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Ultrasensitive Time-Resolved Fluoroimmunoassay for Saikosaponin a in Chaihu (Bupleuri Radix)

Author

Listed:
  • Zhi Chao
  • Qian Cui
  • Enwei Tian
  • Weiping Zeng
  • Xuan Cai
  • Xiaolei Li
  • Hiroyuki Tanaka
  • Yukihiro Shoyama
  • Yingsong Wu

Abstract

The aim of this study is to establish a time-resolved fluoroimmunoassay (TRFIA) system for quantitative analysis of saikosaponin a (SSa) in the crude drug of Chaihu (Bupleuri Radix). A 96-well microplate coated with rabbit anti-mouse IgG was incubated with the methanol extracts of Chaihu samples and a mouse anti-SSa monoclonal antibody, and a Eu3+-labeled SSa-human serum albumin conjugate was used as the tracer. The established competitive TRFIA showed a good fourth order polynomial fitting from 0.01 to 10.0 μg/mL for standard SSa sample with a detection limit of 0.006 μg/mL. The intra- and inter-assay coefficients of variation of the assay were 7.3% and 8.9%, respectively, and the average SSa recovery was 119.2%. For samples of Chaihu extract, the results of this assay showed a good correlation with those by enzyme-linked immunosorbent assay established previously. This TRFIA system is ultrasensitive for detecting SSa with a wide detection range and a good stability and represents the first attempt of using TRFIA for quality evaluation of the crude drug of Chaihu.

Suggested Citation

  • Zhi Chao & Qian Cui & Enwei Tian & Weiping Zeng & Xuan Cai & Xiaolei Li & Hiroyuki Tanaka & Yukihiro Shoyama & Yingsong Wu, 2016. "Ultrasensitive Time-Resolved Fluoroimmunoassay for Saikosaponin a in Chaihu (Bupleuri Radix)," PLOS ONE, Public Library of Science, vol. 11(3), pages 1-11, March.
    • Handle: RePEc:plo:pone00:0151032
    DOI: 10.1371/journal.pone.0151032
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