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引用本文:于生兰,徐加兵,欧阳臻.基于黄芪甲苷单克隆抗体的ELISA检测方法的建立[J].中国现代应用药学,2019,36(3):322-326.
YU Shenglan,XU Jiabing,OUYANG Zhen.Establishment of Enzyme-linked Immunosorbent Assay Method Based on Monoclonal Antibody Against Astragaloside Ⅳ[J].Chin J Mod Appl Pharm(中国现代应用药学),2019,36(3):322-326.
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基于黄芪甲苷单克隆抗体的ELISA检测方法的建立
于生兰1, 徐加兵2, 欧阳臻3
1.江苏农牧科技职业学院, 江苏 泰州 225300;2.泰州市食品药品检验所, 江苏 泰州 225300;3.江苏大学药学院, 江苏 镇江 212013
摘要:
目的 建立黄芪甲苷的酶联免疫分析(enzyme-linked immunosorbent assay,ELISA)检测方法。方法 以制备的黄芪甲苷单克隆抗体为基础,选择其最佳工作浓度,建立黄芪甲苷间接竞争ELISA法,并用该方法检测黄芪饮片中的黄芪甲苷含量。结果 该方法线性范围为39~1 830 ng·mL-1,组内和组间差异均≤ 10%,平均加样回收率为102.7%(n=6)。采用该方法检测黄芪饮片中黄芪甲苷的含量,所得结果与HPLC法基本一致。结论 建立了黄芪甲苷的ELISA检测方法,可为黄芪饮片及其制品的质量控制提供方法补充。
关键词:  黄芪甲苷  单克隆抗体  ELISA  质量控制
DOI:10.13748/j.cnki.issn1007-7693.2019.03.013
分类号:R915
基金项目:江苏省普通高校研究生科研创新计划(CXLX12_0669)
Establishment of Enzyme-linked Immunosorbent Assay Method Based on Monoclonal Antibody Against Astragaloside Ⅳ
YU Shenglan1, XU Jiabing2, OUYANG Zhen3
1.Jiangsu Agri-animal Husbandry Vocational College, Taizhou 225300, China;2.Taizhou Food and Drug Inspection Institute, Taizhou 225300, China;3.School of Pharmacy, Jiangsu University, Zhenjiang 212013, China
Abstract:
OBJECTIVE To establish an enzyme-linked immunosorbent assay(ELISA) method for the determination of Astragaloside Ⅳ(AST). METHODS Based on the preparation of monoclonal antibody against AST, an indirect competitive enzyme immunoassay method which could detect the content of AST in Radix Astragali was established by selecting the optimum working concentration. RESULTS The calibration curves were linear within the range of 39-1 830 ng·mL-1. The average recovery(n=6) was 102.7%, and the relative standard deviations(RSD) of measurements in the intra-assay and in the inter-assay were ≤ 10%. The analysis result of ELISA method was consistent with that of HPLC test in AST determination of Radix Astragali. CONCLUSION ELISA method for the determination of AST is well established and can be applied to the quality control for Radix Astragali and its products.
Key words:  Astragaloside Ⅳ(AST)  monoclonal antibody  ELISA  quality control
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