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Thin-Layer Chromatographic Identification of Flavonoids and Phenolic Acids

Contained in Cosmetic Raw Materials

Article  in  Journal of Liquid Chromatography & Related Technologies · March 2016

DOI: 10.1080/10826076.2016.1163467

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Thin-layer chromatographic identification of

flavonoids and phenolic acids contained in
cosmetic raw materials

Marta Skorek, Karolina Jurczyk, Mieczysław Sajewicz & Teresa KowaIska

To cite this article: Marta Skorek, Karolina Jurczyk, Mieczysław Sajewicz & Teresa Kowalska
(2016)Thin-|ayer chromatographic identification of flavonoids and phenolic acids contained
in cosmetic raw materials, Journal of Liquid Chromatography & Related Technologies, 39:5-6,
286-291, DOl: 1 0.1 080/'1 0826076,201 6.1 1 63467

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 ]oURNAL oF LlQUlD CHRoN,]AToGRAPHY & RELATED TECHNoLoGIE5
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http://dx,doi,or9l ] 0.1 080/ ] 0826076,2o 16.1163Ą67 Taylor&F.anCis Croup

Thin-laYer chromatographic identification of flavonoids and phenotic acids

contalned in cosmetic raw materials
Marta skoreĘ karolina )urczyk, Mieczysław sajewicą and Teresa kowalska
Department of General Chemistry and Chromatography. lnstitute of Chemistry, University of Si|esia,
Katowice, Poland

ABsTRAcT
KEYWoRDs
PolYPhenols, widespread in the kingdom of plants, are targeted in numerous chemical, biochemical, Botani(al raw materials;
and clinical studies. Flavonoids make a particularly interesting group of polyphenols due to theii cosmetic in9redients;
diverse, biological activitie5. ln both in vivo and in vltro studies, tńe-y strongly demonstrate antioxidant, flavonoids; phenolic acids;
anti-inflammatory, anticancer, and anti-aggregation activities, and they aie also known for tl9ńt"nin9 quercetin; rutin; TLC
\c the caPillary blood vessels. Due to their remarkable biological activities, flavonoids find'a wldó
sPectrum of Practical applications and the botanical materia|s rich in flavonoids are omnipresent in
N traditional medicines, phytotherapy, pharmacotherapy, and cosmetics, particularly attractive
d biological properties and, hence, considerable valor' of the cosmetics are attribuńd to such
flavonoids, as rutin, quercetin, naringenin, catechin, catechin hydrate, and apigenin, ln our study, we
\c
selected a simple and cost-frie_ndly thin-layer chromatographiŹ technique to identify the aforeńen-
cn tioned flavonoids, and also_caffeic, ferulic,9allic, and cóumaric acids, in the commórcially available
,{ cosmetic raw materials (such as extracts derived from red wine, grape skins, pomegranate peel5 and
juice, green tea, etc.) Quercetin and rutin proved to be thó two flavonoid5 mo5t fiequently
Ci
encountered in the examined cosmetic raw materials. The results of such investigations provide the
d
? Phenolics-oriented evaluation of the quality of the cosmetic raw materials ind, hence, better
o judgment oh the applicability thereof, e.9,/'a5 components of specific cosmetics prepared to
treat
rJ) various skin problems.

