生脉注射液超高效液相色谱指纹图谱的建立及特征成分分析

目的: 采用超高效液相色谱建立生脉注射液指纹图谱,并应用超高效液相色谱-离子阱质谱联用技术对主要色谱峰进行指认。 方法: 色谱分离采用Acquity UPLC HSS(2.1 mm×100 mm,1.8 μm)色谱柱,以乙腈-10 mmol·L^-1醋酸铵为流动相梯度洗脱,流速0.4 mL·min^-1;质谱检测,雾化器压力207 kPa,干燥气流12 mL·min^-1,干燥气温度350 ℃,碰撞电压1.0 V,正负离子扫描模式。 结果: 建立的超高效液相指纹图谱,标定了18个共有峰,8个生产企业的相似度除2个企业外均高于0.9;利用二级质谱信息,结合对照品及文献报道,确定18个共有峰化合物的可能结构。 结论: 超高效液相指纹图谱可快速有效地评价生脉注射液的产品质量。同时,通过超高效液相色谱-离子阱质谱联用技术,为中药注射液的化学成分研究提供了一种快速、灵敏、高效的分析方法。

Objective: To establish the fingerprint of Shengmai injection and analyze its chemical constituents by ultra performance liquid chromatography (UPLC)-ion trap mass spectrometry. Methods: Chromatographic separation was performed on an Acquity UPLC HSS C₁₈(2.1 mm×100 mm,1.8 μm) column.The mobile phase consisting of acetonitrile and 10 mmol·L^-1 ammonium acetate was used in linear gradient elution with a flow rate of 0.4 mL·min^-1.Agilent 632 ion trap LC/MS was employed for analysis under the following conditions:spray gas pressure was 207 kPa,drying gas(N₂) flow was 12 mL·min^-1,drying gas temperature was 350 ℃,capillary voltage was 1.0 V,the sample was analyzed in the positive-ion mode. Results: Eighteen common peaks from the fingerprint of Shengmai injection were determined,the similarity of testing samples(except two of the totally eight enterprises) were all above 0.9.Based on MS/MS information,potential structures of eighteen chemical constituents were identified by comparing with reference standards and reference data. Conclusion: The method provides a reference for the quality control of Shengmai injection.The UPLC-MS/MS method is a rapid,sensitive and efficient method for chemical constituent study of traditional Chinese medicine injection.

1 ZHENG Xiao-wei(郑笑为),WANG Qi(汪祺),YU Jian-dong(于建东),et al.Establishment of HPLC fingerprint of Shengmai injection(生脉注射液HPLC指纹图谱的建立).Chin J Pharm Anal(药物分析杂志),2012,32(8):1471
2 LIU Dan(刘丹),PU She-ban(濮社班),QIAN Shi-hui(钱士辉),et al.Chemical constituents of Chinese red ginseng(中国红参化学成分的研究).Chin J Chin Mater Med(中国中药杂志),2011,36(4):462
3 HU Chun-xiu(胡春秀),KONG Hong-wei(孔宏伟),ZHU Chao(朱超),et al.An ultra performance liquid chromatography-time-of-flight-mass spectrometric method for fast analysis of ginsenosides in Panax ginseng root(超高效液相色谱-飞行时间质谱快速分析人参根中的皂甙化合物).Chin J Chromatogr(色谱),2011,29(6):488
4 MA Xiao-qiong(马小琼),XU Qing(徐青),LIANG Xin-miao(梁鑫淼).Analysis of ginsenosides in Ginseng by liquid chromatography-atmospheric pressure chemical ionization mass spectrometry(液相色谱-大气压化学电离质谱法分析人参中的人参皂甙).Chin J Chromatogr(色谱),2005,23(4):389
5 Zhang HJ,Wu YJ,Cheng YY.Analysis of 'SHENGMAI' injection by HPLC/MS/MS.J Pharm Biomed Anal,2003,31(1):175
6 Zheng CN,Hao HP,Wang X,et al.Diagnostic fragment-ion-based extension strategy for rapid screening and identification of serial components of homologous families contained in traditional Chinese medicine perscription using high-resolution LC-ESI-IT-TOF/MS:Shengmai injection as an example.J Mass Spectrom,2009,44(2):230
7 Sollorz G.Quality evaluation of ginseng roots.Quantitative HPLC determination of ginsenosides.Dtsch Apoth Ztg,1985,125(41):2052
8 YANG Liu(杨柳),LIU Yi-ming(刘奕明),ZENG Xing(曾星),et al.HPLC/MS/MS determination ginsenoside Rg₁ in Shenmai injection and in human plasma(HPLC/MS/MS法测定参麦注射液及人血浆中的人参皂苷Rg₁).Chin J Pharm Anal(药物分析杂志),2005,25(8):905
9 ZHANG Ping(张萍),ZHANG Nan-ping(张南平),XIAO Xin-yue(肖新月),et al.Chemical analysis of saponins in Panax ginseng(人参皂苷类成分的化学分析).Chin J Pharm Anal(药物分析杂志),2004,24(3):229
10 BI Fu-jun(毕福钧),ZHONG Shun-hao(钟顺好),GU Li-hong(顾利红).Comparison between RRLC and HPLC for determination of ginsenoside Rg₁,Re and Rb₁ in Radix et Rhizoma Ginseng Rubra and Radix Panacis Quinuefolii(RRLC法与HPLC法在红参和西洋参人参皂苷含量测定中的分析比较).Chin J Pharm Anal(药物分析杂志),2010,30(9):1720