spectrophotometric determination of nicotine in

www.wjpps.com Vol 3, Issue 8, 2014.

1327

Omara et al. World Journal of Pharmacy and Pharmaceutical Sciences

SPECTROPHOTOMETRIC DETERMINATION OF NICOTINE IN

CIGARETTE TOBACCO AND BIOLOGICAL SAMPLES OF

SMOKERS

Hany A. Omara* and Salma M. M. Attaf

Chemistry Department, Faculty of Science, Sirte University, Sirte, Libya.

ABSTRACT

A new simple, accurate, sensitive and economical procedure for the

estimation of nicotine (indirect) methods (A and B) for the

determination of nicotine in bulk sample and in biological samples are

described. The first method is based on the oxidation of nicotine by N-

bromosuccinimide (NBS) and determination of the unreacted NBS by

measurement of the decrease in absorbance of methyl orange dye

(MO) at a suitable λmax = 507 nm. The absorbance concentration plot is

linear over the range (0.1-4.8 µg/ml). The second method is based on

oxidation of nicotine by cerric sulphate in acidic medium, and

determination of the unreacted oxidant by measuring the decrease in absorbance using

amaranth dye; (AM) at a suitable λmax (530 nm), respectively. Regression analysis of Beer's

law plots showed good correlation in the concentration ranges (0.3-5.9 µg/ml), respectively.

The limits of detection as well as quantification are reported. Sex replicate analyses (n=6) of

solutions containing three different concentrations of nicotine was carried out. The percent

error and the RSD values have been reported. The proposed methods were applied to the

determination of nicotine in cigarette tobacco present in Libya and in biological samples

(spiked human plasma and urine) and the results demonstrate that the method is equally

accurate and precise as found from the t- and F-values. The reliability of the method was

established by recovery studies using standard-addition technique.

Key Words: Nicotine; Spectrophotometry; Oxidation reaction; N-bromosuccinimide; Cerric

sulphate, Cigarette tobacco in Libyan markets; Spiked human plasma and Urine.

WWOORRLLDD JJOOUURRNNAALL OOFF PPHHAARRMMAACCYY AANNDD PPHHAARRMMAACCEEUUTTIICCAALL SSCCIIEENNCCEESS SSJJIIFF IImmppaacctt FFaaccttoorr 22..778866

VVoolluummee 33,, IIssssuuee 88,, 11332277--11334400.. RReesseeaarrcchh AArrttiiccllee IISSSSNN 2278 – 4357

Article Received on 17 May 2014, Revised on 12 June 2014, Accepted on 23 July 2014

*Correspondence for Author

Dr. Hany A. Omara

Chemistry Department, Faculty

of Science, Sirte University,

Sirte, Libya


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INTRODUCTION

Nicotine (pyridine, 3-(1-methyl-2-pyrrolidinyl), (S) is one of the highly toxic tobacco

alkaloids present in tobacco leaves and cigarette smoke [1]. Nicotine appears to be a

promising tracer for environmental tobacco smoke (ETS) because of its specificity for

tobacco [2]. It is also a systemic and contact insecticide and is also used as a drug and in

chemical [3]. The threshold limit value reported for nicotine is 0.05 mg/m3 [1]. The most

important symptoms of exposure to nicotine are bronchitis, emphysema, cyanosis, and

exenation of the central nervous system. Excessive smoking has been implicated in lung

cancer, bladder cancer, and cancer of the larynx and oesophagus [4].

The determination of total nicotine alkaloids is of particular importance to the tobacco

industry and in the area of toxicology. Various analytical methods such as GC [5], TLC [6],

HPLC [7-9], ASS [10], capillary electrophoresis [11], radio immuno assay [12], GC-MS [13] and

circular dichroism spectropolarimetry [14] have been reported. Most spectrophotometric

methods reported are based on cleavage of the pyridine ring with cyanogen bromide or

chloride and condensation of the resulting compound with p-amino benzoic acid [15] and

diethyl thiobarbituric acid [16] and other spectrophotometric methods [17-24], determination of

outdoor Tobacco smoke exposure by distance From a smoking source [25 ], an

electrochemically [26].