o
ą

.+ lntroduction
o witlr diverse chemical structures and biological properties.
O
Flal,onoids and plrenolic acids are popular constituents of raw Anrong pharn-racological properties of phenolic acids, one
a
U materials utilized in pharmaccutical, alimentary, and cosmetic can name their anti-inflammatory, antimutagenic, antipyretic,
o antirheumatic, antibactcrial, antiviral, antifungal, cholagogic,
O industry. Flavonoids are occasionally named flavones or bio-
flavorroids, and due to tlreir plrarmacological activity, in the and antihepatotoxic properties.t5-7] Flavonoids and phenolic
6
past they were also referred to as vitamin P.|'] The discovery acids find applications in cosmetics both as active substances
J
of flavonoids dates back to the l930s of the past century, when which directly affect human skin and as antioxidants w]rich
-c
.o the Nobel Prize winner Albert Szent-Gyórgyi first identified extcnd the shelf life of cosmetic products.[8]
o
.o In the context of skin and beauty, the main role of llavo-
flavonoids in citrus fruits. Moreover, he also pointed out to
their positive role in lolvering the permeability of blood ves- noids and phenolic acids is caused by their antioxidant
sels, tvhich rvas reflected in the vitamin's name, and where p
properties. Flavonoids directly reduce the reactive oxygen
a
c stands for permeability.I2] Flavonoids are omnipresent in species to more stable and inactive forms.[n] Moreover, flavo-
rroids hamper the oxidation of vitamin C responsible ior
fruits, seeds, leaves, and flowers ofplants. So far, over 4000 fla-
vonoids havc been discovered and described. In plant material, maintaining the tightness of the blood vessel wails; so in
they function as natural pigments and their color spectrum
an indirect way, they are also responsible for the tightness
reaches lrom yellow in citrus fruits (due to thc presence of fla-
of tlre wall.t9'10] Certain flavonoids characterize with anti-
allergic properties. Astragalin, fisetin, kaempferol, myricetin,
vonols, flavones, chalkones, and aurones) to navy blue irr blue-
quercetin, and rutin obstruct the formation of the inl]amma-
berrics and somc other berries (due to the presence of
anthocyanins).t'] Flavonoids exert a vast number of physio-
toIy state nrcdiators, i.e., histamine and proinllammatory
cytokines, TNF-z, and intcrleukins-1 /i,6, and 8. It was pro-
logical effects, such as antioxidant, antifungal, anti-inflamma-
tory, ven t]rat flavones (luteolin arrd apigenin) present in plants
anti-atherosclerotic, spasmolytic, anti-aggIegation,
obstruct potentially pathogenic functions of lymphocytes T,
diuretic, detoxicant, anti-arrhlthmic, hlpoterrsic, tightening
the capillary blood vessels, anticancer, improving tlre eyesight
which can become active in the case of autoimmune
diseases.["'"] Owing to the hampering effect exerted on
sharpness, etc.['l'5] Phenolic acids belong to thc group of poly-
the activity of selected enzymes (hyaluronidase, lvhich cata-
phenols and abundantly occur in different plant tissues, They
lyses depolinrerization of hyalurorric acid, and elastase),
belorrg to the group of secondary metabo]ites and characterize

coNTAcT Teresa Kowalska @ teresa,kowa|ska6us.edu.pl l?ł lnstitute of Chemistry, University of silesia, 9 Szkolna Street, 40-006 Katowice, Poland
Color versions of one or morJof the figures in ńis articl'e cli be found online on at www,tandfonline.com/ljlc.
() 20']6 Taylor & Francis

