基于指纹图谱的女金丸质量一致性评价

Quality consistency evaluation of Nyujin Pills based on fingerprint analysis

目的分别建立女金丸的高效液相色谱(HPLC)和气相色谱(GC)指纹图谱,客观评价女金丸的产品质量一致性。方法采用Agilent Eclipse plus C18(4.6 mm×250 mm,5μm)色谱柱,乙腈(A)-0.2%甲酸溶液(B)为流动相,梯度洗脱(0~25 min,10%A^35%A;25~37 min,35%A^59%A;37~40 min,59%A^75%A),柱温和流速分别为30℃和1 mL·min-1,波长切换,0~20 min,230 nm;20~40 min,270 nm,建立女金丸的高效液相色谱指纹图谱;采用HP-5毛细管柱(30 m×250μm×0.25μm),升温程序为初始温度50℃,保持1 min,以每分钟20℃的速率升温至130℃,保持10 min;再以每分钟2℃的升温速率升至180℃,保持1 min;进样口温度230℃,检测器温度250℃;分流进样,分流比10∶1,建立女金丸的气相色谱指纹图谱。采用高分辨质谱和对照品对指纹图谱中的特征峰进行确认,并通过《中药色谱指纹图谱相似度评价软件》(2012年版)对指纹图谱进行相似度计算,来评价女金丸的产品质量一致性。结果共确定女金丸高效液相色谱指纹图谱15个共有峰,通过对照品比较指认其中12个成分;共确定女金丸气相色谱指纹图谱30个共有峰,通过高分辨质谱和对照品比较指认其中28个成分。不同企业之间和同一企业内部产品质量一致性较差。结论所建立的女金丸高效液相色谱和气相色谱指纹图谱信息较为全面,可用于女金丸的产品质量一致性评价。

Objective To establish a method of HPLC and GC fingerprint analysis for quality consistency evaluation of Nyujin Pills.Methods The HPLC fingerprint analysis was carried out on a column of Agilent Eclipse plus C18(4.6 mm×250 mm,5 μm),using a mobile phase of acetonitrile(A)-0.2% formic acid solution(B) by a gradient eluted program(0~25 min,10%A^35%A;25~37 min,35%A^59%A;37~40 min,59%A^75%A)with a flow rate of 1 mL·min-1 at 30 ℃,the detection wavelength was set at 230 nm in first 20 min,and then changed to 270 nm between 20 and 40 min;The GC fingerprint analysis was carried out on a column of HP-5 capillary column(30 m×250 μm×0.25 μm),temperature programming:initial temperature 50 ℃,keep 1 minutes,heating up to 130 ℃ with a rate of 20 ℃ per minute,keep 10 minutes,and then heating up to 180 ℃ with a rate of 2 ℃ per minute,keep 10 minutes,introduction port temperature 230 ℃,detector temperature 250 ℃,split sampling,split ratio 10∶1.The marker peaks were identified by comparison with high resolution mass spectrometry and reference substance,the similarity of specific chromatogram were calculated through similarity evaluation software(2012)and the quality consistency evaluation of Nyujin Pills,can be evaluated.Results The common mode of the HPLC fingerprint was set up with 15 common peaks,and 12 of them were identified by comparison with the reference;the common mode of the GC fingerprint was set up with 30 common peaks,and 28 of them were identified by comparison with high resolution mass spectrometry and reference.The consistency of product quality between different enterprises and the same enterprise was poor.Conclusion The established HPLC and GC fingerprint information of Nyujin Pills was relatively comprehensive and can be used to evaluate the product quality consistency of Nyujin Pills.