IndustrialCropsandProducts60(2014)239–246
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Industrial
Crops
and
Products
j ou rn a l h o m epa g e :w w w . e l s e v i e r . c o m / l o c a t e / i n d c r o p
HPTLC
simultaneous
quantification
of
triterpene
acids
for
quality
control
of
Plantago
major
L.
and
evaluation
of
their
cytotoxic
and
antioxidant
activities
Kartini
a,b,∗,
S.
Piyaviriyakul
c,
P.
Siripong
c,
O.
Vallisuta
aaFacultyofPharmacy,MahidolUniversity,447Sri-AyudhayaRoad,Ratchathewi,Bangkok10400,Thailand bFacultyofPharmacy,UniversityofSurabaya,RayaKalirungkutRoad,Surabaya60293,Indonesia cNaturalProductsResearchSection,ResearchDivision,NationalCancerInstitute,Bangkok10400,Thailand
a
r
t
i
c
l
e
i
n
f
o
Articlehistory: Received12March2014
Receivedinrevisedform11June2014 Accepted13June2014
Keywords: Cytotoxic Antioxidant Ursolicacid Oleanolicacid HPTLC
Simultaneousanalysis
a
b
s
t
r
a
c
t
Avalidatedhighperformancethin-layerchromatography(HPTLC)methodwasdevelopedfor simul-taneousdeterminationofursolicacid(UA)andoleanolicacid(OA)contentsinPlantagomajorwhich werecollectedfromseveralplantationareasinIndonesia.Thecytotoxiceffectagainsttwocancercell lines,SiHaandHepG2,andantioxidantactivitywereevaluatedusingtheMTTandDPPH-radical scav-engingassay,respectively.ThetestsamplesincludedvariousextractsofP.majorfromdifferentplant partsusingmethanolandwaterasextractingsolventsandpurecompoundsderivedfromthisplant. Theresultsshowedthatbothplantpartsandextractingsolventsaffectedthechemicalcontentsand theirbiologicalactivities.ThecontentsofUAandOAvariedaccordingtotheorgansandprovenancesof plant.ThehighestcontentofUA(0.22–0.48%dryweight)andOA(0.17–0.33%dryweight)werefound inthemethanolextractofseed.Thisextractalsoexhibitedthehighestcytotoxicactivity(IC50value:
174.42–246.38g/ml),whereasthestrongestfreeradicalscavengingactivitywasobtainedfromtheleaf
methanolextract(IC50value:263.57g/ml).ThedevelopedHPTLCmethodcanbeusedforroutine
anal-ysisandstandardizationofP.majorcrudedrugs,extracts,and/orfinishedproductsusingUAandOAas appropriatemarkersforanticancerproducts.
©2014ElsevierB.V.Allrightsreserved.
1. Introduction
PlantagomajorL.isamemberofPlantagogenus(Plantaginaceae family)havingmorethan256species(Galvez etal.,2005).This perennialplantisgloballydistributedandfoundinremarkably dif-ferentlocationsandclimaticconditions,fromsealevelupto3300m altitude.Itisgenerallyacceptedthatenvironmentaffectschemicals composition,whichthenaffectthebiologicalactivitiesof medic-inalplants(Zhangetal.,2009).ThedecoctionofaerialpartsofP. major,itsleavesorseeds,areusedtraditionallyasdiuretic,laxative, anti-diabetes,andforskindiseases.Therootisempiricallyused asanti-cough(dePaduaandBunyapraphatsara,1999).Atpresent, severalherbalmedicineindustriesinIndonesiacommercializethis plantintomoderndosageformssuchascapsulesandtablets with-outanystandardizationoftheproducts.Onlytheaerialpartsare
∗ Correspondingauthorat:FacultyofPharmacy,MahidolUniversity,447 Sri-AyudhayaRoad,Ratchathewi,Bangkok10400,Thailand.Tel.:+66026448677.
E-mailaddresses:[email protected],[email protected](Kartini).
beingused,whereastherootsbecomewastewithoutanyusage. Furtherstudiesarerequiredtomaximizeitsutilization.
Plantagomajorcontainsvariousclassesofcompoundsuchas flavonoids (e.g., baicalein), iridoid glycosides (e.g.,aucubin and catalpol), triterpeneacids (e.g.,ursolicacidand oleanolicacid), caffeic acidderivatives (e.g., caffeic acidand chlorogenicacid), andphenoliccompounds(e.g.,vanillicacidandp-coumaricacid) (Samuelsen,2000).Ursolicacid(UA)andoleanolicacid(OA)(Fig.1.) arehighlyinterestingduetotheirbroadspectrumofactivities;i.e. COX-2andcholinesteraseinhibitor(Kolaketal.,2011;Ringbom etal.,1998;Stenholmetal.,2013),immunomodulator(Chiangetal., 2003c),andcancerpreventionandtreatment(Chiangetal.,2003a; Kolaket al.,2011;Lee etal.,2005;Liu, 1995,2005;Pollierand Goossens,2012;Ringbom etal.,1998;Shanmugametal.,2013; Stenholm etal., 2013).Variousdetermination methodssuchas GC–MS(Caligianietal.,2013;Janicsáketal.,2003),HPLC(Wang etal.,2011a;Wangetal.,2008;Wójciak-Kosioretal.,2013), LC-(UV)-APCI–MS(Tarvainenetal.,2010),and2DNMR(Kontogianni etal.,2009)wereappliedtoquantifytheUAandOAcontentinsome plants,includingP.major.Thesemethods,however,arehighcost, time-consuming,andspecializedexpertisesareneeded.Forthese
