

964

00

1378

0672

03015

64[41]

(+)-120572-Tocop

herol

acetate

Silicag

el60

toluene

With

outu

sing

visualizingreagent

33796

591

115373

1024

0740

03011

00[41]

(+)-120572-Tocop

herol

acetate

Rhod

amineB

26031

455

153033

0386

0960

04515

11[41]

(+)-120572-Tocop

herol

acetate

Gentia

nviolet

2944

1545

105450

0392

0849

07518

84[41]

(+)-120572-Tocop

herol

acetate

Methylgreen

30226

564

115127

0366

0827

0753911

[41]

(+)-120572-Tocop

herol

acetate

Janu

sblue

30125

565

115136

0365

0830

0753750

[41]

Ibup

rofen

Silicag

el60F 2

54With

outu

sing

visualizingreagent

5564

444

526

17115

0342

044

90623325

[42]

Ibup

rofen

chloroform

+methano

l(50

14vv)

Methylene

violet

53696

348

3599

430123

046

612

544

38[42]

Stearic

acid

Silicag

el60

n-hexane

+aceton

e(4

1vv)

Rhod

amineB

6567

8231

264

50017

432

125900

[45]

Stearic

acid

Methylene

blue

30918

368

1426286

3155

092

007245

[45]

Stearic

acid

Janu

sblue

36175

462

1142000

2741

079

012604

[45]

Stearylalcoh

olSilicag

el60

n-hexane

+ethylacetate+

methano

l(46

3505vvv)

Rhod

amineB

3605

100

1376

920019

749

148234

[45]

Stearylalcoh

olMalachitegreen

12378

333

747571

0553

218

032382

[45]

Stearylalcoh

olJanu

sblue

10963

269

1320692

0264

246

032214

[45]


Table 3 Detection results of selected corticosteroids [46]

Corticosteroid 119877119865

Reagent 1 Reagent 2Color of

corticosteroid spotLimit of detection

(120583g)Color of


(120583g)Prednisolone 040 Blue-green 0400 Violet 0150Dexamethasone 045 Bray-violet 0300 Violet 0100Triamcinolone 029 Grayish 0200 Violet 0075Beclomethasone 038 Blue 0060 Violet 0040Prednisone 050 Brown-yellow 0300 Violet 0100Betamethasone 038 Blue 0050 Violet 0030Hydrocortisone 039 Greenish 0050 Violet 0040

Table 4 Characteristics of densitometric bands of stigmasterol after dipping and spraying with the use of particular visualizing reagents [47]

10 120583g of stigmasterol 5 120583g of stigmasterolArea (AU) Height (AU) Angle 120572 (∘) 120582max (nm) Area (AU) Height (AU) Angle 120572 (∘) 120582max (nm)

Without using visualizing reagent 7111 198 9 200 4400 126 9 200Rhodamine B

Dipping 2264 86 32 200 1040 373 12 591Spraying 4753 1433 9 200 2721 906 14 200

Gentian violetDipping 2685 958 76 200 712 301 34 693Spraying 5844 1795 75 601 4655 1377 8 602

Methylene violetDipping 3792 119 135 200 1177 582 12 200Spraying 8988 2512 7 200 6122 1766 8 200

Methyl greenDipping 3514 934 73 200 2237 557 39 200Spraying 7857 216 7 200 5447 1583 12 200

Malachite greenDipping 2350 844 735 200 1052 373 37 200Spraying 7740 221 10 638 4942 1516 13 636

Janus blueDipping 2334 826 66 200 1429 475 36 663Spraying 4024 1473 95 674 2302 862 115 674

bromocresol green brilliant cresyl blue bromophenol blueand neutral red were applied for visualizing cholesterol andits derivatives in adsorption thin-layer chromatography (onsilica gel and aluminum oxide) and in argentation thin-layerchromatography (on silica gel impregnated with an aqueoussolution of silver nitrate (5)) [49] Visualizing effects ofdetection of cholesterol derivative depend on several factorsincluding the chemical structure of the visualizing reagentsthe structure of the detected substance and the type ofchromatographic sorbent Cholesteryl acetate was revealedwith all visualizing reagents after separation on silica gelCholesteryl acetanilide cholesteryl acetate and cholesterylarachidonate could be determined on silica gel by use of allthe reagents except bromocresol greenOnneutral aluminumoxide 03 120583g cholesterol could be revealed with bromophe-nol blue neutral red and aniline blue 03 120583g cholesteryl

acetanilide could be visualized with bromophenol blue andalkaline blue 03 120583g cholesteryl acetate could be detectedwithbromophenol blue neutral red helasol red helasol greenand alkaline blue 01 120583g cholesteryl arachidonate could bevisualized with bromophenol blue The worst results wereobtained after argentation TLC the compounds studiedcould be detected only by the use of phenol red helasol greenbromophenol blue neutral red aniline blue and alkalineblue The best and most universal visualizing reagents forcholesterol and the derivatives investigated in adsorptionand argentation TLC were aniline blue bromophenol bluehelasol green and alkaline blue [49]

Wardas and Jędrzejczak [50] separated the selected freeand conjugated bile acids by NP-TLC Eleven visualizingagents were applied for detection of these investigated bileacidsThe best results of detection of bile acids were obtained


Table 5 Characteristics of densitometric bands of 120573-sitosterol after dipping and spraying with the use of particular visualizing reagents [47]

10 120583g of 120573-sitosterol 5120583g of 120573-sitosterolArea (AU) Height (AU) Angle 120572 (∘) 120582max (nm) Area (AU) Height (AU) Angle 120572 (∘) 120582max (nm)

Without using visualizing reagent 4797 1445 15 200 3243 826 10 200

Rhodamine BDipping 646 34 110 200 112 94 21 595

Spraying 3807 1158 12 200 1770 596 225 584

Gentian violetDipping 7161 3123 45 510 1418 478 5 517

Spraying 5148 1469 95 200 4354 1124 19 606

Methylene violetDipping 3017 110 19 200 1490 49 95 200

Spraying 7561 2139 85 200 4056 1263 10 200

Methyl greenDipping 1415 533 105 200 538 248 47 473

Spraying 5966 1674 9 200 4560 1035 16 200

Malachite greenDipping 2235 827 51 640 966 407 42 643

Spraying 5338 1555 8 643 3444 1114 10 649

Janus blueDipping 1029 49 134 678 53 62 415 685

Spraying 2474 691 24 667 1772 501 30 662

with bromocresol blue [50] Bile acids can be also visualizedby dipping plates into phosphomolybdic acid in ethanoland then heating for 10min at 105ndash120∘C [1] or by sprayingthe plates with a 10 solution of phosphomolybdic acidin methanol and then heating for 20min at 50ndash80∘C [51]Chromatographic bands of bile acids on the densitogramafter the use of spray solution of phosphomolybdic acidin methanol were irregular [51] Therefore this way of thedetection of bile acids cannot be recommended Regularchromatographic bands of bile acids on the densitogramwereobtained after the use of dipping water solution of sulphuricacid [52]

312 Detection of Antiepileptic Anticonvulsant and Psy-chotropic Drugs Phenytoin was detected by 1 aqueousmercuric nitrate reagent [1] 2 ethanolic mercuric chloridefollowed by 02 ethanolic diphenylcarbazone [1 53] and001 carbazone followed by 1 ferrous sulfate [1] Howeverthese reagents with phenytoin give a similar reaction tothe barbiturates Kulkarni et al [54] reported a selectiveand sensitive chromogenic reagent namely 01 brominein carbon tetrachloride followed by 01 o-tolidine in 05acetic acid for detection and determination of phenytoinin biological samples investigated by TLC Phenytoin withthis reagent gives blue spot on a white background ofchromatogram Color of blue spot is stable for an hour Thisreagent does not give any color reaction with barbituratesHowever this reagent gives a similar color reaction withthe benzodiazepines Sensitivity for phenytoin alprazolam

diazepam lorazepam nitrazepam and oxazepam is equal to5 120583g 20120583g 30 120583g 5 120583g 30 120583g and 5 120583g respectively [54]A single and sensitive chromogenic chlorination with theuse of o-tolidine was used for detection of diazepam phe-nobarbitone and saccharin [55] Diazepam phenobarbitoneand saccharin solution were spotted on silica gel G platesand developed by the use of one from three mobile phasesnamely n-hexane-acetone-methanol (8 3 05 vvv) n-hexane-acetone-butanol (12 8 05 vvv) and chloroform-acetic acid (9 1 vv) After development plates were driedin air and were placed for ca 5min in a chamber con-taining chloride gas excess chlorine was removed fromthe plates and the next plates were sprayed with o-tolidinereagent (50mg o-tolidine in 100mL 10 acetic acid) Whena faint blue color appeared the plates were next sprayedwith 1 phosphomolybdic acid for stabilization The limitsof detection for diazepam phenobarbitone and saccharinwith o-tolidine reagent were 05 120583g per spot 03 120583g perspot and 01 120583g per spot respectively [55] Cyclodol anddiprazin were separated on silica gel by the use of one fromsix mobile phases toluene-acetone-ethanol-25 NH

4OH

(45 45 75 25 vvvv) hexane-ethyl acetate (15 5 vv)chloroform-heptene-25 NH

4OH (16 3 3 vvv) ethyl

acetate-hexane (1 1 vv) acetonitrile-methanol (1 1 vv)or heptene-chloroform-ethanol-25 NH

4OH (5 10 3 1

vvvv) Iodine vapours UV light at 254 nm and freshnormal plasma (FNP) can be applied as visualizing reagentsfor cyclodol and diprazin FNP gives colored spot withdiprazin [56]


313 Detection of Antibiotics Quintens et al [57] describedthe procedure which enables the identification of thirtycephalosporins by TLC on silanized silica gel F

254 The

obtained results with seven mobile phases were reportedResults of some reactions with all cephalosporins were alsopresented The following reagents were applied (1) sulfuricacid (2) sulfuric acid-formaldehyde (1mL of a 37 mmsolution of formaldehyde mixed with 50mL of concentratedsulfuric acid) (3) sulfuric acid-nitric acid (1mL of concen-trated nitric acid mixed with 99mL of an 80 vv solutionof sulfuric acid) [57] The reagents (1) and (2) were usedby mixing the cephalosporin (2mg) with 005mL water andadding the reagent (2mL) The reagent (3) was administeredby mixing the cephalosporin (2mg) directly with 025mLof reagent (3) The details of results obtained with the colorreactions were presented in Quintens paper [57]The sulfuricacid-formaldehyde reagent (2) gives the most discriminatorycolors TLC on silanized silica gel combined with sim-ple color reactions enables identification of all investigatedcephalosporins [57]

Bacitracin is a mixture of related cyclic polypeptides pro-duced byBacillus licheniformis andBacillus subtilis organismsand is used in human medicine as a polypeptide antibioticBacitracin with dabsyl chloride gives a complex and can bedetected by densitometric measurements at 460 nm [58]

314 Detection of Essential Oil Compounds Pyka et al [59]applied eighteen new visualizing reagents for the visualiza-tion of nineteen standard essential oil components (hydro-carbons aliphatic and monocyclic alcohols phenols ethersketones and aldehydes) after chromatography on silica geland benzene asmobile phaseThe visualizing effects obtainedwere compared with detection of these compounds by usingiodine vapor and 5 solution potassium dichromate in 40sulfuric acid These investigations possess quality characterfor the detection substances which occur in various essentialoils Quality identification was based on the ground of the119877119865values of investigated compounds and on the color of

detected compounds in TLCThe substances belonging to thesame class of compounds investigated bymeans of adsorptionthin-layer chromatography may have slight differences in119877119865values For example hydrocarbons investigated by Pyka

et al [59] camphene (R)-(+) limonene and p-cymenealcohols farnesol menthol and borneol phenols eugenoland guaiacol ethers coumarin cineole and carvone andcinnamic aldehyde as well as camphor and (1R)-(minus) fen-chone with regard to their incomplete separation by meansof these techniques (small differences in 119877

119865values) can

be identified by means of diversified colours of spots onchromatogram These results can be used in the practicefor qualitative investigation of essential oils of plants Forexample thymol linalool and borneol are presented inthymeherbHerbaThymiThedifference in119877

119865values between

thymol and linalool was notable and was equal to Δ119877119865=

0265 while between linalool and borneol was small andwas equal to Δ119877

119865= 0043 That is why the confirmation

of appearance particularly linalool and borneol in mixture

can be by diversified colours of their chromatographic spotswith visualizing reagents 01 aqueous HCl + 1 aqueousNi(NO

3)2 01 aqueous HCl + 2 aqueous CuSO

4 and

01 aqueous Na2CO3+ 2 aqueous CuSO

4 Furthermore

linalool and geraniol are the main components of corianderfruit (Fructus Coriandri)The difference in119877


these compounds was equal to 0085The different colours ofchromatographic spots of linalool and geraniol were obtainedwith visualizing reagents 01 aqueous HCl + 1 aqueousNi(NO


4 01

aqueous HCl + 2 aqueous CuSO4+ saturated aqueous solu-

tion of variamine blue hydrochloride 01 aqueous HCl +saturated aqueous solution of variamine blue hydrochloridesaturated aqueous solution of variamine blue hydrochloride01 aqueous Na

2CO3+ saturated aqueous solution of

variamine blue hydrochloride and 5 solution ofK2Cr2O7in

40 H2SO4 Fennel oil (Fructus Foeniculi) contains among

others trans-anethole fenchone and limonene Limonenewas visualized only by means of visualizing reagent 01aqueous HCl + 2 aqueous CuSO

4+ saturated aqueous

solution of variamine blue hydrochloride whilst this agentdoes not give the reaction with fenchone and trans-anetholeOnly the visualizing reagent 01 aqueous Na

2CO3gives

different colour of chromatographic spots with fenchoneand trans-anethole Anise oil (Oleum Anisi) contains amongothers trans-anethole fenchone and p-cymene p-Cymeneand trans-anethole give chromatographic bands with variouscolours with visualizing reagent 01 aqueous Na

2CO3+ 1

aqueousNi(NO3)2+ saturated aqueous solution of variamine

blue hydrochloride whilst fenchone could not be detectedby means of this reagent To detect fenchone it is necessaryto make a second chromatographic analysis and to applyfor example visualizing reagent saturated aqueous solutionof variamine blue hydrochloride for fenchone detectionThis reagent does not detect trans-anethole but with p-cymene gives band with orange-yellow colour whilst thespot has light grey colour with fenchone The results canbe also applied for quick estimation of purity of somecompounds obtained synthetically The natural vanillin ispresent in vanilla fruits (Vanilla planifolia) but syntheticallycan be obtained from eugenol Vanillin (119877

119865= 0138) and

eugenol (119877119865= 0364) can be analysed qualitatively on

chromatogram and the presence of eugenol can also bechecked in the product The diversified colours of spots ofthese compounds with visualizing reagents 01 aqueousHCl + 1 aqueous Ni(NO

3)2+ saturated aqueous solution

of variamine blue hydrochloride 01 aqueous HCl + 2aqueous CuSO

4+ saturated aqueous solution of variamine

blue hydrochloride 01 aqueous HCl + saturated aqueoussolution of variamine blue hydrochloride saturated aqueoussolution of variamine blue hydrochloride 01 aqueousNa2CO3+ 2 aqueous CuSO

4+ saturated aqueous solution

of variamine blue hydrochloride 01 aqueous Na2CO3+

saturated aqueous solution of variamine blue hydrochloride01 aqueous Na

2CO3+ 1 aqueous Ni(NO

3)2+ saturated

aqueous solution of variamine blue hydrochloride and 5solution of K

2Cr2O7in 40 H

2SO4 can be useful for

interpretation of the results obtained [59]


315 Detection of Quinones Quinones are a class of aro-matic yellow compounds that are biologically important ascoenzymes or acceptors or vitamins Kocjan [60] investigatedeleven new visualizing reagents for the detection of fivequinones Quinones were separated on silica gel 60 using amixture of dichloromethane-n-hexane (8 2 vv) as mobilephase Variamine blue hydrochloride hydrochloric aciddithizone 8-hydroxyquinoline and different salts [Ni(NO

3)2

Na2CO3 Co(NO

3)2] were used to prepare the solutions of

visualizing reagents The 119877119865values and different colors of

spots obtained for quinones enabled unequivocal identifica-tion of all quinonesThe detection limit of detected quinoneswas from 01120583g to 5 120583g [60]

316 Detection of Phenolic Drugs and Flavonoids Wardaset al [61] examined alkacymetric indicators for detec-tion of adrenaline dopamine phenylephrine metaraminolfenoterol and bithionol after TLC separation on silica gelpolyamide 11F

254 and a mixture of silica gel and kieselguhr

by using glacial acetic acid-n-butanol-water (1 + 4 + 1 vv)as mobile phase They investigated the following indicatorsas visualizing reagents phenol red thymol blue bromoth-ymol blue bromophenol blue cresol green erythrosine Beriochrome black T brilliant cresyl blue eosin yellow titanyellow and helasol green dissolved in 5 NaOH as well asbromocresol green aniline blue alkaline blue brilliant cresylblue and brilliant green dissolved in water Additionallydimethyl yellow and thymolphthalein dissolved in methanolwere applied but before used the chromatographic plate wassprayed with 5 NaOH In each case the concentration ofthe solutions was 05mgmL The visualizing effects werecharacterized by the detectability index which was definedby Gregorowicz and Sliwiok [39 40] using (1) Charac-teristics of the visualizing effect for adrenaline dopaminephenylephrine metaraminol fenoterol and bithionol withthe best visualizing reagents investigated on Kieselgel 60F

254

Kieselgel 60kieselguhr F254

and Polyamide 11F254

are listedin Table 6 However phenylephrine and bithionol were notdetected on polyamide 11F

254with none of the studied

visualizing reagents [61] Pyka et al [62] detected silica gel60 (E Merck 105721) following phenolic drugs bamethaneethamivan hexachlorophene salicylanilide pyrocatechinethymol pentazocine phloroglucinol eugenol niclosamideterbutaline methyldopa and norepinephrine Plates withmethyldopa norepinephrine terbutaline bamethane andethamivan were developed with a mixture of glacial aceticacid-n-butanol-water (1 4 1 vv) as mobile phase Plateswith phloroglucinol pentazocine hexachlorophene pyrocat-echine niclosamide salicylanilide and thymol were devel-oped with a mixture of chloroform-methanol (9 1 vv) Theplate with eugenol was developed with benzene as mobilephase Thirteen visualizing reagents were used to detect theabove-mentioned phenolic drugs Alkaline blue aniline blueneutral red and brilliant green were used as 50mg100mLsolutions in water Bromophenol blue bromothymol bluebrilliant cresyl blue thymol blue phenol red bromocresolgreen and helasol green were applied as 50mg100mLsolutions in 2 aqueous sodium hydroxide solution Bro-mophenol blue solution was prepared directly before use

Plates were evaluated after spraying and heated at 100∘C for15min By the means of the following visualizing reagentsalkaline blue aniline blue bromophenol blue bromothymolblue alkaline solution of brilliant cresyl blue bromocresolgreen helasol green and aqueous solution of brilliant cresylblue it is possible to detect all of the drugs investigated inthe amount of 100 120583g The results with the best visualizingreagents are also presented in Table 6

Smolarz et al [63] visualized phenolic acid in the petiolesof Rheum undulatum and Rheum rhaponticum Separationand identification of phenolic acids were performed oncellulose plates and by using the mobile phases benzene-methanol-acetic acid-acetonitrile (80 10 5 5 vvvv) inthe first direction and sodium formate-formic acid-water(10 1 200 wvv) in the second direction After drying thechromatograms were sprayed with diazotized sulfanilic acidin 20 sodium carbonate solution or with 2 aqueous ferricchloride The limits of detection for gallic protocatechuiccaffeic p-hydroxybenzoic p-coumaric syringic vanillic andferulic acids were equal to 13 ng 10 ng 12 ng 16 ng 60 ng60 ng 11 ng 10 ng and 64 ng respectively [63]

Phenolic acids and flavonoids were also detected in someCroatian Stachys Taxa Analysis was performed on silica gel60F254

HPTLC plates using ethyl acetate-acetic acid-formicacid-water (100 11 11 26 vvvv) as mobile phase Afterdrying the plates were sprayed with the natural productsreagent polyethylene glycol After spraying chromatogramswere observed at 254 nm and 366 nm Fluorescence colorsof phenolic acid and flavonoid standards were intense lightblue (for caffeic acid chlorogenic acid) dark orange (forhyperoside isoquercitrin luteolin luteolin 7-O-glucosiderutin quercetin and quercitrin) and light orange for vitexin[64] Selected flavonoids (rutin narcissin nicotiflorin andisoquercitrin) were isolated and identified in the extractfrom Caragana spinosa [65] TLC was performed on silicagel plates using ethyl acetate-12-dichloroethane-acetic acid-85 formic acid-water (10 25 1 1 08 vvvvv) as mobilephase First the being development distance was 6 cm Nextplates were dried and developed using the same conditionsAfter dried the investigated flavonoids were detected usingvisualizing reagent (05 2-aminoethyl diphenylborinatesolution in ethyl acetate5 polyethylene glycol solution in12-dichloroethane) [65 66]The detection limits were 44004421 2728 and 2129 ng for rutin narcissin nicotiflorin andisoquercitrin respectively

Głowniak et al [67] analyzed phenolic compounds inthe flowers of Lavatera trimestris L (Malvaceae) Analysiswas performed on silica gel 60 using different mobile phasesCaffeic p-coumaric ferulic protocatechuic gentisic chloro-genic and gallic acids in UV light give blue light blueblue violet blue blue and violet spots respectively Caffeicp-coumaric ferulic protocatechuic gentisic chlorogenicand gallic acids in UV light after treatment with ammoniavapor give blue blue blue violet yellow yellow-green andviolet spots respectively Caffeic p-coumaric ferulic proto-catechuic gentisic chlorogenic isovanillic gallic syringicvanillic and p-hydroxybenzoic acids after spraying withdiazotized sulfanilic acid in 20 sodium carbonate solutiongive brown red violet brown gray light brow orange


