基于HPLC及GC-MS技术的血尿胶囊融合指纹图谱

目的：建立血尿胶囊的融合指纹图谱。方法：采用高效液相色谱（HPLC），使用C₁₈色谱柱（4.6 mm×250 mm，5 μm），以乙腈-0.1%磷酸水为流动相，进行梯度洗脱，流速1.0 mL·min^-1，检测波长280 nm，进样量10 μL。采用气相色谱-质谱联用（GC-MS），使用HP-5MS色谱柱（30 m×250 μm×0.25 μm），载气为氦气，体积流量为1.0 mL·min^-1，进样口温度250℃，柱温采用程序升温（初始温度45℃，保持2 min，以5℃·min^-1程序升温至180℃，保持10 min，后以10℃·min^-1程序升温至250℃，保持10 min），进样量1 μL，分流比20：1；电离方式EI，电子能量70 eV，离子源温度230℃，四极杆温度150℃，质量范围m/z 50~550。将HPLC与GC-MS的色谱数据进行归一化处理，并将数据导入国家药典委员会制定的《中药色谱指纹图谱相似度评价系统2.0版》进行指纹图谱融合。结果：在血尿胶囊融合指纹谱获得34个共有峰，HPLC指纹图谱得到22个共有峰，并指认了原儿茶酸、儿茶素、绿原酸、咖啡酸、虎杖苷、落新妇苷、黄杞苷、白藜芦醇、槲皮素9个成分，GC-MS指纹图谱鉴定了棕榈酸甲酯、棕榈酸乙酯、9，12-十八碳二烯酸甲酯、（E）-9-十八烯酸甲酯、硬脂酸甲酯、亚油酸乙酯、油酸乙酯、7，10-十八碳二烯酸甲酯、硬脂酸乙酯、11-二十烯酸甲酯、二十酸甲酯、二十二酸甲酯12个成分。结论：建立的融合指纹图谱能够全面反映血尿胶囊的整体特征，克服了单一成分含量测定及单一指纹图谱的不全面性，为复方中药制剂质量控制方法研究提供依据。

Objective:To establish fusion-fingerprint of Xueniao capsules.Methods:HPLC analysis was performed on a C₁₈ column (4.6 mm×250 mm,5 μm) with mobile phase consisting of acetonitrile-0.1% phosphoric acid with gradient elution at a flow rate of 1.0 mL·min^-1.The detection wavelength was 280 nm and the injection volume was 10 μL.GC-MS analysis was performed on an HP-5MS column (30 m×250 μm×0.25 μm) and carrier gas was helium (He) at a flow rate of 1.0 mL·min^-1.The inlet temperature was 250℃.The column temperature was held at 45℃ for 2 min and then programmed at 5℃·min^-1 to 180℃ and held for 10 min,then at 10℃·min^-1 to 250℃ and held for 10 min.The injection volume was 1 μL and the split ratio was 20:1.The ionization mode was EI and the electron energy was 70 eV.The ion source temperature was 230℃ and the quadrupole temperature was 150℃.The scan range was m/z 50-550.The fusion-fingerprint was originated by similarity evaluation software based on the normalized data which acquired from HPLC and GC-MS.Results:34 common peaks were found in the fusion-fingerprint of Xueniao capsules. 22 peaks were from HPLC and 9 of them were identified as protocatechuic acid,catechins,chlorogenic acid,caffeic acid,polydatin,astilbin,engeletin,resveratrol and quercetin. 12 peaks were from GC-MS and identified as hexadecanoic acid,methyl ester;hexadecanoic acid,ethyl ester;9,12-octadecadienoic acid,methyl ester;(E)-9-octadecenoic acid,methyl ester;octadecanoic acid,methyl ester;linoleic acid ethyl ester;ethyl oleate;7,10-octadecadienoic acid,methyl ester;octadecanoic acid,ethyl ester;11-eicosenoic acid,methyl ester;eicosanoic acid,methyl ester;docosanoic acid and methyl ester.Conclusion:Superior to single component quantification and single fingerprint,the proposed fusion fingerprint could reflect the comprehensive characteristics of Xueniao capsules,thus providing reference for quality control of traditional Chinese medicine preparations.