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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 prepare the artificial antigen of chlorogenic acid (CGA), which is a traditional Chinese medicine ingredient, and to optimize the synthesis reaction conditions, so as to provide the material basis and technical basis for the in-depth study of adverse reactions of traditional Chinese medicine injections. METHODS: The complete antigen (CGA-BSA) was prepared by coupling CGA with the carrier protein bovine serum albumin (BSA) using the carbodiimide method. The orthogonal reaction method was used to optimize the reaction conditions. Matrix-assisted laser desorption/time-of-flight mass spectrometry was used. (MALDI-TOF-MS) and UV absorption spectrum (UV) were used to determine the coupling ratio (Mr). RESULTS: The MR of artificial antigen CGA-BSA synthesized by MALDI-TOF-MS and UV was 20 and 18.8, respectively; the optimal reaction conditions were NHS, EDC, BSA and CGA in a ratio of 4:1,1:1,1. : 200, pH is 9. Conclusion: The chlorogenic acid total antigen (CGA-BSA) was successfully synthesized, which provided the material basis for the further study of the allergenicity and correlation of chlorogenic acid.