【摘要】 目的:本文通过不同纯度绿原酸的致敏试验、通过平衡透析除去大分子后不同纯度绿原酸的致敏性试验、透析获得的大分子物质的致敏性试验、通过LC-MS分离鉴定致敏透析物以及比较分析国内外有关绿原酸致敏研究的文献,综合研究并论证了绿原酸是否具有致敏原性。方法:以豚鼠和SD大鼠为受试动物,分别采用了主动和被动过敏试验验证不同纯度绿原酸提取物、注射用绿原酸及经平衡透析后的透析内液和外液、过期和降解后的注射用绿原酸等受试药物;采用LC-MS法(Kromasil C-18 150×4.6mm,5μm,300A;流动相为0.1%甲酸:乙腈=95:5(v/v);流速:1μl/min;进样量:2μl。质谱条件:电喷雾离子源,Esi+;喷雾电压:2.8kv)鉴定了透析内液中1000道尔顿以上的大分子残留物的分子量范围;对比分析了从60年代初期至今绿原酸致敏性研究的相关文献。结果:纯度高于92%的绿原酸提取物、注射剂、过期注射剂、降解后注射剂、透析外液和透析内液均未发生过敏反应;纯度低于40%的绿原酸提取物及其透析内液均被LC-MS鉴定出大分子化合物并且发生明显的过敏反应。结论:试验结果证明了绿原酸没有致敏原性,与文献最后结论绿原酸无致敏性相吻合,同阐明引起致敏性的主要原因与提取物中存在的大分子物质密切相关。更多还原

【Abstract】 Objective:To evaluate and demonstrate whether chlorogenic acid (CHA) is the characteristic of allergen,the sensitization test (STT) of different purity of CHA extractives,STT of different purity of CHA without of macromolecules by equilibrium dialysis,STT of macromolecules obtained from the equilibrium dialysis were carried out,sensitized dialysates were separated and identified by LC-MS,and domestic and overseas literatures were compared and analyzed about allergy of CHA.Methods:Guinea pigs and SD rats were used as the animal subjects.The active systemic anaphylaxis(ASA) and passive cutaneous anaphylaxis(PCA) were applied respectively to validate different purity of CHA extract,chlorogenic acid(CHA) for injection,dialysis diffusible solutions and nondiffusible solution,overdue CHA for injection and degradative CHA for injection.The range of molecular weight above 1 000 dalton of macromolecule residue was identified by LC-MS(Kromasil C-18 150×4.6mm,5μm,300A;mobile phase 0.1% formic acid:acetonitrile=95:5(v/v);flow rate 1μl/min.ion source Esi+;V 2.8kv).Also,the pertinent literatures of anaphylaxis study of CHA were contrasted and analyzed.Results:No anaphylaxis occurred in the purity more than 92% of CHA extracts,injection,overdue injection,degradative injection.dialysis internal solution and external solution.There were macromolecular compounds which identified by LC-MS in the purity of CHA less than 40% of extracts and dialysis internal solutions and anaphylaxis occurred apparently.Conclusion:CHA is not an allergen demonstrated by the experiments,which agrees with the results of the literatures.Besides,anaphylaxis is closely correlated with the macromolecular contaminants in the low purity of CHA extract.更多还原