certain flavorroids (c.g., kacmpferol) strengthen the connec-
tive tissues and tighten.the blood vessels. As a result, such
flavonoids contributc to the minimization of internal bleed-
ings, inllammation cases, and swellings. At the same time, an
improvement is observed in the blood flow of the capillary
vessels which results in more effective nourishment and aer-
atiorr of the tissues, and in improved moisture and elasticity
of skin. Quercetin hampers the activity of the histidine
decarboxylase, lowers the histamine levels in the organism,
arrd, in effect, exerts antiswelling, anti-inflamnratory, and
pairr-killil-rg activities. Isoflavonoids, (e g , genisteir-r) also
knorvn as phytoestrogens, exert a retardation effect on skin
aging, due to their ability to bind to certain rcceptor groups,
un uttructive component of antiwrinkle
.lf,i'.i,'.'r".,tlflr,
External use of the phytohormones in cosmetics shields
the ccl]ular membranes from oxidation and damage, and
cl cxerts a retardation effect on the activity o[ collagenase (an
enzyme responsible for thc dcgradation of the collagen
ż
6
structure) and e]astase (an enzynre which degrades elastine).
An additional positive effect exerted by the phytohormones
cJ
is the strengthening of the capillary walls and the better
A
nourishment of the skin. Increased amounts of the hyaluro-
d nic acid (due to hampering an activity of hyaluronidase
which destroys its structure) promotes increased elasticity
E
! Preparation of phytochemical standard and cosmetic
O and smoothncss of the skin. Apart from delaying skin aging,
a raw materiąl solutions
plrytohornrorres perfectly tend to the skin which 1oses
o.
o its lirnrness. Contrary to the estrogens of animal origin,
o externally applied herbal phytohormones only 1imit the
..- sphere of action to the application sitc, and do not provoke
o
5! any unwanted side effects.t1] Numerous investigations
O
c have confirmed the effect of flavonoids in canccr prevention.
U Such activity is attributed, e.g., to quercetin, kaempferol,
o
genistcin, 5,7-dimethoxyflavone, 5,7,4'-trimethoxyflavone,
0
apigenin, and tricinc.L''-'']
d Flavonoids and phcrrolic acids present in plants are not
J
easy to obtain synthetically and such syntheses are fairly
-c
cor-nplicated.t"] In the pharmaceutical and cosmetic indus-
o
-o trics, one takes advantage of the natural plant rarv materiais
o known for being rich in flavonoids. This trend is enhanced
by contemporary fashion promoting phytochemistry, eco-
o
cosmetics, and healthy diet. In order to attract customels'
attention and cnhance their own prestige, manufacturers
of cosmetics and cosmetic raw materials emphasize botan-
ical cornposition of their products and the contents of
herbal antioxidants therein. However, their specifications
rel-er mostly to the total 1lavonoid or phenolic acid content,
,lvithout specifying individual chernical compounds. Tlrus,
the ain of this study lvas to identify selected flavonoids
and phcnolic acids playing a particularly important role in
skin care, which are contained in commercially available
cosmetic rarv materials of herbal origin. Identification of
sclected examples and quantification of thcse compounds
rvere carried out using thin-layer chromatography (TLC)
due to the sinrp}icity and rapidity of this particular tech-
nique, cost-eft'ectiveness, possibility to ana|yze severai
products in parallel, arrd to a wealth of helpful ]iterature
relerences on the analysis of flavonoids ar-rd phenolic acids
in natura] samples.
JOURNAL OF L|QUlD CHROMATOGRAPHY & RELATED TECHNOLOGIES Q Zal
Experimental
Apparatus, reagents, sdmples
In the experiments, ethyl acetate, methanol, ethanol, toluene,
formic acid, and acetic acid of analytical purity (PPH POCh,
Gliwice, Poland) wele used. Phytochemical standards, i.e.,
flavonoids (rutin, quercetin, naringenin, catechin, cateclrin
lrydrate, and apigenin) and plrenolic acids (caffeic, ferulic,
gallic, and coumaric acid) of analyical purity were purchased
fronr Sigmir-Aldrich (St. Louis, MO, USA). Water lvas
de-ionized and doub]e-distilled in our laboratory by means
of the Elix Advantage model, Millipore system (Millipore,
Molsheim, France).
Thin-layer chromatographic analyses were carried out rvith
use of the precoated silica gel F2§4 TLC plates (20 x 20 cm; cat.
# I.057 47 ; Merck, Darmstadt, Germany).
The commercial solid (iyophilized) samples of the cosmctic
raw materials of botanic origin were purchased from the
online stores. The producer/distributor names were Natur
Vit Chem and Kosmetyki Naturalne. During the course of
our investigations, all samples of phytochemical standards
and the cosmetic raw materials (known for their photosensi-
tivity) were kept in a cool and dark place.
For tlre purpose of the TLC arralysis, the flavonoid and
pherrolic acicl standards were dissolved in methanol and
disso]ution was enhanced by ultrasonication. Solutions of
phytochemical standards were prepared at the concentration
of 1.00nrgmL l, Due to high costs of these standards, the
calibration curves based on different vo]ume amounts thereof
(instead of the recommended series of standards at diffcrent
concentration) and each calibration curve was built on five
different microgram amounts of a given phytochemical stan-
dard per one chromatographic spot. Based on the calibration
curves obtained, the contents of flavonoids and phenolic
acids in the cosmetic raw materials were assessed. It is note-
worthy that the standard solutions were prepaled immcdi-
ately before use.
Each solution of the phytochemical standard used for tlre
calibration curve was spotted on one and the same chroma-
tographic plate in the aiiquots depending on the standard.
The chromatographic peak heights (i.e., signal intensities in
mAV) were plotted against the microgram amounts of the
standards applied to the chromatographic plate. The limit
of detection (LOD) and tlre 1imit of quantification (LOQ)
were calculated using the formulas LOD :3.3 x SD/n, and
LOQ : 10 x SD/a, rcspectively, where SD is the standard
deviation of tlre pcak height taken as a measurc ol noise,
and a is the slope of the corresponding calibration curve
(y: r*+ b). Then, the quantification of the flavonoid and
phenolic acid contents in the cosmetic raw materials was
performed.
The commercial lyophilized herbal extracts used for the
preparation of cosmetics rvere also dissolved in methanol to
obtain 1% (lv/v) solutions and dissolution was supported by
ultrasonication. Although mcthanoi is not a 'green' solvent,
 288 O M. sKoREK ET AL.

