Vous êtes sur la page 1sur 10

j. Cosmet.

sci., 58, 35-44 (January/February


2007)

High-performance liquid chromatographic


determinationof
arbutinin skin-whiteningcreamsand medicinal
plant extracts

WISANU THONGCHAI, BOONSOM LIAWRUANGRATH,


and SAISUNEE LIAWRUANGRATH, Department of
PharmaceuticalSciences,
Facultyof Pharmacy(W.T., B.L.), and
Departmentof Chemistry,
FacultyofScience
(S.L.), Chiang
University,
ChiangMai, 50200, Thailand.

Accepted
for publication
November
1, 2006.

Synopsis
A high-performance
liquid chromatographic methodwasdeveloped for quantitativeanalysis
of arbutin.The
arbutinwasseparated on an ODS Hypersil© %8 columnwith a mobilephaseof water:methanol:0.1 M
hydrochloric
acid(89:10:1,v/v/v).The levelof arbutinwasmeasuredby meansof UV detectionat 222 nm.
The optimum conditionsfor arbutin quantitativeanalysiswere investigated.The calibrationcurvewas
foundto be linearup to 1,000[tg/ml-• of arbutinconcentration,
andtheworkingcalibrationcurvefor
arbutindetermination overthe range0.5-30.0 lxg/ml-• of arbutin(r2 = 0.9999)wasestablished.
The
relativestandarddeviationsfor intradayand interdaywerefoundto be 0.98% and 1.15%, respectively.
A
detection
limit (3cr)andquantitation
limit(10cr)of0.02pg/ml-• and0.212g/ml
-•, respectively,
anda mean
percentagerecoveryof the spikedarbutinof 99.88 -+1.12% wereobtained.The proposed
methodhasbeen
appliedto the determination of arbutinin commercial skin-whitening
creams(Arbuwhite© cream,Super
Whitening
© cream,andShiseido
© cream)with average
contents
of 7.60, 5.30,and57.90mg/g-•, respec-
tively. It wasalsoappliedto the determinationof arbutin in medicinalplant extractsfrom Betu/aa/noides
Buch. Ham., Clerodendrum petasites
S. Moore, Curculigo latifo/ia Dryand. Var. latifolia, and HeJperethusa
crenulata
(Roxb.)Roem,levels
ofwhichwerefoundto be3.50,1.50,1.10,and0.12ing/g
-• , respectively
(no
articlereportedin the literatureaboutarbutinanalysis).
The proposed
HPLC methodis rapid,simple,and
selectivefor routineanalysis.

INTRODUCTION

Skin-whiteningproductshavebecomeincreasingly in demandin the pastfew years.The


main purposefor skin-lighteningproductsis to lighten the skin aswell as to evenout
skin toneor to treatpigmentationdisorders suchasfreckles,melasma,pregnancymarks,
and age spots(1). The most successful recentand natural skin-whiteningagentsare
arbutin, vitamin C, kojic acid, licoriceextract,burnet root extract,scutellariaextract,

Addressall correspondence
to BoonsomLiawruangrath.