Table6Ch

aracteris

ticso

fthe

visualizingeffectfor

selected

phenoliccompo

unds

Detecteddrug

Stationary

phase

Mob

ileph

ase

Visualizingreagent

Detectio

nlim

it(120583g)

Detectability

index

(120583gmm

2 )Color

ofspot

Backgrou

ndcoloro

fchromatogram

Reference

Adrenalin

eSilicag

el60F 2

54Glacialaceticacid

+n-bu

tano

l+water

(14

1vvv)

Brilliant

cresylblue

in5

NaO

H010

01010

Brow

nLightviolet

[61]

Silicag

el60kieselguh

rF254

Brom

ocresolgreen

in5

NaO

H025

02547

Lightb

rown

Seagreen

Polyam

ide11F

254

Eosin

yello

win

5NaO

H250

25030

Lightb

rown

Lightyellow

Dop

amine

Silicag

el60F 2

54Glacialaceticacid

+n-bu

tano

l+water

(14

1vvv)

Brom

ocresolgreen

in5

NaO

H010

01015

Orange

Lightb

lue

[61]

Silicag

el60kieselguh

rF254

025

02510

Brow

norange

Seagreen

Polyam

ide11F

254

Eosin

yello

win

5NaO

H075

07517

Lightb

rown

Lightyellow

Metaram

inol

Silicag

el60F 2

54Glacialaceticacid

+n-bu

tano

l+water

(14

1vvv)

Brom

ocresolgreen

in5

NaO

H050

05015

Lemon

Lightb

lue

[61]

Silicag

el60kieselguh

rF254

Eriochromeb

lack

Tin

5NaO

H050

05035

Brow

nLightb

rown

Polyam

ide11F

254

Anilin

eblueinwater

505042

Lightyellow

Blue

Feno

terol

Silicag

el60F 2

54Glacialaceticacid

+n-bu

tano

l+water

(14

1vvv)

Brom

ocresolgreen

in5

NaO

H010

01013

Brow

norange

Lightb

lue

[61]

Silicag

el60kieselguh

rF254

Eriochromeb

lack

Tin

5NaO

H050

05038

Brow

nLightb

rown

Polyam

ide11F

254

Eosin

yello

win

5NaO

H50

515

Yello

wLightyellow

Phenylephrine

Silicag

el60F 2

54Glacialaceticacid

+n-bu

tano

l+water

(14

1vvv)

Brom

ocresolgreen

in5

NaO

H025

02521

Lemon

Lightb

lue

[61]

Silicag

el60kieselguh

rF254

Eriochromeb

lack

Tin

5NaO

H050

05041

Brow

nLightb

rown

Bithiono

lSilicag

el60F 2

54Glacialaceticacid

+n-bu

tano

l+water

(14

1vvv)

Brilliant

cresylblue

in5

NaO

H050

05077

Brow

nLightviolet

[61]

Silicag

el60kieselguh

rF254

Brilliant

cresylblue

in5

NaO

H250

25068

Brow

nLightb

rown

Methyldop

a

Silicag

el60

Glacialaceticacid

+n-bu

tano

l+water

(14

1vvv)

Thym

olblue

in2

NaO

H32

327

Brow

nblack

Beige

[62]

Norepinephrine

Brom

othymolblue

in2

NaO

H06

0628

Yello

wbrown

Lightb

lue

Terbutaline

Brilliant

cresylblue

in2

NaO

H20

2017

Lightb

rown

Lightb

lue

Bamethane

Anilin

ebluein2

NaO

H10

1025

Blue

Lightb

lue

Etam

ivan

Anilin

ebluein2

NaO

H50

5018

Whiteblue

Lightb

lue

Phloroglucinol

Silicag

el60

Chloroform

+methano

l(9

1vv)

Brilliant

cresylblue

in2

NaO

H20

2023

Lightb

rown

Lightb

lue

[62]

Pentazocine

Anilin

ebluein2

NaO

H05

0568

White

Lightb

lue

Hexachlorop

hene

Thym

olblue

in2

NaO

H50

5037

Yellowbeige

Beige

Pyrocatechine

Brilliant

greenin

2NaO

H03

0317

Light

orangered

Lightg

reen

Niclosamide

Brilliant

greenin

2NaO

H08

0818

Green

Lightg

reen

Salicylanilide

Brilliant

greenin

2NaO

H08

0816

Green

Lightg

reen

Thym

olBrilliant

greenin

2NaO

H20

2042

Yello

wgreen

Lightg

reen

Eugeno

lSilicag

el60

Benzene

Brom

othymolblue

in2

NaO

H50

5037

Lightyellow

Lightb

lue

[62]


light brown dark red orange and yellow spots respectivelyCaffeic p-coumaric ferulic protocatechuic gentisic chloro-genic gallic syringic vanillic and p-hydroxybenzoic acidsafter spraying with diazotized p-nitroaniline give brownbrown-blue blue red-brown gray-green lemon green-redblue violet and red spots respectively Caffeic ferulicprotocatechuic gentisic chlorogenic isovanillic gallic andvanillic acids after spraying with a 2 aqueous solution offerric chloride give brown-green red light-orange brown-blue navy blue gray-green gray and brown spots respec-tively [67]

Bhujbal and Sandhya [68] investigated the extracts fromtwo commercial formulations of Vidangarista Analysis wasperformed on silica gel 60F

254plates The presence of gallic

acid and conessine in extracts was observed using ethylacetate-toluene-methanol-formic acid (3 3 03 08 vvvv)and toluene-ethyl acetate-diethyl amine (65 25 10 vvv)as mobile phases respectively After development gallic acidand conessine on chromatographic plate were visualizedusing Dragendorff rsquos reagent Densitometric analysis wasperformed at 120582max 280 nm for gallic acid and at 120582max 520 nmfor conessine

317 Detection of Fatty Acids Fatty Alcohols Amide of FattyAcids and Esters of Fatty Acids The fatty acids fatty alcoholsand esters of higher fatty acids have great biochemicalmedical and pharmaceutical significance The alkacymetricindicators namely thymol blue bromophenol blue andbromothymol blue were also used for the detection ofdifferent classes including the compounds with biologicalsignificance namely acids (myristic acid palmitic acidand stearic acid) glycerides (glycerol trioleate glyceroltripalmitate) alcohols (oleyl alcohol stearyl alcohol) andamides (stearamide palmitamide) on silica gel 60F

254after

separation using the n-hexane + acetone (8 2 vv) asmobile phase The colors of spots were durable and sufficientcontrast between spots and background was observedThe detection limits with these visualizing reagents werefrom 05 120583g to 10 120583g for acids from 10 120583g to 20120583g forglycerides from 10 120583g to 20120583g for alcohols and 10 120583gfor amides [48] Aniline blue alkaline blue bromothymolblue thymol blue bromophenol blue phenol red helasolgreen bromocresol green and brilliant cresyl blue were alsoapplied to the detection of unsaturated fatty acids (cis-6-octadecenoic cis-9-octadecenoic cis-11-octadecenoic cis-12-octadecenoic cis-13-octadecenoic trans-9-octadecenoictrans-11-octadecenoic cis-6912-octadecatrienoic and cis-91215-octadecatrienoic) chromatographed on silica gel60F254

and on mixture of silica gel and kieselguhr F254

[69]The best detectability (2120583g or 3 120583g) of unsaturated fatty acidswas obtained with aniline blue alkaline blue bromothymolblue thymol blue and bromophenol blue [69] Niestroj et al[70] separated palmitic acid 120572-hydroxypalmitic acid stearicacid and 12-hydroxystearic acid by normal phase thin-layerchromatography (NP-TLC) using a mixture of n-hexaneand acetone in volume composition 7 3 (vv) as mobilephase This group of compounds is not active in the rangeUV light Therefore suitable visualizing reagent was appliednamely aqueous solution of rhodamine B which activates

the detected chromatographic band [70] The linearity rangeof stearic acid using rhodamine B has been established tobe from 2120583g to 20120583g [70] However short-chain fatty acidsfrom ethanoic to octanoic were chromatographed on silicagel with n-hexane-acetone (4 1 vv) and acetone-water-chloroform-ethanol-aqueous ammonia (30 1 3 5 1 vv) asmobile phases [71] New visualizing reagents (aniline bluealkaline blue neutral red brilliant green bromothymol bluebrilliant cresyl blue thymol blue bromophenol blue phenolred bromocresol green and helasol green) were used forthe detection of the free fatty acids and their ammoniumsalts The results obtained for the free fatty acids and theirammonium salts by the use of the individual visualizingreagents depend on method of applicationmdashspraying ordipping Dipping of chromatographic plates into solutions ofthe visualizing reagents leads to chromatographic spots ofthe fatty acids with better contrast than the spots obtained bydirect spraying of the plates Generally better visualizationwas achieved with ammonium salt of the fatty acids thanwith the free acids Of the reagents described in thescientific literature free fatty acids from ethanoic to octanoiccan be visualized only by the use of bromocresol greenbromophenol blue potassium permanganate and methylred Among the visualizing reagents investigated dipping ofchromatographic plates into an aqueous solution of alkalineblue enables the detection of the free acids from propanoic tooctanoic The ammonium salts of fatty acids from ethanoicto octanoic can be detected by spraying or dipping usingthe visualizing reagents investigated except bromophenolblue [71] Eighteen new reagents (alkaline blue aniline bluebrilliant green neutral red bromothymol blue thymol bluephenol red helasol green bromocresol green brilliant cresylblue bromophenol blue eriochrome black erythrosin Beosin yellow spands thymolphthalein and Congo red) wereapplied for visualizing eight esters of higher fatty acids afterchromatography on silica gel on a mixture of silica gel andkieselguhr and on neutral aluminum oxide [72] For all theesters investigated the best detectability was obtained on themixture of silica gel 60 and kieselguhr F

254 it was worse

on silica gel 60F254

and the worst on neutral aluminumoxide Bromophenol blue was the best and most universalvisualizing reagent for all the esters investigated on all thechromatographic supports [72]

318 Detection of Quinolones The quinolones were detectedby the Dragendorff Forrest and Folin-Ciocalteu reagents aswell as iron (III) chloride in hydrochloric acid iodic reagentphosphomolybdic acid in sulphuric (VI) acid [73 74] andterbium (III) and europium (III) ions [75] Wardas et al [76]also visualized quinolones using different indicators (eosinyellow thymol blue bromothymol blue thymolphthaleinhelasol green spands and titan yellow) For all the quinolonecompounds studied whether chromatographed on silica gelor aluminum oxide the best detectability was obtained withhelasol green [76] Bober [77] showed the possibility of use-fulness of the visualizing reagents such as Janus blue methy-lene violet gentian violet methyl green cresol red Rho-damine B malachite green methylene blue eosin yellow and


metanil yellow in the detection of selected quinolones andfluoroquinolones The cinoxacin pipemidic acid ofloxacinand pefloxacin were separated by thin-layer chromatography(TLC) and high performance thin-layer chromatography(HPTLC) on silica gel 60 using acetonitrile + water + aceticacid (6 40 4 vvv) as mobile phase The visualizing effectswere evaluated 3 times namely directly after dipping insolutions of visualizing reagents after drying at a roomtemperature during 24 hours and after dipping in solutionsof visualizing reagents and drying at 120∘Cduring 10minutesCresol redwas the best visualizing reagent for the detection ofofloxacin and pefloxacin Janus blue was the best visualizingreagent for the detection of cinoxacin and pipemidic acidThe detection limits for ofloxacin pefloxacin cinoxacin andpipemidic acid were 050 075 010 and 010 120583g respectively[77] However ciprofloxacinHCl in coated tablets can be alsodirectly obtained on silica gel plate 60F

254after scanning in

absorbency mode at 278 nm [22]

319 Detection of Hypnotic Compounds Zolpidem and zopi-clone are hypnotic reagents and can be used as potentialdrugs Pushpalatha et al [78] applied 05 solution ofchloranilic acid in 14-dioxane for the detection of zolpidemand zopiclone on silica gel 60 Colored complex spots ofzolpidem and zopiclone with chloranilic acid are formed innonaqueous medium Zolpidem with chloranilic acid givesdark pink spot with sensitivity of method equal to 2 120583gZopiclone with chloranilic acid gives light pink spot withsensitivity of method equal to 5120583g

3110 Detection of Saponins Saponins have anti-inflamma-tory antifungal hemolytic cytotoxicity molluscicidal anti-tumor antiviral and immunomodulatory properties Sap-onins were detected on silica gel 60 TLC and HPTLC plateswith 10 sulfuric acid in ethanol or anisaldehyde-sulfuricacid reagent and heated at 110∘C in an oven for 5min and thenevaluated in visible andUV light at 120582 = 365 nm Saponins canbe also visualized by using blood reagent [79]

3111 Detection of Glucosamine Glucosamine is an aminosugar a prominent precursor in the biochemical synthesis ofglycosylated proteins and lipids aswell as dietary supplementsused by adults On heating the amino plate the glucosaminereacts to form a compound which strongly absorbs lightbetween 305 nm and 330 nm with weak fluorescence Thedetection limit in absorption mode is equal to 25 ng per spot[80] Ninhydrin has usually been applied in combinationwith acetic acid pyridine and collodine for the detectionof glucosamine Ammonium bisulfate anisaldehyde andElson-Morgan reagents were also used for the detectionof glucosamine The detection limit of glucosamine withninhydrin is equal to 1000 ng per spot [80]

Esters et al [81] separated glucosamine from the plantextracts on a silica gel 60F

254HPTLC plates using 2-

propanol-ethyl acetate-ammonia solution (8) in volumecomposition 10 10 10 as mobile phase For visualization theplate was dipped into a modified anisaldehyde (detectionreagent solution consisting of diethyl ether-glacial acetic

acid-anisaldehyde-sulphuric acid in volume composition(136 91 1 20)) for exactly 2 s and heated at 120∘C for 30minin a drying oven Glucosamine gives brownish-red chromato-graphic spots Densitometric quantificationwas performed at415 nm [81]

3112 Detection of Hypoglycemic Drug Pioglitazone is ahypoglycemic (antihyperglycemic antidiabetic) drug used toimprove glucose control in adults over the age of 18 withtype 2 diabetes Mohamed et al [82] elaborated new sensitiveand selective high performance thin-layer chromatography(HPTLC) coupled with densitometry for determination ofpioglitazone hydrochloride in pharmaceutical formulationsAnalysis was performed on silica gel 60 with methanol-chloroform (10 1 vv) asmobile phase Chromogenic reagentas visualizing reagent namely o-phthalaldehyde gives thepink colored product with pioglitazone hydrochloride onchromatographic plate

3113 Detection of Selected Compounds in Extract from Dif-ferent Plants Nowak et al [83] investigated the extracts fromAsteraceae family (Centaurea bella Dugaldia hoopesii Inulaaschersoniana Serratula wolffii Stizolophus balsamita andZoegea baldschuanica) using silica gel 60 platesThreemobilephases were used Methylene chloride-acetonemobile phasesin different volume compositions (8 1 7 1 5 1 3 1 2 11 1) were used for the separation of sesquiterpenes Howevermethylene chloride-acetone mobile phase in volume com-position 1 2 and methylene chloride-methanol in volumecomposition 5 1 were applied for the separation of ecdysonesSesquiterpenes and ecdysones were detected on thin-layerusing anisaldehyde reagent (anisaldehyde 05mL acetic acid100mL methanol 85mL and sulphuric acid 45mL) andheated at 103∘C for 3min The spots of isolated com-pounds have different colors namely mauve (for stizolin)violet (for 9120572-hydroxyparthenolide) pink (for partheno-lide and cynaropicrin) orange (for hymenoratin B) yellowfor hymenoratin B 2-O-120573-D-(61015840-O-acetyl)-glucopyranosidedark violet (for kandavanolide) dark blue (for cebellin O)cherry-red (for limonin A) grey (for acetylhymenograndin)and blue (for polypodine B) [83] Anisaldehyde reagent(15mL p-anisaldehyde 25mL sulphuric acid and 1mLacetic acid in 37mL ethanol) andor Natural Product Reagent(1 g diphenylborinic acid aminoethyl ester in 200mL of ethylacetate) were applied to the identification of thin-layer of thecompounds from the extracts of the constituent plants (Urticadioica L Arctostaphylos uva-ursi L Helichrysum italicumRoth Ribes nigrum L Citrus limon L Centaurium erythraeaRafn Melissa officinalis L and Arctium lappa L) [84]

Podolak et al [85] elaborated HPTLC-densitometricmethod for quantitative determination of triterpene saponinsin various parts of eight Lysimachia Lspecies (roots stemsleaves fruits and flowers of Lysimachia nemorum L Lysi-machia nummularia L Lysimachia vulgaris L Lysimachiapunctate L Lysimachia thyrsiflora L Lysimachia ephemerumL Lysimachia ciliate L and Lysimachia clethroides) Plateswere developed with two mobile phases First mobile phasewas chloroform-methanol-water (8 7 1 vv) for L ciliata


L punctata and L nemorum Second mobile phase was n-butanol-acetic acid-water (6 1 3 vvv) for L ephemerumL vulgaris L thyrsiflora and L nummularia Plates withsaponins after the separation were detected using 25 solu-tion of sulphuric acid in methanol and heated at 105∘Cfor 5min Saponin spots showed violet color Quantitativedetermination of saponins was performed by scanning den-sitometry at 545 nm

Yadav andGupta [86] analyzedPremna integrifolia whichis general component of herbal formulation ldquoDashmoolrdquo 10-O-trans-p-Coumaroylcatalpol 410158401015840-hydroxy-E-globularininand premnosidic acid are major iridoid glycosides presentin Premna integrifolia These compounds were separatedon silica gel 60F

254using ethyl acetate-methanol-water-

acetic acid (80 12 6 2 vvvv) as mobile phase Afterdevelopment the plates were dried and dipped into vanillin-sulphuric acid reagent (5 g vanillin + 475mL ethanol + 25mLsulphuric acid) and next were heated for 3min at 110∘CQuantitative determination of investigated compounds wasperformed by scanning densitometry at 510 nm

Maurya and Srivastava [87] elaborated new HPTLC-densitometric method for the simultaneous quantitationof four bioactive markers ursolic acid betulinic acid 120573-sitosterol and lupeol in the stem and root barks of Alstoniascholaris Analysis was performed on silica gel 60F

254plates

using chloroform-methanol (99 1 vv) as mobile phaseUrsolic acid betulinic acid 120573-sitosterol and lupeol werevisualized by vanillin-sulphuric acid reagent Quantitativedetermination of ursolic acid betulinic acid 120573-sitosterol andlupeol was performed by scanning densitometry at 680 nm

Alkaloids occurring in leaves of Lobelia cardinalis (Cam-panulaceae) on thin-layer were visualized with Dragendorff rsquosreagent [88]

3114 Detection of Vitamins Hydrophobic and hydrophilicvitamins were detected by many visualizing reagents whichwere described in review [29]

32 Detection by Photocatalysis of Different Drugs Makowskiet al [89] applied the photocatalytic reactions to visu-alize drugs (antibiotics antirheumatic anti-inflammatoryanalgesic antitussive broncholytic spasmolytic anesthetichypnotic sympathomimetic and vitamin) The drugs wereseparated on silica gel using a mixture of butanol anhy-drous acetic and water (6 2 2 vvv) as mobile phaseAfter development and air-drying plates were sprayed withthe four visualizing reagents namely (a) 10mL potassiumpermanganate with 025 g titanium dioxide powder andafter spraying plates were illuminated for 10min (b) 10mLpotassium iodide with 025 g titanium dioxide powder andafter spraying plates were illuminated for 5min and nextwere sprayed again with 2 aqueous starch solution (c)10mL potassium bromide with 025 g titanium dioxide pow-der (d) 10mL potassium chloride with 025 g titaniumdioxide powder After spraying with (c) and (d) reagentsthe plates were illuminated for 10min Next these plateswere sprayed with silver nitrate (01molsdotLminus1) and illumi-nated again for 3min The most sensitive detections were

obtained with a reagent containing titanium dioxide powderin potassium bromide solutionThe detection limits with thisreagent were 05 120583g for benzyl-penicillin procaine 02120583g forbenzyl-penicillin potassium 02120583g for penicillic acid 05 120583gfor tetracycline hydrochloride 05120583g for oxytetracyclinehydrochloride 05 120583g for chlortetracycline hydrochloride02 120583g for penicillamine 03 120583g for metamizole sodium02 120583g for aminophenazone 03 120583g for salicylamide 3 120583gfor codeine phosphate 1120583g for aminophylline 07 120583g forpapaverine hydrochloride 10 120583g for phenazone 02 120583g forprocaine hydrochloride 02 120583g for phenobarbital 05 120583g forephedrine hydrochloride and 05 120583g for ascorbic acid [89]

33 Microbiological Detection (Bioautography Detection)Sensitive selective and rapid methods are presented fordetecting inhibition of microbial growth by various com-pounds Bioautography is a special method of the detectionand is suitable for investigating antimicrobial activity bythe application of thin-layer chromatography [90] Bioauto-graphy can be realized in three versions namely contactimmersion and direct [91]Many antibacterial and antifungalcompounds can be detected by bioautography method [90ndash92] Many applications of bioautography detection are pre-sented and discussed in review articles [90ndash92]

