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Determination of Iriflophenone 3‐ C ‐β‐ d ‐Glucoside From Aquilaria spp. by an Indirect Competitive Enzyme‐linked Immunosorbent Assay Using a Specific Polyclonal Antibody
Author(s) -
Putalun Waraporn,
Yusakul Gorawit,
Saensom Paritad,
Sritularak Boonchoo,
Tanaka Hiroyuki
Publication year - 2013
Publication title -
journal of food science
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.772
H-Index - 150
eISSN - 1750-3841
pISSN - 0022-1147
DOI - 10.1111/1750-3841.12225
Subject(s) - polyclonal antibodies , antibody , chemistry , enzyme , cross reactivity , microbiology and biotechnology , chromatography , biology , biochemistry , cross reactions , immunology
Polyclonal antibody against iriflophenone 3‐ C ‐β‐ d ‐glucoside (IP3G), a major compound from the leaves of Aquilaria spp., was produced for the development of an enzyme‐linked immunosorbent assay (ELISA). The results showed that the antibodies were specific for IP3G. The produced antibody has low cross reactivity with iriflophenone 3,5‐ C ‐β‐ d ‐diglucopyranoside (13%), genkwanin 5‐ O ‐β‐primeveroside (3.55%) and no cross reactivity found in other compounds. The range of ELISA assay extends from 100 to 1560 ng/mL with coefficient of variation (CV) 1.19% to 2.07% for intra‐assay and 3.76% to 7.15% for inter‐assay precision levels. The recovery rates of IP3G in the leaves of Aquilaria spp. were in the range of 96.0% to 99.0% with CV 4.50% to 5.32%. A correlation between ELISA and high‐performance liquid chromatography methods was obtained when analysis of IP3G in the plant samples ( R 2 = 0.9321). These results suggest that the developed ELISA method can be applied to determine IP3G content with high specificity, rapidity, and simplicity. The developed immunosorbent assay in this study provides a useful tool for the analysis of IP3G in plant samples and products.