35
36 JOURNAL OF COSMETIC SCIENCE

andmulberry.Theseagentsareall tyrosinase inhibitors,whichinactivatetyrosinase


(the
enzymeresponsible for skin pigmentation)by chelatingwith its vital copperion and
suppressing tautomerizationfrom dopachrome to DHICA. Normal skin coloris formed
by melanin,a naturalpigment that alsodetermineshair and eyecolor.In the skin,the
enzymetyrosinase biochemicallyconvertsthe aminoacidtyrosineinto melanin.Hyper-
pigmentationoccurswhentoomuchmelaninis producedandformsdepositsin the skin.
Hyperpigmentationis not a medicallyharmfulcondition.However,it is alwaysadvis-
ableto havenewbrownspotscheckedby a dermatologist to makesurethey arenot skin
cancers(2). Arbutin is a naturallyoccurringglycosideof hydroquinone(Figure 1).
Arbutin is foundin the barkandleavesof variousplants,usuallyoccurringtogetherwith
methylarbutin.Naturally occurringarbutinwasfirst characterized by Kawalier(3), who
obtainedit from bearberryleaves.It is alsofoundin the leavesof blueberry,cranberry,
and pear. Syntheticarbutin was first reportedby Mannich (4), and later by others.
Commercialarbutin is almostalwayssyntheticin origin. Becauseof its antibacterial
properties,arbutinis a constituentof the traditionalmedicine•va •rsi, and it is widely
usedin a varietyof formulations(5). The ability of arbutin to inhibit humanmelanin
synthesishas given rise to its wide use in many cosmeticformulations(6). Arbutin
protectsthe skinagainstdamagecausedby freeradicals.It is a skin-whiteningagentthat
is verypopularin JapanandAsiancountriesfor skin depigmentation. Arbutin inhibits
the formationof melaninpigmentby inhibitingtyrosinase activity(7). Backin the 18th
centuryarbutinwasfirst usedin medicalareasasan anti-inflammatory andantibacterial
agent.It wasusedparticularlyfor cystiris,urethritis,andpyelitis.Theseuseshavebeen
applied until today, when natural medicineusesonly natural ingredientsto treat any
disease.It may be usedto repressthe virulenceof bacterialpathogensand to repress
contaminatingbacteria.It is alsousedfor treatingallergicinflammationof the skin.It
can be usedto whiten the skin, to prevent liver spotsand freckles,to treat sunburn
marks,and to regulatemelanogenesis (8).
Arbutin is a very safe skin-whiteningagent for external use, which doesnot have
toxicity,a stimulatingeffect,an unpleasantodor,or sideeffectssuchas hydroquinone
(Figure la). Hydrophilicarbutin can be incorporatedin lipophilic media by encapsu-
lation. Arbutin hasthreemain properties:a whiteningeffect,an anti-agingeffect,and
a UVB/UVC filter (9). Arbutin is determinedby manymethods:the spectrophotometric
method (10-12), capillary zone electrophoresis(13), and thin-layer chromatography-
densitometry(14). The proposedHPLC method for determiningarbutin in skin-
whitening cosmeticproductsand somemedicinalplants is more sensitive,precise,and
lesstime-consumingthan the previousHPLC methodsdescribedin the literature (15-

OH

• OH
CH20
H OH

Hydroquinone Arbutin
(a) (b)
Figure 1. Chemicalstructuresof hydroquinone
(a) and arbutin (b).
HPLC DETERMINATION OF ARBUTIN 37

18). Furthermore,there is no data in the literature about the isolationand quantitative


analysisof arbutin in BetMaalnoides Buch.Ham., Clerodendrz/m petasites
S. Moore,C•r-
cMigolatifoliaDryand.Var. latifolia,andHesperethz•sacrenzdata
(Roxb.)Roem.Therefore,
it is interestingto investigatethe arbutin content in thesemedicinalplants, because
naturallyoccurringarbutin is very safeskin-whiteningagent.

EXPERIMENTAL

APPARATUS

HPLC analyseswere carriedout with Hewlett PackardModel 1100 liquid chromato-


graph with autosampler,thermostaticcolumn compartment,online degasser,and a
UV-visibledetectormodelG 1313 A. The columnusedwasODS Hypersil© C•s (125
mm x 4 mm, 5.0 t•m [Chromtech,
Stockholm,
Sweden])with a Lichrosphere
© 100
RP-18 (4 mm x 4 mm, 5.0 t•m) guard column. The mobile phasewas a mixture
containingvaryingratiosof methanol,water, and 0.1 M hydrochloricacid, vacuum-
filtered through 0.45-t•m nylon membranes(Germany)beforeuse.The following in-
strumentswerealsoused:a simultaneous spectrophotometer
(Spekol1200), a pH-meter
(Model pH 900, Precisa,Switzerland),a water bath and shaker(Model SB-200-10,
Thailand), an ultrasonicator(Model 889, Cole Parmer,USA), and a rotary evaporator
(EYELA N-N series).

REAGENTS

The followingstandardreagents
wereused:arbutinHPLC grade98% (Sigma,St Louis,
MO) and resorcinol98% (Fluka). The following reagentswere used:hydrochloricacid
(AR) (FarmitaliaCarlo Erba, Italy), glacial aceticacid (AR) (FarmitaliaCarlo Erba),
acetonitrile(HPLC grade) (Lab-ScanAnalytical Sciences,Ireland), ethyl acetate(AR)
(BDH laboratorysupplies,England),methanol(HPLC grade)(Lab-Scan AnalyticalSci-
ences),and ether (AR) (Lab-ScanAnalytical Sciences). De-ionizeddistilled water was
usedthroughout.