Amphotericin B [93] and Brazilian medicinal and fruitbearing plants [94] were visualized with Candida albicansShiitake mushroom was detected with Micrococcus luteus[95] Two-dimensional thin-layer chromatography (2D-TLC)bioautography with C fragariae was used for the detectionof agricultural fungicides and antifungal drugs [96] VitaminB12can be detected using Escherichia coli [97] Pseudomonas

savastanoi pv phaseolicola was applied for the detection ofaflatoxin B

1 aflatoxin B

2 aflatoxin G

1 aflatoxin G

2[98ndash

100] trans-resveratrol [101ndash104] antibacterial compounds inboth red and white wine extracts [104] and Chelidoniummajus L alkaloids [105] Doxycycline [106] flumequine [106107] cefacetrile [108] enrofloxacin [109 110] ciprofloxacin[109 110] and Hypericum brasiliense polyphenols [111] canbe detected with Bacillus subtilis Essential oils and extractsfrom different plants can be visualized with Bacillus subtilisB cereus Vibrio fischeri C sphaerospermum (Penzig) C cla-dosporioides (Fresen) de Vries Staphylococcus aureusMicro-coccus luteus Staphylococcus epidermidis Escherichia coliPseudomonas aeruginosa Candida albicans Xanthomonascampestris pv vesicatoria Pseudomonas syringae pv phaseoli-cola Enterobacter cloacae andHumulus lupulus [91 112ndash119]However Scaligeria tripartite essential oils can be detectedwith Colletotrichum acutatum [120] Erythrina vogelii rootwas visualized with Cladosporium cucumerinum [121] Gladi-olus dalenii van Geel (Iridaceae) bulb extracts were detectedwith Aspergillus niger [122] The antibacterial compoundsfrom extract from Urginea sanguinea bulbs were visualizedwith Staphylococcus aureus [123] The rapid TLC autobio-graphic methods were also elaborated for the detection ofacetylcholinesterase and butyrylcholinesterase inhibitors invarious plants [124 125]


4 Conclusion

(i) In this review known indicators and dyes applied asnew visualizing reagents and various visualizing sys-tems as well as photocatalytic reactions andmicrobio-logical detection (bioautography detection) describedshould serve as a supplement to those used previouslyfor the detection of selected drugs

(ii) The visualizing effect depends on the chemical struc-ture of the visualizing reagent the structure of thesubstance detected and the chromatographic adsor-bent applied

(iii) Particular application will have those visualizingreagents which with substances present in analyzedmixtures will give diversified colors of chromato-graphic spots

(iv) The visualizing results of detected drugs depend onmethod of applicationmdashspraying or dipping

(v) Broadening index detection index characteristics ofdensitometric band modified contrast index limitof detection densitometric visualizing index andlinearity range of detected compounds can be usedfor the evaluation of visualizing effects of appliedvisualizing reagents

(vi) Presented data indicate the detection progress ofselected drugs investigated by thin-layer chromatog-raphy

Conflict of Interests

The authors declare that there is no conflict of interestsregarding the publication of this paper

Acknowledgment

This research was financed by the Medical University of Sile-sia as part of statutory research project KNW-1-013N30

References

[1] H Jork W Funk W Fischer and H Wimmer Dunnschicht-Chromatographie Reagenzien Und NachweismethodenPhysicalische Und Chemische Nachweismethoden GrUndlagenReagenzien vol 1 Wiley-VCH Weinheim Germany 1989

[2] H Jork W Funk W Fischer and H Wimmer Thin-LayerChromatography Reagents and DetectionMethods Physical andChemical Detection Methods Activation Reactions ReagentsSequences Reagents II vol 1Wiley-VCHWeinheim Germany1994

[3] J Sherma and B Fried Handbook of Thin-Layer Chromatogra-phy Marcel Dekker New York NY USA 3rd edition 2003

[4] A Pyka ldquoPhenolic drugs new visualizing reagents for detectionin TLCrdquo in Encyclopedia of Chromatography J Cazes Ed pp1ndash8 Marcel Dekker New York NY USA 2nd edition 2004

[5] A Pyka ldquoDrugs new visualizingreagents for detection in TLCrdquoin Encyclopedia of Chromatography J Cazes Ed pp 1267ndash1273Taylor amp Francis New York NY USA 2nd edition 2005

[6] A Pyka ldquoPhenolic drugs new visualizing reagents for detectionin TLCrdquo in Encyclopedia of Chromatography J Cazes Ed vol1 pp 1ndash9 Taylor amp Francis New York NY USA 2nd edition2007

[7] A Pyka ldquoPhenolic drugs TLC detectionrdquo in Encyclopedia ofChromatography J Cazes Ed pp 1777ndash1789 Taylor amp Francis3rd edition 2010

[8] EMerckDyeing Reagents forThin Layer and Paper Chromatog-raphy Merck Darmstadt Germany 1980

[9] A Pyka ldquoLipophilic vitaminsrdquo in Handbook of Thin-LayerChromatography J Sherma and B Fried Eds pp 671ndash696Marcel Dekker New York NY USA 3rd edition 2003

[10] A Pyka ldquoVitamins hydrophobic TLC analysisrdquo in Encyclope-dia of Chromatography J Cazes Ed pp 2415ndash2425 Taylor ampFrancis 3rd edition 2010

[11] VDMody K K PandyaMC Satia I AModi R IModi andT P Gandhi ldquoHigh performance thin-layer chromatographicmethod for the determination of sparfloxacin in human plasmaand its use in pharmacokinetic studiesrdquo Journal of Pharmaceu-tical and Biomedical Analysis vol 16 no 8 pp 1289ndash1294 1998

[12] C F Poole ldquoPlanar chromatography at the turn of the centuryrdquoJournal of Chromatography A vol 856 no 1-2 pp 399ndash4271999

[13] J Krzek and A Kwiecien ldquoApplication of densitometry fordetermination of beta-adrenergic-blocking agents in pharma-ceutical preparationsrdquo Journal of Planar Chromatography vol18 no 104 pp 308ndash313 2005

[14] S De and P Nariya ldquoA new rapid online-HPTLC methodfor evaluation of DPPH reduction with ascorbic acidrdquo JournalPlanar Chromatography vol 26 pp 21ndash25 2013

[15] J Sherma ldquoThin-layer chromatography in food and agriculturalanalysisrdquo Journal of Chromatography A vol 880 no 1-2 pp129ndash147 2000

[16] D Widiretani I Luailia and G Indrayanto ldquoSimultaneousdensitometric determination of hydrocortisone acetate and 2-phenoxyethanolrdquo Journal Planar Chromatography vol 26 pp37ndash42 2013

[17] B N Patel B N Suhagia C N Patel and H J Panchal ldquoAsimple and sensitive HPTLC method for quantitative analysisof darunavir ethanolate tabletsrdquo Journal of Planar Chromatog-raphy vol 24 no 3 pp 232ndash235 2011

[18] A Yadav R M Singh S C Mathur P K Saini and G NSingh ldquoA simple and sensitiveHPTLCmethod for simultaneousanalysis of domperidone and paracetamol in tablet dosageformsrdquo Journal of Planar Chromatography vol 22 no 6 pp421ndash424 2009

[19] B Spangenberg A Seigel J Kempf andWWeinmann ldquoForen-sic drug analysis by means of diode-array HPTLC using RF andUV library searchrdquo Journal of Planar Chromatography vol 18no 105 pp 336ndash343 2005

[20] VMirakor V Vaidya SMenon P Champanerker andA LaudldquoHPTLC method for determination of colchicine in a pharma-ceutical formulationrdquo Journal of PlanarChromatography vol 21no 3 pp 187ndash189 2008

[21] A Maslanka J Krzek and M Stolarczyk ldquoSimultaneousanalysis of hydrochlorothiazide triamterene furosemide andspironolactone by densitometric TLCrdquo Journal of Planar Chro-matography vol 22 no 6 pp 405ndash410 2009

[22] J Novakovic K Nesmerak H Nova and K Filka ldquoAnHPTLCmethod for the determination and the purity control ofciprofloxacin HCl in coated tabletsrdquo Journal of Pharmaceuticaland Biomedical Analysis vol 25 no 5-6 pp 957ndash964 2001


[23] M Khakpour A Jamshidi A A Entezami and H MirzadehldquoHPTLC procedure for determination of levonorgestrel in thedrug-release media of an in-situ-forming delivery systemrdquoJournal of Planar Chromatography vol 18 no 104 pp 326ndash3292005

[24] M Starek andM Rejdych ldquoDensitometric analysis of celecoxibetoricoxib and valdecoxib in pharmaceutical preparationsrdquoJournal of Planar Chromatography vol 22 no 6 pp 399ndash4032009

[25] W Parys and A Pyka ldquoUse of tlc and densitometry to evaluatethe chemical stability of nicotinic acid and its esters on silicagelrdquo Journal of Liquid Chromatography amp Related Technologiesvol 33 no 7-8 pp 1038ndash1046 2010

[26] A Pyka E Wiatr K Kwiska and D Gurak ldquoValidation thinlayer chromatography for the determination of naproxen intablets and comparison with a pharmacopeil methodrdquo Journalof Liquid Chromatography amp Related Technologies vol 34 no10-11 pp 829ndash847 2011

[27] P Bochenska and A Pyka ldquoDetermination of acetylsalicylicacid in pharmaceutical drugs by TLC with densitometricdetection in UVrdquo Journal of Liquid Chromatography amp RelatedTechnology vol 35 pp 1346ndash1363 2012

[28] A Pyka K Bober and A Stolarczyk ldquoDensitometric determi-nation of arbutin in cowberry leaves (Vaccinium vitis idaeae)rdquoActa Poloniae Pharmaceutica vol 64 no 5 pp 395ndash400 2007

[29] A Pyka ldquoTLC of vitamins including nicotinic acid derivativesrdquoinThin Layer Chromatography in Drug Analysis L Komsta andMWaksmundzka-Hajnos Eds CRCPress Taylor and Francis2013

[30] M Del Bubba L Checchini L Ciofi S Furlanetto and L LeprildquoDirect resolution and quantitative analysis of flurbiprofenenantiomers using microcrystalline cellulose triacetate platesapplications to the enantiomeric purity control and opticalisomer determination in widely consumed drugsrdquo BiomedicalChromatography 2013

[31] R M Youssef H M Maher E L El-Kimary E M Hassanand M H Barary ldquoValidated stability-indicating methods forthe simultaneous determination of amiloride hydrochlorideatenolol and chlorthalidone using HPTLC and HPLC withphotodiode array detectorrdquo Journal of AOAC International vol96 pp 313ndash323 2013

[32] G Mustafa A Ahuja S Baboota and J Ali ldquoBox-Behnkensupported validation of stability-indicating high performancethin-layer chromatography (HPTLC)method an application indegradation kinetic profiling of ropinirolerdquo Saudi Pharmaceu-tical Journal vol 21 pp 93ndash102 2013

[33] N Perisic-Janjic R Kaliszan NMilosevic G Uscumlic andNBanjac ldquoChromatographic retention parameters in correlationanalysis with in silico biological descriptors of a novel series ofN-phenyl-3-methyl succinimide derivativesrdquo Journal of Phar-maceutical and Biomedical Analysis vol 72 pp 65ndash73 2013

[34] Y Chen and W Schwack ldquoPlanar chromatography mediatedscreening of tetracycline and fluoroquinolone antibiotics inmilk by fluorescence and mass selective detectionrdquo Journal ofChromatography A vol 1312 pp 143ndash151 2013

[35] N W Ali M Gamal and M Abdelkawy ldquoChromatographicmethods for simultaneous determination of diiodohydrox-yquinoline and Metronidazole in their binary mixturerdquo Pak-istan Journal of Pharmaceutical Sciences vol 26 pp 865ndash8712013

[36] M Kaminska and I Choma ldquoThe influence of perchlorate ionconcentration on the retention of fluoroquinolones on cyano-silica platesrdquo Journal of Liquid Chromatography amp RelatedTechnologies vol 35 pp 1306ndash1313 2012

[37] D H Shewiyo E Kaale P G Risha et al ldquoAccuracy profilesassessing the validaty for routine use of high-performance thin-layer chromatographic assays for drug formulationsrdquo Journal ofChromatography A vol 1293 no 42 pp 159ndash169 2013

[38] A Pyka M Budzisz and M Dołowy ldquoValidation thin layerchromatography for the determination of acetaminophen intablets and comparison with a pharmacopeial methodrdquo BioMedResearch International vol 2013 Article ID 545703 10 pages2013

[39] J Sliwiok ldquoThe application of fuchsine dyes in the detection ofhigher fatty acids by thin-layer chromatographyrdquoMicrochemicalJournal vol 13 no 1 pp 108ndash110 1968

[40] Z Gregorowicz and J Sliwiok ldquoIndexes for estimation of devel-oping reagents in thin-layer chromatographyrdquo MicrochemicalJournal vol 15 no 1 pp 60ndash63 1970

[41] A Pyka ldquoAnalytical evaluation of visualizing reagents used todetect tocopherol and tocopherol acetate on thin layerrdquo Journalof Liquid Chromatography amp Related Technologies vol 32 no 2pp 312ndash330 2009

[42] A Pyka ldquoAnalytical evaluation of visualizing reagents used todetect ibuprofen on thin layersrdquo Journal of Liquid Chromatog-raphy amp Related Technologies vol 32 no 4 pp 578ndash588 2009

[43] A Pyka W Klimczok and D Gurak ldquoEvaluation of visualizingreagents for estradiol on thin layer by densitometric methodrdquoJournal of Liquid Chromatography amp Related Technologies vol31 no 4 pp 555ndash566 2008

[44] A Pyka ldquoThe application of densitometry to evaluate thevisualizing effects of salicylanilide using brilliant greenrdquo Journalof Liquid Chromatography amp Related Technologies vol 31 no 13pp 1943ndash1958 2008

[45] A Pyka and W Klimczok ldquoAnalytical and densitometric eval-uation of visualizing reagents of selected aliphatic compoundson thin layerrdquo Journal of Liquid Chromatography amp RelatedTechnologies vol 31 no 10 pp 1492ndash1510 2008

[46] O Huetos T Reuvers and J J Sanchez ldquoComparative studyof the thin-layer chromatographic detection of different corti-costeroidsrdquo Journal of Planar Chromatography vol 11 no 4 pp305ndash308 1998

[47] L Grząbą and A Pyka Densitometric evaluation of the stabilityof the chromatographic bands of selected substances in thin-layerchromatography [PhD thesis] Sosnowiec Poland 2011

[48] W Wardas and A Pyka ldquoNew visualizing agents for selectedpolybasic phenols and chlorophenols in thin-layer chromatog-raphyrdquo Journal of Planar Chromatography vol 6 pp 320ndash3221993

[49] W Wardas and A Pyka ldquoVisualizing agents for cholesterolderivatives in TLCrdquo Journal of Planar Chromatography vol 11no 1 pp 70ndash73 1998

[50] W Wardas and M Jędrzejczak ldquoNew visualizing agents forselected bile acid in TLCrdquo Chemia Analityczna vol 40 pp 73ndash79 1995

[51] P K ZarzyckiM A Bartoszuk andA I Radziwon ldquoOptimiza-tion of TLC detection by phosphomolybdic acid staining forrobust quantification of cholesterol and bile acidsrdquo Journal ofPlanar Chromatography vol 19 no 107 pp 52ndash57 2006

[52] A Pyka ldquoTLC of selected bile acids detection and separationrdquoJournal of Liquid Chromatography amp Related Technologies vol31 no 9 pp 1373ndash1385 2008


[53] I Sunshine E Rose and J Le Beau ldquoBarbiturate detection usingthin-layer chromatographyrdquo Clinical Chemistry vol 9 pp 312ndash316 1963

[54] R R Kulkarni V B Patil A G Bhoi and S Knandode ldquoAnew spray reagent for detection of phenytoin by thin-layerchromatographyrdquo Journal of Planar Chromatography vol 11 no4 pp 309ndash310 1998

[55] B D Mali D S Rathod and M V Garad ldquoThin-layer chro-matographic determination of diazepam phenobarbitone andsaccharin in toddy samplesrdquo Journal of Planar Chromatographyvol 18 no 104 pp 330ndash332 2005

[56] R Makharadze L Adeishvili T Chelidze N Imnadze andN Nizharadze ldquoSimultaneous determination of cyclodol anddiprazin by thin layer chromatography and high performanceliquid chromatographyrdquo Georgian Medical News vol 176 pp66ndash71 2009

[57] IQuintens J Eykens E Roets and JHoogmartens ldquoIdentifica-tion of cephalosporins by thin layer chromatography and colorreactionsrdquo Journal of PlanarChromatography vol 6 pp 181ndash1861993

[58] J Krzek J Piotrowska B Zuromska-Witek U Hubicka and EŁyson ldquoValidation of the method of identification and quan-titative determination of bacitracin in the form of bacitracinderivative with dabsyl chloride by thin-layer chromatographyand densitometryrdquo Journal Planar Chromatography vol 26 pp67ndash72 2013

[59] A Pyka D Gurak K Bober and A Niestroj ldquoApplication ofnew visualizing agents for selected essential oil components inTLCrdquo Chemia Analityczna vol 47 no 5 pp 691ndash699 2002

[60] B Kocjan ldquoDetection and identification of quinones in TLCrdquoJournal of Planar Chromatography vol 13 no 5 pp 396ndash3972000

[61] W Wardas I Lipska and K Bober ldquoTLC fractionation andvisualization of selected phenolic compounds applied as drugsrdquoActa Poloniae Pharmaceutica vol 57 no 1 pp 15ndash21 2000

[62] A Pyka D Gurak and K Bober ldquoNew visualizing reagents forselected phenolic drugs investigated by thin layer chromatogra-phyrdquo Journal of Liquid Chromatography amp Related Technologiesvol 25 no 10-11 pp 1483ndash1495 2002

[63] HD Smolarz EMedynska andGMatysik ldquoDetermination ofemodin and phenolic acids in the petioles of Rheumundulatumand Rheum rhaponticumrdquo Journal of Planar Chromatographyvol 18 no 104 pp 319ndash322 2005

[64] V Bilusic Vundac Z Males M Plazibat P Golja and B Cetina-Cizmek ldquoHPTLC determination of flavonoids and phenolicacids in some Croatian Stachys taxardquo Journal of Planar Chro-matography vol 18 no 104 pp 269ndash273 2005

[65] D N Olennikov and V V Partilkhaev ldquoIsolation and densit-ometric HPTLC analysis of rutin narcissin nicotiflorin andisoquercitrin in Caragana spinosa shootsrdquo Journal of PlanarChromatography vol 25 no 1 pp 30ndash35 2012

[66] HWagner and S BladtPlantDrugAnalysis LondonUK 2008[67] K Głowniak K Skalicka A Ludwiczuk and K Jop ldquoPhenolic

compounds in the flowers ofLavatera trimestrisL (Malvaceae)rdquoJournal of Planar Chromatography vol 18 no 104 pp 264ndash2682005

[68] P Bhujbal and P Sandhya ldquoChromatographic analysis ofbioactive markers in Vidangarista an ayurvedic polyherbalformulationrdquo Journal of Planar Chromatography vol 25 no 1pp 42ndash47 2012

[69] WWardas andA Pyka ldquoNewvisualizing agents for unsaturatedhigher fatty acids in TLCrdquo Journal of Planar Chromatographyvol 10 no 1 pp 63ndash67 1997

[70] A Niestroj M Stefaniak and J Sliwiok ldquoApplication of Rho-damine B in densitometric determination of long chain fattyacids in thin layer chromatographyrdquo inChemistry and Biochem-istry in the Agricultural Production Environmental ProtectionHuman and Animal Health H Gorecki Z Dobrzanski and PKafarski Eds pp 1ndash4 Czech-Pol-Trade Prague Brussels 2006

[71] A Pyka and K Bober ldquoVisualizing agents for short-chain fattyacids in TLCrdquo Journal of Planar Chromatography vol 18 no 2pp 141ndash146 2005

[72] W Wardas and A Pyka ldquoVisualizing agents for esters of higherfatty acidsrdquo Journal of Planar Chromatography vol 14 no 1 pp8ndash15 2001

[73] D Kowalczuk and H Hopkala ldquoSeparation of fluoroquinoloneantibiotics by TLC on silica gel cellulose and silanized layersrdquoJournal of Planar Chromatography vol 19 no 109 pp 216ndash2222006

[74] S Thangadurai S K Shukla and Y Anjaneyulu ldquoSeparationand detection of certain beta-lactam and fluoroquinoloneantibiotic drugs by thin layer chromatographyrdquo AnalyticalSciences vol 18 no 1 pp 97ndash100 2002

[75] M Juhel-Gaugain and J P Abjean ldquoScreening of quinoloneresidues in pig muscle by planar chromatographyrdquo Chro-matographia vol 47 no 1-2 pp 101ndash104 1998

[76] W Wardas A Pyka and M Jędrzejczak ldquoNew visualizingagents for quinoline isoquinoline and selected methylquino-lines in TLCrdquo Journal of Planar Chromatography vol 6 pp 238ndash241 1993

[77] K Bober ldquoDetermination of selected quinolones and fluoro-quinolones by use of TLCrdquo Analytical Letters vol 41 no 10 pp1909ndash1913 2008

[78] P Pushpalatha R K Sarin M A Rao and T R Baggi ldquoA newthin-layer chromatographic method for analysis of zolpidemand zopiclonerdquo Journal of Planar Chromatography vol 22 no6 pp 449ndash451 2009

[79] M Glensk MWłodarczyk M Radom andW Cisowski ldquoTLCas a rapid and convenient method for saponin investigationrdquoJournal of Planar Chromatography vol 18 no 2 pp 167ndash1702005

