quality control and standardization of quercetin in herbal

Kagawad et al. Futur J Pharm Sci (2021) 7:176 https://doi.org/10.1186/s43094-021-00327-y

REVIEW

Quality control and standardization of Quercetin in herbal medicines by spectroscopic and chromatographic techniquesPooja Kagawad, Shankar Gharge, Kadambari Jivaje, Sushmita I. Hiremath and Shailendra S. Suryawanshi*

Abstract

Background: Herbal medicines and their preparations have been mostly used since from thousands of years in all developing and developed countries in the primary health care of society and community. Flavonoids are the class of polyphenolic compounds, which are mainly distributed throughout the plant kingdom. Quercetin is a flavonoid which shows major pharmacological activities and effectively used for the management and treatment of various diseases and disorders. Many herbal medicines and their formulations containing Quercetin are available in market and hence quality control of Quercetin in is very important and essential in manufacturing industries

Main body of the abstract: We have reviewed various scientific research published on quality control analysis and standardization of Quercetin in its isolated form, extract or any other herbal or polyherbal preparation. We have mainly focused on the spectroscopic and chromatographic methods for qualitative and quantitative analysis of Quercetin and they were comprehensively presented in the present review work.

Short conclusion: The present review concludes that the spectroscopic and chromatographic methods play great role in the quality control and standardization of Quercetin in its isolated form, extract and in its herbal and polyherbal preparation.

Keywords: Quality control, Standardization, Herbal medicines, Quercetin, Flavonoids

© The Author(s) 2021. Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/.

BackgroundHerbal medicine and most of their preparations have been widely used from the thousands of years in all devel-oping and developed countries in the primary health care of society and community. They have great demand due its safety, efficacy with minimum side or adverse effects. India is considered as well recorded as well practiced knowledge country of traditional herbal medicine. The Indian systems of medicine mainly composed of Ayur-veda, Siddha and Unani system of medicines. Traditional

herbal medicines are generally naturally occurring and derived from plants with very less or no requirement of industrial methodology in its final production and used to treat illness, dieses or disorders in local or regional healing practices [1].

Many herbal medicines and phytomedicines are introduced into the market for primary health care of communities without any mandatory safety or toxi-cological evaluation and quality control parameters regarding the effect of drug. Many countries who are dealing with manufacturing and production of herbal medicines are lacking with availability of effective machineries, laboratories for effective quality control testing to regulate manufacturing practices and quality

Open Access

Future Journal ofPharmaceutical Sciences

*Correspondence: [email protected] of Pharmaceutical Chemistry, KLE College of Pharmacy Belagavi, KLE Academy of Higher Education and Research, Belagavi, Karnataka 560010, India

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of 12Kagawad et al. Futur J Pharm Sci (2021) 7:176

standards of the herbal preparations [2]. Quality con-trol is one of the very important and essential steps in the manufacturing of herbal preparations as quality of product affects the safety and efficacy of medicines. Quality control is mainly applied for both raw materials along with excipients used and finished product. The quality of the raw materials and other material used as excipients used in the manufacturing of phytomedi-cines need to be tested and evaluated before production of its finished product. Modern analytical instruments play great role in the quality control and standardiza-tion of Herbal Medicines. Especially the spectroscopic and chromatographic methods play essential role in quality control and analytical validation of herbal prep-arations [3].

Flavonoids are the class of polyphenolic compounds, which are mainly distributed throughout the plant kingdom. There are number of flavonoids are known. Flavonoids, as a major active constituent, shows best various pharmacological activities Rutin and logical activities like anti-inflammatory, anti-allergic anti-oxi-dant effects. Quercetin possesses antioxidant activity and reduces low density lipoproteins oxidation level. Quercetin and rutin are the some important flavonoids known for its, anti-inflammatory, anti-allergic, anti-thrombic, anti-spasmodic anti-cancer, and hepatopro-tective properties [4]. Main flavonoid constituents like quercetin (Fig. 1) quercetin is a flavonoid chemically known as 5.7,3′-4′ tetra hydroxy flavanol, which shows a major pharmacological activity like hepatoprotective activity, anti-spasmodic, anti-inflammatory, etc. And mainly belongs to polyphenolic derivatives [5].

Main textMany herbal medicines and their formulations con-taining Quercetin are available in market. We have reviewed various scientific research published on qual-ity control analysis and standardization of Querce-tin in its isolated form, extract or any other herbal or polyherbal preparation. In the present review arti-cle, we have mainly focused on the spectroscopic and

chromatographic methods. The details of methodology and interpretation of results were presented as below.

Spectroscopic techniquesV.C. Yeligar et al., have reported the validation of UV spectrophotometric method for simultaneous estima-tion of Melatonin and Quercetin in liposome formula-tion by calibration curve method using methanol as a solvent. The UV spectra run in the range of 200–400 nm. The λmax of standard solution of melatonin (10 µg/ml) and Quercetin (10 µg/ml) was found to be 276 nm and 372 nm. The absorption and absorptivity coefficient were found by simultaneous equation method [6].

Singh Upendra et al., have validated UV spectrometric methods for the simultaneous estimation of Quercetin and Silymarin using double beam UV–Vis spectropho-tometer by preparing the standard solution of 0.01 g of each drug in methanol scanned in the range of 200–400 nm. The maximum absorption is found at 256 and 288 nm for quercetin and silymarin respectively [7].

Ginpreet Aneja et al., have validated the method for simultaneous estimation of Piperine, Quercetin, and Curcumin by UV spectrometry by making standard stock solution of 1000 ppm of each drug in methanol and scanned in the range of 200–400 nm and λmax of querce-tin, curcumin and piperine were found to be 371.31 nm, 424.68 nm, 343.76 nm respectively [8].

A Viswanath et al., have reported the UV spectrom-etry method for the estimation of Quercetin from Ipo-moea sepiaria Koenig. The two-extraction scanned for the absorption maxima. Quercetin standard absorbs at 350 nm, acetone extract at 328.2 nm and methanol extracts absorbs at 323.4 nm [9].

Marzanna Kurzawa et al., have determined the querce-tin and rutin in selected herbs and pharmaceutical prepa-rations by UV–Vis double beam spectrometer using 1 cm quartz cell. The standard solution of quercetin and rutin was prepared in methanol and ethanol. The absorbance at maximum absorbance of 425 nm for quercetin and 362 for rutin [10].

The detailed summery of spectroscopic standardization of Quercetin is presented in Table 1.

Chromatographic techniquesSunita Shailajan et al., have reported the validated method for estimation of Quercetin content from Cus-cuta reflexa Roxb by the HPLC using 0.025 M NaH2PO4: ACN as mobile phase. The analysis was performed using C18 column (150 mm × 4.6 mm × 5 µm) peaks were recorded at 378 nm [11].

Asmaˈa Ai-Rifai et al., have describes the HPLC method for analysis of Quercetin and Kaempferol of alcoholic extraction of convolvulus pilosellifolius by

O

OOH

OH

OH

HO

OHFig. 1 Chemical structure of Quercetin


using isocratic mixture of methanol and water contain-ing 0.1% v/v formic acid (80:20) using BETASIL C18 col-umn(150 mm × 4.6 mm,3 µm) and peaks were found at 258 nm [12].

Lee Fung Ang et al., have describes the quantita-tive detection of quercetin and curcuminoids (dimeth-oxycurcumin, bis-demethoxycurcumin, curcumin) in traditional Chinese medicines by HPLC. The analy-sis is carried out by using thermo ersil Gold column (250 mm × 4.6 mm ID.:5 µm) and a C-18 cartridge guard column(12.5 mm × 4.6mmID.:5 mm) with a mobile phase system of Acetonitrile and 2%v/v acetic acid (40:60) at the detection wavelength 370 nm.In this method the simul-taneous estimation of quercetin, bisdemethoxycurcumin, demethoxycurcumin and curcumin for the concentration range of 0.00488-200 µg/ml, 0.625-320 µg/ml, 0.07813-320 µg/ml and 0.03906-320 µg/ml respectively [13].