SAMPLES

Skin whitening
prodz/cts.
Arbuwhite© cream(Nature Best Health ProductCo., Ltd.,
Thailand),SuperWhitening© cream(Aunyamanee Herbs,Thailand),and Shiseido
©
cream(ShiseidoCo., Ltd., Tokyo,Japan)were used.
Plant materialand location.The bark of BetMaalnoideswas collectedat Bah Bae Village,
Mae Dtang District, ChiangMai Province,Thailand,in October2003 and wasiden-
tified by W. Thongchai1. Voucherspecimens havebeendepositedat CMU Herbarium,
ChiangMai University,ChiangMai, Thailand. The rootsof Clerodendrz/m petasites
S.
Moorewerecollectedfrom ChiangDow, ChaingMai, Thailand.The tubersof Curculigo
latij$liaDryand.Var. latifoliawerecollectedfrom Papae,Maetang,ChiangMai, Thai-
land. The trunk of HeJ;Oerethmacremdata(Roxb.) Roere. was collectedfrom Mae Sai,
Chiang Rai, Thailand.
38 JOURNAL OF COSMETIC SCIENCE

PROCEDURES

Samplepreparation
(skin-whitening
cream).About 0.5 g of each whitening cream was
accuratelyweighed and transferredinto three separate25-ml volumetricflasksand
dissolved
in methanol.
To eachflask50 t•g/ml-• of resorcinol
wasaddedasaninternal
standard.The solutionwassonicatedvigorouslyfor 30 min, centrifugedat 4000 rpm for
30 min, and filtered on a Millipore membrane(0.45 t•m) to obtain a transparent
solution.The supernatantliquid wasusedfor chromatographic analysis.
Extraction
of medicinal plants.The dried medicinalplantswerepowdered.Then 6 kg of
the powderwasextractedwith two successive portionsof 5.0 1 of de-ionizedwater and
methanol.They wereshakenin a wrist-actionshakerfor five hoursandfiltered.Then the
solventof the filtrate couldbe removedeither by usinga spray-driedtechnique(tem-
perature
100øCandflowrate1.0 ml/min-•) to givea brownpowder,
or by usinga
rotatoryevaporatorto give a dark browncruderesidue.
Preparation
ofstandard
soluions.
A 1,000tag/ml
-• stocksolution
ofarbutinstandard
was
preparedin methanol.A seriesof eachstandardsolutioncontaining0.5, 1.0, 3.0, 5.0,
10.0,and30.0lag/ml
-• wasprepared
fromthestockstandard
solution.
Preparationof samplesoluions.
Three setsof medicinalcrudeextracts(5 g) and cosmetic
samples(0.5 g) of eachset were extractedunderrefluxwith 100 ml of 75% methanol
for 30 min andfiltered.The flitrate wasevaporated
to about12 ml andtransferred into
a 250-ml separatorfollowedby addition of 50 ml of water. The mixture was then
extractedwith ether(2 x 30 ml). The combinedaqueouslayerwasextractedwith ethyl
acetate(3 x 50 ml). The combinedethyl acetateextractwasthen evaporated to dryness
and dissolved in 10 ml of methanol.

Preliminaryinvestigation.
A preliminary investigationwas carriedout to separatesome
chemicalconstituentsby TLC. The crude extract was extractedwith 75% methanol
underrefluxfor 30 min andthen filtered.The flitrate wasevaporated to about12 ml and
transferredto a 250-ml separatingfunnel togetherwith 50 ml of water.This solution
wasextractedthree times with 50 ml of ethyl acetate,and the combinedethyl acetate
extractswere evaporatedto drynessand the residuesdissolvedin 10% methanol.The
samplesolutionandthe standardsolutionswereseparated on a silicagel GF254(20 x 20
cm) glassplate, using ethyl acetate:methanol(9:1) as a developingsolvent.The crude
extractgavefive well-definedspots.The Rf valueof eachspotwasexactlythe sameas
that obtainedfrom eachspotof standard.
Optimizationof experimentalconditions
for RP-HPLC. RP-HPLC was performedunder
isocraticconditions.All experimentalconditionswereoptimizedby meansof a univari-
ate method as follows:
Analyticalwavelength.Optimum absorbance
of eachstandardsolutionwasdetermined
byinjection
ofthesame
amount
ofmixedstandard
solutions
(5.0lag/ml
-•) at different
wavelengths
from 200 nm to 400 nm. The mobilephasewasa mixtureconsisting
of
water:methanol
(80:20v/v)witha flowrateof 1.0ml/min-•.Astheoptimum
to obtain
the bestsensitivity,•maxwaschosen.
Mobilephase.Varioussolventsystems weretestedasthe mobilephasefor the separation
of arbutinin the samples,
e.g.,water:acetonitrile:0.1
M hydrochloricacid(94:5:1,v/v/v),
water:methanol:0.1M hydrochloricacid (89:10:1, v/v/v), and methanol:100mM phos-
phatebuffer,pH 2.1 (10:90 v/v).
HPLC DETERMINATION OF ARBUTIN 39