[80] M Bleichert H-S Eckhardt K-F Klein and B SpangenbergldquoA simple and reliablemethod for quantification of glucosaminein nutritional supplementsrdquo Journal of Planar Chromatographyvol 21 no 1 pp 55ndash59 2008

[81] V Esters L Angenot V Brandt et al ldquoValidation of a high-performance thin-layer chromatographydensitometry methodfor the quantitative determination of glucosamine in a herbaldietary supplementrdquo Journal of Chromatography A vol 1112 no1-2 pp 156ndash164 2006

[82] A M I Mohamed F A F Mohamed S A R Ahmed andY A S Mohamed ldquoNew and selective HPTLC-densitometricmethod for determination of piolitazone hydrochloriderdquo Jour-nal of Planar Chromatography vol 26 pp 209ndash214 2013

[83] G Nowak M Urbanska J Nawrot M K Bernard and RDawid-Pac ldquoColor and chemical reactions of selected sesquiter-pene lactones and ecdysones from Asteraceae on TLC platesrdquoJournal of Planar Chromatography vol 26 pp 289ndash293 2013

[84] M Nicoletti Ch Toniolo F R Gallo GMultari and G Palazz-ino ldquoTraceability in multi-ingredient botanicals by HPTLCfingerprint approachrdquo Journal of Planar Chromatography vol26 pp 243ndash248 2013


[85] I Podolak U Hubicka B Zuromska-Witek Z Janeczko andJ Krzek ldquoQuantification of saponin in different plant partsof Lysimachia L species by validated HPTLC-densitometricmethodrdquo Journal of Planar Chromatography vol 26 pp 248ndash253 2013

[86] D Yadav and M Gupta ldquoIsolation and HPTLC analysis ofiridoids in Premna integrifolia an important ingredient ofayurvedic drug Dashmoolrdquo Journal of Planar Chromatographyvol 26 pp 260ndash266 2013

[87] A Maurya and S K Srivastava ldquoA simple and reliable HPTLCmethod for the determination of four marker components inthe quality control of Alstonia scholarisrdquo Journal of PlanarChromatography vol 26 pp 254ndash259 2013

[88] C B Oppel D E Dussourd and U Garimella ldquoVisualizinga plant defense and insect counterploy alkaloid distributionin Lobelia leaves trenched by a plusiine caterpillarrdquo Journal ofChemical Ecology vol 35 no 6 pp 625ndash634 2009

[89] A Makowski E Adamek andW Baran ldquoUse of photocatalyticreactions to visualize drugs in TLCrdquo Journal of Planar Chro-matography vol 23 no 1 pp 84ndash86 2010

[90] L Botz S Nagy and B Kocsis ldquoDetection of microbiologicallyactive compoundsrdquo in Planar Chromatography A RetrospectiveView for the Third Millennium Sz Nyiredy Ed SpringerBudapest Hungary 2001

[91] I M Choma and E M Grzelak ldquoBioautography detection inthin-layer chromatographyrdquo Journal of Chromatography A vol1218 no 19 pp 2684ndash2691 2011

[92] Z Cheng and T Wu ldquoTLC bioautography high throughputtechnique for screening of bioactive of bioactive natural prod-uctsrdquo Combinatorial Chemistry amp High Throughput Screeningvol 16 pp 531ndash549 2013

[93] A Fittler B Kocsis Z Matus and L Botz ldquoA sensitive methodfor thin-layer chromatographic detection of amphotericin BrdquoJournal of Planar Chromatography vol 23 no 1 pp 18ndash22 2010

[94] G Schmourlo R R Mendonca-Filho C S Alviano andS S Costa ldquoScreening of antifungal agents using ethanolprecipitation and bioautography of medicinal and food plantsrdquoJournal of Ethnopharmacology vol 96 no 3 pp 563ndash568 2005

[95] V Molnar F Billes E Tyihak and P G Ott ldquoBioautographicdetection of antimicrobial compounds in the edible Shiitakemushroom (Lentinus edodes)rdquo Journal of Planar Chromatogra-phy vol 21 no 6 pp 423ndash426 2008

[96] D E Wedge and D G Nagle ldquoA new 2D-TLC bioautographymethod for the discovery of novel antifungal agents to controlplant pathogensrdquo Journal of Natural Products vol 63 no 8 pp1050ndash1054 2000

[97] K Ueta M Nishioka Y Yabuta and F Watanabe ldquoTLC-bioautography analysis of vitamin B12 compound from theshort-necked clam (Ruditapes Philipponarum) extract usedas a flavoringrdquo Journal of Liquid Chromatography amp RelatedTechnologies vol 33 no 7-8 pp 972ndash979 2010

[98] A M Moricz E Horvath P G Ott and E Tyihak ldquoRamanspectroscopic evaluation of the influence of Pseudomonas bac-teria on aflatoxin B1 in the BioArena complex bioautographicsystemrdquo Journal of Raman Spectroscopy vol 39 no 10 pp 1332ndash1337 2008

[99] R M Baosic D M Milojkovia-Opsenica and Z L Tesic ldquoTheeffect of the substituents of 120573-ketoiminato ligand of copper(II)and nickel(II) complexes on their retention on thin layers ofpolyacrylonitrilerdquo Journal of Planar Chromatography vol 16no 6 pp 412ndash416 2003

[100] A Moricz K H-Otta P Ott and E Tyihak ldquoSeparation anddetection of aflatoxins using overpressured-layer chromatogra-phy and bioautographyrdquo Journal of Planar Chromatography vol16 no 6 pp 417ndash420 2003

[101] E Tyihak P Ott A MMoricz G Katay and Z Kiraly-VeghelyldquoAntibiosis antibiotics and the formaldehyde cycle the uniqueimportance of planar chromatographic techniques to progressin these fieldsrdquo Journal Planar Chromatography vol 17 pp 84ndash88 2004

[102] E Tyihak L Botz P Ott et al ldquoThe combination of theoverpressured layer chromatography and bioautography andits applications to the analysis of molecules influencing cellproliferationrdquo Chemia Analityczna vol 48 no 3 pp 543ndash5532003

[103] E Mincsovics G Katay P G Ott Z Kiraly-Veghely A MMoricz and E Tyihak ldquoIsolation of some antimicrobial com-pounds of red wine by OPLC flowing eluent wall techniquerdquoChromatographia vol 62 no 13 pp S51ndashS56 2005

[104] Z Kiraly-Veghely A M Moricz P G Ott G Katay I Belaiand E Tyihak ldquoComparison of components from red andwhite wines for antimicrobial activity by biodetection afterOPLC separationrdquo Journal of Liquid Chromatography amp RelatedTechnologies vol 32 pp 1259ndash1272 2009

[105] A Sarkozi A M Moricz P G Ott E Tyihak and A KeryldquoInvestigation of Chelidonium alkaloids by use of a complexbioautographic systemrdquo Journal PlanarChromatography vol 19pp 267ndash272 2006

[106] I Choma A Choma and K Staszczuk ldquoDirect bioautography-thin-layer chromatography of flumequine and doxycycline inmilkrdquo Journal of Planar Chromatography vol 15 no 3 pp 187ndash191 2002

[107] I Choma ldquoThin-layer chromatography-direct bioautography offlumequine residues inmilkrdquo Journal of Liquid Chromatographyamp Related Technologies vol 29 no 14 pp 2083ndash2093 2006

[108] I M Choma C Kowalski R Lodkowski A Burmanczukand I Komaniecka ldquoTLC-DB as an alternative to the HPLCmethod in the determination of cefacetril residues in cowrsquosmilkrdquo Journal of LiquidChromatographyampRelated Technologiesvol 31 no 13 pp 1903ndash1912 2008

[109] I M Choma ldquoScreening of enrofloxacin and ciprofloxacinresidues in milk by HPLC and by TLC with direct bioautog-raphyrdquo Journal of Planar Chromatography vol 19 no 108 pp104ndash108 2006

[110] IMChoma and I Komaniecka ldquoMatrix solid-phase dispersioncombined with thin-layer chromatographymdashdirect bioauto-graphy for determination of enrofloxacin and ciprofloxacinresidues in milkrdquo Journal of Liquid Chromatography amp RelatedTechnologies vol 28 no 16 pp 2467ndash2478 2005

[111] L Rocha A Marston O Potterat M A Kaplan H Stoeckli-Evans and K Hostettmann ldquoAntibacterial phloroglucinols andflavonoids fromHypericum brasilienserdquo Phytochemistry vol 40no 5 pp 1447ndash1452 1995

[112] D Tasdemir A A Donmez I Calis and P Ruedi ldquoEvaluationof biological activity of Turkish plants Rapid screening for theantimicrobial antioxidant and acetylcholinesterase inhibitorypotential by TLC bioautographic methodsrdquo PharmaceuticalBiology vol 42 no 4-5 pp 374ndash383 2004

[113] K Polatoglu F Demirci B Demirci N Goren and K H CBaser ldquoAntibacterial activity and the variation of Tanacetumparthenium (L) schultz bip essential oils from Turkeyrdquo Journalof Oleo Science vol 59 no 4 pp 177ndash184 2010


[114] K Polatoglu F Demirci B Demirci N Goren and K HCan Baser ldquoAntimicrobial activity and essential oil compositionof a new T argyrophyllum (C koch) Tvzel var argyrophyllumchemotyperdquo Journal of Oleo Science vol 59 no 6 pp 307ndash3132010

[115] K Polatoglu F Demirci B Demirci N Goren and K HC Baser ldquoEssential oil composition and antibacterial activityof Tanacetum argenteum (Lam) willd ssp argenteum and Tdensum (Lab) Schultz Bip ssp AmaniHeywood from TurkeyrdquoJournal of Oleo Science vol 59 no 7 pp 361ndash367 2010

[116] G Horvath N Jambor A Vegh et al ldquoAntimicrobial activity ofessential oils the possibilities of TLC-bioautographyrdquo Flavourand Fragrance Journal vol 25 no 3 pp 178ndash182 2010

[117] A D A Morandim A R Pin N A S Pietro et al ldquoComposi-tion and screening of antifungal activity against Cladosporiumsphaerospermum and Cladosporium cladosporioides of essentialoils of leaves and fruits of Piper speciesrdquo African Journal ofBiotechnology vol 9 no 37 pp 6135ndash6139 2010

[118] A R Shahverdi F Abdolpour H R Monsef-Esfahani andH Farsam ldquoA TLC bioautographic assay for the detectionof nitrofurantoin resistance reversal compoundrdquo Journal ofChromatography B vol 850 no 1-2 pp 528ndash530 2007

[119] G Horvath L G Szabo E Lemberkovics L Botz and BKocsis ldquoCharacterization and TLCmdashbioautographic detectionof essential oils from sone Thymus taxa Determination ofthe activity of the oils and their components against plantpathogenic bacteriardquo Journal of Planar Chromatography vol 17no 4 pp 300ndash304 2004

[120] N Tabanca B Demirci K H C Baser et al ldquoCharacterizationof volatile constituents of Scaligeria tripartita and studies onthe antifungal activity against phytopathogenic fungirdquo Journalof Chromatography B vol 850 no 1-2 pp 221ndash229 2007

[121] E F Queiroz J-L Wolfender K K Atindehou D Traoreand K Hostettmann ldquoOn-line identification of the antifungalconstituents of Erythrina vogelii by liquid chromatography withtandem mass spectrometry ultraviolet absorbance detectionand nuclear magnetic resonance spectrometry combined withliquid chromatographic micro-fractionationrdquo Journal of Chro-matography A vol 974 no 1-2 pp 123ndash134 2002

[122] J A Odhiambo G M Siboe C W Lukhoba and S F DossajildquoAntifungal activity of crude extracts of Gladiolus dalenii vangeel (Iridaceae)rdquoAfrican Journal of Traditional Complementaryand Alternative Medicines vol 7 no 1 pp 53ndash58 2010

[123] V Naidoo D R Katerere G E Swan and J N Eloff ldquoPre-treatment of Urginea sanguinea bulbs used in ethnoveterinarymedicine influences chemical composition and biological activ-ityrdquo Pharmaceutical Biology vol 42 no 7 pp 529ndash533 2004

[124] Z-D Yang Z-W Song J Ren M-J Yang and S Li ldquoImprovedthin-layer chromatography bioautographic assay for the detec-tion of actylcholinesterase inhibitors in plantsrdquo PhytochemicalAnalysis vol 22 no 6 pp 509ndash515 2011

[125] A Marston J Kissling and K Hostettmann ldquoA rapid TLCbioautographicmethod for the detection of acetylcholinesteraseand butyrylcholinesterase inhibitors in plantsrdquo PhytochemicalAnalysis vol 13 no 1 pp 51ndash54 2002

Submit your manuscripts athttpwwwhindawicom

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Hindawi Publishing Corporationhttpwwwhindawicom

Volume 2014

ToxinsJournal of

VaccinesJournal of

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AntibioticsInternational Journal of

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StrokeResearch and TreatmentHindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Drug DeliveryJournal of



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Emergency Medicine InternationalHindawi Publishing Corporationhttpwwwhindawicom Volume 2014


Autoimmune Diseases


AnesthesiologyResearch and Practice

ScientificaHindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Journal of


Pharmaceutics


MEDIATORSINFLAMMATION

of


these types of compounds for which GC or HPLC is not suit-able The above-mentioned arguments explain enough whythe TLC-densitometry is proposed in many drug analyses asan alternative method to the pharmacopoeias methods

Important step of thin-layer analysis of compounds is thedetection of investigated substances including drugsThe sep-arated substances by thin-layer chromatography (TLC) canbe detected by the following methods physical (individualcolour of substance or fluorescence of substance in UV light)chemical (coloured reactions of separated substances withvisualizing reagents) physicochemical (eg the applicationof isotopes as visualizing reagent) biological (the applicationof biodetectors) [1ndash10] Many drugs are active in the rangeof UV light and can be directly detected and determinedon chromatographic plate for example by densitometricanalysis in short or long wavelength UV light [11ndash39] Thevisualizing reagents have the special significance to detectseparated compounds on thin layers In view of the detectionmechanism of the compound the visualizing reagents canbe sorted as follows conservative reagents which do notdestroy separated substances destructive reagents whichdestroy or change the structure of separated substances Cur-rently the most important field of application of thin-layerchromatography is pharmacy The number of publications inthe field of pharmacy steadily increases It results from thefact that contemporary thin-layer chromatography is a fullyinstrumentalized and automated technique [1ndash3 29]

The applications of many reagents for the detection oforganic compounds including drugs after their separationon thin-layer were presented in books [1 2] on the basisof manuscripts published up to 1992 The objectives of thisreview are the presentation and discussion of the significanceof known indicators and dyes as new visualizing reagentsand various visualizing systems as well as photocatalytic reac-tions and bioautography method for detection of bioactivecompounds including drugs and compounds isolated fromherbal extracts in TLC investigations that were described inthe selected manuscripts from 1993

2 Parameters Describingthe Visualizing Effects

The estimation of visualizing effects of the detected com-pounds must be objective Therefore Gregorowicz and Sli-wiok [39 40] introduced the indexes for estimation ofrhodamine B new fuchsine basic fuchsine and crystal violetfor detection of cholesterol and higher fatty acids To comparethe developing effects of the above-mentioned dyes it wassuggested [39 40] to estimate the chromatographic spotsobtained by means of indexes given bellow

(i) detection index which indicates the ratio of the mini-mal number of micrograms of the substance detectedto the area of the chromatographic spot in squaremillimeters obtained by the planimetric method

119868det =1198981

1199011

[120583g

mm2] (1)

where 1198981is the smallest quantity of substances

detected [120583g] with the visualizing reagent (limit ofdetection) and 119901

1is the spot area of the substance

[mm2] at the limit of detection of the substance(ii) developing index which gives the numerical value of

the part of the chromatographic spot area relating to1 120583g

(iii) contrast index which represents two independentvalues namely the angle between tangents drawn tothe arms of the densitometric band formulated inthe degrees and the densitometric band height incentimeters

(iv) durability of the spot in order to formulate the dura-bility of chromatographic spot colors The followingrelative scale was proposed spot of minor durabilityvisible up to 2 hours spot of mean durability visiblefrom 2 to 6 hours and durable spot durability visiblefor over 6 hours

Today the area of chromatographic spot is determinedby densitometric analysis Therefore Pyka et al modifiedthe above-mentioned indexes and also proposed new indexeswhich characterized the visualizing effects [41ndash45] Thelimit of detection (detectability) detection index broadeningindex modified contrast index densitometric visualizingindex and linearity range were determined and tested forestradiol tocopherol tocopherol acetate ibuprofen stearicacid stearyl alcohol and salicylanilide detected by the useof visualizing reagents The broadening index [41 42] wasdefined as

119868broad =119886

1199012

times 1000 [120583gAU] (2)

where 119886 is the mass [120583g] of the detected substance and 1199012is

the spot area [AU] of 119886 120583g of this analyzed substanceThe detection index was defined as

119868det =1198981

1199011

[120583gAU] (3)

where 1198981is the smallest quantity of substance detected [120583g]

(limit of detection) and 1199011is the spot area of substance [AU]

at the limit of detection of substanceThe modified contrast index [43] was defined as

119868contr(mod if) =ℎ

120572[AU∘] (4)

where ℎ is the height of the densitometric band [AU] of119886 120583g of analyzed substance and 120572 is the angle [∘] betweenthe tangents at the inflection points to the curves of thedensitometric band of substance

The densitometric visualizing index (DVI) [44] wasproposed and defined as

DVI =1199012

1198981times 120572times 10minus4[AU120583g sdot∘] (5)

where 1198981is the limit of detection of the analyzed substance

[120583g] 1199012is the spot area [AU] of 119886 120583g of analyzed substance





150F254





minum oxide 150F254













Table1Fu

ndam

entalabsorptionband

ofdrugscoloro

fspo

tofd


ndcoloro

fchrom

atogramlim

itof

detection

andlin

earityrange

Detecteddrug

Stationary

phase

Detectio

nway

Fund

amental

absorptio

nband

ofdrug120582max(nm)

Color

ofspot

ofdetected

drug

Backgrou

ndcoloro

fchromatogram

Limitof

detection

(120583g)

Linearity

range

(120583gspo

tminus1 )

(rcorrelatio

ncoeffi

cient)

Reference

Estradiol

Neutralalum

inum

oxide6

0F254

toluene+

ethylacetate(11vv)

With

outu

sing

visualizingreagent

205

Lack

ofcoloredspot

invisib

lelight

White

032

110divide2500

(119903=09951)

[43]

Estradiol

Bartonrsquosreagent

233

Darkbluegreen

Willow

-green

032

053divide1024

(119903=09975)

[43]

Estradiol

Sulphu

ricacid

278

Lighto

range

Lightb

eige

010

088divide1280

(119903=09974)

[43]

Estradiol

Gentia

nviolet

226

Violetblue

Violet

032

053divide2500

(119903=09943)

[43]

Estradiol

Methylene

violet

227

Greyblue

Lightg

rey

032

088divide336

(119903=09975)

[43]

Estradiol

Neutralalum

inum

oxide150F 2

54toluene+

ethylacetate(11vv)

With

outu

sing

visualizingreagent

207

Lack

ofcoloredspot

invisib

lelight

White

032

137divide2000

(119903=09972)

[43]

Estradiol

Gentia

nviolet

598

Darkviolet

Violet

032

088divide2000

(119903=09879)

[43]

Estradiol

Methylene

violet

204

Greyblue

Lightg

rey

053

137divide2500

(119903=09983)

[43]

Salicylanilide

Silicag

el60F 2

54(10

5744

)chloroform

With

outu

sing

visualizingreagent

307

Lack

ofcoloredspot

invisib

lelight

White

007

500divide3000

(119903=09802)

[44]

Salicylanilide

Brilliant

green

597

Darkgreen

Green

070

300divide3000

(119903=09929)

[44]


herol

Silicag

el60

toluene

With

outu

sing

visualizingreagent

270

Lack

ofcoloredspot

invisib

lelight

White

045

346divide2000

(119903=09894)

[41]


herol


n(III)chloride

524

Raspberryred

Lightb

eige

208

576divide3000

(119903=09927)

[41]


herol

Methylen

eviolet

270

Azure

with

white

border

Lightg

rey-blue

030

075divide576

(119903=09949)

[41]


herol

Methylgreen

270

Lightg

reen

Green

030

075divide576

(119903=09863)

[41]

(+)-120572-Tocop

herol

acetate

Silicag

el60

toluene

With

outu

sing

visualizingreagent

203

Lack

ofcoloredspot

invisib

lelight

White

030

075divide2000

(119903=09978)

[41]

(+)-120572-Tocop

herol

acetate

Rhod

amineB

203

Darkpink

Pink

045

125divide1600

(119903=09936)

[41]

(+)-120572-Tocop

herol

acetate

Gentia

nviolet

203

Darkviolet

Violet

075

208divide2500

(119903=09908)

[41]

(+)-120572-Tocop

herol

acetate

Methylgreen

203

Green

(smallcon

trast

with

backgrou

ndcolor)

Green

075

125divide2500

(119903=09943)

[41]

(+)-120572-Tocop

herol

acetate

Janu

sblue

203

Darkblue

Blue

075

125divide2500

(119903=09938)

[41]


Table1Con

tinued

Detecteddrug

Stationary

phase

Detectio

nway

Fund

amental

absorptio

nband

ofdrug120582max(nm)

Color

ofspot

ofdetected

drug

Backgrou

ndcoloro

fchromatogram

Limitof

detection

(120583g)

Linearity

range

(120583gspo

tminus1 )

(rcorrelatio

ncoeffi

cient)

Reference

Ibup

rofen

Silicag

el60F 2

54chloroform

+methano

l(50

14

vv)