it dissolved phytochenrical standards and the lyophilized c ś

.9
plant extracts (known for their photosensitivity) faster than ,9

ethano], and in this sense it considerably outperlormed the .!

latter one. =
Ń6:
o
o
o
^c-
Development of chromatograms o _* c^
ą
'I'he 5-60-pLL aliquots of herbal extfact solutions were
Eą
applicd to the chromatographic plates with use of the cali- :E o
o >999o o o o oF
E
c
@
brated microcapillary pipettcs (Sigma-Aldrich). Different c o
o c 9
o
@
@
a ,-o o !
=
! -^ =
E
a
aliquots of the herbal extract solutions were needed due
to the diff'erent natural concentrations of flavonoids and o
E
o .!
ś
^
E
c -E o o ś =ś,:'ó-ś_
*.^:-'nć^=

,9 H=ĘY19
J
!-
!
a
a ś
śś o Ec
=
c \9 ; F 6 @>]j:oó
plrenolic acids wit}r diff'crcnt investigated species. Tlrirteen =
ooś
_ H -o 6, 9 = Pl . !,9 o
samples rvere applied per one żOx2Ocm plate in the 15- o
^;-E^E,^ co aJ
mm distance from one another and in the 10-mm distance
ó !! > =^i:E=-::
t>t9c_9E=tsa]o<otr=
*6xax3xixpEbbsr
c9c _ c_ c9EU!-! 9c9 @
o o
from the plate bottom and the side edges. To each plate, = =i ': *'a
N łŹ N rc N ru N oN o c = c ń
d-.&tróó
o o
- -,- - q
\c two to four samples of the flavonoid and/or phenoiic acid >>(9
f
c.l standards were applied (in the aliquots ranging from l to E 6c
aś 30 ptl, depending on a given standard) and thc remaining .E
!o
! o€
samples were those of the cosmetic raw materials. Chroma- c t
o c!o cĄ
c
tograms were developed to tlre distance of 15cm in stan- E
c.l
dard (vertical) chromatographic chambers (CAMAG,
c
,9
!',
ń o=
@U
:ś-
'' oU
6@=
Muttenz, Switzerland), for 20 min presaturated with mobile .E
.|! E =o -.E
śj phase. Due to a variety of cosmetic raw materials and phy- }+i- c -- c
9?E
=
o 6<;"łt
o
E tochcmlcal standards investigated in this study, a number = X-o<In
ś z--
of n-robile phases and visualization methods earlier
,9
o o
a described ir-r the literature were tested ('I'able 1). The -E
a-
o developed chromatogranrs were densitotnetrically scanned a!- :-=
::- =
- :
E
:
-Y =
Ę = =
-
a in the rcflectancc or lluoresccnce mode rvith use of thc Łc
'óq oOO o
N
O
N
O
N
o
N
O
N
ó
N
=
O
N
;o Desaga (Heidelberg, Germany) model CD 60 densitonreter
-QN
Nńń ń ń ń ń ń ń ń
o a6
equipped with the Windows-compatiblc ProQuant software, o
o o
and the working wavelengths were selected depending on
€ ;o
o-6 h o N ń h @
a 6\c9 Ę 9 !,l 9 \ \
U the traced flavonoid or phenolic acid (Tab]e 1), For each OOO Q O O O O O
o ś
o cosmetic raw material, the analyses were repeated six times o
E 6
.a: (n :6). The analogous number of analytical repetitions c
,9 =a
od
J
6
were applicd to phytochemical standards (flavonoids and ,! 1ruć--
_: :_.j-!
phcr-rolic acids), ivhich moreover lvere analyzed at five ! 9 f '- 1
:--ć';a=ó!p= =
-o
diflerent aliquots each.
=
-c =. ; E : H H
o oi =ć
9ó
!@,=,=.=ń
F = ś 9 ,9
!o
6
E oz,
=6 ą
< Uó 6
U 9 Uo Io U6
Visualization of chromatograms
}
c A fairly wide selection of stationary and mobile phases : ń
were tested in our study. Eventually, we decided to use sil- F= e1 + +
ica gel as stationary phase and the mobile phases were o
E
Ti
c
taken from the literature (Table 1), lvhich performed the ,9
y- ,9 9
,9
o
best in our pilot experiments. In this tab]e, wc also sum- o
ą
marize tlre basic details on densitometric scanning of tlre ę
o
a E
o
ć/ó
;l
-Y
irj aj
ańE
TLCs (the wavelengths, the reflection, or fluorescence o
--
9ŃH
lnode), atrd on chcmical visualization thereof with use of @ =
ś
.9
ąo
P y_
ś
visualizing reagents (assessment of the chromatograms in E
.!!
=ś @

