苦木注射液高效液相色谱指纹图谱研究及多成分定量分析

Fingerprint research and multi-component quantitative analysis of Kumu injection by HPLC

目的:建立苦木注射液的HPLC指纹图谱,并同时测定3种咔巴啉类生物碱的含量,全面评价其内在质量。方法:采用Phenomenex Gemini C18色谱柱(4.6 mm×250 mm,5μm),甲醇-30 mmol.L-1乙酸铵(冰乙酸调pH4.5)溶液梯度洗脱,用国家药典委员会"中药色谱指纹图谱相似度评价系统2004 A版"处理分析,建立苦木注射液的HPLC指纹图谱共有模式。结果:在指纹图谱研究中,标定了16个共有峰,指认了7个化合物,分析的11批苦木注射液与共有模式之间相似性良好,相似度均在0.9以上;3种咔巴啉类生物碱在含量分析测定条件下,各组分分离良好,相关系数均为0.999 9,线性范围分别为0.020 0～0.300 0,0.102 0～1.530 0,0.015 2～0.228 0μg,加样回收率在99.5%～102%。结论:本法灵敏度高、专属性强,HPLC指纹图谱结合多成分含量测定为苦木注射液的质量控制提供科学依据。

Objective： To establish the HPLC chromatographic fingerprint of Kumu injection and to simultaneously determine the contents of three β-carboline alkaloids,comprehensively evaluating the immanent quality of Kumu injection.Method： The chromatographic analysis was performed on a Phenomenex Gemini C18（4.6 mm×250 mm,5 μm） column with the gradient elution solvent system composed of methanol and 30 mmol·L-1 aqueous ammonium acetate（adjusted with glacial acetic acid to pH 4.5）.Similarity evaluation system for chromatographic fingerprint of traditional Chinese medicine（2004 A） was used in data analysis.Result： Sixteen co-possessing peaks were selected as the fingerprints of Kumu injection,and 7 peaks were identified by chemical reference substances.There were good similarities between the standard fingerprint chromatogram and each fingerprint chromatogram from the eleven samples for their similarity coefficients were not less than 0.9.Three kinds of β-carboline alkaloids were separated well.The correlation coefficients were 0.999 9.The linear ranges of three components were 0.020 0-0.300 0,0.102 0-1.530 0,0.015 2-0.228 0 μg,respectively,and the average recoveries ranged were from 99.5% to 102%.Conclusion： The method of fingerprint combined with quantitative analysis is sensitive,selective,and provide scientific basis for quality control of Kumu Injection.