With

outu

sing

visualizingreagent

200

Lack

ofcoloredspot

invisib

lelight

White

062

125divide1000

(119903=09958)

[42]

Ibup

rofen

Methylene

violet

200

White-bluew

ithdark

blue

border

Light

125

500divide2500

(119903=099549)

[42]

Stearic

acid

Silicag

el60

n-hexane

+aceton

e(4

1vv)

Rhod

amineB

585

Darkpink

Pink

125

156divide2840

(119903=09913)

[45]

Stearic

acid

Methylene

blue

687

Blue

with

light

border

Blue

007

041divide744

(119903=09975)

[45]

Stearic

acid

Janu

sblue

617

Navyblue

with

white

border

Blue

012

156divide744

(119903=09928)

[45]

Stearylalcoh

olSilicag

el60

n-hexane

+ethylacetate+

methano

l(46

3505vvv)

Rhod

amineB

586

Darkpink

Pink

148

290divide2700

(119903=09964)

[45]

Stearylalcoh

olSilicag

el60

n-hexane

+ethylacetate+

methano

l(46

3505vvv)

Malachitegreen

644

Green

with

light

border

Green

032

089divide885

(119903=09959)

[45]

Stearylalcoh

olSilicag

el60

n-hexane

+ethylacetate+

methano

l(46

3505vvv)

Janu

sblue

655

Navyblue

Blue

032

053divide708

(119903=09995)

[45]


Table2Ch

aracteris

ticso

fdensitom

etric

band

ofdetected

drugsmod

ified


metric

visualizingindexmod

ified

broadening


Detecteddrug

Stationary

phasemobile

phase

Detectio

nway

Densitom

etric

band

characteris

ticof

detected

drug

Mod

ified

contrast

index(AU∘ )

Densitom

etric

visualizing


Mod

ified

broadening

index(120583gAU

)

Detectio

nindex

(120583gAU

)Re

ference

Area

(AU)

Height

(AU)120572(∘ )

Estradiol

Neutralalum

inum

oxide

60F 2

54toluene+

ethylacetate

(11vv)

With

outu

sing

visualizingreagent

1304

8246

145

16966

mdash19

16032495

[43]

Estradiol

Bartonrsquosreagent

23786

255

1319615

mdash10

5103210

70[43]

Estradiol

Sulphu

ricacid

44709

497

1338231

mdash0559

010850

[43]

Estradiol

Gentia

nviolet

11348

220

1514667

mdash2203

032760

[43]

Estradiol

Methylene

violet

13622

221

1713000

mdash18

35032705

[43]

Estradiol

Neutralalum

inum

oxide

150F

254toluene+

ethylacetate

(11vv)

With

outu

sing

visualizingreagent

11424

192

131476

9mdash

2188

032409

[43]

Estradiol

Gentia

nviolet

10906

127

2077

00mdash

2292

032796

[43]

Estradiol

Methylene

violet

8466

154

334667

mdash2953

053490

[43]

Salicylanilide

Silicag

el60F 2

54(10

5744

)chloroform

With

outu

sing

visualizingreagent

72889

707

9785

11570

0686

00715

90[44]

Salicylanilide

Brilliant

green

91842

704

8880

1640

0544

07011

112

[44]


herol

Silicag

el60

toluene

With

outu

sing

visualizingreagent

34519

459

162869

0479

0724

0453100

[41]


herol


iron(III)

chlorid

e60

474

698

97756

0323

0413

2088915

[41]


herol

Methylene

violet

40426

636

87950

1684

0618

0303456

[41]


herol

Methylgreen

37196

576

964

00

1378

0672

03015

64[41]

(+)-120572-Tocop

herol

acetate

Silicag

el60

toluene

With

outu

sing

visualizingreagent

33796

591

115373

1024

0740

03011

00[41]

(+)-120572-Tocop

herol

acetate

Rhod

amineB

26031

455

153033

0386

0960

04515

11[41]

(+)-120572-Tocop

herol

acetate

Gentia

nviolet

2944

1545

105450

0392

0849

07518

84[41]

(+)-120572-Tocop

herol

acetate

Methylgreen

30226

564

115127

0366

0827

0753911

[41]

(+)-120572-Tocop

herol

acetate

Janu

sblue

30125

565

115136

0365

0830

0753750

[41]

Ibup

rofen

Silicag

el60F 2

54With

outu

sing

visualizingreagent

5564

444

526

17115

0342

044

90623325

[42]

Ibup

rofen

chloroform

+methano

l(50

14vv)

Methylene

violet

53696

348

3599

430123

046

612

544

38[42]

Stearic

acid

Silicag

el60

n-hexane

+aceton

e(4

1vv)

Rhod

amineB

6567

8231

264

50017

432

125900

[45]

Stearic

acid

Methylene

blue

30918

368

1426286

3155

092

007245

[45]

Stearic

acid

Janu

sblue

36175

462

1142000

2741

079

012604

[45]

Stearylalcoh

olSilicag

el60

n-hexane

+ethylacetate+

methano

l(46

3505vvv)

Rhod

amineB

3605

100

1376

920019

749

148234

[45]

Stearylalcoh

olMalachitegreen

12378

333

747571

0553

218

032382

[45]

Stearylalcoh

olJanu

sblue

10963

269

1320692

0264

246

032214

[45]






(120583g)Color of




















Spraying 3807 1158 12 200 1770 596 225 584


Spraying 5148 1469 95 200 4354 1124 19 606


Spraying 7561 2139 85 200 4056 1263 10 200


Spraying 5966 1674 9 200 4560 1035 16 200


Spraying 5338 1555 8 643 3444 1114 10 649


Spraying 2474 691 24 667 1772 501 30 662




4OH




4OH (5 10 3 1




254 The






119865values) can









4 and


4 Furthermore





4 01









2CO3gives


2CO3+ 1



119865= 0138) and











3)2+ saturated


2Cr2O7in 40 H





3)2

Na2CO3 Co(NO







254












Table6Ch

aracteris

ticso

fthe


selected

phenoliccompo

unds

Detecteddrug

Stationary

phase

Mob

ileph

ase

Visualizingreagent

Detectio

nlim

it(120583g)

Detectability

index

(120583gmm

2 )Color

ofspot

Backgrou

ndcoloro

fchromatogram

Reference

Adrenalin

eSilicag

el60F 2

54Glacialaceticacid

+n-bu

tano

l+water

(14

1vvv)

Brilliant

cresylblue

in5

NaO

H010

01010

Brow

nLightviolet

[61]

Silicag

el60kieselguh

rF254

Brom

ocresolgreen

in5

NaO

H025

02547

Lightb

rown

Seagreen

Polyam

ide11F

254

Eosin

yello

win

5NaO

H250

25030

Lightb

rown

Lightyellow

Dop

amine

Silicag

el60F 2

54Glacialaceticacid

+n-bu

tano

l+water

(14

1vvv)

Brom

ocresolgreen

in5

NaO

H010

01015

Orange

Lightb

lue

[61]

Silicag

el60kieselguh

rF254

025

02510

Brow

norange

Seagreen

Polyam

ide11F

254

Eosin

yello

win

5NaO

H075

07517

Lightb

rown

Lightyellow

Metaram

inol

Silicag

el60F 2

54Glacialaceticacid

+n-bu

tano

l+water

(14

1vvv)

Brom

ocresolgreen

in5

NaO

H050

05015

Lemon

Lightb

lue

[61]

Silicag

el60kieselguh

rF254

Eriochromeb

lack

Tin

5NaO

H050

05035

Brow

nLightb

rown

Polyam

ide11F

254

Anilin

eblueinwater

505042

Lightyellow

Blue

Feno

terol

Silicag

el60F 2

54Glacialaceticacid

+n-bu

tano

l+water

(14

1vvv)

Brom

ocresolgreen

in5

NaO

H010

01013

Brow

norange

Lightb

lue

[61]

Silicag

el60kieselguh

rF254

Eriochromeb

lack

Tin

5NaO

H050

05038

Brow

nLightb

rown

Polyam

ide11F

254

Eosin

yello

win

5NaO

H50

515

Yello

wLightyellow

Phenylephrine

Silicag

el60F 2

54Glacialaceticacid

+n-bu

tano

l+water

(14

1vvv)

Brom

ocresolgreen

in5

NaO

H025

02521

Lemon

Lightb

lue

[61]

Silicag

el60kieselguh

rF254

Eriochromeb

lack

Tin

5NaO

H050

05041

Brow

nLightb

rown

Bithiono

lSilicag

el60F 2

54Glacialaceticacid

+n-bu

tano

l+water

(14

1vvv)

Brilliant

cresylblue

in5

NaO

H050

05077

Brow

nLightviolet

[61]

Silicag

el60kieselguh

rF254

Brilliant

cresylblue

in5

NaO

H250

25068

Brow

nLightb

rown

Methyldop

a

Silicag

el60

Glacialaceticacid

+n-bu

tano

l+water

(14

1vvv)

Thym

olblue

in2

NaO

H32

327

Brow

nblack

Beige

[62]

Norepinephrine

Brom

othymolblue

in2

NaO

H06

0628

Yello

wbrown

Lightb

lue

Terbutaline

Brilliant

cresylblue

in2

NaO

H20

2017

Lightb

rown

Lightb

lue

Bamethane

Anilin

ebluein2

NaO

H10

1025

Blue

Lightb

lue

Etam

ivan

Anilin

ebluein2

NaO

H50

5018

Whiteblue

Lightb

lue

Phloroglucinol

Silicag

el60

Chloroform

+methano

l(9

1vv)

Brilliant

cresylblue

in2

NaO

H20

2023

Lightb

rown

Lightb

lue

[62]

Pentazocine

Anilin

ebluein2

NaO

H05

0568

White

Lightb

lue

Hexachlorop

hene

Thym

olblue

in2

NaO

H50

5037

Yellowbeige

Beige

Pyrocatechine

Brilliant

greenin

2NaO

H03

0317

Light

orangered

Lightg

reen

Niclosamide

Brilliant

greenin

2NaO

H08

0818

Green

Lightg

reen

Salicylanilide

Brilliant

greenin

2NaO

H08

0816

Green

Lightg

reen

Thym

olBrilliant

greenin

2NaO

H20

2042

Yello

wgreen

Lightg

reen

Eugeno

lSilicag

el60

Benzene

Brom

othymolblue

in2

NaO

H50

5037

Lightyellow

Lightb

lue

[62]







254after





254 it was worse
























254plates









1 aflatoxin B

2 aflatoxin G

1 aflatoxin G

2[98ndash



4 Conclusion









Acknowledgment


References






































































































































Volume 2014

ToxinsJournal of

VaccinesJournal of




















Autoimmune Diseases




Journal of


Pharmaceutics



of





150F254





minum oxide 150F254













Table1Fu

ndam

entalabsorptionband

ofdrugscoloro

fspo

tofd


ndcoloro

fchrom

atogramlim

itof

detection

andlin

earityrange

Detecteddrug

Stationary

phase

Detectio

nway

Fund

amental

absorptio

nband

ofdrug120582max(nm)

Color

ofspot

ofdetected

drug

Backgrou

ndcoloro

fchromatogram

Limitof

detection

(120583g)

Linearity

range

(120583gspo

tminus1 )

(rcorrelatio

ncoeffi

cient)

Reference

Estradiol

Neutralalum

inum

oxide6

0F254

toluene+

ethylacetate(11vv)

With

outu

sing

visualizingreagent

205

Lack

ofcoloredspot

invisib

lelight

White

032

110divide2500

(119903=09951)

[43]

Estradiol

Bartonrsquosreagent

233

Darkbluegreen

Willow

-green

032

053divide1024

(119903=09975)

[43]

Estradiol

Sulphu

ricacid

278

Lighto

range

Lightb

eige

010

088divide1280

(119903=09974)

[43]

Estradiol

Gentia

nviolet

226

Violetblue

Violet

032

053divide2500

(119903=09943)

[43]

Estradiol

Methylene

violet

227

Greyblue

Lightg

rey

032

088divide336

(119903=09975)

[43]

Estradiol

Neutralalum

inum

oxide150F 2

54toluene+

ethylacetate(11vv)

With

outu

sing

visualizingreagent

207

Lack

ofcoloredspot

invisib

lelight

White

032

137divide2000

(119903=09972)

[43]

Estradiol

Gentia

nviolet

598

Darkviolet

Violet

032

088divide2000

(119903=09879)

[43]

Estradiol

Methylene

violet

204

Greyblue

Lightg

rey

053

137divide2500

(119903=09983)

[43]

Salicylanilide

Silicag

el60F 2

54(10

5744

)chloroform

With

outu

sing

visualizingreagent

307

Lack

ofcoloredspot

invisib

lelight

White

007

500divide3000

(119903=09802)

[44]

Salicylanilide

Brilliant

green

597

Darkgreen

Green

070

300divide3000

(119903=09929)

[44]


herol

Silicag

el60

toluene

With

outu

sing

visualizingreagent

270

Lack

ofcoloredspot

invisib

lelight

White

045

346divide2000

(119903=09894)

[41]


herol


n(III)chloride

524

Raspberryred

Lightb

eige

208

576divide3000

(119903=09927)

[41]


herol

Methylen

eviolet

270

Azure

with

white

border

Lightg

rey-blue

030

075divide576

(119903=09949)

[41]


herol

Methylgreen

270

Lightg

reen

Green

030

075divide576

(119903=09863)

[41]

(+)-120572-Tocop

herol

acetate

Silicag

el60

toluene

With

outu

sing

visualizingreagent

203

Lack

ofcoloredspot

invisib

lelight

White

030

075divide2000

(119903=09978)

[41]

(+)-120572-Tocop

herol

acetate

Rhod

amineB

203

Darkpink

Pink

045

125divide1600

(119903=09936)

[41]

(+)-120572-Tocop

herol

acetate

Gentia

nviolet

203

Darkviolet

Violet

075

208divide2500

(119903=09908)

[41]

(+)-120572-Tocop

herol

acetate

Methylgreen

203

Green

(smallcon

trast

with

backgrou

ndcolor)

Green

075

125divide2500

(119903=09943)

[41]

(+)-120572-Tocop

herol

acetate

Janu

sblue

203

Darkblue

Blue

075

125divide2500

(119903=09938)

[41]


Table1Con

tinued

Detecteddrug

Stationary

phase

Detectio

nway

Fund

amental

absorptio

nband

ofdrug120582max(nm)

Color

ofspot

ofdetected

drug

Backgrou

ndcoloro

fchromatogram

Limitof

detection

(120583g)

Linearity

range

(120583gspo

tminus1 )

(rcorrelatio

ncoeffi

cient)

Reference

Ibup

rofen

Silicag

el60F 2

54chloroform

+methano

l(50

14

vv)

With

outu

sing

visualizingreagent

200

Lack

ofcoloredspot

invisib

lelight

White

062

125divide1000

(119903=09958)

[42]

Ibup

rofen

Methylene

violet

200

White-bluew

ithdark

blue

border

Light

125

500divide2500

(119903=099549)

[42]

Stearic

acid

Silicag

el60

n-hexane

+aceton

e(4

1vv)

Rhod

amineB

585

Darkpink

Pink

125

156divide2840

(119903=09913)

[45]

Stearic

acid

Methylene

blue

687

Blue

with

light

border

Blue

007

041divide744

(119903=09975)

[45]

Stearic

acid

Janu

sblue

617

Navyblue

with

white

border

Blue

012

156divide744

(119903=09928)

[45]

Stearylalcoh

olSilicag

el60

n-hexane

+ethylacetate+

methano

l(46

3505vvv)

Rhod

amineB

586

Darkpink

Pink

148

290divide2700

(119903=09964)

[45]

Stearylalcoh

olSilicag

el60

n-hexane

+ethylacetate+

methano

l(46

3505vvv)

Malachitegreen

644

Green

with

light

border

Green

032

089divide885

(119903=09959)

[45]

Stearylalcoh

olSilicag

el60

n-hexane

+ethylacetate+

methano

l(46

3505vvv)

Janu

sblue

655

Navyblue

Blue

032

053divide708

(119903=09995)

[45]


Table2Ch

aracteris

ticso

fdensitom

etric

band

ofdetected

drugsmod

ified


metric

visualizingindexmod

ified

broadening


Detecteddrug

Stationary

phasemobile

phase

Detectio

nway

Densitom

etric

band

characteris

ticof

detected

drug

Mod

ified

contrast

index(AU∘ )

Densitom

etric

visualizing


Mod

ified

broadening

index(120583gAU

)

Detectio

nindex

(120583gAU

)Re

ference

Area

(AU)

Height

(AU)120572(∘ )

Estradiol

Neutralalum

inum

oxide

60F 2

54toluene+

ethylacetate

(11vv)

With

outu

sing

visualizingreagent

1304

8246

145

16966

mdash19

16032495

[43]

Estradiol

Bartonrsquosreagent

23786

255

1319615

mdash10

5103210

70[43]

Estradiol

Sulphu

ricacid

44709

497

1338231

mdash0559

010850

[43]

Estradiol

Gentia

nviolet

11348

220

1514667

mdash2203

032760

[43]

Estradiol

Methylene

violet

13622

221

1713000

mdash18

35032705

[43]

Estradiol

Neutralalum

inum

oxide

150F

254toluene+

ethylacetate

(11vv)

With

outu

sing

visualizingreagent

11424

192

131476

9mdash

2188

032409

[43]

Estradiol

Gentia

nviolet

10906

127

2077

00mdash

2292

032796

[43]

Estradiol

Methylene

violet

8466

154

334667

mdash2953

053490

[43]

Salicylanilide

Silicag

el60F 2

54(10

5744

)chloroform

With

outu

sing

visualizingreagent

72889

707

9785

11570

0686

00715

90[44]

Salicylanilide

Brilliant

green

91842

704

8880

1640

0544

07011

112

[44]


herol

Silicag

el60

toluene

With

outu

sing

visualizingreagent

34519

459

162869

0479

0724

0453100

[41]


herol


iron(III)

chlorid

e60

474

698

97756

0323

0413

2088915

[41]


herol

Methylene

violet

40426

636

87950

1684

0618

0303456

[41]


herol

Methylgreen

37196

576

964

00

1378

0672

03015

64[41]

(+)-120572-Tocop

herol

acetate

Silicag

el60

toluene

With

outu

sing

visualizingreagent

33796

591

115373

1024

0740

03011

00[41]

(+)-120572-Tocop

herol

acetate

Rhod

amineB

26031

455

153033

0386

0960

04515

11[41]

(+)-120572-Tocop

herol

acetate

Gentia

nviolet

2944

1545

105450

0392

0849

07518

84[41]

(+)-120572-Tocop

herol

acetate

Methylgreen

30226

564

115127

0366

0827

0753911

[41]

(+)-120572-Tocop

herol

acetate

Janu

sblue

30125

565

115136

0365

0830

0753750

[41]

Ibup

rofen

Silicag

el60F 2

54With

outu

sing

visualizingreagent

5564

444

526

17115

0342

044

90623325

[42]

Ibup

rofen

chloroform

+methano

l(50

14vv)

Methylene

violet

53696

348

3599

430123

046

612

544

38[42]

Stearic

acid

Silicag

el60

n-hexane

+aceton

e(4

1vv)

Rhod

amineB

6567

8231

264

50017

432

125900

[45]

Stearic

acid

Methylene

blue

30918

368

1426286

3155

092

007245

[45]

Stearic

acid

Janu

sblue

36175

462

1142000

2741

079

012604

[45]

Stearylalcoh

olSilicag

el60

n-hexane

+ethylacetate+

methano

l(46

3505vvv)

Rhod

amineB

3605

100

1376

920019

749

148234

[45]

Stearylalcoh

olMalachitegreen

12378

333

747571

0553

218

032382

[45]

Stearylalcoh

olJanu

sblue

10963

269

1320692

0264

246

032214

[45]






(120583g)Color of




















Spraying 3807 1158 12 200 1770 596 225 584


Spraying 5148 1469 95 200 4354 1124 19 606


Spraying 7561 2139 85 200 4056 1263 10 200


Spraying 5966 1674 9 200 4560 1035 16 200


Spraying 5338 1555 8 643 3444 1114 10 649


Spraying 2474 691 24 667 1772 501 30 662




4OH




4OH (5 10 3 1




254 The






119865values) can









4 and


4 Furthermore





4 01









2CO3gives


2CO3+ 1



119865= 0138) and











3)2+ saturated


2Cr2O7in 40 H





3)2

Na2CO3 Co(NO







254












Table6Ch

aracteris

ticso

fthe


selected

phenoliccompo

unds

Detecteddrug

Stationary

phase

Mob

ileph

ase

Visualizingreagent

Detectio

nlim

it(120583g)

Detectability

index

(120583gmm

2 )Color

ofspot

Backgrou

ndcoloro

fchromatogram

Reference

Adrenalin

eSilicag

el60F 2

54Glacialaceticacid

+n-bu

tano

l+water

(14

1vvv)

Brilliant

cresylblue

in5

NaO

H010

01010

Brow

nLightviolet

[61]

Silicag

el60kieselguh

rF254

Brom

ocresolgreen

in5

NaO

H025

02547

Lightb

rown

Seagreen

Polyam

ide11F

254

Eosin

yello

win

5NaO

H250

25030

Lightb

rown

Lightyellow

Dop

amine

Silicag

el60F 2

54Glacialaceticacid

+n-bu

tano

l+water

(14

1vvv)

Brom

ocresolgreen

in5

NaO

H010

01015

Orange

Lightb

lue

[61]

Silicag

el60kieselguh

rF254

025

02510

Brow

norange

Seagreen

Polyam

ide11F

254

Eosin

yello

win

5NaO

H075

07517

Lightb

rown

Lightyellow

Metaram

inol

Silicag

el60F 2

54Glacialaceticacid

+n-bu

tano

l+water

(14

1vvv)