Ujjwala supe et al., have reported the HPLC method for analysis of quercetin in Momordica charanta. Merck C-18 Bondapack was used which is maintained at a tem-perature of 27℃ and methanol:ACN:water (60:20:20v/v) were used as mobile phase. At 260 and 262 nm wave-length the investigation carried out for flavonoid, phenols and Quercetin [14].

A. Srinivasa Rao et al., have accessed the HPLC method for the simultaneous determination of Rutin, Quercetin and Kaempferol in Catharanthus roseus by using Athena C18 column and phosphate buffer (pH = 5.8) and ACN as mobile phase 55:45 Ratio and maximum absorbance were measured at 254 nm. The retention time of Rutin, Quercetin, Kaempferol in the extract was found 2.403, 6.143, 8.903 respectively [15].

Deepak Mundkinajeddu et al., have developed a method for estimation of the flavonoid glycoside in

Withenia somnifera by HPLC using Phenomenex Luna C18 column (5µ, 250 × 4.6 mm) and the mobile sys-tem consist of potassium dihydrogen orthophosphate (0.136 g) dissolved in 900 ml of HPLC grade water to that 0.5 ml of orthophosphoric acid added volume made up to 1000 ml. the method for the estimation of 3 flavo-noid glycosides that are quercetin-3-orobinoside-7-O-glucoside (1), quercetin- 3-O-rutinoside-7-O-glucoside (2), kaempferol 3-O-robinobioside-7-O-glucoside [16].

Aline Augusti Boligon et al., Havevalidate the HPLC method for the estimation of flavonoid in gel of Scutia buxifolia using C18 column and mixture of Acetoni-trile: water (70:30, v/v) as a mobile phase at 356 nm of maximum absorbance wavelength the quercetin and rutin are quantified.In this method the concentration of quercetin and rutin were found to be 28.15 ± 0.04 and 59.73 ± 0.13 mg/g respectively [17].

Gomathy Subramanian et al., have reported the development and validation of HPLC method for the simultaneous estimation of quercetin and rutin in Aga-nosma dicotoma. The separation is achieved on C18 column (150 × 4.6 mm) and mobile phase containing acetonitrile: ammonium acetate (40:60v/v) at the flow rate 1 ml/min. The analysis monitored at 259 nm [4].

Haritha Krishna prasad et al., havedeveloped and validated the RP-HPLC method for simultaneous estimation of Resveratrol and Quercetin in bulk and pharmaceutical dosage form. The estimation is per-formed by using Sunfire C18 (150 × 3.0 nm I.D5µm particle size) column having Rheodyne injector using Methanol:Water:Formic acid: Triethylaminein the ratio 10:70:15:5 as mobile phase at the wavelength 277 nm.resveratrol and Quercetin eluted at retention time 1.24 and 2.14 respectively [18].

Table 1 Spectroscopic standardization of Quercetin in herbal medicines

Sl. no Author name Title of the work Description of analysis

1 V.C. Yeligar et al. Development and Validation of UV Spectrophotometric Method for Simultaneous Estimation of Melatonin and Quercetin in Liposome Formulation

The UV spectra run in the range of 200–400 nm. The λmax of standard solution of melatonin (10 µg/ml) and Quercetin (10 µg/ml) was found to be 276 nm and 372 nm

2 Singh Upendra et al. Simultaneous Estimation of Quercetin and Silymarin: Method Development and Validation

The UV spectrum run in the range of 400–200 nm. The maximum absorption found at 256 and 288 for querce-tin and silymarin

3 Ginpreet Aneja et al. Simultaneous Estimation of Piperine, Quercetin, and Curcumin in A Mixture using UV–Visible spectropho-tometer and Method Validation

The standard stock solution of 1000 ppm of each drug in methanol and scanned in the range of 200–400 nm and λmax of quercetin, curcumin and piperine was found to be 371.31 nm, 424.68 nm, 343.76 nm respectively

4 A Viswanath et al. UV spectrometry method for the estimation of Quercetin from Ipomoea sepiariaKoenig

The two-extraction scanned for the absorption maxima. Quercetin standard absorbs at 350 nm, acetone extract at 328.2 nm and methanol extracts absorbs at 323.4 nm

5 Marzanna Kurzawa et al. Determination of Quercetin and Rutin in Selected Herbs and Pharmaceutical Preparations

The standard solution of quercetin and rutin was prepared in methanol and ethanol respectively. The absorbance was found at 425 nm for quercetin and 362 for rutin


Tabl

e 2

Chr

omat

ogra

phic

sta

ndar

diza

tion

of Q

uerc

etin

in h

erba

l med

icin

es

Sl. n

oA

utho

r nam

eTi

tle o

f the

wor

kD

escr

iptio

n of

ana

lysi

s

1Su

nita

sha

ilaja

n et

al.

A c

ompa

rativ

e es

timat

ion

of q

uerc

etin

con

tent

from

Cus

cuta

refle

xa R

oxbu

s-in

g va

lidat

ed H

PTLC

and

HPL

C te

chni

ques

By H

PLC

met

hod

usin

g 0.

025

M N

aH2P

O4:

AC

N a

s m

obile

pha

se. T

he a

naly

sis

was

per

form

ed u

sing

C18

col

umn

(150

mm

× 4

.6 m

m ×

5 µ

m) p

eaks

wer

e re

cord

ed a

t 378

nm

2A

smaˈ

aAi-R

ifai e

t al.

Ana

lysi

s of

Que

rcet

in a

nd K

aem

pfer

ol in

an

Alc

ohol

ic E

xtra

ct o

f Con

volv

ulus

pi

lose

llifo

lius

usin

g H

PLC

HPL

C m

etho

d us

ing

isoc

ratic

mix

ture

of m

etha

nol a

nd w

ater

con

tain

ing

0.1%

v/

v fo

rmic

aci

d (8

0:20

) usi

ng B

ETA

SIL

C18

col

umn

(150

mm

× 4

.6 m

m,3

µm

) an

d pe

aks

wer

e fo

und

at 2

58 n

m

3Le

e Fu

ng A

ng e

t al.

HPL

C M

etho

d fo

r Sim

ulta

neou

s Q

uant

itativ

e D

etec

tion

of Q

uerc

etin

and

Cu

rcum

inoi

ds in

Tra

ditio

nal C

hine

se M

edic

ines

Qua

ntita

tive

dete

ctio

n of

que

rcet

in a

nd c

urcu

min

oids

(dim

etho

xy c

urcu

min

, bi

s de

met

hoxy

cur

cum

in, c

urcu

min

) in

trad

ition

al C

hine

se m

edic

ines

by

HPL

C u

sing

ther

mo

ersi

l Gol

d co

lum

n (2

50 m

m ×

4.6

mm

ID.:5

µm

) and

a

C-1

8 ca

rtrid

ge g

uard

col

umn

(12.

5 m

m ×

4.6

mm

ID.:5

mm

) with

a m

obile

ph

ase

syst

em o

f Ace

toni

trile

and

2%

v/v

acet

ic a

cid

(40:

60) a

t the

det

ectio

n w

avel

engt

h 37

0 nm

4U

jjwal

a su

pe e

t al.

Prel

imin

ary

Phyt

oche

mic

al A

naly

sis

and

Qua

ntita

tive

Ana

lysi

s of

Que

rcet

in

by H

PLC

of M

omor

dica

Cha

rant

iaH

PLC

met

hod

perf

orm

ed u

sing

Mer

ck C

-18

Bond

apac

k w

hich

is m

aint

aine

d at

a

tem

pera

ture

of 2

7℃ a

nd m

etha

nol:A

CN

:wat

er (6

0:20

:20v

/v) w

ere

used

as

mob

ile p

hase

. At 2

60 a

nd 2

62 n

m w

avel

engt

h th

e in

vest

igat

ion

carr

ied

out

for fl

avon

oid,

phe

nols

and

Que

rcet

in [1

4]

5A

. Srin

ivas

a Ra

o et

al.