Mobile phaseflow rate.The optimum flow rate of the mobilephaseshouldprovidegood


separation,high sensitivity,and short analysistime. In this work, after the optimal
wavelengthwasselected,optimizationof the flow rate wascarriedout by injecting the
same
concentration
ofmixedstandard
solutions
atvarying
flowratesfrom0.5ml/min•
to 1.0 ml/min -•.

Recommended
RP-HPLC procedure.
A sampleand/orstandardsolutioncontainingarbutin
wasseparated
ona reverse-phase
ODSHypersil
© C•8column(125 mm x 4 mm, 5.0 pm)
and detectedat 222 nm. An aliquot of 100 pl of a seriesof arbutin standardsolutions
and 100 pl of sampleextractwasinjectedinto the LC systemand elutedwith the mobile
phase,water:methanol:0.1M hydrochloricacid (89:10:1, v/v/v) (flow rate = 1.0 ml/
min •). Theareaof thearbutinpeakwasmeasured.
Arbutinconcentration
in theplant
extractwas determinedby referenceto the calibrationcurvepreparedunder identical
experimentalconditions.

RESULTS AND DISCUSSION

A high-performanceliquid chromatographicmethodfor the determinationof arbutinin


skin-whiteningcreamsand medicinalplant extractscontainingarbutin wasdeveloped.
The experimentalconditionswere investigatedand the proposedmethodwasalsovali-
dated.

OPTIMIZATION OF RP-HPLC CONDITIONS

The optimal conditionsof HPLC for determiningarbutin were carriedout under iso-
cratic conditions.Variousmobile phasesystemswith differentcompositions were in-
vestigated.First, the optimal wavelengthfor the detectionof arbutin and other com-
pounds,as mentionedearlier,wasinvestigated,and the UV spectrumof eachstandard
compoundshowedthe absorptionmaximaat 222 nm. A wavelengthof 222 nm showed
the highestsensitivityfor arbutin. Second,amongthe mobile phasesstudied,a mixture
consistingof water:methanol:0.1 M hydrochloricacid (89:10:1, v/v/v) wasusedas the
mobilephase,and it wasfoundthat this mobile phasewasthe most suitablebecauseit
resultedin goodretentiontimes, resolution,and satisfactory peak profiles(Figure2).
Finally,theoptimumflowratewas1.0 ml/min-•, asit gavea goodresolution,
high
sensitivity,a shortanalysistime, etc. In the RP-HPLC analysis,arbutin and resorcinol
(internal standard)showedsinglesymmetricalpeaks(retentiontime 5.7 min and 10.7
min), respectively.

METHOD VALIDATION

The proposedmethod wasvalidated accordingto U.S. Pharmacopoeia;


USP (19).
Sensitivity.
The sensitivityof the assaywasdeterminedin termsof the detectionlimit
(LOD) and the quantitationlimit (LOQ). Detection limits and quantitationlimits were
estimatedfor eachof the examinedcompounds.The valueswere calculatedfrom the
standarddeviation(S.D.) of response and the slopeof the curve(S) by meansof the
equations:LOD = 3.3 (S.D./S)and LOQ = 10 (S.D./S).Low LOD and LOQ valueswere
40 JOURNAL OF COSMETIC SCIENCE

mAU Resorcinol

2o.i

Arbutm
8O

4O

2 4 6 8 10 12 14

Figure2. HPLCchromatogram
of thearbutinstandard
(5 pg/ml •) andtheresorcinol
internalstandard
(5
pg/ml •).

obtainedas shownin Table I, which indicatesthe goodsensitivityof the proposedLC


method.