Many of these methods involve a prolonged extraction procedure prior to the determination

of nicotine and the use of highly toxic cyanide [15, 16]. These methods were sophisticated to

perform and/or time consuming. The oxidation reaction between NBS or Cerric in acidic

medium and nicotine have not been investigated yet but for other drugs [27-37].

EXPERIMENTAL

Apparatus

All the spectral measurement were made using double-beam UV/Vis spectrophotometer

(Biotech Engineering Ltd., UK), with wavelength range 190 –1100 nm, spectral bandwidth

2.0 nm, with scanning speed 400 nm/min, equipped with 10 mm matched quartz cells. A

thermostat water bath, Buchi 461 water bath, Schwiz (France) was used to carry out the

temperature studies and Magnetic Mix. 100, Thermo Scientific, UK.


1329


Material and reagents

All chemicals used were of analytical reagent grade of the best available quality. and all

solutions were freshly prepared in doubly distilled water.

Standard nicotine solution (Merck) was prepared by dissolving 0.01 g of pure nicotine in 50

ml of bi-distilled water and complete to 100 ml with bi-distilled water to obtain the working

standard solution of 100 µg/ml.

A stock solution of 1.0 x 10-3 M NBS (Aldrich Co., Ltd., Gillingham-Dorst, Germany) was

freshly prepared by dissolving 0.0177 g of NBS in a least amount of warm water in a 100 ml

measuring flask and then diluted with distilled water to the mark.

A stock solution of 1.0 x 10-3 M cerric sulphate (Aldrich Co., Ltd., Gillingham-Dorst,

Germany) was freshly prepared by dissolving appropriate weight of Ce (VI) in a least amount

of warm water in a 100 ml measuring flask and then diluted with distilled water to the mark.

A 2.0 M of HCl was prepared by adding exact volume from stock concentrated acid to

bidistilled water in 500 ml measuring flask (Merck, Darmstadt, Germany, sp. gr. 1.18%,

37%) to 100 ml with distilled water.

A solution of 2.0 M H2SO4, was prepared by adding exact volume from stock (98%)

concentrated acid to bidistilled water in 500 ml measuring flask, and standardized as recorded [38].

Aqueous solutions of dyes (1.0 x 10-3 M) methyl orange (MO) (Merck), (1.0 x 10-3 M)

amaranth (AM) (Merck) were prepared by dissolving in an appropriate weight in 100 ml

bidistilled water.

General procedure

For the first method, (method A) depends on oxidation of nicotine by addition of 0.01-4. 8 ml

(100 µg/ml) to 1.8 ml of 1.0 x 10-3 M NBS containing 1.1 ml HCl, 2.0 M. The solution was

heated in a water bath at 80±1 oC for 4.0 min. The solution was cooled and 1.7 ml (1.0 x 10-3

M) of MO was added, the volume was completed to 10 ml with bidistilled water. For the

second method (method B) depends on oxidation of nicotine by addition of 0.01-5.9 ml

nicotine (100 µg/ml) to 1.5 ml of 1.0 x 10-3 M Ce (VI) containing 1.2 ml of 2.0 M H2SO4 was

added. The solution was heated in a water bath at 80±1 oC for 5.0 min, the mixture of acidic

solution was cooled and 1.6 ml (1.0 x 10-3 M) of AM was added, the volume was completed

to 10 ml with bidistilled water. The decrease in color intensity of dye was measured

spectrophotometrically against a blank solution containing the same constituent except drug

treated similarly, at their corresponding λmax 507 (method A), 530 nm (method B),


1330


respectively. The concentration range was determined in each case by plotting the

concentration of nicotine against absorbance at the corresponding maximum wavelengths.