Sim

ulta

neou

s de

term

inat

ion

of p

heno

lic c

ompo

unds

in C

atha

rant

hus

rose

us

leav

es b

y H

PLC

met

hod

HPL

C m

etho

d us

ing

Ath

ena

C18

col

umn

and

phos

phat

e bu

ffer (

pH =

5.8

) and

A

CN

as

mob

ile p

hase

55:

45 R

atio

and

max

imum

abs

orba

nce

wer

e m

easu

red

at 2

54 n

m. T

he re

tent

ion

time

of R

utin

, Que

rcet

in, K

aem

pfer

ol w

as fo

und

2.40

3, 6

.143

, 8.9

03 re

spec

tivel

y

6D

eepa

k M

undk

inaj

eddu

et a

l.D

evel

opm

ent a

nd V

alid

atio

n of

Hig

h Pe

rfor

man

ce L

iqui

d C

hrom

atog

raph

y M

etho

d fo

r Sim

ulta

neou

s Es

timat

ion

of F

lavo

noid

Gly

cosi

des

in W

ithan

ia

som

nife

ra A

eria

l Par

ts

HPL

C m

etho

d by

usi

ng P

heno

men

ex L

una

C18

col

umn

(5µ,

250

× 4

.6 m

m)

and

the

mob

ile s

yste

m c

onsi

st o

f pot

assi

um d

ihyd

roge

n or

thop

hos-

phat

e (0

.136

g) d

isso

lved

in 9

00 m

l of H

PLC

gra

de w

ater

to th

at 0

.5 m

l of

orth

opho

spho

ric a

cid

adde

d vo

lum

e m

ade

up to

100

0 m

l, fo

r the

est

imat

ion

of 3

flav

onoi

d gl

ycos

ides

that

are

que

rcet

in-3

-oro

bino

side

-7-O

-glu

cosi

de(1

), qu

erce

tin- 3

-O-r

utin

osid

e-7-

O-g

luco

side

(2),

kaem

pfer

ol 3

-O-r

obin

obio

-si

de-7

-O-g

luco

side

(3)

7A

line

Aug

usti

Bolig

on e

t al.

Dev

elop

men

t and

Val

idat

ion

of a

n H

PLC

–DA

D A

naly

sis

for F

lavo

noid

s in

the

gel o

f Scu

tia b

uxifo

liaTh

ey h

ave

valid

ated

the

HPL

C u

sing

C18

col

umn

and

mix

ture

of A

ceto

nitr

ile:

wat

er (7

0:30

, v/v

) as

a m

obile

pha

se a

t 356

nm

of m

axim

um a

bsor

banc

e w

avel

engt

h th

e qu

erce

tin a

nd ru

tin a

re q

uant

ified

8G

omat

hy S

ubra

man

ian

et a

l.D

evel

opm

ent a

nd V

alid

atio

n of

HPL

C M

etho

d fo

r the

Sim

ulta

neou

s Es

tima-

tion

of Q

uerc

etin

and

Rut

in in

Aga

nosm

a D

icho

tom

a [R

oth]

K. S

chum

For H

PLC

met

hod

sepa

ratio

n is

ach

ieve

d us

ing

C18

col

umn

(150

× 4

.6 m

m)

and

mob

ile p

hase

con

tain

ing

acet

onitr

ile: a

mm

oniu

m a

ceta

te (4

0:60

v/v)

at

the

flow

rate

1 m

l/min

.the

anal

ysis

mon

itore

d at

259

nm

9H

arith

a Kr

ishn

a pr

asad

et a

l.M

etho

d D

evel

opm

ent a

nd V

alid

atio

n fo

r the

Sim

ulta

neou

s Es

timat

ion

of

Resv

erat

rol a

nd Q

uerc

etin

in B

ulk

and

Phar

mac

eutic

al D

osag

e Fo

rm b

y RP

-HPL

C

RP-H

PLC

met

hod

for t

he e

stim

atio

n is

per

form

ed b

y us

ing

Sunfi

re C

18

(150

× 3

.0nm

I.D5µ

m p

artic

le s

ize)

col

umn

havi

ng R

heod

yne

inje

ctor

usi

ng

Met

hano

l:Wat

er:F

orm

ic a

cid:

Trie

thyl

amin

e in

the

ratio

10:

70:1

5:5

as m

obile

ph

ase

at th

e w

avel

engt

h 27

7 nm

.resv

erat

rol a

nd Q

uerc

etin

elu

ted

at re

ten-

tion

time

1.24

and

2.1

4 re

spec

tivel

y


Tabl

e 2

(con

tinue

d)

Sl. n

oA

utho

r nam

eTi

tle o

f the

wor

kD

escr

iptio

n of

ana

lysi

s

10Vi

shal

Sha

rad

Cha

udha

ri et

al.

Ana

lytic

al m

etho

d de

velo

pmen

t and

val

idat

ion

of re

vers

e-ph

ase

high

-pe

rfor

man

ce li

quid

chr

omat

ogra

phy

(RP-

HPL

C) m

etho

d fo

r sim

ulta

neou

s qu

antifi

catio

ns o

f que

rcet

in a

nd p

iper

ine

in d

ual-d

rug

load

ed n

anos

truc

-tu

red

lipid

car

riers

RP-H

PLC

met

hod

for T

he e

stim

atio

n w

as c

arrie

d ou

t by

usin

g H

yper

sil g

old

C-1

8 co

lum

n (1

50 m

m ×

4.6

mm

, 5 µ

m p

artic

le s

ize

OD

S) a

nd m

ixtu

re o

f ac

eton

itrile

and

HPL

C g

rade

wat

er (p

H 2

.6 a

djus

ted

with

2%

v/v

glac

ial a

cetic

ac

id) a

s a

mob

ile p

hase

wav

elen

gth

346

nm Q

uerc

etin

and

Pip

erin

e is

elu

ted

at re

tent

ion

time

2.80

min

and

10.

36 m

in re

spec

tivel

y

11A

shok

Kum

ar e

t al.

Estim

atio

n of

Gal

lic a

cid,

Rut

in a

nd Q

uerc

etin

in T

erm

enel

ia c

hebu

laby

HPT

LCH

PTLC

met

hod

by u

sing

pre

coat

ed s

ilica

gel

GF2

54 a

s st

atio

nary

pha

se a

nd

Tolu

ene:

ace

tone

: gla

cial

ace

tic a

cid

(3:2

:1 v

/v/v

/v/v

) as

mob

ile p

hase

for o

f ta

nnin

s, an

d et

hyl a

ceta

te: d

ichl

orom

etha

ne: f

orm

ic a

cid:

gla

cial

ace

tic a

cid:

w

ater

(10:

2.5:

1:1:

0.1v

/v/v

/v/v

) the

qua

ntifi

catio

n is

car

ried

out f

or R

utin

and

Q

uerc

etin

at 3

66 n

m a

nd fo

r gal

lic a

cid

at 2

54 n

m d

ensi

tom

etric

ally

and

Rf

valu

e of

gal

lic a

cid,

rutin

, and

que

rcet

in a

re 0

.30,

0.1

3, 0

.93

resp

ectiv

ely

12Sa

chin

U. R

akes

h et

al.