Li,earity. To determinelinearity, five different concentrations


of the arbutin standard
wereusedin a workingrangeof0.5-30.0Hg/ml-• . Thelinearregression
equations
and
thecorrelation
coefficient
(r2) values
forarbutinandtheinternalstandard
aregivenin
TableI. Ther2 values
showgoodlinearityin theexamined concentration
range.

PRECISION AND ACCURACY

The intradayreproducibilitystudywasperformedduring a period of three days.The


resultsobtainedshowedthat the arbutin peak areavariabilitiesfor standardsolutions
werewithin 0.00-0.02% R.S.D. For interdayreproducibility,five replicateinjectionsof
variousconcentrationsof arbutin weremadewithin a day. The resultsobtainedshowed
that the arbutin peak area variabilities for the standardsolutionswere within 0.00-
0.02% R.S.D. The resultsare shownin Table II. The R.S.D. valuesdemonstrated good
precision.

Table I
LinearRegression
Analysis(n = 3) and Limits of Detection(S/N = 3) and Quantitation(S/N = 10)

Linearityrange Equation r2 LOD LOQ


Standard (Hg/ml -•) Y = SX + C Mean(+S.D.) (Hg/ml1) (Hg/ml-•)
Arbutin 0.5-30 Y = 132.84 (_+0.36)X 0.9999 5.07 x 10 .3 1.01 x 10 2
- 9.32 (+1.23) (-+5.77x 10 5)
Resorcinol* 0.5-30 Y = 257.03 (_+0.91)X 0.9999 5.04 x 10 • 1.00 x 10 2
+ 7.34 (_+0.04) (+1.49 x 10-s)
* Internal standard.
HPLC DETERMINATION OF ARBUTIN 41

Table II
Intraday and Interday PrecisionAnalysisof Arbutin StandardSolutions

Within-day variability (n - 5) Between-dayvariability (n = 5)


(concentration found) (concentrationfound)
Concentration added
(pg/ml•) Meanñ S.D.(pg/ml•) %R.S.D. Mean_+S.D.(pg/ml•) %R.S.D.
1.00 0.99 + 0.02 0.02 1.01 _+0.02 0.02
5.00 5.00 + 0.02 0.00 5.00 _+0.02 0.00
10.00 10.09 + 0.07 0.01 10.06 _+0.11 0.01
15.00 15.02 _+0.08 0.00 15.02 _+0.08 0.00
20.00 19.88 + 0.28 0.01 19.88 _+0.28 0.01

The accuracyof the proposedmethod was determinedby analyzingsampleextract


solutionsspiked with three different concentrationsof standardarbutin solution (3.0,
5.0,and10.0pg/ml-•, respectively),
usingtheproposed
LCmethod.
Therecoveries
of
the arbutinstandardsolutionswerealsoanalyzed.The resultsarepresentedin Table III.
The excellentrecoveries
of the standardadditionamountssuggestthat the method is
accurate.

APPLICATION TO COSMETIC AND MEDICINAL PLANTS

The proposedRP-HPLC methodwasappliedto the determinationof arbutin in whit-


enlug creamsand medicinal plant extractsafter TLC extraction.The sampleswere
separated on a silicagel GF254(20 cm x 20 cm) glassplate usingethyl acetate:metha-
nol:water(85:17:13, v/v/v) as mobilephase.After visualization,eachsampleshoweda
blue spotwith an Rf valueof 0.36, whichcorresponded to that of the standard.Arbutin
in eachsamplewas then determinedby the proposedRP-HPLC method(Figure 3). A
comparativedeterminationof arbutin in the originalskin-whiteningcreamsampleswas
also carried out using the UV spectrophotometric method. The resultsare shown in
TableIV. It is indicatedthat the amountsof arbutinfoundby both methodsarequite
identical.Excellentcorrelationbetweenthe two methodswasobtained.The performance
of the methodwasassessed by calculationof the Studentt-test comparedwith the UV
spectrophotometricmethod. It was evident that the t-value for arbutin determination,
comparedto the resultsobtained by the UV spectrophotometric method, was 1.38,
which did not exceedthe theoreticalvalue (2.44) at the 95% confidencelimit for the six
degreesof freedom.Theseresultsindicatedthat the 95% confidencelimit betweenthe
proposedHPLC methodand the UV spectrophotometric method for the arbutin assay
doesnot differ significantly.Fourmedicinalplant sampleswerecollected:BetzdaM, oides
Buch. Ham, Clerodendr•m petasites
S. Moore., Cz•rc•ligo
latifolia Dryand. Var. latifolia,

