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DOI: 10.1055/s-0036-1597057
An enzyme-linked immunosorbant assay for genistein 7-O-[α-rhamnopyranosyl-(1→6)]-β-glucopyranoside determination in Derris scandens by using a polyclonal antibody
Publication History
Publication Date:
14 December 2016 (online)
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 [1]. There are many commercial products of D. scandens available in Thailand. To evaluate the quality of products and raw material from the plants, we developed an immunoassay for GTG determination in D. scandens and its products by using a polyclonal antibody. An immunogen was synthesized by conjugating GTG with a carrier protein. The polyclonal antibody against GTG was produced in New Zealand white rabbits. The ELISA method was validated for specificity, sensitivity, accuracy, precision and correlation with HPLC. The prepared GTG-PAb was specific to GTG and genistin. The developed ELISA for GTG determination was performed 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 D. scandens extracts ranged from 100.7 – 107.0% with a coefficient of determination less than 7.0%. The variation of intra- and inter-assay were less than 5.0%. The developed ELISA and HPLC methods showed good correlation for GTG determination in samples with a coefficient of determination (r 2) of 0.9880. The developed ELISA could be used for quality control of GTG content in D. scandens and its products with simplicity, speed and reliability.
Acknowledgements: Financial support from Faculty of Pharmaceutical Sciences, Khon Kaen University to Mrs. Kamonthip Jutathis is acknowledged.
Keywords: Derris scandens, ELISA, genistein 7-O-[α-rhamnopyranosyl-(1→6)]-β-glucopyranoside, polyclonal antibody.
References:
[1] Laupattarakasem P, Houghton PJ, Hoult JRS. Anti-inflammatory isoflavonoids from the stem of Derris scandens. Planta Med 2004; 70: 496 – 501
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No conflict of interest has been declared by the author(s).