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建立间接竞争酶联免疫吸附测定法检测桔梗中桔梗皂苷D的含量

Establishment of indirect competitive enzyme-linked immunosorbent assay for the detection of platycodin D in Radix Platycodonis.

作者信息

Luo Shuai, He Yanfei, Sun Hongxiang

机构信息

College of Animal Sciences, Zhejiang University, Hangzhou, Zhejiang, 310058, China.

出版信息

J Food Drug Anal. 2020 Sep 15;28(3):399-406. doi: 10.38212/2224-6614.1009.

Abstract

Platycodin D (PD) has been used as the quality control marker of Radix Platycodonis for its high content and various pharmacological properties. In this study, a specific polyclonal antibody against PD (PD-pAb) was developed, and PD-pAb-based indirect competitive enzyme-linked immunosorbent assay (icELISA) was established for the detection of PD in Radix Platycodonis. The 50% inhibition concentration (IC) of PD was 2.70 μg/mL and the linearity range for PD was from 0.032 μg/mL to 100 μg/mL. No cross reactivity with PD-pAb was found in five PD analogs except for platycodin D2 (PD2, 0.93%). The average recovery of PD by icELISA was 97.14% (RSD = 1.17%). The icELISA was used for the detection of PD in different Radix Platycodonis samples and the results were confirmed by high performance liquid chromatography (HPLC). The correlation coefficient between the two assays was 0.9654. Taken together, the established icELISA might be a simple, cheap, rapid, sensitive, reliable and high-throughput method for determining the contents of PD in Radix Platycodonis.

摘要

桔梗皂苷D(PD)因其含量高且具有多种药理特性,一直被用作桔梗的质量控制指标。本研究制备了一种针对PD的特异性多克隆抗体(PD-pAb),并建立了基于PD-pAb的间接竞争酶联免疫吸附测定法(icELISA)用于检测桔梗中的PD。PD的50%抑制浓度(IC)为2.70μg/mL,PD的线性范围为0.032μg/mL至100μg/mL。除桔梗皂苷D2(PD2,0.93%)外,5种PD类似物与PD-pAb均无交叉反应。icELISA法测定PD的平均回收率为97.14%(RSD = 1.17%)。icELISA法用于检测不同桔梗样品中的PD,结果经高效液相色谱法(HPLC)验证。两种检测方法的相关系数为0.9654。综上所述,所建立的icELISA法可能是一种简单、廉价、快速、灵敏、可靠且高通量的测定桔梗中PD含量的方法。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/33c4/9261796/a06ee7034dab/jfda-28-03-399f1.jpg

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