Extraction and determination of nicotine in cigarette tobacco [39]

Weigh 10 g of cigarettes leaves in beaker. Add 100 ml NaOH solution and stir very well for

15 min, filter in Buchner using glass wool and press the cigarettes very well by using other

beaker, transfer the cigarettes again to beaker, add 30 ml DW and stir and filter again, collect

the filtrate together, (if there is any impurities re-filter), transfer the filtrate to the SF and

extract by 25 ml ether, repeat the extraction 3 times, gather the 4 filtrates in conical flask, dry

by using 1.0 teaspoon anhydrous potassium carbonate, filter. evaporate ether on water bath,

(avoid extra heat because nicotine is hydrolyzed by extreme heating), after evaporation of

ether add 4.0 ml methanol to dissolve the resulted oil, to the filtrate 14 ml of 1.0 mol/L HCl

was added and the solution was made up to 100 ml with water. An aliquot of this solution

was taken and analyzed as described above. The results are shown in Table 2.

Procedure for spiked human urine

A 50 ml volume of nicotine – free human urine taken in a 125 ml separating funnel was

spiked with 2.5 mg nicotine. Ten milliliters of 1.0 M NaOH was added, mixed and kept aside

for 3.0 min. Then, 25 ml of ethyl acetate was added, shaken well for about 15 min and the

upper organic layer was collected in a beaker containing anhydrous sodium sulphate. The

water-free organic layer was transferred into a dry beaker and solvent removed by

evaporation on a hot water bath. The dry residue was dissolved in glacial acetic acid and

transferred into a 25 ml calibrated flask, and diluted to the mark with the same acid. The

resulting solution equivalent to 100 µg/ml nicotine was diluted with the same acid and

analyzed by following the procedure described above.

Procedure for spiked human plasma samples

Aliquots of 1.0 ml of plasma were spiked with different concentration levels of nicotine. The

spiked plasma samples were treated with 0.1 ml of 70% perchloric acid and vortexes for 1.0

min. The samples were centrifuged for 20 min at 13000 rpm. The supernatants were

transferred into test tubes and neutralized with 1.0 M NaOH solution.

Stoichiometric relationship

The stoichiometry of the reaction between nicotine and the oxidants at the selected conditions

were established by the molar ratio method. In this method 1.0 ml of 1.0 x 10-3 M NBS


1331


(method A) 1.0 x 10-3 M Ce (VI) (method B) is kept constant and variable concentrations

(0.1-6.0 ml) of nicotine (1.0 x 10-3 M) were added. The absorbance was measured at λmax

against blank solution prepared in the same manner. The absorbance values were then plotted

against the molar ratio [D]/[O] as shown in Figure 6.

RESULTS AND DISCUSSION

First Method (A)

The proposed spectrophotometric methods are based on the reaction between nicotine and

measured excess of NBS and subsequent determination of the latter by reacting it with a fixed

amount of MO dye, and measuring the absorbance at 507 nm. The reaction takes place

completely after 4.0 min., was heated in a thermostat water bath at 80±1 oC. This method

make use of the bleaching action of NBS on the dye, the decolorization being caused by the

oxidative destruction of the dye. nicotine when added in increasing concentrations to a fixed

concentration of NBS consumes the latter and there will be a concomitant decrease in the

concentration of NBS. When a fixed concentration of dye is added to decreasing

concentrations of NBS, a concomitant increase in the concentration of dye is obtained.

Consequently, a proportional increase in the absorbance at the respective λmax is observed

with increasing concentrations of nicotine, the color remains constant for at least 48 h.

Second Method (B)

The method involves two steps namely:

Oxidation of nicotine with Ce(SO4)2 in acidic medium by heating in water bath 80±1 oC.

Determination of unreacted oxidant by measuring the decrease in absorbance of dye at a

suitable λmax.

Nicotine + Ce(SO4)2 42. SOHDil Oxidized products + Ce (III) + Ce(SO4)2

(Unreacted)

Unreacted Ce(SO4)2 + Amaranth dye → Ce (III) + Unreacted Amaranth dye

Pink colour, λmax 530 nm

The influence of each of the following variables on the reaction was tested.

Effect of NBS concentration

The influence of NBS concentration was studied in the range from 10-4 - 10-2 M, as final

concentration. The optimum results were obtained with 1.8 ml of 1.0 x 10-3 M.