the dayliglrt and with use of the CAMAG UV cabinet, o

o 9Pi,;
-Ęfi j ó.j
c
at the rvavelengths 254 and 366 nm). Pictures of the selec- E o
o
->! -
6
ted TLCs provide examples of differcnces in visibility of śś @ ts

chromatographic spots prior and after visualization .=

(Figure 1). Flavonoids and phenolic acids found in the TC Ec

p
investigated herbal raw materials are listed in Table 2.
E
F !
c
In T'able 3, the results of exenrplary quantification of +3
oo
-_ą
,9
6c
otre flavor-roid (quercetin) and one phenolic acid (caffeic ś
L
acid) are given.
 JoURNAL oF LlQUtD cHRoMATOGRAPHY & RELATED TEcHNoLoGlEs @ zss

Results and discussion

Ja
a6
J
* Samples scrutinized in this study were prepared from the com-
§
mercially available cosmetic lrerbal raw materials (Table 2).
9o "ł
ta dl The main criterion for the selection of raw materials was an
o
cą
EE
ą
F expected high content of phenolics, and in the first instance,
of highiy appreciated antioxidants (flavonoids and phenolic
acids). Working samples of raw materials were prepared as
the 17o methanol (w/v) solutions, which guaranteed good
solubility of thesc substances and their concentrations anal-
ogous to those applied in cosmetic preparations. Selection of
the flavonoid and phenolic acid standards was based on their
particular importance for the skin well-being and care.
Currently, information on the contents of individual
flavonoids and plrenolic acids in tlre cosmetic ralv materials
is scarcely available, as manufacturers prefer to characterize
the total contents of the phenolics without itemizing individ-
ual compounds. Arr emerging interest in a more detailed
ól characterization of the cosmetic raw materials is due to a
(B
new ecological trend in the cosmetics. C]ients who buy rather
\o expensive cosmetics based on natural active substances expect
c! Łęffi',Ę their free-radical scavenging action, according to the manufac-
,-i turer's declaration. Instead, they often become the recipients of
incomplete information on a composition of a given cosmetic
ś
Ń
in form of laconic statements, e.g., that 'the cream contains
Ę polyphenols', 'high contents of flavonoids', or 'free-radical
scavenging activity due to phenols'.
a
Identification and exemplary quantification of the cosmetic
o.
o raw tnaterials targeting selected llavonoids and phenolic acids
(presented in tlris study) were based on the TLC results. Aller
O
havirrg tested a wide selection of the TLC systems proposed in
50
o the literature, certain systems were selected as thc best per-
o Figure 1. Selected examples of the Vi5ualiżed thinlayer chromatograms (a) forming orr.s.|'u-'o] These systems included the chromato-
U Determination of rutin in the extract from skin of red grapes (sample no, 1); graphic plates precoated with silica gel F25a and one of the
ó/
mobile phase: ethyl acetate: toluene: formic acid, 7:3:1(v/v); VisUalization in the three different mobile phases. A11 the details regarding the
o
CAMAG UV cabinet at 254 nm and in the day|ight, (b) Determination of coumaric
acid in the extract from red wine (sample no, 3) and the extract from young applied mobile phase, the densitometric visualization rnode
J barley §ample no.']1); mobile phase: toluene: ethyl acetate: acetic acid, (reflectance or fluorescence), the applied wavelength deper'd-
34:15:1 (v/v); visualization in the CAi\4AG UV cabinet at 254nm.
,.o
ing on the targeted compound, and the visualization effect
-ó
o are sumntarized in Table 1. Two working examples of the
ci visualized chromatograms (valid for rutin and coumaric acid)
o
are given in Figure 1. The results obtained confirm the

