นโยบายงานวิจัย /จรรยาบรรณนักวิจัย /ระดับคุณภาพบทความวิจัยตีพิมพ์ /ระดับคุณภาพผลงานวิชาการ /แหล่งทุน /ดาวน์โหลด /ฐานข้อมูลวิจัย /วิเคราะห์-สังเคราะห์งานวิจัย /ลิขสิทธิ์ /ข่าว


Highly selective and sensitive determination of deoxymiroestrol using apolyclonal antibody-based enzyme-linked immunosorbent assay

Author

- Gorawit Yusakul, Orapin Udomsin,Thaweesak Juengwatanatrakul, Hiroyuki Tanaka, Chaiyo Chaichantipyuth, Waraporn Putalun


Journal

- Talanta

Volume

- 114

Year

- 2013

Publication type

- Research article (Inter)

Page list

- 73-78

Abstract

    Puerariacandollei-associatedproductsareofinteresttoworldwideconsumersfortheirrejuvenatingand
cosmeticpurposes.Inaddition,clinicaltrialshavesupportedthebeneficialeffectsof P.candollei onthe
alleviationofmenopausalsymptoms.Deoxymiroestrol,whichwasreportedasthemostpotent
phytoestrogeninthetuberousrootof P.candollei,exhibitedpotentialasabiomarkerof P.candollei-
derivedsamplesandproducts.Apolyclonalantibody-basedenzyme-linkedimmunosorbentassay
(ELISA)wasdevelopedfordeoxymiroestroldetermination.Theraisedantibodyboundspecificallyto
deoxymiroestrolwithverylowcrossreactivitiesof1.26and0.42%tostructurallyrelatedmiroestroland
isomiroestrol,respectively.Thelinearrangewas0.73–3000.00ngmL−1
,andthecoefficientsofvariation
forboththeintra-andinter-platedeterminationswerelessthan5%.Insamplesspikedwithaknown
amountofdeoxymiroestrol,therecoverieswere99.82–102.58%in P.candollei samplesand98.07–
106.33%initsproductssamples.Incomparisonwithotheranalyticalmethods,thevalidatedELISAwas
comparabletopublishedHPLC-UVmethodsforsampleswithhighdeoxymiroestrolcontent
(R2
¼0.9993).Furthermore,ELISAcanbeusedforsampleswithdeoxymiroestrolconcentrationsthat
aretoosmalltodetectbyHPLCandforconditionswhenotherchemicalscauseinterferencewith
chromatographicanalysis.Forthe P.candollei-derivedproducts,thepreparationscontained0.154–
10.998 mgg−1
drywt.OurELISAsuccessfullymeasureddeoxymiroestrolcontentwithhighsensitivity,
selectivity,accuracyandrapidity.Therefore,thisELISAshowedpotentialfordosagestandardizationof P.
candollei-associatedsamples.


Keywords

    Puerariacandollei
Deoxymiroestrol
Enzyme-linked immunosorbent assay
Phytoestrogen
Polyclonal antibody