Article ID | Journal | Published Year | Pages | File Type |
---|---|---|---|---|
7626133 | Journal of Pharmaceutical and Biomedical Analysis | 2018 | 7 Pages |
Abstract
In this paper, a simple, rapid and high-throughput fluorescence polarization immunoassay (FPIA) based on polyclonal antibodies (PAb) is described for the determination of glycocholic acid (GCA) in human urine. Three fluorescein-labeled GCA (tracers) with different structures and spacer bridges were synthesized and purified by thin-layer chromatography (TLC). The structure effect of tracers on the assay was investigated and the sensitivity of best tracer in the optimized FPIA demonstrated an IC50 value of 306â¯ng/mL. The working range of FPIA was 36 â¼ 2 600â¯ng/mL and the limit of detection (LOD) was 9â¯ng/mL. The developed FPIA was time-saving that could be completed within 10â¯min. Human urine samples spiked with GCA were analyzed by this method, followed by confirmation with commercial enzyme immunoassay analysis (EIA). Excellent recoveries and correlation between these two methods were observed (R2â¯=â¯0.996), suggesting the developed FPIA could be applied to screening of GCA in human urine samples without complicated cleanup.
Keywords
Related Topics
Physical Sciences and Engineering
Chemistry
Analytical Chemistry
Authors
Qiyi He, Xiping Cui, Ding Shen, Yingshan Chen, Zhengyun Jiang, Rui Lv, Sergei A. Eremin, Suqing Zhao,