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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

Authors

  • Waraporn Putalun,

    Corresponding author
    1. Research Group for Pharmaceutical Activities of Natural Products using Pharmaceutical Biotechnology (PANPB), Natl. Research Univ.-Khon Kaen Univ., Thailand
    • Faculty of Pharmaceutical Sciences, Khon Kaen Univ., Thailand
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  • Gorawit Yusakul,

    1. Faculty of Pharmaceutical Sciences, Khon Kaen Univ., Thailand
    2. Research Group for Pharmaceutical Activities of Natural Products using Pharmaceutical Biotechnology (PANPB), Natl. Research Univ.-Khon Kaen Univ., Thailand
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  • Paritad Saensom,

    1. Faculty of Pharmaceutical Sciences, Khon Kaen Univ., Thailand
    2. Research Group for Pharmaceutical Activities of Natural Products using Pharmaceutical Biotechnology (PANPB), Natl. Research Univ.-Khon Kaen Univ., Thailand
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  • Boonchoo Sritularak,

    1. Faculty of Pharmaceutical Sciences, Chulalongkorn Univ., Thailand
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  • Hiroyuki Tanaka

    1. Graduate School of Pharmaceutical Sciences, Kyushu Univ., Japan
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Direct inquiries to author Putalun (E-mail: waraporn@kku.ac.th).

Abstract

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 (R2 = 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.

Practical Application

Agarwood leaves are consumed as health drink due to their proposed anti-inflammatory, anti-diabetic properties, and as a laxative. An indirect competitive enzyme-linked immunosorbent assay method for determination of iriflophenone 3-C-β-d-glucoside showed potential assay for quality control of agarwood leaves raw material and product.

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