1332


0.0

0.2

0.4

0.6

0.8

0.0 0.5 1.0 1.5 2.0 2.5 3.0

ml added of NBS

Abs

orba

nce

MO

0.0

0.2

0.4

0.6

0.8

1.0

0.0 0.5 1.0 1.5 2.0 2.5 3.0 3.5

ml added of Ce(VI)

Abs

orba

nce

AM

Effect of cerric sulphate concentration

The influence of Ce (VI) concentration was studied in the range from 10-4 - 10-2 M, as final

concentration. The optimum results were obtained with 1.5 ml of 1.0 x 10-3 M; higher

concentration of Ce(SO4)2 caused the color to disturbed.

Effect of acid medium

The different types of acid were examined (HCl, HClO4, H2SO4, H3PO4, CH3COOH and

HNO3). The most suitable acid to achieve maximum yield of redox reaction was found to be

1.1 ml of 2.0 M HCl for (method A) and 1.2 of 2.0 M H2SO4 was added on using AM, for

(method B) Figure 3.

Fig. 1. Effect of ml added of NBS (1 x 10-3 M);

nicotine (4.0 µg/ml) 1.1 ml of 2.0 M

hydrochloric acid, reaction temperature

80±1 °C; reaction time: 4.0 min

Fig. 2. Effect of ml added of Ce (VI) (1 x 10-3 M)

(10 M) on 4.0 µg/ml nicotine, 1.2 ml of 2.0 M

sulphoric acid; reaction temperature 80±1 oC;

reaction time: 5.0 min

Effect of temperature and time

The reaction takes place completely after 20 min at room temperature 25±1 oC. The oxidation

process of nicotine with NBS is catalyzed by heating in a thermostat water bath at 80±1 oC

for 4.0 min. using MO dye. The oxidation process of nicotine for the (method B) Ce(SO4)2 in

acidic medium was catalyzed by heating in a thermostat water bath of 80±1 oC for 5.0 min,

using AM. After oxidation process, the solution must be cooled at least for 2.0 min before

addition of dye.

Effect of sequence of additions

The effect of sequence of additions on the oxidation process of nicotine was studied by

measuring the absorbance of solution prepared by different sequence of additions against a


1333


blank solution prepared in the same manner. Experiments showed that (Oxidant-Acid-Drug)

for both methods, gave the best results.

Effect of dye concentration

The optimum volume of dye used for production of maximum color intensity was 1.7 ml of

1.0 x 10-3 M (MO) for (method A), 1.6 ml of 1.0 x 10-3 M (AM) for (method B) Figure 4. The

effect of time after the addition of dye indicated that shaking for 1.0 min was sufficient to

give reliable results for all dyes. The color remains constant for at least 24 h.

Stoichiometric ratio

The stoichiometry of the reaction between nicotine and the oxidant at the selected conditions

was established by the molar ratio method. In this method 1.0 ml of 1.0 x 10-3 M NBS for

(method A), 1.0 ml of 1.0 x 10-3 M Ce(VI) for (method B) is kept constant and variable

concentrations (0.1 - 6.0 ml) of nicotine (1.0 x 10-3 M) were added using micropipette. The

absorbance was measured at λmax against blank solution prepared in the same manner. The

absorbance values were then plotted against the molar ratio [D]/[O]. The stoichiometry of

[D]/[O] at the selected conditions showed that the inflection of the two straight lines at 0.63

for (method A) 0.71 AM for (method B), Figure 6.

Reproducibility of the method

The reproducibility of the method was assessed by carrying out seven replicate analyses of a

solution containing 3.0 µg/ml of nicotine in a final solution volume of 25 ml. The standard

deviation and relative standard deviation of absorbance values were found to be ±0.011 and

0.56 %, respectively.

Analytical data

Beer's law was verified up to (0.1-4.8 µg/ml) for method A, and (0.3-5.9 µg/ml) for method B

as shown in (Figure 5) and Ringbom limits, molar absorptivities, Sandell sensitivities,

regression equations and correlation coefficients were calculated and recorded in (Table 1).

The limits of detection (K=3) and quantitation (K=10) were established according to IUPAC

definitions [40] are recorded in (Table 1).

In order to determine the accuracy and precision of the methods, solution containing three

different concentrations of nicotine were prepared and analyzed in six replicates (Table 2).


