Production of polyclonal antibody against madecassoside and development of immunoassay methods for analysis of triterpene glycosides in Centella asiatica

Patcharin Tassanawat, Waraporn Putalun, Gorawit Yusakul, Boonchoo Sritularak, Thaweesak Juengwatanatrakul, Hiroyuki Tanaka

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Introduction Centella asiatica (L.) Urban consists of two major triterpene glycosides, asiaticoside (AS) and madecassoside (MA), as active components used for wound healing and enhancing memory. Objective To produce a polyclonal antibody against madecassoside (MA-PAb) and develop enzyme-linked immunosorbent assay (ELISA) and Eastern blotting methods for quantitative analysis of triterpene glycosides in Centella asiatica. Methods An ELISA method was developed using polyclonal antibody against MA. An Eastern blotting method on the PES membrane was established for determination of MA and AS. The immunoassays were validated for sensitivity, precision, specificity and accuracy. Results The prepared MA-PAb shows specificity to MA and AS. The measuring range of triterpene glycosides was 0.39-50 μg/mL using the ELISA method. An Eastern blotting method was developed for determining individual MA and AS, which could be detected in the range of 62.5-500 ng. The limit of detection for MA and AS was 31.25 ng. The two methods developed showed good specificity, precision, and accuracy, and also correlated with high-performance liquid chromatography. Conclusion These immunoassays have several advantages that include high sensitivity as well as being rapid and facile for determination of the triterpene glycosides in C. asiatica.

Original languageEnglish
Pages (from-to)256-262
Number of pages7
JournalPhytochemical Analysis
Issue number3
Publication statusPublished - May 1 2013


All Science Journal Classification (ASJC) codes

  • Analytical Chemistry
  • Food Science
  • Biochemistry
  • Molecular Medicine
  • Plant Science
  • Drug Discovery
  • Complementary and alternative medicine

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