Brom

ocresolgreen

in5

NaO

H050

05015

Lemon

Lightb

lue

[61]

Silicag

el60kieselguh

rF254

Eriochromeb

lack

Tin

5NaO

H050

05035

Brow

nLightb

rown

Polyam

ide11F

254

Anilin

eblueinwater

505042

Lightyellow

Blue

Feno

terol

Silicag

el60F 2

54Glacialaceticacid

+n-bu

tano

l+water

(14

1vvv)

Brom

ocresolgreen

in5

NaO

H010

01013

Brow

norange

Lightb

lue

[61]

Silicag

el60kieselguh

rF254

Eriochromeb

lack

Tin

5NaO

H050

05038

Brow

nLightb

rown

Polyam

ide11F

254

Eosin

yello

win

5NaO

H50

515

Yello

wLightyellow

Phenylephrine

Silicag

el60F 2

54Glacialaceticacid

+n-bu

tano

l+water

(14

1vvv)

Brom

ocresolgreen

in5

NaO

H025

02521

Lemon

Lightb

lue

[61]

Silicag

el60kieselguh

rF254

Eriochromeb

lack

Tin

5NaO

H050

05041

Brow

nLightb

rown

Bithiono

lSilicag

el60F 2

54Glacialaceticacid

+n-bu

tano

l+water

(14

1vvv)

Brilliant

cresylblue

in5

NaO

H050

05077

Brow

nLightviolet

[61]

Silicag

el60kieselguh

rF254

Brilliant

cresylblue

in5

NaO

H250

25068

Brow

nLightb

rown

Methyldop

a

Silicag

el60

Glacialaceticacid

+n-bu

tano

l+water

(14

1vvv)

Thym

olblue

in2

NaO

H32

327

Brow

nblack

Beige

[62]

Norepinephrine

Brom

othymolblue

in2

NaO

H06

0628

Yello

wbrown

Lightb

lue

Terbutaline

Brilliant

cresylblue

in2

NaO

H20

2017

Lightb

rown

Lightb

lue

Bamethane

Anilin

ebluein2

NaO

H10

1025

Blue

Lightb

lue

Etam

ivan

Anilin

ebluein2

NaO

H50

5018

Whiteblue

Lightb

lue

Phloroglucinol

Silicag

el60

Chloroform

+methano

l(9

1vv)

Brilliant

cresylblue

in2

NaO

H20

2023

Lightb

rown

Lightb

lue

[62]

Pentazocine

Anilin

ebluein2

NaO

H05

0568

White

Lightb

lue

Hexachlorop

hene

Thym

olblue

in2

NaO

H50

5037

Yellowbeige

Beige

Pyrocatechine

Brilliant

greenin

2NaO

H03

0317

Light

orangered

Lightg

reen

Niclosamide

Brilliant

greenin

2NaO

H08

0818

Green

Lightg

reen

Salicylanilide

Brilliant

greenin

2NaO

H08

0816

Green

Lightg

reen

Thym

olBrilliant

greenin

2NaO

H20

2042

Yello

wgreen

Lightg

reen

Eugeno

lSilicag

el60

Benzene

Brom

othymolblue

in2

NaO

H50

5037

Lightyellow

Lightb

lue

[62]







254after





254 it was worse
























254plates









1 aflatoxin B

2 aflatoxin G

1 aflatoxin G

2[98ndash



4 Conclusion









Acknowledgment


References






































































































































Volume 2014

ToxinsJournal of

VaccinesJournal of




















Autoimmune Diseases




Journal of


Pharmaceutics



of


Table1Fu

ndam

entalabsorptionband

ofdrugscoloro

fspo

tofd


ndcoloro

fchrom

atogramlim

itof

detection

andlin

earityrange

Detecteddrug

Stationary

phase

Detectio

nway

Fund

amental

absorptio

nband

ofdrug120582max(nm)

Color

ofspot

ofdetected

drug

Backgrou

ndcoloro

fchromatogram

Limitof

detection

(120583g)

Linearity

range

(120583gspo

tminus1 )

(rcorrelatio

ncoeffi

cient)

Reference

Estradiol

Neutralalum

inum

oxide6

0F254

toluene+

ethylacetate(11vv)

With

outu

sing

visualizingreagent

205

Lack

ofcoloredspot

invisib

lelight

White

032

110divide2500

(119903=09951)

[43]

Estradiol

Bartonrsquosreagent

233

Darkbluegreen

Willow

-green

032

053divide1024

(119903=09975)

[43]

Estradiol

Sulphu

ricacid

278

Lighto

range

Lightb

eige

010

088divide1280

(119903=09974)

[43]

Estradiol

Gentia

nviolet

226

Violetblue

Violet

032

053divide2500

(119903=09943)

[43]

Estradiol

Methylene

violet

227

Greyblue

Lightg

rey

032

088divide336

(119903=09975)

[43]

Estradiol

Neutralalum

inum

oxide150F 2

54toluene+

ethylacetate(11vv)

With

outu

sing

visualizingreagent

207

Lack

ofcoloredspot

invisib

lelight

White

032

137divide2000

(119903=09972)

[43]

Estradiol

Gentia

nviolet

598

Darkviolet

Violet

032

088divide2000

(119903=09879)

[43]

Estradiol

Methylene

violet

204

Greyblue

Lightg

rey

053

137divide2500

(119903=09983)

[43]

Salicylanilide

Silicag

el60F 2

54(10

5744

)chloroform

With

outu

sing

visualizingreagent

307

Lack

ofcoloredspot

invisib

lelight

White

007

500divide3000

(119903=09802)

[44]

Salicylanilide

Brilliant

green

597

Darkgreen

Green

070

300divide3000

(119903=09929)

[44]


herol

Silicag

el60

toluene

With

outu

sing

visualizingreagent

270

Lack

ofcoloredspot

invisib

lelight

White

045

346divide2000

(119903=09894)

[41]


herol


n(III)chloride

524

Raspberryred

Lightb

eige

208

576divide3000

(119903=09927)

[41]


herol

Methylen

eviolet

270

Azure

with

white

border

Lightg

rey-blue

030

075divide576

(119903=09949)

[41]


herol

Methylgreen

270

Lightg

reen

Green

030

075divide576

(119903=09863)

[41]

(+)-120572-Tocop

herol

acetate

Silicag

el60

toluene

With

outu

sing

visualizingreagent

203

Lack

ofcoloredspot

invisib

lelight

White

030

075divide2000

(119903=09978)

[41]

(+)-120572-Tocop

herol

acetate

Rhod

amineB

203

Darkpink

Pink

045

125divide1600

(119903=09936)

[41]

(+)-120572-Tocop

herol

acetate

Gentia

nviolet

203

Darkviolet

Violet

075

208divide2500

(119903=09908)

[41]

(+)-120572-Tocop

herol

acetate

Methylgreen

203

Green

(smallcon

trast

with

backgrou

ndcolor)

Green

075

125divide2500

(119903=09943)

[41]

(+)-120572-Tocop

herol

acetate

Janu

sblue

203

Darkblue

Blue

075

125divide2500

(119903=09938)

[41]


Table1Con

tinued

Detecteddrug

Stationary

phase

Detectio

nway

Fund

amental

absorptio

nband

ofdrug120582max(nm)

Color

ofspot

ofdetected

drug

Backgrou

ndcoloro

fchromatogram

Limitof

detection

(120583g)

Linearity

range

(120583gspo

tminus1 )

(rcorrelatio

ncoeffi

cient)

Reference

Ibup

rofen

Silicag

el60F 2

54chloroform

+methano

l(50

14

vv)

With

outu

sing

visualizingreagent

200

Lack

ofcoloredspot

invisib

lelight

White

062

125divide1000

(119903=09958)

[42]

Ibup

rofen

Methylene

violet

200

White-bluew

ithdark

blue

border

Light

125

500divide2500

(119903=099549)

[42]

Stearic

acid

Silicag

el60

n-hexane

+aceton

e(4

1vv)

Rhod

amineB

585

Darkpink

Pink

125

156divide2840

(119903=09913)

[45]

Stearic

acid

Methylene

blue

687

Blue

with

light

border

Blue

007

041divide744

(119903=09975)

[45]

Stearic

acid

Janu

sblue

617

Navyblue

with

white

border

Blue

012

156divide744

(119903=09928)

[45]

Stearylalcoh

olSilicag

el60

n-hexane

+ethylacetate+

methano

l(46

3505vvv)

Rhod

amineB

586

Darkpink

Pink

148

290divide2700

(119903=09964)

[45]

Stearylalcoh

olSilicag

el60

n-hexane

+ethylacetate+

methano

l(46

3505vvv)

Malachitegreen

644

Green

with

light

border

Green

032

089divide885

(119903=09959)

[45]

Stearylalcoh

olSilicag

el60

n-hexane

+ethylacetate+

methano

l(46

3505vvv)

Janu

sblue

655

Navyblue

Blue

032

053divide708

(119903=09995)

[45]


Table2Ch

aracteris

ticso

fdensitom

etric

band

ofdetected

drugsmod

ified


metric

visualizingindexmod

ified

broadening


Detecteddrug

Stationary

phasemobile

phase

Detectio

nway

Densitom

etric

band

characteris

ticof

detected

drug

Mod

ified

contrast

index(AU∘ )

Densitom

etric

visualizing


Mod

ified

broadening

index(120583gAU

)

Detectio

nindex

(120583gAU

)Re

ference

Area

(AU)

Height

(AU)120572(∘ )

Estradiol

Neutralalum

inum

oxide

60F 2

54toluene+

ethylacetate

(11vv)

With

outu

sing

visualizingreagent

1304

8246

145

16966

mdash19

16032495

[43]

Estradiol

Bartonrsquosreagent

23786

255

1319615

mdash10

5103210

70[43]

Estradiol

Sulphu

ricacid

44709

497

1338231

mdash0559

010850

[43]

Estradiol

Gentia

nviolet

11348

220

1514667

mdash2203

032760

[43]

Estradiol

Methylene

violet

13622

221

1713000

mdash18

35032705

[43]

Estradiol

Neutralalum

inum

oxide

150F

254toluene+

ethylacetate

(11vv)

With

outu

sing

visualizingreagent

11424

192

131476

9mdash

2188

032409

[43]

Estradiol

Gentia

nviolet

10906

127

2077

00mdash

2292

032796

[43]

Estradiol

Methylene

violet

8466

154

334667

mdash2953

053490

[43]

Salicylanilide

Silicag

el60F 2

54(10

5744

)chloroform

With

outu

sing

visualizingreagent

72889

707

9785

11570

0686

00715

90[44]

Salicylanilide

Brilliant

green

91842

704

8880

1640

0544

07011

112

[44]


herol

Silicag

el60

toluene

With

outu

sing

visualizingreagent

34519

459

162869

0479

0724

0453100

[41]


herol


iron(III)

chlorid

e60

474

698

97756

0323

0413

2088915

[41]


herol

Methylene

violet

40426

636

87950

1684

0618

0303456

[41]


herol

Methylgreen

37196

576

964

00

1378

0672

03015

64[41]

(+)-120572-Tocop

herol

acetate

Silicag

el60

toluene

With

outu

sing

visualizingreagent

33796

591

115373

1024

0740

03011

00[41]

(+)-120572-Tocop

herol

acetate

Rhod

amineB

26031

455

153033

0386

0960

04515

11[41]

(+)-120572-Tocop

herol

acetate

Gentia

nviolet

2944

1545

105450

0392

0849

07518

84[41]

(+)-120572-Tocop

herol

acetate

Methylgreen

30226

564

115127

0366

0827

0753911

[41]

(+)-120572-Tocop

herol

acetate

Janu

sblue

30125

565

115136

0365

0830

0753750

[41]

Ibup

rofen

Silicag

el60F 2

54With

outu

sing

visualizingreagent

5564

444

526

17115

0342

044

90623325

[42]

Ibup

rofen

chloroform

+methano

l(50

14vv)

Methylene

violet

53696

348

3599

430123

046

612

544

38[42]

Stearic

acid

Silicag

el60

n-hexane

+aceton

e(4

1vv)

Rhod

amineB

6567

8231

264

50017

432

125900

[45]

Stearic

acid

Methylene

blue

30918

368

1426286

3155

092

007245

[45]

Stearic

acid

Janu

sblue

36175

462

1142000

2741

079

012604

[45]

Stearylalcoh

olSilicag

el60

n-hexane

+ethylacetate+

methano

l(46

3505vvv)

Rhod

amineB

3605

100

1376

920019

749

148234

[45]

Stearylalcoh

olMalachitegreen

12378

333

747571

0553

218

032382

[45]

Stearylalcoh

olJanu

sblue

10963

269

1320692

0264

246

032214

[45]






(120583g)Color of




















Spraying 3807 1158 12 200 1770 596 225 584


Spraying 5148 1469 95 200 4354 1124 19 606


Spraying 7561 2139 85 200 4056 1263 10 200


Spraying 5966 1674 9 200 4560 1035 16 200


Spraying 5338 1555 8 643 3444 1114 10 649


Spraying 2474 691 24 667 1772 501 30 662




4OH




4OH (5 10 3 1




254 The






119865values) can









4 and


4 Furthermore





4 01









2CO3gives


2CO3+ 1



119865= 0138) and











3)2+ saturated


2Cr2O7in 40 H





3)2

Na2CO3 Co(NO







254












Table6Ch

aracteris

ticso

fthe


selected

phenoliccompo

unds

Detecteddrug

Stationary

phase

Mob

ileph

ase

Visualizingreagent

Detectio

nlim

it(120583g)

Detectability

index

(120583gmm

2 )Color

ofspot

Backgrou

ndcoloro

fchromatogram

Reference

Adrenalin

eSilicag

el60F 2

54Glacialaceticacid

+n-bu

tano

l+water

(14

1vvv)

Brilliant

cresylblue

in5

NaO

H010

01010

Brow

nLightviolet

[61]

Silicag

el60kieselguh

rF254

Brom

ocresolgreen

in5

NaO

H025

02547

Lightb

rown

Seagreen

Polyam

ide11F

254

Eosin

yello

win

5NaO

H250

25030

Lightb

rown

Lightyellow

Dop

amine

Silicag

el60F 2

54Glacialaceticacid

+n-bu

tano

l+water

(14

1vvv)

Brom

ocresolgreen

in5

NaO

H010

01015

Orange

Lightb

lue

[61]

Silicag

el60kieselguh

rF254

025

02510

Brow

norange

Seagreen

Polyam

ide11F

254

Eosin

yello

win

5NaO

H075

07517

Lightb

rown

Lightyellow

Metaram

inol

Silicag

el60F 2

54Glacialaceticacid

+n-bu

tano

l+water

(14

1vvv)

Brom

ocresolgreen

in5

NaO

H050

05015

Lemon

Lightb

lue

[61]

Silicag

el60kieselguh

rF254

Eriochromeb

lack

Tin

5NaO

H050

05035

Brow

nLightb

rown

Polyam

ide11F

254

Anilin

eblueinwater

505042

Lightyellow

Blue

Feno

terol

Silicag

el60F 2

54Glacialaceticacid

+n-bu

tano

l+water

(14

1vvv)

Brom

ocresolgreen

in5

NaO

H010

01013

Brow

norange

Lightb

lue

[61]

Silicag

el60kieselguh

rF254

Eriochromeb

lack

Tin

5NaO

H050

05038

Brow

nLightb

rown

Polyam

ide11F

254

Eosin

yello

win

5NaO

H50

515

Yello

wLightyellow

Phenylephrine

Silicag

el60F 2

54Glacialaceticacid

+n-bu

tano

l+water

(14

1vvv)

Brom

ocresolgreen

in5

NaO

H025

02521

Lemon

Lightb

lue

[61]

Silicag

el60kieselguh

rF254

Eriochromeb

lack

Tin

5NaO

H050

05041

Brow

nLightb

rown

Bithiono

lSilicag

el60F 2

54Glacialaceticacid

+n-bu

tano

l+water

(14

1vvv)

Brilliant

cresylblue

in5

NaO

H050

05077

Brow

nLightviolet

[61]

Silicag

el60kieselguh

rF254

Brilliant

cresylblue

in5

NaO

H250

25068

Brow

nLightb

rown

Methyldop

a

Silicag

el60

Glacialaceticacid

+n-bu

tano

l+water

(14

1vvv)

Thym

olblue

in2

NaO

H32

327

Brow

nblack

Beige

[62]

Norepinephrine

Brom

othymolblue

in2

NaO

H06

0628

Yello

wbrown

Lightb

lue

Terbutaline

Brilliant

cresylblue

in2

NaO

H20

2017

Lightb

rown

Lightb

lue

Bamethane

Anilin

ebluein2

NaO

H10

1025

Blue

Lightb

lue

Etam

ivan

Anilin

ebluein2

NaO

H50

5018

Whiteblue

Lightb

lue

Phloroglucinol

Silicag

el60

Chloroform

+methano

l(9

1vv)

Brilliant

cresylblue

in2

NaO

H20

2023

Lightb

rown

Lightb

lue

[62]

Pentazocine

Anilin

ebluein2

NaO

H05

0568

White

Lightb

lue

Hexachlorop

hene

Thym

olblue

in2

NaO

H50

5037

Yellowbeige

Beige

Pyrocatechine

Brilliant

greenin

2NaO

H03

0317

Light

orangered

Lightg

reen

Niclosamide

Brilliant

greenin

2NaO

H08

0818

Green

Lightg

reen

Salicylanilide

Brilliant

greenin

2NaO

H08

0816

Green

Lightg

reen

Thym

olBrilliant

greenin

2NaO

H20

2042

Yello

wgreen

Lightg

reen

Eugeno

lSilicag

el60

Benzene

Brom

othymolblue

in2

NaO

H50

5037

Lightyellow

Lightb

lue

[62]







254after





254 it was worse
























254plates









1 aflatoxin B

2 aflatoxin G

1 aflatoxin G

2[98ndash



4 Conclusion









Acknowledgment


References






































































































































Volume 2014

ToxinsJournal of

VaccinesJournal of




















Autoimmune Diseases




Journal of


Pharmaceutics



of


Table1Con

tinued

Detecteddrug

Stationary

phase

Detectio

nway

Fund

amental

absorptio

nband

ofdrug120582max(nm)

Color

ofspot

ofdetected

drug

Backgrou

ndcoloro

fchromatogram

Limitof

detection

(120583g)

Linearity

range

(120583gspo

tminus1 )

(rcorrelatio

ncoeffi

cient)

Reference

Ibup

rofen

Silicag

el60F 2

54chloroform

+methano

l(50

14

vv)

With

outu

sing

visualizingreagent

200

Lack

ofcoloredspot

invisib

lelight

White

062

125divide1000

(119903=09958)

[42]

Ibup

rofen

Methylene

violet

200

White-bluew

ithdark

blue

border

Light

125

500divide2500

(119903=099549)

[42]

Stearic

acid

Silicag

el60

n-hexane

+aceton

e(4

1vv)

Rhod

amineB

585

Darkpink

Pink

125

156divide2840

(119903=09913)

[45]

Stearic

acid

Methylene

blue

687

Blue

with

light

border

Blue

007

041divide744

(119903=09975)

[45]

Stearic

acid

Janu

sblue

617

Navyblue

with

white

border

Blue

012

156divide744

(119903=09928)

[45]

Stearylalcoh

olSilicag

el60

n-hexane

+ethylacetate+

methano

l(46

3505vvv)

Rhod

amineB

586

Darkpink

Pink

148

290divide2700

(119903=09964)

[45]

Stearylalcoh

olSilicag

el60

n-hexane

+ethylacetate+

methano

l(46

3505vvv)

Malachitegreen

644

Green

with

light

border

Green

032

089divide885

(119903=09959)

[45]

Stearylalcoh

olSilicag

el60

n-hexane

+ethylacetate+

methano

l(46

3505vvv)

Janu

sblue

655

Navyblue

Blue

032

053divide708

(119903=09995)

[45]


Table2Ch

aracteris

ticso

fdensitom

etric

band

ofdetected

drugsmod

ified


metric

visualizingindexmod

ified

broadening


Detecteddrug

Stationary

phasemobile

phase

Detectio

nway

Densitom

etric

band

characteris

ticof

detected

drug

Mod

ified

contrast

index(AU∘ )

Densitom

etric

visualizing


Mod

ified

broadening

index(120583gAU

)

Detectio

nindex

(120583gAU

)Re

ference

Area

(AU)

Height

(AU)120572(∘ )

Estradiol

Neutralalum

inum

oxide

60F 2

54toluene+

ethylacetate

(11vv)

With

outu

sing

visualizingreagent

1304

8246

145

16966

mdash19

16032495

[43]

Estradiol

Bartonrsquosreagent

23786

255

1319615

mdash10

5103210

70[43]

Estradiol

Sulphu

ricacid

44709

497

1338231

mdash0559

010850

[43]

Estradiol

Gentia

nviolet

11348

220

1514667

mdash2203

032760

[43]

Estradiol

Methylene

violet

13622

221

1713000

mdash18

35032705

[43]

Estradiol

Neutralalum

inum

oxide

150F

254toluene+

ethylacetate

(11vv)

With

outu

sing

visualizingreagent

11424

192

131476

9mdash

2188

032409

[43]

Estradiol

Gentia

nviolet

10906

127

2077

00mdash

2292

032796

[43]

Estradiol

Methylene

violet

8466

154

334667

mdash2953

053490

[43]

Salicylanilide

Silicag

el60F 2

54(10

5744

)chloroform

With

outu

sing

visualizingreagent

72889

707

9785

11570

0686

00715

90[44]