1334


0.0

0.1

0.2

0.3

0.4

0.5

0.6

0.7

0.8

0.9

0.0 0.5 1.0 1.5 2.0 2.5 3.0

ml added of acid

Abs

orba

nce

NBSCerric

0.0

0.1

0.2

0.3

0.4

0.5

0.6

0.7

0.8

0.9

0.0 0.5 1.0 1.5 2.0 2.5 3.0

ml add of dyes

Abs

orpa

nce

MOAM

Fig. 3. Effect of ml added of acid (2.0 M HCl

using NBS, 2.0 M H2SO4 using Ce(VI));

nicotine (4.0 µg/ml), 1.1 ml of (1 x 10-3 M)

NBS, reaction temperature 80±1 °C; reaction

time: 4.0 min

Fig. 4. . Effect of ml added of dyes on

4.0 µg/ml nicotine, (MO in case of NBS,

AM in case of Ce(VI)), reaction

temperature 80±1 °C; reaction time:

4.0 min

Stability of colour

The pink or red colour formed was stable for at least 24 h under optimum conditions.

Validation method

The proposed method was successfully applied to determine nicotine in cigarettes tobacco

and in biological samples (spiked human plasma and urine). The accuracy of the proposed

methods is evaluated by applying standard addition technique, in which variable amounts of

the nicotine were added to the previously analyzed portion of extracted nicotine from

cigarettes tobacco and in biological samples (spiked human plasma and urine). The results

recorded in (Table 3), The results are in good agreement with those obtained by the reported

method [17-19], by Student’s t-test (for accuracy), and variance ratio F-test (for precision) [41],

at 95% confidence level as recorded in (Table 4). The results showed that the t- and F- values

were lower than the critical values indicating that there was no significant difference between

the proposed and reported methods. The proposed method was more accurate with high

recoveries.


1335


0.0

0.2

0.4

0.6

0.8

1.0

1.2

0.0 1.0 2.0 3.0 4.0 5.0 6.0 7.0

[D] µg ml-1

Abs

orba

nce

NBSCerric

0.0

0.2

0.4

0.6

0.8

1.0

1.2

1.4

0.0 0.5 1.0 1.5 2.0 2.5 3.0

[D]/[O]

Abs

orba

nce

NBSCe(VI)

Fig. 5. Calibration curve of nicotine µg/ml,

using NBS and Ce(VI) (1.0 x 10-3 M) and

dyes, MO and AM (1.0 x 10-3 M), reaction

temperature 80±1 °C; reaction time: 4.0 min

Fig. 6. Molar ratio method for nicotine

(1.0 x 10-3 M) using NBS and Ce(VI)

(1.0 x 10-3 M) and dyes, MO and AM

(1.0 x 10-3 M), reaction temperature

80±1 °C; reaction time: 4.0 min

Table 1. Optical and regression characteristics of nicotine for the proposed method.

Parameters

Proposed method NBS Ce(SO4)2

λmax (nm) 507 530 Stability / h 24 48 Beer’s law limits (µg/ml) 0.1 – 4.8 0.3 – 5.9 Ringbom limits (µg/ml) 0.3 – 4.5 0.6 – 6.3 Molar absorptivity (L/ mol.cm) 2.76 x 104 3.03 x 104 Sandell sensitivity (ng/cm2) 5.88 5.35 Detection limits (µg/ml) 0.058 0.052 Quantitation limits (µg/ml) 0.193 0.173 Regression equation a: Slope (b)

0.170

0.187

Intercept (a) 8.4 x 10-4 8.1 x 10-4 Correlation coefficient (r) 0.9996 0.9998 RSD b % 0.59 0.46 Calculated t-values (2.57) c 0.49 0.77 Calculated F-test (5.05) c 2.27 1.84 Stiochiometric ratio [D]/[O] 1.0 : 0.63 1.0 : 0.71

a A = a + bC where C is concentration of nicotine in µg/ml and A is absorbance.

b Relative standard deviation for six determinations. c Values in parentheses are the theoretical values for t- and F values at 95% confidence limits

and five degrees of freedom.