Table III
Accuracyof the ProposedLC Method (n = 7)

Concentration added Concentration found:


Standard (pg/ml
-•) mean+_S.D.(pg/ml1) Recovery
(%) Relative
error(%)
Arbutin 3.00 2.96 _+0.006 98.71 + 0.22 0.04
a .[....:• 5.00 q •2 • •2 •• •/, • t• r• r•v
Arbutin 10.00 10.15 + 0.009 101.53 _+0.09 0.15
42 JOURNAL OF COSMETIC SCIENCE

mAU

_L
Figure 3. HPLC chromatogram
of a samplecontainingarbutinfrom skin-whiteningcream.

Table IV
Comparisonof Two Methodsfor MeasuringArbutin in Samplesof CommercialWhitening Cream

Amountsof arbutin found(g%)

Cosmeticsample Spectrophotometric
(Arbuwhite© cream) RP-HPLC method• method• (20) Calculatedt-values

0.75 0.77
0.75 0.78
0.77 0.80
0.76 0.78 1.38
0.76 0.79
0.75 0.78
0.77 0.80
Average 0.76 0.79

a Each value is the averageof 7 determinations.


s Calculated
t-valueforp = 005 andsixdegrees
of freedom
is 2.44.

and HeJ•perethusa
crenu/ata
(Roxb.) M. Roem.The sampleswereextractedwith water and
methanol. Then the amounts of arbutin in the sampleswere determined by the
RP-HPLC method. The amountsof arbutin found in the aqueousextract were 3.50,
1.50,1.10,and0.12pg/g-•,respectively.
Theamounts
ofarbutinfoundin themethanol
extractwere0.77,0.0002,and0.0012pg/g-•,respectively.
Themethod wassuccess-
fully applied to the determination of arbutin in three commercialskin-whitening
creams.The amountsof arbutin found in the samples(Arbuwhite© cream,Super
Whitening
© cream,andShiseido
© cream)were0.76, 0.58, and5.79 mg/g-•, respec-
tively.

CONCLUSION

The reversed-phase
high-performance liquid chromatographic
procedurehas beende-
velopedfor determiningarbutin in commercialskin-whiteningcreamsand someme-
HPLC DETERMINATION OF ARBUTIN 43

dicinalplant extracts,respectively.
The methodwasalsovalidatedfor limit of detection,
limit of quantitation,repeatability,reproducibility,and accuracy.The optimum con-
ditionsand analyticalcharacteristicsfor RP-HPLC determinationof arbutin exhibited
goodresolution,shortanalysistime, andratherhigh sensitivity.In the proposedmethod
for determiningarbutin in skin-whiteningcreams,the workingcalibrationcurvesover
theranges
of 0.5-30.0pg/ml-• wereestablished.
Themethod
wassuccessfully
applied
to the determinationof arbutinin threecommercialskin-whiteningcreams.The content
of arbutinfoundin the samples
(Arbuwhite
© cream,SuperWhitening© cream,and
Shiseido
© cream)
were0.76,0.58,and5.79mg/g-•, respectively.
Themethod
wasalso
appliedto the determinationof arbutin in somemedicinalplant extracts.The amounts
of arbutin in Bet•la alnoidesBuch. Ham., Clerodendr•m petasites
S. Moore., C•rc•/igo
latifoliaDryand.Var. latifolia,andHesperethma
cren•lata(Roxb.).Roemin the aqueous
extracts
were3.50, 1.50, 1.10,and 0.12 l•g/g-•, respectively.
Arbutinfromthese
medicinalplants can be used for the productionof skin-whiteningcosmetics.The
benefitsof the proposedmethodare simplicity, convenience,
rapidity, sensitivity,good
precision,and accuracy.The methodis suitablefor routineanalysisof arbutinin com-
mercial cosmeticsand raw plant materials.

ACKNOWLEDGMENTS

The authorsexpresstheir sincerethanksto the GraduateSchooland the Facultyof


Pharmacy, Chiang Mai University, for financial and chemical support. Saisunee
Liawruangrathexpresses
her sincerethanksto the Postgraduate
Educationand Research
Programin Chemistry(PERCH) for partial support.

