吐根酊特征图谱研究与质量评价方法建立

Study on HPLC specific chromatogram and establishment of quality evaluation method of ipecac tincture

目的:建立吐根酊的HPLC特征图谱,对主要成分进行鉴别,并评价其质量。方法:采用Inertsil ODS-SP(250 mm×4.6mm,5μm)色谱柱,以乙腈-0.4%甲酸水溶液为流动相,梯度洗脱,流速为1 m L·min-1,检测波长210 nm,柱温35℃。利用LC-MS/MS方法对特征图谱中的主要特征峰进行推测,对其中的主要未知峰进行液相制备纯化,并用NMR试验进行结构鉴定。结果:建立了40批吐根酊的HPLC特征图谱,推测其中的6个特征峰,对3个含量较大的色谱峰进行准确指认,分别为吐根酚碱、吐根碱和吐根苷。判断其中36批吐根酊样品为合格样品,4批为劣质吐根酊样品。结论:建立的吐根酊HPLC特征图谱专属性强,方法验证表明其可用于该产品的质量评价。

Objective：To establish the HPLC specific chromatogram,identify the main constituents and evaluate the quality of ipecac tincture.Methods：The HPLC specific chromatogram was performed on an Inertsil ODS-SP column（250 mm × 4.6 mm,5 μm） with a gradient mobile phase of acetonitrile-0.4% formic acid aqueous solution system at a flow rate of 1 m L·min-1.The detection wavelength was 210 nm and the column temperature was 35 ℃.The main peaks in the chromatogram were presumed with LC-MS/MS.The main unknown peak was obtained by prepared HPLC and purified,and its structure was identified by NMR test.Results：The HPLC specific chromatograms of 40 batches of samples were obtained,in which six alkaloids were characterized and three main constituents were obtained and identified as cephaeline,emetine and ipecoside.Thirty-six samples were high qualified,while four samples were low qualified.Conclusion：The method validation shows that the established methods have high specificity and can be used for the quality evaluation of ipecac tincture.