Table 2.Samples of plant extracts used in cosmetic preparations (5olUtion concentrations in methanol, 17o
(w/v)) the names of online prodUcers/distributors,
presence of the investigated phenolics in the
flavonoid, and phenoIic acids found in the investigated samples, and references to the titerature which confirms the
samples of botanlcal
Producer/di5tri butor Flavonoids and/or phenolic acids found References
Sample (17o 5olution; w/v)
,]
Extra(t from skin of red grapes Natur vit chem Rutin, coumaric acid I20]

2 Extra(t from grape seeds Kosmetyki Naturalne Rutin, naringenin, coumaric acid, ferulic acid {21]

3 Extract from red wine Kosmetyki NatUralne Rutin, api9enin, qUercetin, coUmari( a(id, I22)
feruIic acid
jui(e of pomegranate Kosmetyki Naturalne Quercetin t23]
4 EXtraCt from
5 Extract from peel of pomegranate Kosmetyki Naturalne Rutin, gatic acid, coumaric acid, ferulic acid t23]
Ko5metyki NatUralne Rutin L24,26l
6 Extract from raspberry fruit
7 Extract from strawberry,juice Ko5metyki Naturalne Quercetin, coumaric acid |25,ż6)
Ferul]c acid I27)
8 Extra(t from blueberry juice Kosmetyki NaturaIne
n.f,' |25]
9 Extract from banana julce Kosmetyki Naturalne
,]0
Extract from mandarin juice Kosmetyki Naturalne n,f.t 128]

RUtin, apigenin, catechin, coumaric acid,

rrql
1,1 Extract from young barley Kosmetyki NatUralne
ferulic acid
12 Extract from green coffee Natur vit chem Rutin, Catechin, cate(hin hydrate, caffeic acid I30]