HPT

LC M

etho

d Fo

r Qua

ntita

tive

dete

rmin

atio

n of

Que

rcet

in in

Hyd

roal

co-

holic

Ext

ract

Of D

ried

Flow

er o

f Nym

phae

a St

ella

ta W

illd

In H

PTLC

met

hodt

he d

etec

tion

and

quan

tifica

tion

wer

e ca

rrie

d ou

t by

silic

a ge

l 60

F254

pla

tes

and

Tolu

ene:

eth

yl a

ceta

te: f

orm

ic a

cid

in th

e ra

tio 5

:4:0

.2

(v/v

/v) a

s m

obile

pha

se. A

t 380

nm

a s

harp

and

wel

l-defi

ned

peak

foun

d at

Rf

= 0

.29

13A

nsul

Sha

kya

et a

l.A

Rap

id H

igh-

Perf

orm

ance

Thi

n-La

yer C

hrom

atog

raph

ic M

etho

d to

Est

imat

e Q

uerc

etin

in B

enin

cas

ahisp

ida

(Thu

nb.)

Cogn

. Fru

it Pu

lpH

PTLC

for t

he e

stim

atio

n us

ing

alum

ina

plat

es w

ith s

ilica

gel

60

F254

wer

e us

ed w

ith T

olue

ne: e

thyl

ace

tate

: for

mic

aci

d (5

:4:0

.2v/

v) a

s m

obile

pha

se a

t ab

sorb

ance

wav

elen

gth

262n

mth

e Rf

val

ue o

f Que

rcet

in fo

und

Rf =

0.3

92

14Ba

rik L

axm

i Dha

r et a

l.H

PTLC

met

hod

for Q

uant

itativ

e es

timat

ion

of Q

uerc

etin

in a

pol

yher

bal

com

poun

d fo

r uro

lithi

asis

By H

PTLC

met

hod

extr

act c

hrom

atog

ram

med

on

silic

a ge

l 60F

254

alum

iniu

m

plat

es a

nd m

ixtu

re o

f chl

orof

orm

: met

hano

l: fo

rmic

aci

d us

ed a

s m

obile

ph

ase

in th

e ra

tio 7

.5:1

.5:1

v/v

/v. T

he R

f val

ue w

as 0

.51

anal

ysed

at w

ave-

leng

th o

f 254

nm

15Bi

ndu

A R

et a

l.H

igh

Perf

orm

ance

Thi

n La

yer C

hrom

atog

raph

ic M

etho

d fo

r Qua

ntita

tive

Det

erm

inat

ion

of Q

uerc

etin

in T

ende

r Lea

ves

of P

sidi

um g

uaja

vaH

PTLC

met

hod

caar

ied

out o

n si

lica

gel 6

0F25

4 TL

C p

late

s us

ing

tolu

ene:

ac

eton

e: fo

rmic

aci

d (3

0:10

:5) a

s m

obile

pha

se. T

he d

etec

tion

and

quan

tifica

-tio

n do

ne a

t 364

nm

and

Rf v

alue

of a

ceto

ne e

xtra

ct w

as 0

.45

16Va

idev

i Set

hura

m e

t al.

Com

bina

toria

l ana

lysi

s of

que

rcet

in a

nd re

sver

atro

l by

HPT

LC in

Ses

bani

a gr

andi

flora

/ph

yto

base

d na

nofo

rmul

atio

nsBy

HPT

LC m

etho

d In

this

inve

stig

atio

n se

para

tion

achi

eved

by

usin

g m

obile

ph

ase

syst

em o

f tol

uene

: chl

orof

orm

: eth

yl a

ceta

te: f

orm

ic a

cid

(3:2

:4.9

:0.1

%

v/v)

. The

den

sito

met

ric s

cann

ing

at 2

80 n

m w

as p

erfo

rmed

and

Rf v

alue

was

0.

40 (Q

uerc

etin

) and

0.5

0(Re

sver

atro

l)

17M

ange

sh S

. Kha

rate

et a

l.Es

timat

ion

of Q

uerc

etin

from

Cru

de L

eaf E

xtra

ct, M

imus

ops E

leng

i L. B

y H

PTLC

The

estim

atio

n of

Que

rcet

in c

arrie

d ou

t on

prec

oate

d si

lica

gel p

late

s F

254

(10

cm ×

10

cm) a

nd m

ixtu

re o

f tol

uene

: eth

yl a

ceta

te: f

orm

ic a

cid

in th

e ra

tio 5

:4:1

was

use

d as

mob

ile p

hase

. The

det

ectio

n an

d qu

antifi

catio

n do

ne

at 3

65 n

m

18V

Patil

et a

l.Re

cent

pro

gres

s in

sim

ulta

neou

s es

timat

ion

of ru

tin, q

uerc

etin

and

liqu

iritin

in

Coc

culu

s hirs

utus

by

HPT

LCTh

e si

mul

tane

ous

estim

atio

n ca

rrie

d on

sili

ca g

el 6

0F25

4 an

d m

ixtu

re o

f n-

buta

nol:

acet

ic a

cid:

wat

er: f

orm

ic a

cid

(7:1

:1:0

.25)

as

mob

ile p

hase

sys

tem

. Th

e Rf

val

ue fo

r Rut

in, Q

uerc

etin

and

Liq

uirit

in w

as 0

.047

± 0

.03,

0.0

63 ±

0.0

3 an

d 0.

82 ±

0.0

2 re

spec

tivel

yat t

he w

avel

engt

h 25

4 nm

19O

mi L

aila

et a

l.D

evel

opm

ent a

nd V

alid

atio

n of

HPT

LC M

etho

d fo

r Sim

ulta

neou

s Es

tima-

tion

of D

iosg

enin

and

Que

rcet

in in

Fen

ugre

ek S

eeds

(Trig

onel

la fo

enum

-gr

aceu

m)

HPT

LC m

etho

d fo

r val

idat

ion

carr

ied

out u

sing

pre

coat

ed a

lum

iniu

m p

late

s w

ith s

ilica

gel

G60

F254

and

mix

ture

of t

olue

ne: e

thyl

ace

tate

: for

mic

aci

d (5

:4:1

v/v

/v) a

s m

obile

pha

se. T

he s

cann

ing

of p

late

s at

275

nm

and

the

Rf

valu

e w

as 0

.69 ±

0.0

2 (D

iosg

enin

) and

0.5

7 ±

0.0

2(Q

uerc

etin

)


Tabl

e 2

(con

tinue

d)

Sl. n

oA

utho

r nam

eTi

tle o

f the

wor

kD

escr

iptio

n of

ana

lysi

s

20A

. Srin

ivas

Rao

et a

l.Si

mul

tane

ous

estim

atio

n of

que

rcet

in a

nd ru

tin in

eth

anol

ic e

xtra

ct o

f Mel

ia

azed

arac

h. L

inn

leav

es b

y H

PTLC

met

hod

HPT

LC m

etho

d fo

rthe

ana

lysi

s is

car

ried

out w

ith p

reco

ated

sili

ca 6

0F25

4 as

st

atio

nary

pha

se a

nd m

ixtu

re o

f tol

uene

: eth

yl a

ceta

te: m

etha

nol (

5:3:

2) a

s m

obile

pha

se. A

t 254

nm

the

Rf v

alue

s of

Que

rcet

in a

nd R

utin

was

0.6

5 an

d 0.

15 re

spec

tivel

y

21G

undu

Sin

dhu

Cha

krab

orty

et a

l.D

eter

min

atio

n of

Que

rcet

in b

y H

PTLC

in “C

alen

dula

Offi

cina

lis” E

xtra

ctH

PTLC

met

hod

usin

g pr

ecoa

ted

alum

iniu

m p

late

s w

ith s

ilica

gel

60G

F as

sta

-tio

nary

pha

se a

nd c

hlor

ofor

m: m

etha

nol (

9.5:

0.5)

mix

ture

as

mob

ile p

hase

so

lven

t sys

tem

. The

Rf v

alue

was

0.4

3 an

d sc

anni

ng fo

r est

imat

ion

was

car

-rie

d ou

t at 3

66 n

m w

avel

engt

h

22M

d. S

arfa

raj H

ussa

in e

t al.

