title: Development of an enzyme-linked immunosorbent assay to determine puerarin and its aglycone daidzein.
abstract: Pueraria candollei (White Kwao Khuer) is a medicinal plant containing puerarin (8-C-glucosyl-4',7-dihydroxyisoflavone) and its aglycone, daidzein, as major isoflavonoids used for its rejuvenating and estrogenic effects. In order to analyze these isoflavonoids, an enzyme-linked immunosorbent assay (ELISA) was developed using polyclonal antibody (PAb) against puerarin. Puerarin was conjugated with bovine serum albumin (BSA) to be used as an immunogen for production of PAb from a rabbit. The PAb has been shown to be specific to puerarin and its aglycone, daidzein. An ELISA was then performed using anti-puerarin PAb in the range of 6.10 ng/ml-1.56 μg/ml of puerarin. Based on validation analysis, this analytical method by ELISA is a precise, accurate, and sensitive method for determining puerarin and daidzein in plant samples.
authors.affiliation: Faculty of Pharmaceutical Sciences, Khon Kaen University, Khon Kaen 40002, Thailand.
authors.forename: Benyakarn
authors.initials: B
authors.lastname: Pongkitwitoon
authors.forename: Seiichi
authors.initials: S
authors.lastname: Sakamoto
authors.forename: Hiroyuki
authors.initials: H
authors.lastname: Tanaka
authors.forename: Ryota
authors.initials: R
authors.lastname: Tsuchihashi
authors.forename: Junei
authors.initials: J
authors.lastname: Kinjo
authors.forename: Satoshi
authors.initials: S
authors.lastname: Morimoto
authors.forename: Waraporn
authors.initials: W
authors.lastname: Putalun
mesh_terms: D000818:Animals; D002851:Chromatography, High Pressure Liquid; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D005260:Female; D007529:Isoflavones / Q000032:analysis*; D011817:Rabbits; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
chemical_list: D007529:Isoflavones; C004742:daidzein; C033607:puerarin