Ultrasensitive time-resolved fluoroimmunoassay for saikosaponin a in Chaihu (Bupleuri Radix)

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

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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.

Original languageEnglish
Article numbere0151032
JournalPloS one
Volume11
Issue number3
DOIs
Publication statusPublished - Mar 2016

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All Science Journal Classification (ASJC) codes

  • Biochemistry, Genetics and Molecular Biology(all)
  • Agricultural and Biological Sciences(all)
  • General

Cite this

Chao, Z., Cui, Q., Tian, E., Zeng, W., Cai, X., Li, X., ... Wu, Y. (2016). Ultrasensitive time-resolved fluoroimmunoassay for saikosaponin a in Chaihu (Bupleuri Radix). PloS one, 11(3), [e0151032]. https://doi.org/10.1371/journal.pone.0151032