1336


Table 2. Evaluation of the accuracy and precision of the proposed procedures for

nicotine.

Reagent Taken µg/ml

Found µg/ml

Recovery %

RSD a %

RE b

% Confidence Limitsc

NBS

1.0 2.0 4.0

1.02 1.98 4.01

102.0 99.00 100.25

0.55 0.69 0.76

1.23 1.60 0.82

1.02 ± 0.0125 1.98 ± 0.0316 4.01 ± 0.0329

Ce(SO4)2

1.0 3.0 5.0

0.97 2.98 5.01

97.0 99.33 100.2

0.73 0.84 0.91

1.39 1.07 1.02

0.97 ± 0.0135 2.98 ± 0.0318 5.01 ± 0.0510

a Relative standard deviation for six determinations. b Relative error.

c 95% confidence limits and five degrees of freedom

Table 3. Determination of nicotine in cigarette tobacco in Libyan markets and biological

samples using standard addition technique.

Sample

Taken µg/ml

NBS Ce(VI) Added µg/ml

Found* µg/ml

Recovery %

Added µg/ml

Found* µg/ml

Recovery %

American Legend a

**

1.0

0.0 1.03 103.00 0.0 0.96 96.00 1.0 2.05 102.50 1.0 2.01 100.5 2.0 2.97 99.00 2.0 3.02 100.66 3.0 4.01 100.25 3.0 3.96 99.00

Marlloborro b **

1.0

0.0 0.97 97.00 0.0 0.98 98.00 1.0 2.05 101.00 1.0 2.07 103.5 2.0 2.99 99.00 2.0 2.96 98.66 3.0 3.96 99.50 3.0 3.99 99.75

Urine

1.0

0.0 0.98 98.00 0.0 1.01 101.0 1.0 1.98 99.00 1.0 1.96 98.00 2.0 2.97 99.00 2.0 2.95 98.33 4.0 4.03 100.75 4.0 4.07 101.75

Spiked human plasma

1.0

0.0 0.98 98.00 0.0 0.95 95.00 1.0 2.07 103.50 1.0 1.96 98.00 2.0 2.96 98.66 2.0 2.98 99.33 4.0 4.03 100.75 4.0 4.06 101.5

* Average of six determinations. ** Amount of sample: 0.1 gm. After treatment diluted to 50 ml, nicotine found in µg/ml of the

final solution. a Cigarette smoke, made in EU, under the authority of the Trade mark Owners, by KTG, INC.


1337


b. Cigarette smoke, Karelia light, Karelia tobacco company INC, made in Greece, nicotine

content o.7 mg.

Table 4. Determination of nicotine in cigarette tobacco in Libyan markets and in

biological samples using the proposed and reported methods.

Samples

NBS Ce(VI) Reported method[17] Recovery

% t-

test F-

value Recovery

% t-

test F-

value American Legend a * 98.80 1.22 2.49 97.77 0.82 2.33 96.55 Marlloborro b 99.17 0.89 1.99 98.97 0.99 1.87 98.71 Urine 97.89 1.08 2.07 98.10 1.16 2.24 97.07 Spiked human plasma 97.19 0.96 2.55 96.11 0.78 2.19 95.06

Theoretical value for t- and F- values for five degrees of freedom and 95% confidence limits

are 2.57 and 5.05, respectively.

* Amount of sample: 0.1 gm. After treatment diluted to 50 ml, nicotine found in µg/ml of the

final solution. a Cigarette smoke, made in EU, under the authority of the Trade mark Owners, by KTG, INC. b Cigarette smoke, Karelia light, Karelia tobacco company INC, made in Greece, nicotine

content o.7 mg.

CONCLUSION

The proposed method was advantageous over other reported visible spectrophotometric and

colorimetric methods, related to their high reproducibility, high sensitivity, less time

consuming and using simple and inexpensive reagents. Moreover, these methods allowed the

determination of nicotine up to 0.1 µg/ml, in addition to simplicity, rapidity, precision and

stability of colored species for more than 24 h.

REFERENCES

1. Rodricks, JV. Calculated Risks; Cambridge University Press: Cambridge, 1992; 51.

2. Jones FE. Toxic Organic Vapors in the Work Place; Lewis Publishers: London, CRC

Press Inc.: New York, 1994.

