Development and Application of an Enzyme-linked Immunosorbent Assay for Specific Detection of Mangiferin Content in Various Cultivars of Mangifera indica Leaves Using Anti-mangiferin Polyclonal Antibody

Authors

  • Gorawit Yusakul,

    1. Authors Gorawit Yusakul, Wongsathorn Kitirattrakarn, Narunat Tanwanichkul, and Waraporn Putalun are with Faculty of Pharmaceutical Sciences, Khon Kaen Univ., Khon Kaen 40002, Thailand.
    2. Authors Gorawit Yusakul and Waraporn Putalun are also with Research Group for Pharmaceutical Activities of Natural Products using Pharmaceutical Biotechnology (PANPB).
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  • Wongsathorn Kitirattrakarn,

    1. Authors Gorawit Yusakul, Wongsathorn Kitirattrakarn, Narunat Tanwanichkul, and Waraporn Putalun are with Faculty of Pharmaceutical Sciences, Khon Kaen Univ., Khon Kaen 40002, Thailand.
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  • Narunat Tanwanichkul,

    1. Authors Gorawit Yusakul, Wongsathorn Kitirattrakarn, Narunat Tanwanichkul, and Waraporn Putalun are with Faculty of Pharmaceutical Sciences, Khon Kaen Univ., Khon Kaen 40002, Thailand.
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  • Hiroyuki Tanaka,

    1. Author Hiroyuki Tanaka is with Graduate School of Pharmaceutical Sciences, Kyushu Univ., Fukuoka 812–8582, Japan. Direct inquiries to author Waraporn Putalun (E-mail: waraporn@kku.ac.th).
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  • Waraporn Putalun

    1. Authors Gorawit Yusakul, Wongsathorn Kitirattrakarn, Narunat Tanwanichkul, and Waraporn Putalun are with Faculty of Pharmaceutical Sciences, Khon Kaen Univ., Khon Kaen 40002, Thailand.
    2. Authors Gorawit Yusakul and Waraporn Putalun are also with Research Group for Pharmaceutical Activities of Natural Products using Pharmaceutical Biotechnology (PANPB).
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Abstract

Abstract:  An enzyme-linked immunosorbent assay (ELISA) was developed for determining mangiferin content in plant samples using a polyclonal antibody (PAb) against mangiferin. The developed ELISA showed a full measurement range from 0.12 to 31.25 μg/mL mangiferin with a relative standard deviation (RSD) less than 6% for both intra- and interassay precision levels. The accuracy was determined by a percent recovery experiment at three concentration levels and it showed 97.8%–103.7% recovery in Mangifera indica leaf samples. The developed ELISA exhibited a high correlation value (R2= 0.992) with the standard high-performance liquid chromatography (HPLC) method in various mangiferin-containing plant samples. Our results suggest that the validated ELISA methodology using a PAb against mangiferin can be applied to determine mangiferin content with high specificity, rapidity and simplicity in various mangiferin-containing plant samples. The mangiferin content in the mature leaves of fifty M. indica cultivars were determined using the developed ELISA. The mangiferin contents ranged from 1.94 ± 0.13% to 13.79 ± 0.84% dry wt. The Thawai cultivar leaves contained the highest level of mangiferin (13.79 ± 0.84% dry wt), but it is a rare cultivar. The Namdokmai, which is more commonly cultivated in Thailand, contain 12.41 ± 0.60% dry wt mangiferin; therefore, this cultivar leaf was recommended as the source of raw material for the pharmaceutical, nutraceutical and cosmetic product industries.

Practical Application:  Currently, natural heath products are accepted worldwide for healthcare. Mangiferin-containing plants and products exhibit health benefits against oxidative stress-related diseases, such as cardiovascular diseases, cancer, dyslipidemia and diabetes. We have developed an ELISA with high specificity, rapidity and simplicity for the quality control of mangiferin-derived product production. Moreover, we found that the Namdokmai leaf, a Thai M. indica cultivar, was recommended as the source of raw material for the pharmaceutical, nutraceutical and cosmetic product industry because of its high mangiferin content and natural prevalence.

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