13 Extract from 9reen tea Kosmetyki Naturalne QUercetin, Cate(hin, catechin hydrate, 9alic [30,3,1 ]

acid, coumaric acid, ferulic acid, caffeic acid

'n.f., not found.
 29o o M, sKoREK ET AL,

presence ofthe compounds from the groups offlavonoids and
phenolic acids in the botanic raw materials commonly utilized
in cosmetic industry (Tablc 2). The highest numbers of differ-
ent scrutinized phenolics were found in the extracts from
green tea (scven compounds) and red rvine (five compounds).
These results seem to convincingly support a widesprcad
popularity of these two extracts in cosmetics. None of the used
standard phenolics rvas found in the extracts from nrandarin
juice or banana juice.
Quantilication of quercetin and caffeic acid (as the selected
specimens of flavonoid and phenolic acid) was done based on
the respective calibration curvcs prepared with use of the quer-
cetin and caft'eic acid standards. Calibration equation valid for
quercetin was equa] to y:24,96*+74,żż (correlation coei:
ficient, r:0.988 and standard deviation, SD:0.675). The
respective LOD and LOQ values valid for quercetin were equal
\c l.
to 0.101 and 0.306 pg spot Thc analogous equation valid for
N caffeic acid was equal Lo y:136.9,a716,6 (corre\ation coef_
ficient, r:0.984 arrd standard deviation, SD:0.887). The
respective LOD and LOQ values va]id for caffeic acid were
\c
equal to 0.021 and 0.065 pg spot-l. Then the quantification
c]
of quercetin and caffeic acid (as two examples of the discussed
,{
phenolics) was performed for a selection of the investigated
cosmetic raw materia]s and the results obtained are summar-
6
ized in Table 3. As it comes out from tlrese results, thc highest
Ę nolic acids, lvhile in this study fcrulic acid alonc was identificd.
irmounts of quercetin were found in the extracts from peel
a and juice of pomegranate, whereas quercetin was not lound
O in the extract from green coffee. Calfeic acid was found in
the extracts from grecn tea and green coffee only, and not in
the remaining scrutinized samples.
o
oo Quantitative results presented in this study cannot
o
directly comparcd with those taken from the literature, as no
U sources are available on the TLC quantification of flavonoids
o
or phenolic acids contained in the cosmetic rar,v materials.
o
ó Tlrus, the results presented in this study can only be compared
J rvith the data obtained for the juices and/or extracts derived
from natural products. A roughly comparable exampie is furn-
-o ished in a study on detertnination of quercetin in the dried
o
leaves of the Yerba mate tea.[32] The established content of
d
quercetin in these leaves was 2.2mgg-\, which is double the
amount of the samc flavonoid in the lyophilized extract from
green tea (yet the nragnitude order of quercerin in the samples
of the two plants is the same; Table 3). Additionally, one has to
takc into the account that the two specimcns are botanically
different and the quantified products have been differently
pretreated (dried leaves vs. 1yophilized herbal extract).
App}ication of more sophisticated instrumenta] chromato-
graphic techniques (in the first instance, HPLC-DAD or
LC-MS) allows identification of considerably more flavonoids
and phenolic acids than TLC, certainly due to higher sensi-
tivity of the former techniques than tlre latter one. Now lct
us give several practical examples to confirm this statement.
By means of TLC, in the extract from grape skins we found
rutin and feru]ic acid only, lvhereas with use of HPLC rvith dif-
ferent detectors, additionally gallic acid and catechin were
found.t2"'3'] In the extract from raspberry fruit, we managed
to find rutin, whereas the RP-HPLC analysis of tlre raspberry
fruit revcaled caffeic, coumaric, and fcrulic acids, but in fact no
rutin, Moreover, another HPLC-DAD analysis of raspberry
fruit revca]ed an additional plesence of quercctin.[2o'"] At tl-ri.
point, it is quite difficuit to comment on a reai source of this
discrepancy between our results and those taken from the
Iiterature. As next example, iet us consider the pomegranate
juice. By means of TLC, we managed to identify in this pro-
duct quercetin only, whereas another soulce reports on an
additionaI presence of catechin, gallic acid, and yet some other
phenolic acids.i']] According to the litcrature report based on
the HPLC results,[27] extract from blueberries contains ferulic,
caffeic, and coumaric acids, quercetin and yet some other phe-
Nevełtheless, a final statement can be made tl-rat identification
of flavonoids and phenolic acids made by use of TLC pre-
sented in this study remains in reasonable agreement lvith
tlre results obtained with use of the other chromatographic
be techniques (as shown in Table 2).
conclusions
Based on the identification and quantification results obtained
by means of TLC and discussed in this study which refer to the
presence of flavonoids and phenolic acids in the cosnretic raw
materials of botanical origin, and on the validity thereof
confirmed by a comparison with the data obtained from the
iitcrature, a justified statement can be madc that the proposed
TLC protocols can effectively be used for the screening and
quar-rtification of flavonoids and phenoiic acids in a rapid con-
trol of the aforementioned cosmetic raw materials.

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