中国中药杂志

2009, v.34(15) 1906-1909

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绿原酸人工抗原的合成及反应条件的优化
Synthesis and optimize reaction conditions of chlorogenic acid artificial antigen

林辉;刘屏;王建勋;李斌;穆丽华;胡园;
LIN Hui,LIU Ping,WANG Jianxun,LI Bin,MU Lihua,HU Yuan(Dept of Clinical Pharmacology,General Hospital of People Liberation Army,Beijing 100853,China)

摘要(Abstract):

目的:制备中药成分绿原酸(CGA)的人工抗原,并优化合成反应条件,为中药注射剂不良反应的深入研究提供物质基础和技术基础。方法:采用碳二亚胺法,将CGA与载体蛋白牛血清白蛋白(BSA)偶联起来制得完全抗原(CGA-BSA);正交法优化合成反应条件;经基质辅助激光解吸飞行时间质谱(MALDI-TOF-MS)和紫外吸收光谱(UV)测定其偶联率(Mr)。结果:MALDI-TOF-MS和UV测得合成的人工抗原CGA-BSA的Mr分别为20和18.8;反应最佳条件为NHS,EDC,BSA与CGA的比率为4:1,1:1,1:200,pH为9。结论:成功合成了绿原酸全抗原(CGA-BSA),为进一步深入研究绿原酸的过敏原性及其相关性研究提供物质基础。
Objective:To synthesize artificial antigen of the chlorogenic acid(CGA),a constituent in Traditional Chinese Medicine,then optimize the reaction conditions,so that to supply the basis in material and technique for study on Chinese medicine injection of adverse reactions in-depth.Method:The complete antigen of chlorogenic acid(CGA-BSA)was synthesized by CGA coupled to carrier protein bovine serum albumi(BSA) using carbodiimide method.We used the orthogonal design test to optimize reaction conditions and used the MALDI-TOF-MS and UV motheds to determine the coupling rate of the CGA-BSA.Result:The coupling rate were 20 and 18.8,counting by MALDI-TOF-MS and UV,respectively;while the best reaction conditions were NHS:CGA,EDC:CGA,CGA:BSA were 4:1,1:1,200:1,respectively,with the pH 9.Conclusion:The complete antigen is preparated successfully.Which supplied the basis to advanced study of correlation.

关键词(KeyWords): 绿原酸;基质辅助激光解吸飞行时间质谱;完全抗原
chlorogenic acid;MALDI-TOF-MS;complete antigen

Abstract:

Keywords:

基金项目(Foundation): 国家自然科学基金项目(90709040)

作者(Authors): 林辉;刘屏;王建勋;李斌;穆丽华;胡园;
LIN Hui,LIU Ping,WANG Jianxun,LI Bin,MU Lihua,HU Yuan(Dept of Clinical Pharmacology,General Hospital of People Liberation Army,Beijing 100853,China)

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