Valid

atio

n of

the

met

hod

for t

he s

imul

tane

ous

estim

atio

n of

bio

activ

e m

arke

r gal

lic a

cid

and

quer

cetin

in A

butil

on in

dicu

m b

y H

PTLC

The

anal

ysis

per

form

ed o

n al

umin

ium

foil-

back

ed s

ilica

gel

60F

254

HPT

LC

plat

es u

sing

sol

vent

sys

tem

of t

olue

ne: e

thyl

ace

tate

: for

mic

aci

d (5

:4:1

v/v

) as

mob

ile p

hase

. At a

bsor

banc

e w

avel

engt

h 27

0 nm

and

Rf v

alue

was

0.3

1 (g

allic

aci

d), 0

.50

(Que

rcet

in)

23Jin

an H

ussa

in e

t al.

Qua

litat

ive

and

quan

titat

ive

com

paris

on o

f rut

in, q

uerc

etin

and

gal

lic a

cid

conc

entr

atio

ns in

Syr

ian

Capp

aris

spin

osa.

L L

eave

sTh

e an

alys

is p

erfo

rmed

on

silic

a ge

l 60F

254

and

mix

ture

of e

thyl

ace

tate

: gla

-ci

al a

cetic

aci

d: fo

rmic

aci

d: d

istil

led

wat

er (1

00:1

1:11

:25)

as

mob

ile p

hase

. At

the

scan

ning

wav

elen

gth

of 3

66 n

m th

e Rf

val

ue o

btai

ned

are

0.39

(Rut

in),

0.79

(Que

rcet

in) a

nd a

t 254

nm

the

Rf v

alue

was

0.8

1 (g

allic

aci

d)

24A

bhay

Pra

kash

Mis

hra

et a

l.A

Dev

elop

ed a

nd V

alid

ated

Hig

h-Pe

rfor

man

ce T

hin-

Laye

r Chr

omat

ogra

phic

M

etho

d fo

r the

Qua

ntita

tive

Det

erm

inat

ion

of Q

uerc

etin

in S

atyr

ium

ne

pale

nse

Tube

rs

HPT

LC a

naly

sis

carr

ied

with

sili

ca g

el 6

0F25

4 as

sta

tiona

ry p

hase

usi

ng to

lu-

ene:

eth

yl a

ceta

te: f

orm

ic a

cid

(7:5

:1 v

/v) a

s so

lven

t sys

tem

at 3

66 n

m

25Sh

iv K

Yad

av e

t al.

Dev

elop

men

t and

val

idat

ion

of a

HPT

LC m

etho

d fo

r sim

ulta

neou

s es

tima-

tion

of q

uerc

etin

, Chl

orog

enic

aci

d an

d tr

igon

ellin

e in

pol

yher

bal a

ntib

ac-

teria

l for

mul

atio

n

HPT

LC a

naly

sis

with

alu

min

ium

pla

tes

60F2

54 u

sing

sol

vent

sys

tem

chl

o-ro

form

: eth

yl a

ceta

te:m

etha

nol:

form

ic a

cid

(5:3

:1.5

:0.5

v/v/

v/v)

as

mob

ile

phas

e. T

he w

ell s

epar

ated

spo

ts w

ere

obta

ined

with

Rf v

alue

s of

0.1

3, 0

.24,

0.

62 fo

r trig

onel

line,

chl

orog

enic

aci

d, a

nd q

uerc

etin

resp

ectiv

ely

26H

iteks

ha P

anch

al e

t al.

Dev

elop

men

t of V

alid

ated

Hig

h-pe

rfor

man

ce T

hin-

laye

r Chr

omat

ogra

phy

Met

hod

for S

imul

tane

ous

Det

erm

inat

ion

of Q

uerc

etin

and

Kae

mpf

erol

in

Thes

pesia

pop

ulne

a

Ana

lysi

s pe

rfor

med

On

alum

iniu

m p

late

s pr

ecoa

ted

with

sili

ca g

el 6

0F25

4 us

ing

tolu

ene:

eth

yl a

ceta

te: f

orm

ic a

cid

(6:4

:0.3

v/v

/v).

At t

he s

cann

ing

abso

rban

ce w

avel

engt

h 36

6 nm

, the

Rf v

alue

obt

aine

d ar

e 0.

39 fo

r Que

rce-

tin a

nd 0

.50

for k

aem

pfer

ol

27Kh

an D

ures

hahw

ar e

t al.

Qua

ntifi

catio

n of

Que

rcet

in O

btai

ned

from

Alli

um c

epa

Lam

. Lea

ves

and

its

Effec

ts o

n St

rept

ozot

ocin

-indu

ced

Dia

betic

Neu

ropa

thy

The

quan

tifica

tion

carr

ied

out w

ith p

reco

ated

sili

ca g

el G

F254

pla

tes

and

usin

g m

obile

pha

se s

yste

m to

luen

e: e

thyl

ace

tate

: for

mic

aci

d (5

:4:1

) and

the

scan

ning

abs

orba

nce

at 2

54 n

m

28Su

priy

a S.

Jirg

e et

al.

Sim

ulta

neou

s Es

timat

ion

of K

aem

pfer

ol, R

utin

, and

Que

rcet

in in

Var

ious

Pla

nt

Prod

ucts

and

Diff

eren

t Dos

age

Form

s of

Bhu

iam

la a

nd A

mla

The

anal

ysis

per

form

ed w

ith p

reco

ated

sili

ca g

el a

lum

iniu

m p

late

s 60

F254

(20

cm ×

10

cm w

ith 2

50 µ

m th

ickn

ess)

and

mix

ture

of

chlo

rofo

rm:m

etha

nol:

form

ic a

cid

(8.2

:1.5

:1) u

sed

as m

obile

pha

se a

nd a

t 25

4 nm

the

Rf v

alue

of k

aem

pfer

ol-0

.69,

que

rcet

in-0

.53,

and

rutin

-0.1

2 ar

e re

port

ed

29Sh

ikar

Ver

ma

et a

l.H

PTLC

Ana

lysi

s fo

r the

Sim

ulta

neou

s Q

uant

ifica

tion

of G

allic

Aci

d, V

anill

ic

Aci

d, P

roto

cate

chui

c A

cid,

and

Que

rcet

in in

the

Met

hano

lic F

ract

ion

of

Lim

onia

aci

diss

ima

L. F

ruits

HPT

LC m

etho

d us

ing

silic

a ge

l 60F

254

plat

es. T

he m

ixtu

re o

f sol

vent

s to

luen

e:et

hyl a

ceta

te: f

orm

ic a

cid

(5:4

:1) u

sed

as m

obile

pha

se a

nd th

e de

tect

ion

carr

ied

out a

t 254

nm

and

repo

rted

Rf v

alue

s ar

e 0.

3 ±

0.0

0 (g

allic

aci

d), 0

.47 ±

0.0

05 (v

anill

ic a

cid)

, 0.3

7 ±

0.0

0 (p

roto

cate

chui

c ac

id),

0.42

± 0

.005

(que

rcet

in)


Tabl

e 2

(con

tinue

d)

Sl. n

oA

utho

r nam

eTi

tle o

f the

wor

kD

escr

iptio

n of

ana

lysi

s

30Sn

ehal

B. B

hand

are

et a

l.Si

mul

tane

ous

Qua

ntifi

catio

n of

Kae

mpf

erol

and

Que

rcet

in in

Med

icin

al

Plan

ts u

sing

HPT

LCH

PTLC

met

hod

usin

g to

luen

e: a

ceto

ne: f

orm

ic a

cid

in th

e ra

tio 7

:3:0

.25

as

mob

ile p

hase

on R

P-18

F25

4 pl

ates

. The

Rf v

alue

for K

aem

pfer

ol a

nd Q

uerc

e-tin

wer

e fo

und

0.46

and

0.3

9 re

spec

tivel

y at

the

dete

ctio

n w

avel

engt

h 25

4 nm

31Sh

ikar

Ver

ma

et a

l.G

as C

hrom

atog

raph

ic–M

ass

Spec

trom

etric

Pro

file

of N

on-P

olar

Fra

ctio

n an

d H

igh-

Perf

orm

ance

Thi

n-La

yer C

hrom

atog

raph

ic A

naly

sis

of M

etha

nolic

Fr

actio

n w

ith S

imul

tane

ous

Qua

ntifi

catio

ns o

f Pro

to c

atec

huic

Aci

d an

d Q

uerc

etin

in C

ariss

a ca

rand

as L

. Fru

its

HPT

LC a

naly

sis

usin

g pr

ecoa

ted

plat

es w

ith s

ilica

gel

60F

254

with

sol

vent

sy

stem

tolu

ene:

eth

yl a

ceta

te: f

orm

ic a

cid

(6:3

:1, v

/v).