REFERENCES

(1) H. Zhai and H. I. Maibach,Skin whiteningagents,Cosmet.


Toiletr.,116, 21 (2001).
(2) B. Stefania,C. Emanuela,and P. Mauro, Chemical and instrumentalapproachesto treat hyperpig-
menration,PigmentCell Res.,16, 101 (2003).
(3) A. Kawalier, Ann., 82, 241 (1852).
(4) C. Manich, Arch.Pharm.,250, 547 (1912).
(5} K. Maedaand M. Fukuda,J. Pharmacol. Exp. Ther.,276, 765 (1996).
(6) L. Petit and G. E. Pierard,Skin-lighteningproductsrevisited,Int. J. Cosmet.
Sd., 25, 169 (2003).
(7) M. H. Assaf,A. A. All, M. A. Makboul,J.P. Beck,and R. Anton, Preliminarystudyof phenolic
glycosides from Origamun majorana;Quantitativeestimationof arbutin:Cytotoxicactivity of hydro-
quinone,PlantaMedica,343 (1987).
(8) H. Matusda,M. Higashino,Y. Nakai, I. Iinuma, M. Kubo, and F. A. Lqang,Studiesof cuticledrugs
from naturalsources. IV. Inhibitory effectsof someArctostaphylos
plantson melaninbiosynthesis,Bio.
Pharm. Bull., 19, 153 (1996).
(9) J. A. Parrish,R. R. Anderson,F. Urbach,and D. Pitts, UVA: Biological
Efj•ctsof Ultraviolet
Radiation
with Emphasis on HumanResponses to LongWaveUltraviolet(Plenum Press,New York and London,
1978), pp. 15 1.
(10) Lj. Kraus and E. Stahl,Chromatography and photomerryof arbutin,StateInst. Contr.Drug., 17, 252
(1968).
(11) B. Franciszek,D.-Z. Elzbieta,andC. Malgorzata,Spectrophotometric determination of arbutinin herb
raw materials usingcerium(IV) and arsenazo III as reagents,Chem.AnaL, 36, 13 (1991).
(12) G. Jiri and S. Iveta, Color reactionof arbutin with nitrousacid and its analyticalapplications,Fo/ia
Pharm. Univ. CaroL, 7, 29 (2003).
44 JOURNAL OF COSMETIC SCIENCE

(13) E. Kenndler and C. Schwer,Determination of arbutin in uvae-ursifolium (bearberryleaves)by


capillaryzoneelectrophoresis, J. Chromatogr.,514, 383 (1990).
(14) M. VanhaelenandR. Vanhaelen-Fastre, Quantitativedeterminationof biologicallyactiveconstituents
in medicinalplant crudeextractsby thin-layerchromatography-densitometry, J. Chromatogr.,281,
263 (1983).
(15) I. Parejo,F. Viladomat, J. Bastidaand C. Codina,A singleextractionstepin the quantitativeanalysis
of arbutin in bearberry(Arctostaphylos
•va-•rsi) leavesby high performance
liquid chromatography,J.
Phytochem. Anal., 12, 336 (2001).
(16) V. N. Bubenchikova andI. L. Drozdova,
HPLCanalysis
ofphenoliccompounds
in yellowsweet-clover,
J. Pharm.Ch•z., 38, 195 (2004).
(17) M.-O. Masse,V. Duvallet, M. Borremans,and L. Goeyens,Identificationand quantitativeanalysisof
kojic acid and arbutin in skin whitening cosmetics,Int. J. Cosmet.
Sci.,23, 219 (2001).
(18) M.-L. Changand C.-M. Chang,Simultaneous HPLC determinationof hydrophilicwhiteningagents
in cosmeticproducts, J. Pharm.Biomed. Anal., 33, 617 (2003).
(19) UnitedStates
Pharmacopeia, 26th ed. (U.S. Pharmacopeia,
Rockville,MD, 2003).
(20) S.-C. Huang, C.-C. Lin, M.-C. Huang, and K.-C. Wen, Simultaneousdeterminationof magnesium
ascrobylphosphate,ascorbylglucoside,kojic acid, arbutin and hydroquinonein skin whitening
cosmetics,J.Food.Drug. Anal., 12, 13 (2004).

All in-text references underlined in blue are linked to publications on ResearchGate, letting you access and read them immedi

Vous aimerez peut-être aussi