3. Hamm JG. The Handbook of Pest Control; Frederick Fell Publishers Inc.: New York,

1982.

4. Newcomb PA, Storer BE, Marcus PM. Cancer Res. 1995; 55: 4906.


1338


5. Drummer OH., Horomidis SK., Sophie-Syrjanen ML., Tippett P., J. Anal. Toxicol. 1994;

18: 134.

6. Romano G., Caruso G., Masumarra G., Povone D., Guciani G., J. Planer Chromatogr.

1994; 7: 233.

7. Pichini S., Altiere L., Passa AR., Rosa M., Zuccaro P., Pacifici R., J. Chromatogr. A.

1995; 697: 383.

8. Yang X., Changjia W., Xiaopan Z., Haiyan C., Qi Z., Weitao S., Hui W., Qinglei Z. and

Lan D., Fast and selective extraction of nicotine from human plasma based on magnetic

strong cation exchange resin followed by liquid chromatography–tandem mass

spectrometry. Anal. & Bioanal. Chem. 2011; 400(2): 517-26.

9. Adnan MM., Ahmad AG., and Khaled WO., J. Chrom. Scie., 2009; 47: 273-81.

10. Lang HY., Xie ZH., Lei ZH., Li HE., Xi B., Dauxe XB., Ziran Kexueban. 1994; 24: 215.

11. Yang SS., Smetena I., Chromatographia 1995; 40: 375.

12. Pacifi R., Aetieri F., Gandini L., Lenzi A., Passa R., Pichini S., Rosa, M., Zuccaro P.,

Environ. Res. 1995; 69.

13. Koyano MKO., Oike Y., Goto S., Endo OW., Ikuo FK., Matsushita H., J. Toxicol.

Environ. Health. 1996; 42: 263.

14. Atkinson MV., Soon MH., Purdie N., Anal. Chem. 1984; 56: 1947.

15. Kodicek E., Reddi KK., Nature. 1951; 168: 475.

16. Smith CL., Cooke M., Analyst. 1987; 112: 1515.

17. Rai M., Ramachandran KN., Gupta VK., Analyst. 1994; 119: 1883.

18. Li Z., Zhang X., Zhou H., Song H., Fenxi Hauxue. 1986; 14(1): 60.

19. Farrag NM., Said AA., Diab AM., Defrawi EM., J. Pharm. Sci. 1988; 29(1): 439.

20. Yunyou Z., Huapeng Y., Li Z., Hongwei X., Lian W., Junyong S., Lun W., A new

spectrofluorometric method for the determination of nicotine base on the inclusion

interaction of methylene blue and cucurbit[7]uril. Microchimica Acta 2009; 164(1-2): 63-

68.

21. Eduardo CF., Daniela MO., Maria EP., Marco AZ., On-line molecularly imprinted solid-

phase extraction for the selective spectrophotometric determination of nicotine in the

urine of smokers, Analytica Chimica Acta. 2009; 635(1, 2): 102–107.

22. Al-Tamrah SA, Spectrophotometric determination of nicotine, Analytica Chimica Acta,

1999; 379(1–2): 75–80.

23. Shuqin G., Lifu L., , Xilin X., Zhiyuan Z., Nan D., Jiangfeng D., Determination of

nicotine in tobacco with second-order spectra data of charge-transfer complex in ethanol–


1339


water binary solvents processed by parallel factor analysis, Spectrochimica Acta Part A:

Molec. and Biom. Spect., 2010; 75 (5): 1540–45.

24. Anupama A., Rachana R., Sunita G. and Gupta V. K., J. Chin. Chem. Soc., 2004; 51:

949-53.

25. Jihee H., and Kiyoung L., Nicotine & Tobacco Research., 2013; 15 (12): 2045-52.

26. Jichao L., Fengmei H., Yong C., An electrochemical method for high sensitive detection

of nicotine and its interaction with bovine serum albumin, Electrochemistry

Communications., 2012; 24: 93–96.