Salicylanilide

Brilliant

green

91842

704

8880

1640

0544

07011

112

[44]


herol

Silicag

el60

toluene

With

outu

sing

visualizingreagent

34519

459

162869

0479

0724

0453100

[41]


herol


iron(III)

chlorid

e60

474

698

97756

0323

0413

2088915

[41]


herol

Methylene

violet

40426

636

87950

1684

0618

0303456

[41]


herol

Methylgreen

37196

576

964

00

1378

0672

03015

64[41]

(+)-120572-Tocop

herol

acetate

Silicag

el60

toluene

With

outu

sing

visualizingreagent

33796

591

115373

1024

0740

03011

00[41]

(+)-120572-Tocop

herol

acetate

Rhod

amineB

26031

455

153033

0386

0960

04515

11[41]

(+)-120572-Tocop

herol

acetate

Gentia

nviolet

2944

1545

105450

0392

0849

07518

84[41]

(+)-120572-Tocop

herol

acetate

Methylgreen

30226

564

115127

0366

0827

0753911

[41]

(+)-120572-Tocop

herol

acetate

Janu

sblue

30125

565

115136

0365

0830

0753750

[41]

Ibup

rofen

Silicag

el60F 2

54With

outu

sing

visualizingreagent

5564

444

526

17115

0342

044

90623325

[42]

Ibup

rofen

chloroform

+methano

l(50

14vv)

Methylene

violet

53696

348

3599

430123

046

612

544

38[42]

Stearic

acid

Silicag

el60

n-hexane

+aceton

e(4

1vv)

Rhod

amineB

6567

8231

264

50017

432

125900

[45]

Stearic

acid

Methylene

blue

30918

368

1426286

3155

092

007245

[45]

Stearic

acid

Janu

sblue

36175

462

1142000

2741

079

012604

[45]

Stearylalcoh

olSilicag

el60

n-hexane

+ethylacetate+

methano

l(46

3505vvv)

Rhod

amineB

3605

100

1376

920019

749

148234

[45]

Stearylalcoh

olMalachitegreen

12378

333

747571

0553

218

032382

[45]

Stearylalcoh

olJanu

sblue

10963

269

1320692

0264

246

032214

[45]






(120583g)Color of




















Spraying 3807 1158 12 200 1770 596 225 584


Spraying 5148 1469 95 200 4354 1124 19 606


Spraying 7561 2139 85 200 4056 1263 10 200


Spraying 5966 1674 9 200 4560 1035 16 200


Spraying 5338 1555 8 643 3444 1114 10 649


Spraying 2474 691 24 667 1772 501 30 662




4OH




4OH (5 10 3 1




254 The






119865values) can









4 and


4 Furthermore





4 01









2CO3gives


2CO3+ 1



119865= 0138) and











3)2+ saturated


2Cr2O7in 40 H





3)2

Na2CO3 Co(NO







254












Table6Ch

aracteris

ticso

fthe


selected

phenoliccompo

unds

Detecteddrug

Stationary

phase

Mob

ileph

ase

Visualizingreagent

Detectio

nlim

it(120583g)

Detectability

index

(120583gmm

2 )Color

ofspot

Backgrou

ndcoloro

fchromatogram

Reference

Adrenalin

eSilicag

el60F 2

54Glacialaceticacid

+n-bu

tano

l+water

(14

1vvv)

Brilliant

cresylblue

in5

NaO

H010

01010

Brow

nLightviolet

[61]

Silicag

el60kieselguh

rF254

Brom

ocresolgreen

in5

NaO

H025

02547

Lightb

rown

Seagreen

Polyam

ide11F

254

Eosin

yello

win

5NaO

H250

25030

Lightb

rown

Lightyellow

Dop

amine

Silicag

el60F 2

54Glacialaceticacid

+n-bu

tano

l+water

(14

1vvv)

Brom

ocresolgreen

in5

NaO

H010

01015

Orange

Lightb

lue

[61]

Silicag

el60kieselguh

rF254

025

02510

Brow

norange

Seagreen

Polyam

ide11F

254

Eosin

yello

win

5NaO

H075

07517

Lightb

rown

Lightyellow

Metaram

inol

Silicag

el60F 2

54Glacialaceticacid

+n-bu

tano

l+water

(14

1vvv)

Brom

ocresolgreen

in5

NaO

H050

05015

Lemon

Lightb

lue

[61]

Silicag

el60kieselguh

rF254

Eriochromeb

lack

Tin

5NaO

H050

05035

Brow

nLightb

rown

Polyam

ide11F

254

Anilin

eblueinwater

505042

Lightyellow

Blue

Feno

terol

Silicag

el60F 2

54Glacialaceticacid

+n-bu

tano

l+water

(14

1vvv)

Brom

ocresolgreen

in5

NaO

H010

01013

Brow

norange

Lightb

lue

[61]

Silicag

el60kieselguh

rF254

Eriochromeb

lack

Tin

5NaO

H050

05038

Brow

nLightb

rown

Polyam

ide11F

254

Eosin

yello

win

5NaO

H50

515

Yello

wLightyellow

Phenylephrine

Silicag

el60F 2

54Glacialaceticacid

+n-bu

tano

l+water

(14

1vvv)

Brom

ocresolgreen

in5

NaO

H025

02521

Lemon

Lightb

lue

[61]

Silicag

el60kieselguh

rF254

Eriochromeb

lack

Tin

5NaO

H050

05041

Brow

nLightb

rown

Bithiono

lSilicag

el60F 2

54Glacialaceticacid

+n-bu

tano

l+water

(14

1vvv)

Brilliant

cresylblue

in5

NaO

H050

05077

Brow

nLightviolet

[61]

Silicag

el60kieselguh

rF254

Brilliant

cresylblue

in5

NaO

H250

25068

Brow

nLightb

rown

Methyldop

a

Silicag

el60

Glacialaceticacid

+n-bu

tano

l+water

(14

1vvv)

Thym

olblue

in2

NaO

H32

327

Brow

nblack

Beige

[62]

Norepinephrine

Brom

othymolblue

in2

NaO

H06

0628

Yello

wbrown

Lightb

lue

Terbutaline

Brilliant

cresylblue

in2

NaO

H20

2017

Lightb

rown

Lightb

lue

Bamethane

Anilin

ebluein2

NaO

H10

1025

Blue

Lightb

lue

Etam

ivan

Anilin

ebluein2

NaO

H50

5018

Whiteblue

Lightb

lue

Phloroglucinol

Silicag

el60

Chloroform

+methano

l(9

1vv)

Brilliant

cresylblue

in2

NaO

H20

2023

Lightb

rown

Lightb

lue

[62]

Pentazocine

Anilin

ebluein2

NaO

H05

0568

White

Lightb

lue

Hexachlorop

hene

Thym

olblue

in2

NaO

H50

5037

Yellowbeige

Beige

Pyrocatechine

Brilliant

greenin

2NaO

H03

0317

Light

orangered

Lightg

reen

Niclosamide

Brilliant

greenin

2NaO

H08

0818

Green

Lightg

reen

Salicylanilide

Brilliant

greenin

2NaO

H08

0816

Green

Lightg

reen

Thym

olBrilliant

greenin

2NaO

H20

2042

Yello

wgreen

Lightg

reen

Eugeno

lSilicag

el60

Benzene

Brom

othymolblue

in2

NaO

H50

5037

Lightyellow

Lightb

lue

[62]







254after





254 it was worse
























254plates









1 aflatoxin B

2 aflatoxin G

1 aflatoxin G

2[98ndash



4 Conclusion









Acknowledgment


References






































































































































Volume 2014

ToxinsJournal of

VaccinesJournal of




















Autoimmune Diseases




Journal of


Pharmaceutics



of


Table2Ch

aracteris

ticso

fdensitom

etric

band

ofdetected

drugsmod

ified


metric

visualizingindexmod

ified

broadening


Detecteddrug

Stationary

phasemobile

phase

Detectio

nway

Densitom

etric

band

characteris

ticof

detected

drug

Mod

ified

contrast

index(AU∘ )

Densitom

etric

visualizing


Mod

ified

broadening

index(120583gAU

)

Detectio

nindex

(120583gAU

)Re

ference

Area

(AU)

Height

(AU)120572(∘ )

Estradiol

Neutralalum

inum

oxide

60F 2

54toluene+

ethylacetate

(11vv)

With

outu

sing

visualizingreagent

1304

8246

145

16966

mdash19

16032495

[43]

Estradiol

Bartonrsquosreagent

23786

255

1319615

mdash10

5103210

70[43]

Estradiol

Sulphu

ricacid

44709

497

1338231

mdash0559

010850

[43]

Estradiol

Gentia

nviolet

11348

220

1514667

mdash2203

032760

[43]

Estradiol

Methylene

violet

13622

221

1713000

mdash18

35032705

[43]

Estradiol

Neutralalum

inum

oxide

150F

254toluene+

ethylacetate

(11vv)

With

outu

sing

visualizingreagent

11424

192

131476

9mdash

2188

032409

[43]

Estradiol

Gentia

nviolet

10906

127

2077

00mdash

2292

032796

[43]

Estradiol

Methylene

violet

8466

154

334667

mdash2953

053490

[43]

Salicylanilide

Silicag

el60F 2

54(10

5744

)chloroform

With

outu

sing

visualizingreagent

72889

707

9785

11570

0686

00715

90[44]

Salicylanilide

Brilliant

green

91842

704

8880

1640

0544

07011

112

[44]


herol

Silicag

el60

toluene

With

outu

sing

visualizingreagent

34519

459

162869

0479

0724

0453100

[41]


herol


iron(III)

chlorid

e60

474

698

97756

0323

0413

2088915

[41]


herol

Methylene

violet

40426

636

87950

1684

0618

0303456

[41]


herol

Methylgreen

37196

576

964

00

1378

0672

03015

64[41]

(+)-120572-Tocop

herol

acetate

Silicag

el60

toluene

With

outu

sing

visualizingreagent

33796

591

115373

1024

0740

03011

00[41]

(+)-120572-Tocop

herol

acetate

Rhod

amineB

26031

455

153033

0386

0960

04515

11[41]

(+)-120572-Tocop

herol

acetate

Gentia

nviolet

2944

1545

105450

0392

0849

07518

84[41]

(+)-120572-Tocop

herol

acetate

Methylgreen

30226

564

115127

0366

0827

0753911

[41]

(+)-120572-Tocop

herol

acetate

Janu

sblue

30125

565

115136

0365

0830

0753750

[41]

Ibup

rofen

Silicag

el60F 2

54With

outu

sing

visualizingreagent

5564

444

526

17115

0342

044

90623325

[42]

Ibup

rofen

chloroform

+methano

l(50

14vv)

Methylene

violet

53696

348

3599

430123

046

612

544

38[42]

Stearic

acid

Silicag

el60

n-hexane

+aceton

e(4

1vv)

Rhod

amineB

6567

8231

264

50017

432

125900

[45]

Stearic

acid

Methylene

blue

30918

368

1426286

3155

092

007245

[45]

Stearic

acid

Janu

sblue

36175

462

1142000

2741

079

012604

[45]

Stearylalcoh

olSilicag

el60

n-hexane

+ethylacetate+

methano

l(46

3505vvv)

Rhod

amineB

3605

100

1376

920019

749

148234

[45]

Stearylalcoh

olMalachitegreen

12378

333

747571

0553

218

032382

[45]

Stearylalcoh

olJanu

sblue

10963

269

1320692

0264

246

032214

[45]






(120583g)Color of




















Spraying 3807 1158 12 200 1770 596 225 584


Spraying 5148 1469 95 200 4354 1124 19 606


Spraying 7561 2139 85 200 4056 1263 10 200


Spraying 5966 1674 9 200 4560 1035 16 200


Spraying 5338 1555 8 643 3444 1114 10 649


Spraying 2474 691 24 667 1772 501 30 662




4OH




4OH (5 10 3 1




254 The






119865values) can









4 and


4 Furthermore





4 01









2CO3gives


2CO3+ 1



119865= 0138) and











3)2+ saturated


2Cr2O7in 40 H





3)2

Na2CO3 Co(NO







254












Table6Ch

aracteris

ticso

fthe


selected

phenoliccompo

unds

Detecteddrug

Stationary

phase

Mob

ileph

ase

Visualizingreagent

Detectio

nlim

it(120583g)

Detectability

index

(120583gmm

2 )Color

ofspot

Backgrou

ndcoloro

fchromatogram

Reference

Adrenalin

eSilicag

el60F 2

54Glacialaceticacid

+n-bu

tano

l+water

(14

1vvv)

Brilliant

cresylblue

in5

NaO

H010

01010

Brow

nLightviolet

[61]

Silicag

el60kieselguh

rF254

Brom

ocresolgreen

in5

NaO

H025

02547

Lightb

rown

Seagreen

Polyam

ide11F

254

Eosin

yello

win

5NaO

H250

25030

Lightb

rown

Lightyellow

Dop

amine

Silicag

el60F 2

54Glacialaceticacid

+n-bu

tano

l+water

(14

1vvv)

Brom

ocresolgreen

in5

NaO

H010

01015

Orange

Lightb

lue

[61]

Silicag

el60kieselguh

rF254

025

02510

Brow

norange

Seagreen

Polyam

ide11F

254

Eosin

yello

win

5NaO

H075

07517

Lightb

rown

Lightyellow

Metaram

inol

Silicag

el60F 2

54Glacialaceticacid

+n-bu

tano

l+water

(14

1vvv)

Brom

ocresolgreen

in5

NaO

H050

05015

Lemon

Lightb

lue

[61]

Silicag

el60kieselguh

rF254

Eriochromeb

lack

Tin

5NaO

H050

05035

Brow

nLightb

rown

Polyam

ide11F

254

Anilin

eblueinwater

505042

Lightyellow

Blue

Feno

terol

Silicag

el60F 2

54Glacialaceticacid

+n-bu

tano

l+water

(14

1vvv)

Brom

ocresolgreen

in5

NaO

H010

01013

Brow

norange

Lightb

lue

[61]

Silicag

el60kieselguh

rF254

Eriochromeb

lack

Tin

5NaO

H050

05038

Brow

nLightb

rown

Polyam

ide11F

254

Eosin

yello

win

5NaO

H50

515

Yello

wLightyellow

Phenylephrine

Silicag

el60F 2

54Glacialaceticacid

+n-bu

tano

l+water

(14

1vvv)

Brom

ocresolgreen

in5

NaO

H025

02521

Lemon

Lightb

lue

[61]

Silicag

el60kieselguh

rF254

Eriochromeb

lack

Tin

5NaO

H050

05041

Brow

nLightb

rown

Bithiono

lSilicag

el60F 2

54Glacialaceticacid

+n-bu

tano

l+water

(14

1vvv)

Brilliant

cresylblue

in5

NaO

H050

05077

Brow

nLightviolet

[61]

Silicag

el60kieselguh

rF254

Brilliant

cresylblue

in5

NaO

H250

25068

Brow

nLightb

rown

Methyldop

a

Silicag

el60

Glacialaceticacid

+n-bu

tano

l+water

(14

1vvv)

Thym

olblue

in2

NaO

H32

327

Brow

nblack

Beige

[62]

Norepinephrine

Brom

othymolblue

in2

NaO

H06

0628

Yello

wbrown

Lightb

lue

Terbutaline

Brilliant

cresylblue

in2

NaO

H20

2017

Lightb

rown

Lightb

lue

Bamethane

Anilin

ebluein2

NaO

H10

1025

Blue

Lightb

lue

Etam

ivan

Anilin

ebluein2

NaO

H50

5018

Whiteblue

Lightb

lue

Phloroglucinol

Silicag

el60

Chloroform

+methano

l(9

1vv)

Brilliant

cresylblue

in2

NaO

H20

2023

Lightb

rown

Lightb

lue

[62]

Pentazocine

Anilin

ebluein2

NaO

H05

0568

White

Lightb

lue

Hexachlorop

hene

Thym

olblue

in2

NaO

H50

5037

Yellowbeige

Beige

Pyrocatechine

Brilliant

greenin

2NaO

H03

0317

Light

orangered

Lightg

reen

Niclosamide

Brilliant

greenin

2NaO

H08

0818

Green

Lightg

reen

Salicylanilide

Brilliant

greenin

2NaO

H08

0816

Green

Lightg

reen

Thym

olBrilliant

greenin

2NaO

H20

2042

Yello

wgreen

Lightg

reen

Eugeno

lSilicag

el60

Benzene

Brom

othymolblue

in2

NaO

H50

5037

Lightyellow

Lightb

lue

[62]







254after





254 it was worse
























254plates









1 aflatoxin B

2 aflatoxin G

1 aflatoxin G

2[98ndash



4 Conclusion









Acknowledgment


References






































































































































Volume 2014

ToxinsJournal of

VaccinesJournal of




















Autoimmune Diseases




Journal of


Pharmaceutics



of






(120583g)Color of




















Spraying 3807 1158 12 200 1770 596 225 584


Spraying 5148 1469 95 200 4354 1124 19 606


Spraying 7561 2139 85 200 4056 1263 10 200


Spraying 5966 1674 9 200 4560 1035 16 200


Spraying 5338 1555 8 643 3444 1114 10 649


Spraying 2474 691 24 667 1772 501 30 662




4OH




4OH (5 10 3 1




254 The






119865values) can









4 and


4 Furthermore





4 01









2CO3gives


2CO3+ 1



119865= 0138) and











3)2+ saturated


2Cr2O7in 40 H





3)2

Na2CO3 Co(NO







254












Table6Ch

aracteris

ticso

fthe


selected

phenoliccompo

unds

Detecteddrug

Stationary

phase

Mob

ileph

ase

Visualizingreagent

Detectio

nlim

it(120583g)

Detectability

index

(120583gmm

2 )Color

ofspot

Backgrou

ndcoloro

fchromatogram

Reference

Adrenalin

eSilicag

el60F 2

54Glacialaceticacid

+n-bu

tano

l+water

(14

1vvv)

Brilliant

cresylblue

in5

NaO

H010

01010

Brow

nLightviolet

[61]

Silicag

el60kieselguh

rF254

Brom

ocresolgreen

in5

NaO

H025

02547

Lightb

rown

Seagreen

Polyam

ide11F

254

Eosin

yello

win

5NaO

H250

25030

Lightb

rown

Lightyellow

Dop

amine

Silicag

el60F 2

54Glacialaceticacid

+n-bu

tano

l+water

(14

1vvv)

Brom

ocresolgreen

in5

NaO

H010

01015

Orange

Lightb

lue

[61]

Silicag

el60kieselguh

rF254

025

02510

Brow

norange

Seagreen

Polyam

ide11F

254

Eosin

yello

win

5NaO

H075

07517

Lightb

rown

Lightyellow

Metaram

inol

Silicag

el60F 2

54Glacialaceticacid

+n-bu

tano

l+water

(14

1vvv)

Brom

ocresolgreen

in5

NaO

H050

05015

Lemon

Lightb

lue

[61]

Silicag

el60kieselguh

rF254

Eriochromeb

lack

Tin

5NaO

H050

05035

Brow

nLightb

rown

Polyam

ide11F

254

Anilin

eblueinwater

505042

Lightyellow

Blue

Feno

terol

Silicag

el60F 2

54Glacialaceticacid

+n-bu

tano

l+water

(14

1vvv)

Brom

ocresolgreen

in5

NaO

H010

01013

Brow

norange

Lightb

lue

[61]

Silicag

el60kieselguh

rF254

Eriochromeb

lack

Tin

5NaO

H050

05038

Brow

nLightb

rown

Polyam

ide11F

254

Eosin

yello

win

5NaO

H50

515

Yello

wLightyellow

Phenylephrine

Silicag

el60F 2

54Glacialaceticacid

+n-bu

tano

l+water

(14

1vvv)

Brom

ocresolgreen

in5

NaO

H025

02521

Lemon

Lightb

lue

[61]

Silicag

el60kieselguh

rF254

Eriochromeb

lack

Tin

5NaO

H050

05041

Brow

nLightb

rown

Bithiono

lSilicag

el60F 2

54Glacialaceticacid

+n-bu

tano

l+water

(14

1vvv)

Brilliant

cresylblue

in5

NaO

H050

05077

Brow

nLightviolet

[61]

Silicag

el60kieselguh

rF254

Brilliant

cresylblue

in5

NaO

H250

25068

Brow

nLightb

rown

Methyldop

a

Silicag

el60

Glacialaceticacid

+n-bu

tano

l+water

(14

1vvv)

Thym

olblue

in2

NaO

H32

327

Brow

nblack

Beige

[62]

Norepinephrine

Brom

othymolblue

in2

NaO

H06

0628

Yello

wbrown

Lightb

lue

Terbutaline

Brilliant

cresylblue

in2

NaO

H20

2017

Lightb

rown

Lightb

lue

Bamethane

Anilin

ebluein2

NaO

H10

1025

Blue

Lightb

lue

Etam

ivan

Anilin

ebluein2

NaO

H50

5018

Whiteblue

Lightb

lue

Phloroglucinol

Silicag

el60

Chloroform

+methano

l(9

1vv)

Brilliant

cresylblue

in2

NaO

H20

2023

Lightb

rown

Lightb

lue

[62]

Pentazocine

Anilin

ebluein2

NaO

H05

0568

White

Lightb

lue

Hexachlorop

hene

Thym

olblue

in2

NaO

H50

5037

Yellowbeige

Beige

Pyrocatechine

Brilliant

greenin

2NaO

H03

0317

Light

orangered

Lightg

reen

Niclosamide

Brilliant

greenin

2NaO

H08

0818

Green

Lightg

reen

Salicylanilide

Brilliant

greenin

2NaO

H08

0816

Green

Lightg

reen

Thym

olBrilliant

greenin

2NaO

H20

2042

Yello

wgreen

Lightg

reen

Eugeno

lSilicag

el60

Benzene

Brom

othymolblue

in2

NaO

H50

5037

Lightyellow

Lightb

lue

[62]