The

Rf v

alue

obt

aine

d as

0.5

7 an

d 0.

61 fo

r pro

toca

tech

uic

acid

and

que

rcet

in re

spec

tivel

y at

the

max

imum

abs

orba

nce

wav

elen

gth

310

nm

32N

adee

m A

. Sid

diqu

e et

al.

Sim

ulta

neou

s Q

uant

ifica

tion

of U

mbe

llife

rone

and

Que

rcet

in in

Pol

yher

bal

Form

ulat

ions

of A

egle

Mar

mel

os b

y H

PTLC

Ana

lysi

s pe

rfor

med

usi

ng p

reco

ated

alu

min

ium

pla

tes

with

sili

ca g

el 6

0F-2

54

as s

tatio

nary

pha

se. T

he s

olve

nt s

yste

m c

onsi

sts

of to

luen

e: e

thyl

ace

tate

: fo

rmic

aci

d (6

:4:1

v/v/

v) a

s m

obile

pha

se.T

he R

f val

ue w

as re

port

ed a

s 0.

66

and

0.68

for u

mbe

llife

rone

and

que

rcet

in a

t 300

nm

33Pu

shpe

ndra

kum

ar e

t al.

Sim

ulta

neou

s Q

uant

ifica

tion

of Q

uerc

etin

and

Syr

ingi

c A

cid

in M

etha

nolic

Ex

trac

t of L

euca

s lav

andu

lifol

ia b

y us

ing

Valid

ated

HPT

LCD

ensi

tom

etric

M

etho

d

The

anal

ysis

car

ried

out o

n Si

lica

gel 6

0F25

4 pl

ate

usin

g To

luen

e: e

thyl

acet

ate:

fo

rmic

aci

d (7

:2.5

:0.5

v/v

) as

mob

ile p

hase

. The

Rf v

alue

for q

uerc

etin

and

sy

ringi

c ac

id w

as fo

und

to b

e 0.

32 a

nd 0

.41

at th

e de

tect

ion

wav

elen

gth

275–

370

nm

34Th

afsh

ila A

afrin

Am

et a

l.D

eter

min

atio

n qf

Que

rcet

in b

y H

igh

Perf

orm

ance

Thi

n La

yer C

hrom

atog

ra-

phy

Met

hod

in A

chyr

anth

es A

sper

a (L

.) Pl

ant E

xtra

ctTh

e de

term

inat

ion

carr

ied

out o

n si

lica

gel 6

0F25

4 pl

ate

as s

tatio

nary

pha

se

and

mix

ture

of t

olue

ne: e

thyl

ace

tate

: for

mic

aci

d in

the

ratio

of 5

:4:1

. The

Rf

valu

e 0.

60 is

det

ecte

d at

254

nm

35A

vije

et Ja

in e

t al.

Sim

ulta

neou

s es

timat

ion

of q

uerc

etin

and

rutin

in T

ephr

osia

pur

pure

a Pe

rs b

y hi

gh p

erfo

rman

ce th

in la

yer c

hrom

atog

raph

yTh

e es

timat

ion

carr

ied

out o

n pr

ecoa

ted

silic

a ge

l RP-

18 F

254

wer

e us

ed

as s

tatio

nary

pha

se a

nd th

e so

lven

t sys

tem

met

hano

l: w

ater

: for

mic

aci

d (4

0:57

:3v/

v/v)

as

mob

ile p

hase

. At t

he d

etec

tion

wav

elen

gth

254n

mth

e Rf

va

lue

was

0.0

7(qu

erce

tin) a

nd 0

.17

(rutin

)

36G

irme

AS

et a

l.M

etho

d de

velo

pmen

t, op

timiz

atio

n an

d va

lidat

ion

of R

P-U

FLC

met

hod

for

bioa

ctiv

e fla

vono

ids

from

Cas

sia a

uric

ulat

aTh

e an

alys

is is

car

ried

out o

n ac

quity

C18

col

umn

with

ste

pwis

e gr

adie

nt e

lu-

tion

was

car

ried

out w

ith 0

.1%

form

ic a

cid

in w

ater

and

met

hano

l at a

flow

ra

te o

f 0.3

50 m

l/min

. The

max

imum

abs

orba

nce

at 3

50 n

m w

avel

engt

h w

as

dete

cted

for q

uerc

etin

QU

E-3-

O-r

utin

osid

e. T

he re

tent

ion

time

was

foun

d at

3.

95 a

nd 5

.37

for q

uerc

etin

-3-O

-rut

inos

ide

and

quer

cetin

resp

ectiv

ely

37Sh

anm

ugam

R e

t al.

Dev

elop

men

t and

Val

idat

ion

of a

RP-

UFL

C M

etho

d fo

r Sim

ulta

neou

s Es

tima-

tion

of Q

uerc

etin

and

Rut

inTh

e an

alys

is p

erfo

rmed

on

reve

rse

phas

e Sy

stem

C8

colu

mn

(25 ×

4.6

mm

). th

e m

obile

pha

se c

onsi

sts

of m

ixtu

re o

f pot

assi

um d

ihyd

roge

n or

tho

phos

phat

e an

d ac

eton

itrile

the

ratio

of 7

0:30

. The

det

ectio

n w

as d

one

at

230

nm a

nd th

e re

tent

ion

time

was

7.4

and

2.8

min

for q

uerc

etin

and

rutin

re

spec

tivel

y

38Kh

aled

Elg

endy

et a

l.A

naly

sis

of T

otal

Fla

vono

ids

in H

erba

l Dru

gs E

xpre

ssed

as

Que

rcet

in b

y Re

vers

ed P

hase

-UH

PLC

Met

hod

The

anal

ysis

don

e on

col

umn

Phen

omen

ex a

nd m

etha

nol a

nd 0

.5%

pho

s-ph

oric

aci

d (5

0:50

v/v)

as

mob

ile p

hase

with

flow

rate

of 1

.3 m

l/min

. Lin

earit

y of

the

met

hod

is e

stab

lishe

d ov

er th

e co

ncen

trat

ion

rang

e of

240

-960

µg/

ml

for q

uerc

etin

and

rete

ntio

n tim

e w

ere

foun

d 7.

8 m

in

39M

aric

san

tos

et a

l.U

PLC

-MS

for I

dent

ifica

tion

of Q

uerc

etin

Der

ivat

ives

in C

uphe

a gl

utin

osa

Cha

m. a

nd S

chltd

l (Ly

thra

ceae

) and

Eva

luat

ion

of A

ntifu

ngal

Pot

entia

lTh

e an

alys

is p

erfo

rmed

on

fast

C18

col

umn

anal

ytic

al c

olum

n sh

im p

ack

XR-

OD

S co

lum

n (5

0 ×

2 m

m, 2

.1 µ

m).

the

mob

ile p

hase

con

sist

ed o

f mix

ture

of

acet

onitr

ile: m

etha

nol (

4:1v

/v) a

s so

lven

t A a

nd w

ater

con

tain

s 0.

