title: An Enzyme-linked Immunosorbent Assay for Genistein 7-O-[α-rhamnopyranosyl-(1→6)]-β-glucopyranoside Determination in Derris scandens using a Polyclonal Antibody.
abstract: BACKGROUND
  Genistein 7-O-[α-rhamnopyranosyl-(1→6)]-β-glucopyranoside (GTG) is a major bioactive compound in Derris scandens. It is responsible for anti-inflammatory activity by inhibition of cyclooxygenase and lipoxygenase. There are many commercial products of D. scandens available in Thailand.
  
  
  OBJECTIVE
  To develop an enzyme-linked immunosorbent assay (ELISA) for the quantitative analysis of GTG in plant material and derived products using a polyclonal antibody.
  
  
  METHODS
  An immunogen was synthesised by conjugating GTG with a carrier protein. The polyclonal antibody against GTG (GTG-PAb) was produced in New Zealand white rabbits. The ELISA method was validated for specificity, sensitivity, accuracy, precision and correlation with HPLC.
  
  
  RESULTS
  The polyclonal antibody was specific to GTG and genistin within the range of compounds tested. The GTG ELISA was applied in the range 0.04-10.00 μg/mL with a limit of detection of 0.03 μg/mL. The recovery of GTG in spiked Derris scandens extracts ranged from 100.7 to 107.0%, with a coefficient of variation less than 7.0%. The intra- and inter-assay variations were less than 5.0%. The ELISA showed a good correlation with HPLC-UV analysis for GTG determination in samples, with a coefficient of determination (r2) of 0.9880.
  
  
  CONCLUSIONS
  An ELISA was established for GTG determination in Derris scandens. The GTG-PAb can react with GTG and genistin, but genistin has not been found in the plant. Therefore, the ELISA can be used for high throughput quality control of GTG content in D. scandens and its products. Copyright © 2016 John Wiley & Sons, Ltd.
authors.affiliation: Faculty of Pharmaceutical Sciences, Khon Kaen University, Khon Kaen, 40002, Thailand.|Regional Medical Sciences Center 7 Khon Kaen, Khon Kaen, 40000, Thailand.
authors.forename: Kamonthip
authors.initials: K
authors.lastname: Jutathis
authors.affiliation: Faculty of Pharmaceutical Sciences, Khon Kaen University, Khon Kaen, 40002, Thailand.|Research Group for Pharmaceutical Activities of Natural Products using Pharmaceutical Biotechnology (PANPB), National Research University - Khon Kaen University, Khon Kaen, 40002, Thailand.
authors.forename: Tharita
authors.initials: T
authors.lastname: Kitisripanya
authors.affiliation: Faculty of Pharmaceutical Sciences, Khon Kaen University, Khon Kaen, 40002, Thailand.|Research Group for Pharmaceutical Activities of Natural Products using Pharmaceutical Biotechnology (PANPB), National Research University - Khon Kaen University, Khon Kaen, 40002, Thailand.
authors.forename: Orain
authors.initials: O
authors.lastname: Udomsin
authors.affiliation: Faculty of Pharmaceutical Sciences, Khon Kaen University, Khon Kaen, 40002, Thailand.|School of Pharmacy, Eastern Asia University, Pathum Thani, 12110, Thailand.
authors.forename: Chadathorn
authors.initials: C
authors.lastname: Inyai
authors.affiliation: Faculty of Pharmaceutical Sciences, Chulalongkorn University, Bangkok, 10330, Thailand.
authors.forename: Boonchoo
authors.initials: B
authors.lastname: Sritularak
authors.affiliation: Graduate School of Pharmaceutical Sciences, Kyushu University, Fukuoka, 812, -8582, Japan.
authors.forename: Hiroyuki
authors.initials: H
authors.lastname: Tanaka
authors.affiliation: Faculty of Pharmaceutical Sciences, Khon Kaen University, Khon Kaen, 40002, Thailand. waraporn@kku.ac.th.|Research Group for Pharmaceutical Activities of Natural Products using Pharmaceutical Biotechnology (PANPB), National Research University - Khon Kaen University, Khon Kaen, 40002, Thailand. waraporn@kku.ac.th.
authors.forename: Waraporn
authors.initials: W
authors.lastname: Putalun
mesh_terms: D000906:Antibodies / Q000276:immunology*; D002851:Chromatography, High Pressure Liquid; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D019833:Genistein / Q000031:analogs & derivatives* / Q000032:analysis / Q000276:immunology; D013056:Spectrophotometry, Ultraviolet
chemical_list: D000906:Antibodies; D019833:Genistein
keywords: Derris scandens; ELISA; genistein7-O-[α-rhamnopyranosyl-(1→6)]-β-glucopyranoside; genistin; immunoassay; polyclonal antibody