27. El-Didamony AM, Monir ZS, Saleem NO., Spectrophotometric determination of some

analgesic drugs in pharmaceutical formulations using N-bromosuccinimide as an

oxidant., J. Asso. Ara. Univ. Basic Appl. Sci., Article in press (2013).

28. Nafisur R., Manirul SK, Syed NH, Habibur R., Optimized and validated

spectrophotometric methods for the determination of amiodarone hydrochloride in

commercial dosage forms using N-bromosuccinimide and bromothymol blue, J. of Saudi

Chem. Soci., Article in press (2013).

29. Nafisur R, Sayed-Najmul HA, Spectrophotometric method for the determination of

verapamil hydrochloride in pharmaceutical formulations using N-bromosuccinimide as

oxidant, Il Farmaco, 2004; 59(7): 529–36.

30. Amin AS, Ahmed IS, Dessouki HA, Gouda EA., Utility of oxidation–reduction reaction

for the determination of ranitidine hydrochloride in pure form, in dosage forms and in the

presence of its oxidative degradates, Spectrochimica Acta Part A: Mole. and Biom.

Spect., 2003; 59(4): 695–03.

31. Omara HA., Ahmed HA, El-Mahdy AA. and Musbah SA., New spectrophotometric

determination of azithromycin in pure and dosage forms using N-bromosuccinimide and

potassium permanganate as oxidants, world J. of pharm. and pharmace. Scie.., 2014; 3(4):

100-12.

32. Mohamed FA., Mohamed HA., Hussein SA., Ahmed SA., A validated spectrofluorimetric

method for determination of some psychoactive drugs, J. of Pharm. and Biomed. Anal.,

2005; 39(1–2): 139–46.

33. Aniruddha G., Rumpa S., Kakali M., Pintu S., Sumanta KG., Susanta M., Subhendu SB.,

Bidyut S., Rate enhancement via micelle encapsulation for room temperature metal

catalyzed Ce(IV) oxidation of p-chlorobenzaldehyde to p-chlorobenzoic acid in aqueous

medium at atmospheric pressure., J. of Molec. Liq., 2014; 190: 81–93.


1340


34. Hisham EA., Magda ME., El-Sayed HM., Ayad MM., Spectrophotometric and

spectrofluorimetric methods for analysis of acyclovir and acebutolol hydrochloride.,

Spectrochimica Acta Part A: Molec. and Biomol. Spect., 2007; 66(1): 106–10.

35. Kalsang T., Kanakapura B., Hosakere DR., Kanakapura BV., Spectrophotometric

determination of isoxsuprine hydrochloride using 3-methyl-2-benzothiazolinone

hydrazone hydrochloride in spiked human urine and pharmaceuticals. Talanta., 2010;

81(4–5): 1216–23.

36. Yongjun M., Min Z., Xiaoyong J., Ziyu Z., Xiulan T., Hui C., Flow-injection

chemiluminescence assay for ultra-trace determination of DNA using rhodamine B–

Ce(IV)-DNA ternary system in sulfuric acid media., Analytica Chimica Acta, 2004;

501(1, 9): 25–30.

37. Padmarajaiah N., Narayanan A., Anandamurthy S., Nelligere AC., Anantharaman S.,

Kolar VR., Amaranth dye in the evaluation of bleaching of cerium (IV) by antioxidants:

Application in food and medicinal plants, Spectrochimica Acta Part A: Molecular and

Biomolecular Spectroscopy. 2012; 95: 505–510.

38. Basset J, Jeffery GH & Mendham J, Vogel's Text Book of Quantitative Inorganic

Analysis, 1978; 308.

39. Pavia, DL., Lampman GM. and Kriz, GS. JR., Introduction to organic laboratory

technique, W. B. Saunders Co., Philadelphia. 1976; 50-54.

40. IUPAC. "Nomenclature, symbols, units and their usage in spectrochemical analysis II

data interpretation analytical chemistry division", Spectrochim. Acta, B., 1978; 33, 241-

45.

41. Miller JC and Miller JN, "Statistics in Analytical Chemical 3rd Ed. Ellis Horwood

Chichester", 1993.

spectrophotometric determination of nicotine in

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