1% fo

rmic

ac

id a

s so

lven

t B. t

he m

ass

spec

tra

was

reco

rded

by

full

scan

mod

e in

the

rang

e of

m/z

200

–800


Vishal Sharad Chaudhari et al., have reported the simultaneous quantification by RP-HPLC method for Quercetin and Piperine in dual drug loaded nanostruc-tured lipid carriers. The estimation was carried out by using Hypersil gold C-18 column (150 mm × 4.6 mm, 5 µm particle size ODS) and mixture of acetonitrile and HPLC grade water (pH 2.6 adjusted with 2%v/v glacial acetic acid) as a mobile phase at the detection wavelength 346 nm Quercetin and Piperine is eluted at retention time 2.80 min and 10.36 min respectively [19].

Ashok Kumar et al., have investigated the HPTLC method for the estimation of Gallic acid, Rutin and Quercetin in Termenelia chebula. The investigation is carried out by using precoated silica gel GF254 as sta-tionary phase and Toluene: acetone: glacial acetic acid (3:2:1 v/v/v/v/v) as mobile phase for of tannins, and ethyl acetate: dichloromethane: formic acid: glacial acetic acid: water (10:2.5:1:1:0.1v/v/v/v/v) the quantification is car-ried out for Rutin and Quercetin at 366 nm and for gal-lic acid at 254 nm densitometrically and Rf value of gallic acid, rutin, and quercetin are 0.30, 0.13, 0.93 respectively [20].

Sachin U. Rakesh et al., have determined the HPTLC method for the quantitative estimation of Quercetin in Hydrochloric extract of dried flower of Nymphaea stel-late willd. The detection and quantification were carried out by silica gel 60 F254 plates and Toluene: ethyl acetate: formic acid in the ratio 5:4:0.2 (v/v/v) as mobile phase. The densitometric scanning at absorbance wavelength 380 nm was done and a sharp and well-defined peak found at Rf = 0.29 [21].

Ansul Shakya et al., have reported the method to esti-mate Quercetin in Benincasahispida Cong fruit pulp by HPTLC. The estimation is carried out using alumina plates with silica gel 60 F254 were used with Toluene: ethylacetate: formic acid (5:4:0.2v/v) as mobile phase at absorbance wavelength 262nmthe Rf value of Quercetin found Rf = 0.392 [22].

Barik Laxmi Dhar et al., have reported the HPTLC method for the quantitative estimation of Querce-tin in a polyherbal compound for urolithiasis. Cra-taeva nurvala and Bryophyllum pinnatum extracted with methanol and extracts are chromatogrammed on silica gel 60F254 aluminium plates and mixture of chloroform:methanol:formic acid used as mobile phase in the ratio 7.5:1.5:1 v/v/v. The Rf value was 0.51 analysed at wavelength of 254 nm [23].

Bindu A R et al., have described the HPTLC method for the Quantitative determination of Quercetin in ten-der leaves of Psidium guajava on silica gel 60F254 TLC plates using toluene: acetone: formic acid (30:10:5) as mobile phase. The detection and quantification done at 364 nm and Rf value of acetone extract was 0.45 [24].

Vaidevi Sethuram et al., have reported the HPTLC method for combinatorial analysis of quercetin and res-veratrol in Sesbania grandiflora/Phyto based nanopar-ticles. In this investigation separation achieved by using mobile phase system of toluene:chloroform: ethyl acetate: formic acid (3:2:4.9:0.1% v/v). The densitometric scan-ning at 280 nm was performed and Rf value was 0.40 (Quercetin) and 0.50(Resveratrol) [25].

Mangesh S. Kharate et al., have reported the HPTLC method for estimation of quercetin from crude leaf extract Mimusops elengi L. The estimation of Querce-tin carried out on precoated silica gel plates F 254 (10 cm × 10 cm) and mixture of toluene: ethyl acetate: formic acid in the ratio 5:4:1 was used as mobile phase. The detection and quantification done at 365 nm [26].

V Patil et al., have evaluated the simultaneous estima-tion of Rutin, Quercetin and Liquiritin in Cocculus hir-sutus by HPTLC using silica gel 60F254 and mixture of n-butanol: acetic acid: water: formic acid (7:1:1:0.25) as mobile phase system. The Rf value for Rutin, Querce-tin and Liquiritin was 0.047 ± 0.03, 0.063 ± 0.03 and 0.82 ± 0.02 respectively which was scanned at the wave-length 254 nm [27].

Omi Laila et al., have reported the method develop-ment and validation of HPTLC for simultaneous esti-mation of Diosgenin and Quercetin in fenugreek seeds, using precoated aluminium plates with silica gel G60F254 and mixture of toluene: ethylacetate: formic acid (5:4:1 v/v/v) as mobile phase. The densitometric scanning of plates done at 275 nm and the Rf value was 0.69 ± 0.02 (Diosgenin) and 0.57 ± 0.02(Quercetin) [28].

A. Srinivas Rao et al., have described the simultaneous estimation of Quercetin and Rutin in ethanolic extract of Melia azedarach. Linn leaves by HPTLC method. The analysis is carried out with precoated silica 60F254 as stationary phase and mixture of toluene: ethyl acetate: methanol (5:3:2) as mobile phase. The scanning for esti-mation was carried out at 254 nm and the Rf values of Quercetin and Rutin was 0.65 and 0.15 respectively [29].

GunduSindhuChakraborty et al., have described the method for determination of Quercetin by HPTLC in Calendula officinalis extract using precoated alumin-ium plates with silica gel 60GF as stationary phase and chloroform: methanol (9.5:0.5) mixture as mobile phase solvent system. The Rf value was 0.43 and scanning for estimation was carried out at 366 nm wavelength [30].

Md. Sarfaraj Hussain et al., have validated the HPTLC method for simultaneous estimation of bioactive marker Gallic acid and Quercetin in Abutilon indicum. The analysis performed on aluminium foil-backed silica gel 60F254 HPTLC plates using solvent system of toluene: ethyl acetate: formic acid (5:4:1 v/v) as mobile phase. The densitometric scanning performed at absorbance


wavelength 270 nm and Rf value was 0.31 (gallic acid), 0.50 (Quercetin) [31].

Jinan Hussain et al., have reported the qualitative and quantitative comparison of rutin, quercetin and gallic acid concentration in Syrian Capparis Spinosa L leaves using aluminium plates with silica gel 60F254 and mix-ture of ethyl acetate: glacial acetic acid: formic acid: distilled water (100:11:11:25) as mobile phase. At the scanning wavelength of 366 nm the Rf value obtained are 0.39 (Rutin), 0.79 (Quercetin) and at 254 nm the Rf value was 0.81 (gallic acid) [32].

Abhay Prakash Mishra et al., have developed and vali-dated method for the Quantitative determination of Quercetin in Satyrium nepalense tubers by HPTLC with silica gel 60F254 as stationary phase using toluene: ethyl acetate: formic acid (7:5:1 v/v) as solvent system at absorbance wavelength 366 nm [33].

Shiv K Yadav et al., have developed and validated method for simultaneous estimation of Quercetin, chlo-rogenic acid and trigonelline in polyherbal antibacterial formulation by HPTLC with aluminium plates 60F254 using solvent system chloroform: ethyl acetate:methanol: formic acid (5:3:1.5:0.5v/v/v/v) as mobile phase. The well separated spots were obtained with Rf values of 0.13, 0.24, 0.62 for trigonelline, chlorogenic acid, and quercetin respectively [34].

Hiteksha Panchal et al., have developed and validated HPTLC method for simultaneous determination of Quercetin and Kaempferol in Spesia populena. On alu-minium plates precoated with silica gel 60F254 using toluene: ethyl acetate: formic acid (6:4:0.3 v/v/v). At the scanning absorbance wavelength 366 nm, the Rf value obtained are 0.39 for Quercetin and 0.50 for kaempferol [35].