254after





254 it was worse
























254plates









1 aflatoxin B

2 aflatoxin G

1 aflatoxin G

2[98ndash



4 Conclusion









Acknowledgment


References






































































































































Volume 2014

ToxinsJournal of

VaccinesJournal of




















Autoimmune Diseases




Journal of


Pharmaceutics



of






Spraying 3807 1158 12 200 1770 596 225 584


Spraying 5148 1469 95 200 4354 1124 19 606


Spraying 7561 2139 85 200 4056 1263 10 200


Spraying 5966 1674 9 200 4560 1035 16 200


Spraying 5338 1555 8 643 3444 1114 10 649


Spraying 2474 691 24 667 1772 501 30 662




4OH




4OH (5 10 3 1




254 The






119865values) can









4 and


4 Furthermore





4 01









2CO3gives


2CO3+ 1



119865= 0138) and











3)2+ saturated


2Cr2O7in 40 H





3)2

Na2CO3 Co(NO







254












Table6Ch

aracteris

ticso

fthe


selected

phenoliccompo

unds

Detecteddrug

Stationary

phase

Mob

ileph

ase

Visualizingreagent

Detectio

nlim

it(120583g)

Detectability

index

(120583gmm

2 )Color

ofspot

Backgrou

ndcoloro

fchromatogram

Reference

Adrenalin

eSilicag

el60F 2

54Glacialaceticacid

+n-bu

tano

l+water

(14

1vvv)

Brilliant

cresylblue

in5

NaO

H010

01010

Brow

nLightviolet

[61]

Silicag

el60kieselguh

rF254

Brom

ocresolgreen

in5

NaO

H025

02547

Lightb

rown

Seagreen

Polyam

ide11F

254

Eosin

yello

win

5NaO

H250

25030

Lightb

rown

Lightyellow

Dop

amine

Silicag

el60F 2

54Glacialaceticacid

+n-bu

tano

l+water

(14

1vvv)

Brom

ocresolgreen

in5

NaO

H010

01015

Orange

Lightb

lue

[61]

Silicag

el60kieselguh

rF254

025

02510

Brow

norange

Seagreen

Polyam

ide11F

254

Eosin

yello

win

5NaO

H075

07517

Lightb

rown

Lightyellow

Metaram

inol

Silicag

el60F 2

54Glacialaceticacid

+n-bu

tano

l+water

(14

1vvv)

Brom

ocresolgreen

in5

NaO

H050

05015

Lemon

Lightb

lue

[61]

Silicag

el60kieselguh

rF254

Eriochromeb

lack

Tin

5NaO

H050

05035

Brow

nLightb

rown

Polyam

ide11F

254

Anilin

eblueinwater

505042

Lightyellow

Blue

Feno

terol

Silicag

el60F 2

54Glacialaceticacid

+n-bu

tano

l+water

(14

1vvv)

Brom

ocresolgreen

in5

NaO

H010

01013

Brow

norange

Lightb

lue

[61]

Silicag

el60kieselguh

rF254

Eriochromeb

lack

Tin

5NaO

H050

05038

Brow

nLightb

rown

Polyam

ide11F

254

Eosin

yello

win

5NaO

H50

515

Yello

wLightyellow

Phenylephrine

Silicag

el60F 2

54Glacialaceticacid

+n-bu

tano

l+water

(14

1vvv)

Brom

ocresolgreen

in5

NaO

H025

02521

Lemon

Lightb

lue

[61]

Silicag

el60kieselguh

rF254

Eriochromeb

lack

Tin

5NaO

H050

05041

Brow

nLightb

rown

Bithiono

lSilicag

el60F 2

54Glacialaceticacid

+n-bu

tano

l+water

(14

1vvv)

Brilliant

cresylblue

in5

NaO

H050

05077

Brow

nLightviolet

[61]

Silicag

el60kieselguh

rF254

Brilliant

cresylblue

in5

NaO

H250

25068

Brow

nLightb

rown

Methyldop

a

Silicag

el60

Glacialaceticacid

+n-bu

tano

l+water

(14

1vvv)

Thym

olblue

in2

NaO

H32

327

Brow

nblack

Beige

[62]

Norepinephrine

Brom

othymolblue

in2

NaO

H06

0628

Yello

wbrown

Lightb

lue

Terbutaline

Brilliant

cresylblue

in2

NaO

H20

2017

Lightb

rown

Lightb

lue

Bamethane

Anilin

ebluein2

NaO

H10

1025

Blue

Lightb

lue

Etam

ivan

Anilin

ebluein2

NaO

H50

5018

Whiteblue

Lightb

lue

Phloroglucinol

Silicag

el60

Chloroform

+methano

l(9

1vv)

Brilliant

cresylblue

in2

NaO

H20

2023

Lightb

rown

Lightb

lue

[62]

Pentazocine

Anilin

ebluein2

NaO

H05

0568

White

Lightb

lue

Hexachlorop

hene

Thym

olblue

in2

NaO

H50

5037

Yellowbeige

Beige

Pyrocatechine

Brilliant

greenin

2NaO

H03

0317

Light

orangered

Lightg

reen

Niclosamide

Brilliant

greenin

2NaO

H08

0818

Green

Lightg

reen

Salicylanilide

Brilliant

greenin

2NaO

H08

0816

Green

Lightg

reen

Thym

olBrilliant

greenin

2NaO

H20

2042

Yello

wgreen

Lightg

reen

Eugeno

lSilicag

el60

Benzene

Brom

othymolblue

in2

NaO

H50

5037

Lightyellow

Lightb

lue

[62]







254after





254 it was worse
























254plates









1 aflatoxin B

2 aflatoxin G

1 aflatoxin G

2[98ndash



4 Conclusion









Acknowledgment


References






































































































































Volume 2014

ToxinsJournal of

VaccinesJournal of




















Autoimmune Diseases




Journal of


Pharmaceutics



of



254 The






119865values) can









4 and


4 Furthermore





4 01









2CO3gives


2CO3+ 1



119865= 0138) and











3)2+ saturated


2Cr2O7in 40 H





3)2

Na2CO3 Co(NO







254












Table6Ch

aracteris

ticso

fthe


selected

phenoliccompo

unds

Detecteddrug

Stationary

phase

Mob

ileph

ase

Visualizingreagent

Detectio

nlim

it(120583g)

Detectability

index

(120583gmm

2 )Color

ofspot

Backgrou

ndcoloro

fchromatogram

Reference

Adrenalin

eSilicag

el60F 2

54Glacialaceticacid

+n-bu

tano

l+water

(14

1vvv)

Brilliant

cresylblue

in5

NaO

H010

01010

Brow

nLightviolet

[61]

Silicag

el60kieselguh

rF254

Brom

ocresolgreen

in5

NaO

H025

02547

Lightb

rown

Seagreen

Polyam

ide11F

254

Eosin

yello

win

5NaO

H250

25030

Lightb

rown

Lightyellow

Dop

amine

Silicag

el60F 2

54Glacialaceticacid

+n-bu

tano

l+water

(14

1vvv)

Brom

ocresolgreen

in5

NaO

H010

01015

Orange

Lightb

lue

[61]

Silicag

el60kieselguh

rF254

025

02510

Brow

norange

Seagreen

Polyam

ide11F

254

Eosin

yello

win

5NaO

H075

07517

Lightb

rown

Lightyellow

Metaram

inol

Silicag

el60F 2

54Glacialaceticacid

+n-bu

tano

l+water

(14

1vvv)

Brom

ocresolgreen

in5

NaO

H050

05015

Lemon

Lightb

lue

[61]

Silicag

el60kieselguh

rF254

Eriochromeb

lack

Tin

5NaO

H050

05035

Brow

nLightb

rown

Polyam

ide11F

254

Anilin

eblueinwater

505042

Lightyellow

Blue

Feno

terol

Silicag

el60F 2

54Glacialaceticacid

+n-bu

tano

l+water

(14

1vvv)

Brom

ocresolgreen

in5

NaO

H010

01013

Brow

norange

Lightb

lue

[61]

Silicag

el60kieselguh

rF254

Eriochromeb

lack

Tin

5NaO

H050

05038

Brow

nLightb

rown

Polyam

ide11F

254

Eosin

yello

win

5NaO

H50

515

Yello

wLightyellow

Phenylephrine

Silicag

el60F 2

54Glacialaceticacid

+n-bu

tano

l+water

(14

1vvv)

Brom

ocresolgreen

in5

NaO

H025

02521

Lemon

Lightb

lue

[61]

Silicag

el60kieselguh

rF254

Eriochromeb

lack

Tin

5NaO

H050

05041

Brow

nLightb

rown

Bithiono

lSilicag

el60F 2

54Glacialaceticacid

+n-bu

tano

l+water

(14

1vvv)

Brilliant

cresylblue

in5

NaO

H050

05077

Brow

nLightviolet

[61]

Silicag

el60kieselguh

rF254

Brilliant

cresylblue

in5

NaO

H250

25068

Brow

nLightb

rown

Methyldop

a

Silicag

el60

Glacialaceticacid

+n-bu

tano

l+water

(14

1vvv)

Thym

olblue

in2

NaO

H32

327

Brow

nblack

Beige

[62]

Norepinephrine

Brom

othymolblue

in2

NaO

H06

0628

Yello

wbrown

Lightb

lue

Terbutaline

Brilliant

cresylblue

in2

NaO

H20

2017

Lightb

rown

Lightb

lue

Bamethane

Anilin

ebluein2

NaO

H10

1025

Blue

Lightb

lue

Etam

ivan

Anilin

ebluein2

NaO

H50

5018

Whiteblue

Lightb

lue

Phloroglucinol

Silicag

el60

Chloroform

+methano

l(9

1vv)

Brilliant

cresylblue

in2

NaO

H20

2023

Lightb

rown

Lightb

lue

[62]

Pentazocine

Anilin

ebluein2

NaO

H05

0568

White

Lightb

lue

Hexachlorop

hene

Thym

olblue

in2

NaO

H50

5037

Yellowbeige

Beige

Pyrocatechine

Brilliant

greenin

2NaO

H03

0317

Light

orangered

Lightg

reen

Niclosamide

Brilliant

greenin

2NaO

H08

0818

Green

Lightg

reen

Salicylanilide

Brilliant

greenin

2NaO

H08

0816

Green

Lightg

reen

Thym

olBrilliant

greenin

2NaO

H20

2042

Yello

wgreen

Lightg

reen

Eugeno

lSilicag

el60

Benzene

Brom

othymolblue

in2

NaO

H50

5037

Lightyellow

Lightb

lue

[62]







254after





254 it was worse
























254plates









1 aflatoxin B

2 aflatoxin G

1 aflatoxin G

2[98ndash



4 Conclusion









Acknowledgment


References






































































































































Volume 2014

ToxinsJournal of

VaccinesJournal of




















Autoimmune Diseases




Journal of


Pharmaceutics



of



3)2

Na2CO3 Co(NO







254












Table6Ch

aracteris

ticso

fthe


selected

phenoliccompo

unds

Detecteddrug

Stationary

phase

Mob

ileph

ase

Visualizingreagent

Detectio

nlim

it(120583g)

Detectability

index

(120583gmm

2 )Color

ofspot

Backgrou

ndcoloro

fchromatogram

Reference

Adrenalin

eSilicag

el60F 2

54Glacialaceticacid

+n-bu

tano

l+water

(14

1vvv)

Brilliant

cresylblue

in5

NaO

H010

01010

Brow

nLightviolet

[61]

Silicag

el60kieselguh

rF254

Brom

ocresolgreen

in5

NaO

H025

02547

Lightb

rown

Seagreen

Polyam

ide11F

254

Eosin

yello

win

5NaO

H250

25030

Lightb

rown

Lightyellow

Dop

amine

Silicag

el60F 2

54Glacialaceticacid

+n-bu

tano

l+water

(14

1vvv)

Brom

ocresolgreen

in5

NaO

H010

01015

Orange

Lightb

lue

[61]

Silicag

el60kieselguh

rF254

025

02510

Brow

norange

Seagreen

Polyam

ide11F

254

Eosin

yello

win

5NaO

H075

07517

Lightb

rown

Lightyellow

Metaram

inol

Silicag

el60F 2

54Glacialaceticacid

+n-bu

tano

l+water

(14

1vvv)

Brom

ocresolgreen

in5

NaO

H050

05015

Lemon

Lightb

lue

[61]

Silicag

el60kieselguh

rF254

Eriochromeb

lack

Tin

5NaO

H050

05035

Brow

nLightb

rown

Polyam

ide11F

254

Anilin

eblueinwater

505042

Lightyellow

Blue

Feno

terol

Silicag

el60F 2

54Glacialaceticacid

+n-bu

tano

l+water

(14

1vvv)

Brom

ocresolgreen

in5

NaO

H010

01013

Brow

norange

Lightb

lue

[61]

Silicag

el60kieselguh

rF254

Eriochromeb

lack

Tin

5NaO

H050

05038

Brow

nLightb

rown

Polyam

ide11F

254

Eosin

yello

win

5NaO

H50

515

Yello

wLightyellow

Phenylephrine

Silicag

el60F 2

54Glacialaceticacid

+n-bu

tano

l+water

(14

1vvv)

Brom

ocresolgreen

in5

NaO

H025

02521

Lemon

Lightb

lue

[61]

Silicag

el60kieselguh

rF254

Eriochromeb

lack

Tin

5NaO

H050

05041

Brow

nLightb

rown

Bithiono

lSilicag

el60F 2

54Glacialaceticacid

+n-bu

tano

l+water

(14

1vvv)

Brilliant

cresylblue

in5

NaO

H050

05077

Brow

nLightviolet

[61]

Silicag

el60kieselguh

rF254

Brilliant

cresylblue

in5

NaO

H250

25068

Brow

nLightb

rown

Methyldop

a

Silicag

el60

Glacialaceticacid

+n-bu

tano

l+water

(14

1vvv)

Thym

olblue

in2

NaO

H32

327

Brow

nblack

Beige

[62]

Norepinephrine

Brom

othymolblue

in2

NaO

H06

0628

Yello

wbrown

Lightb

lue

Terbutaline

Brilliant

cresylblue

in2

NaO

H20

2017

Lightb

rown

Lightb

lue

Bamethane

Anilin

ebluein2

NaO

H10

1025

Blue

Lightb

lue

Etam

ivan

Anilin

ebluein2

NaO

H50

5018

Whiteblue

Lightb

lue

Phloroglucinol

Silicag

el60

Chloroform

+methano

l(9

1vv)

Brilliant

cresylblue

in2

NaO

H20

2023

Lightb

rown

Lightb

lue

[62]

Pentazocine

Anilin

ebluein2

NaO

H05

0568

White

Lightb

lue

Hexachlorop

hene

Thym

olblue

in2

NaO

H50

5037

Yellowbeige

Beige

Pyrocatechine

Brilliant

greenin

2NaO

H03

0317

Light

orangered

Lightg

reen

Niclosamide

Brilliant

greenin

2NaO

H08

0818

Green

Lightg

reen

Salicylanilide

Brilliant

greenin

2NaO

H08

0816

Green

Lightg

reen

Thym

olBrilliant

greenin

2NaO

H20

2042

Yello

wgreen

Lightg

reen

Eugeno

lSilicag

el60

Benzene

Brom

othymolblue

in2

NaO

H50

5037

Lightyellow

Lightb

lue

[62]







254after





254 it was worse
























254plates









1 aflatoxin B

2 aflatoxin G

1 aflatoxin G

2[98ndash



4 Conclusion









Acknowledgment


References






































































































































Volume 2014

ToxinsJournal of

VaccinesJournal of




















Autoimmune Diseases




Journal of


Pharmaceutics



of


Table6Ch

aracteris

ticso

fthe


selected

phenoliccompo

unds

Detecteddrug

Stationary

phase

Mob

ileph

ase

Visualizingreagent

Detectio

nlim

it(120583g)

Detectability

index

(120583gmm

2 )Color

ofspot

Backgrou

ndcoloro

fchromatogram

Reference

Adrenalin

eSilicag

el60F 2

54Glacialaceticacid

+n-bu

tano

l+water

(14

1vvv)

Brilliant

cresylblue

in5

NaO

H010

01010

Brow

nLightviolet

[61]

Silicag

el60kieselguh

rF254

Brom

ocresolgreen

in5

NaO

H025

02547

Lightb

rown

Seagreen

Polyam

ide11F

254

Eosin

yello

win

5NaO

H250

25030

Lightb

rown

Lightyellow

Dop

amine

Silicag

el60F 2

54Glacialaceticacid

+n-bu

tano

l+water

(14

1vvv)

Brom

ocresolgreen

in5

NaO

H010

01015

Orange

Lightb

lue

[61]

Silicag

el60kieselguh

rF254

025

02510

Brow

norange

Seagreen

Polyam

ide11F

254

Eosin

yello

win

5NaO

H075

07517

Lightb

rown

Lightyellow

Metaram

inol

Silicag

el60F 2

54Glacialaceticacid

+n-bu

tano

l+water

(14

1vvv)

Brom

ocresolgreen

in5

NaO

H050

05015

Lemon

Lightb

lue

[61]

Silicag

el60kieselguh

rF254

Eriochromeb

lack

Tin

5NaO

H050

05035

Brow

nLightb

rown

Polyam

ide11F

254

Anilin

eblueinwater

505042

Lightyellow

Blue

Feno

terol

Silicag

el60F 2

54Glacialaceticacid

+n-bu

tano

l+water

(14

1vvv)

Brom

ocresolgreen

in5

NaO

H010

01013

Brow

norange

Lightb

lue

[61]

Silicag

el60kieselguh

rF254

Eriochromeb

lack

Tin

5NaO

H050

05038

Brow

nLightb

rown

Polyam

ide11F

254

Eosin

yello

win

5NaO

H50

515

Yello

wLightyellow

Phenylephrine

Silicag

el60F 2

54Glacialaceticacid

+n-bu

tano

l+water

(14

1vvv)

Brom

ocresolgreen

in5

NaO

H025

02521

Lemon

Lightb

lue

[61]

Silicag

el60kieselguh

rF254

Eriochromeb

lack

Tin

5NaO

H050

05041

Brow

nLightb

rown

Bithiono

lSilicag

el60F 2

54Glacialaceticacid

+n-bu

tano

l+water

(14

1vvv)

Brilliant

cresylblue

in5

NaO

H050

05077

Brow

nLightviolet

[61]

Silicag

el60kieselguh

rF254

Brilliant

cresylblue

in5

NaO

H250

25068

Brow

nLightb

rown

Methyldop

a

Silicag

el60

Glacialaceticacid

+n-bu

tano

l+water

(14

1vvv)

Thym

olblue

in2

NaO

H32

327

Brow

nblack

Beige

[62]

Norepinephrine

Brom

othymolblue

in2

NaO

H06

0628

Yello

wbrown

Lightb

lue

Terbutaline

Brilliant

cresylblue

in2

NaO

H20

2017

Lightb

rown

Lightb

lue

Bamethane

Anilin

ebluein2

NaO

H10

1025

Blue

Lightb

lue

Etam

ivan

Anilin

ebluein2

NaO

H50

5018

Whiteblue

Lightb

lue

Phloroglucinol

Silicag

el60

Chloroform

+methano

l(9

1vv)

Brilliant

cresylblue

in2

NaO

H20

2023

Lightb

rown

Lightb

lue

[62]

Pentazocine

Anilin

ebluein2

NaO

H05

0568

White

Lightb

lue

Hexachlorop

hene

Thym

olblue

in2

NaO

H50

5037

Yellowbeige

Beige

Pyrocatechine

Brilliant

greenin

2NaO

H03

0317

Light

orangered

Lightg

reen

Niclosamide

Brilliant

greenin

2NaO

H08

0818

Green

Lightg

reen

Salicylanilide

Brilliant

greenin

2NaO

H08

0816

Green

Lightg

reen

Thym

olBrilliant

greenin

2NaO

H20

2042

Yello

wgreen

Lightg

reen

Eugeno

lSilicag

el60

Benzene

Brom

othymolblue

in2

NaO

H50

5037

Lightyellow

Lightb

lue

[62]







254after





254 it was worse
























254plates









1 aflatoxin B

2 aflatoxin G

1 aflatoxin G

2[98ndash



4 Conclusion









Acknowledgment


References






































































































































Volume 2014

ToxinsJournal of

VaccinesJournal of




















Autoimmune Diseases




Journal of


Pharmaceutics



of







254after





254 it was worse
























254plates









1 aflatoxin B

2 aflatoxin G

1 aflatoxin G

2[98ndash



4 Conclusion









Acknowledgment


References






































































































































Volume 2014

ToxinsJournal of

VaccinesJournal of




















Autoimmune Diseases




Journal of


Pharmaceutics



of





















254plates









1 aflatoxin B

2 aflatoxin G

1 aflatoxin G

2[98ndash



4 Conclusion









Acknowledgment


References






































































































































Volume 2014

ToxinsJournal of

VaccinesJournal of




















Autoimmune Diseases




Journal of


Pharmaceutics



of


4 Conclusion









Acknowledgment


References






































































































































Volume 2014

ToxinsJournal of

VaccinesJournal of




















Autoimmune Diseases




Journal of


Pharmaceutics



of
















































































































Volume 2014

ToxinsJournal of

VaccinesJournal of




















Autoimmune Diseases




Journal of


Pharmaceutics



of

detection progress of selected drugs in tlc (thin layer chromatography)

Documents

detection index

gciv tlc

case of tlc

visualizing effect

limit of detection

direct detection

layer chromatography

hplcuvand gc