Khan Dureshahwar et al., have reported method for the quantification of Quercetin from Allium cepa lam. Leaves by HPTLC. The quantification carried out with precoated silica gel GF254 plates and using mobile phase system toluene: ethyl acetate: formic acid (5:4:1) and the scan-ning absorbance at 254 nm [36].

Supriya S. Jirge et al., have discussed the HPTLC method for the simultaneous estimation of kaempferol, rutin, quercetin in various plant products. The analysis performed with precoated silica gel aluminium plates 60F254(20 cm × 10 cm with 250 µm thickness) and mix-ture of chloroform:methanol: formic acid (8.2:1.5:1) used as mobile phase and the absorbance mode kept at 254 nm. Rf value of kaempferol-0.69, quercetin-0.53, and rutin-0.12 are reported [37].

Shikar Verma et al., have reported the simultaneous quantification of gallic acid, vanillic acid, protocatechuic acid and quercetin in Limonia acidissima by HPTLC method using silica gel 60F254 plates. The mixture of

solvents toluene:ethyl acetate: formic acid (5:4:1) used as mobile phase and the detection carried out at 254 nm and reported Rf values are 0.3 ± 0.00 (gallic acid), 0.47 ± 0.005 (vanillic acid), 0.37 ± 0.00 (protocatechuic acid), 0.42 ± 0.005 (quercetin) [38].

Snehal B. Bhandare et al., have estimated the simultane-ous quantification of kaempferol and Quercetin in medic-inal plants by HPTLC method using toluene:acetone: formic acid in the ratio 7:3:0.25 as mobile phaseon RP-18 F254 plates. The Rf value for Kaempferol and Quercetin were found 0.46 and 0.39 respectively at the detection wavelength 254 nm [39].

Shikar Verma et al., have reported the HPTLC method for analysis of methanolic fraction with simultaneous quantification of protocatechuic acid and quercetin in Carissa carandas L. fruits using precoated plates with sil-ica gel 60F254 with solvent system toluene: ethyl acetate: formic acid (6:3:1, v/v). The Rf value obtained as 0.57 and 0.61 for protocatechuic acid and quercetin respectively at the maximum absorbance wavelength 310 nm [40].

Nadeem A. Siddique et al., have discussed the HPTLC method for simultaneous Quantification of umbellifer-one and quercetin in polyherbal formulation of Aegle marmelos by using precoated aluminium plates with sil-ica gel 60F-254 as stationary phase. The solvent system consists of toluene: ethyl acetate: formic acid (6:4:1v/v/v) as mobile phase. The Rf value was reported as 0.66 and 0.68 for umbelliferone and quercetin at the detection wavelength 300 nm [41].

Pushpendra kumar et al., have reported the simultane-ous quantification of quercetin and syringic acid in meth-anolic extract of Leucas lavandulifolia by using validated HPTLC densitometric method.The analysis carried out on Silica gel 60F254 plate using Toluene: ethylacetate: formic acid (7:2.5:0.5 v/v) as mobile phase. The Rf value for quercetin and syringic acid was found to be 0.32 and 0.41 at the detection wavelength 275 to 370 nm [42].

Thafshila Aafrin Am et al., have determined HPTLC method for quercetin in Achyranthes aspera (L) plant extract. The determination carried out on silica gel 60F254 plate as stationary phase and mixture of tolu-ene: ethyl acetate: formic acid in the ratio of 5:4:1. The Rf value 0.60 is detected at 254 nm [43].

Avijeet Jain et al., have reported simultaneous for the estimation of Quercetin and Rutin in Tephrosia purpurea by HPTLC. The estimation carried out on precoated silica gel RP-18 F254 were used as stationary phase and the sol-vent system methanol:water: formic acid (40:57:3v/v/v) as mobile phase. At the detection wavelength 254nmthe Rf value was 0.07(quercetin) and 0.17 (rutin) [44].

Girme AS et al., havediscussed method development, optimization and validation of RP-UFLC method for bio-active flavonoids from Cassia auriculata. The analysis is


carried out on acquity C18 column with stepwise gradi-ent elution was carried out with 0.1% formic acid in water and methanol at a flow rate of 0.350 ml/min. The maxi-mum absorbance at 350 nm wavelength was detected for quercetin QUE-3-O-rutinoside. The retention time was found at 3.95 and 5.37 for quercetin-3-O-rutinoside and quercetin respectively [45].

Shanmugam R et al., have reported the development and validation of RP-UFLC method for simultane-ous estimation of quercetin and rutin. The separation was done by using reverse phase System C8 column (25 × 4.6 mm). the mobile phase consists of mixture of potassium dihydrogen ortho phosphate and acetonitrile the ratio of 70:30. The detection was done at 230 nm and the retention time was 7.4 and 2.8 min for quercetin and rutin respectively [5].

Khaled Elgendy et al., haveanalysed the total flavonoids in herbal drugs expressed as quercetin by reverse phase- UHPLC method. The analysis done on column Phenom-enex and methanol and 0.5% phosphoric acid (50:50v/v) as mobile phase with flow rate of 1.3 ml/min. Linearity of the method is established over the concentration range of 240-960 µg/ml for quercetin and retention time were found 7.8 min [46].

Maric santos et al., have reported the UPLC-MS for identification of quercetin derivative in Cuphea gluti-nosa by using reverse phasesystem and fast C18 col-umn analytical column shim pack XR-ODS column (50 × 2 mm,2.1 µm). the mobile phase consisted of mix-ture of acetonitrile: methanol (4:1v/v) as solvent A and water contains 0.1% formic acid as solvent B. the mass spectra was recorded by full scan mode in the range of m/z 200–800 [47].

The detailed summery of chromatographic standardi-zation of Quercetin is presented in Table 2.

ConclusionsHerbal medicine and their preparations have been widely used from the thousands of years in develop-ing and developed countries in the primary health care of society and community. Quality control is one of the very important and essential steps in the manufacturing of herbal preparations as quality of product affects the safety and efficacy of medicines. Quality control is mainly applied for both raw materials along with excipients used and finished product. Flavonoids are the class of polyphenolic compounds, which are mainly distributed throughout the plant kingdom. Quercetin is a flavonoid which shows major pharmacological activities like anti-cancer, hepatoprotective activity, anti-spasmodic, and anti-inflammatory activity. Many herbal medicines and their formulations containing Quercetin are available in

market. Spectroscopic and Chromatographic methods play great role in the quality control and standardization of Quercetin in its isolated form, extract or any other herbal or polyherbal preparation.

AbbreviationsHPLC: High performance liquid chromatography; HPTLC: High performance thin layer chromatography; RP-UFLC: Reverse phase ultra-fast liquid chroma-tography; UHPLC: Ultra high pressure liquid chromatography.

AcknowledgementsThe authors are very thankful to Principal Dr. S. S. Jalalpure and Vice Principal Dr. M. B. Patil for their support and guidance. Authors are also thankful to the department of pharmaceutical chemistry KLE College of pharmacy.

Authors’ contributionsWe have assured that all authors have read and approved the manuscript. All the authors have equal contribution and participation in this review work. PK has reviewed all manuscripts on quality control and standardization of Quercetin in herbal medicines by spectroscopic and chromatographic techniques. SG helped in paraphrasing and writing the review part on spec-troscopic methods. KJ has collected the data on chromatographic methods for Quercetin. SH has helped in writing and paraphrasing the review on chromatographic methods of Quercetin. SS has selected the objectives and need of study and also helped in writing, correcting and communicating the proposed review article. All authors read and approved the final manuscript.

FundingNot applicable.

Availability of data and materialsThe review work has been carried out by us, and we assure you that it can be provided to you whenever required.

Declarations

Ethics approval and consent to participateNot applicable.

Consent for publicationNot applicable.

Competing interestsThe authors declare that they have no competing interests.

Received: 28 April 2021 Accepted: 22 August 2021

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