硝酸甘油片有关物质方法的辅料扣除及质谱定性研究

目的: 建立液质联用法鉴别硝酸甘油片中的辅料，解决硝酸甘油片质量标准中有关物质辅料峰的判定和扣除问题。方法: 采用Lichro CART C₁₈色谱柱，流动相为乙腈-水(50∶50)，大气压化学电离源，一级质谱全扫描，考察辅料峰的归属。通过重复性、稳定性和耐用性实验，以及水分的测定，考察辅料峰面积的稳定性。结果: 样品色谱图中保留时间2.8min前的色谱峰为辅料峰，且该辅料峰在紫外检测和质谱检测中的响应值均主要源自聚维酮K 29/32。辅料峰的保留时间和峰面积均具有较好的重复性、稳定性和耐用性。扣除辅料峰后，3批硝酸甘油片样品的有关物质结果均符合规定。结论: 本研究建立的液质联用法测定硝酸甘油片中的辅料，操作简单，结果准确，为相关辅料的研究提供了技术依据。根据辅料峰的归属和耐用性考察结果，为中国药典2010年版二部中硝酸甘油片质量标准修订提供了参考。

Objective: To develop a new LC-MS method to determine the excipients in nitroglycerin tablets for the identification and deduction of excipient peaks in related substances. Methods: The Lichro CART C₁₈ column was used with acetonitrile-water as the mobile phase (50∶50).A tandem mass spectrometer equipped with atmospheric pressure chemical ionization source was used as the detector and was operated in full scan to quantify the excipients.An HPLC method and the water assay were used to investigate the stability of excipient area through repeatability,stability and ruggedness tests. Results: The peaks which were obtained before the retention time of 2.8 min were judged as excipients in the chromatograms of samples.The UV and MS response of the excipient peak was mainly sourced from povidone K 29/32.The results of repeatability,stability and ruggedness tests were good in the retention time and peak area of samples.After deduction of the peaks of excipients,the related substance results of three batches of nitroglycerin tablets were all qualified in accordance with related requirements.Conclusion: An accurate and feasible method based on LC-MS was developed to determine the excipients in nitroglycerin tablets.According to the identification of excipients and the results of ruggedness tests,the reference for revising the quality standards of nitroglycerin tablets in the 2010 edition of Chinese Pharmacopoeia was provided.

1 GAO Feng-tong(高凤彤),SUN Zhan-bo(孙湛博),ZHANG Xiao-wen(张晓雯).Research progress in effects and drug - resistant mechanism of nitroglycerin(硝酸甘油作用机制及耐药机制的研究进展).[J] Nat Med Front China(中国医疗前沿),2010,5(8):7
2 YANG Qing-xia(杨清霞),Wang Ling(王伶),Liu Fu-qiang(刘富强).Pharmacologic effects and clinical application of nitroglycerin(硝酸甘油的药理作用及临床应用).[J] Drug evaluat(药品评价),2006,3(3):214
3 ChP(中国药典)[S].2010.Vol Ⅱ(二部):947
4 BP[S].2012.2875
5 USP 35 NF30.-[S]2012.4066
6 JP ⅩⅥ[S].2011.1169
7 LUO Jian-Ming(罗建明),SITU Shao-Jin(司徒少金),LI Chun (黎春),et al.HPLC determination of the related substances of nitroglycerin solution(HPLC法测定硝酸甘油溶液中的有关物质)[J].Jiangsu Pharm Clin Res(江苏药学与临床研究),2005,13(4):17
8 WU Liang-yong(伍良涌),PAN Xi-qiang(潘锡强),WU Jun-cheng(吴均成).RP-HPLC determination of nitroglycerin aerosoland its related substances(反相高效液相色谱法测定硝酸甘油气雾剂有关物质和硝酸甘油含量)[J].Chin J Pharm Anal(药物分析杂志),2012,32(5):895
9 WU Liang-yong(伍良涌),PAN Xi-qiang(潘锡强).Determination of the related substances of nitroglycerin solution and its related substances by RP-HPLC(反相高效液相色谱法测定硝酸甘油溶液及其制剂的有关物质)[J].Chin J Clin Rational Drug Use(临床合理用药),2010,3(20):1
10 LI Hong-shu(李洪书),CHEN Bin(陈斌).Discussion on determination method in Chinese pharmacopoeia of related substances of nitroglycerin Tablets(中国药典中硝酸甘油片有关物质测定方法的探讨)[J].Chin J Pharm(中国医药工业杂志),2012,43(11):946
11 WU Liang-yong(伍良涌),PAN Xi-qiang(潘锡强),LI Zhi-fang(黎志芳),et al.Some suggestions on revision of the quality standards of nitroglycerin in Chinese Pharmacopoeia Volume Ⅱ( 2010 Edition)(中国药典2010年版硝酸甘油系列质量标准修订建议)[J].Chin J Pharm Anal(药物分析杂志),2013,33(5):870
12 Rowe R C,Sheskey P J,Weller P J.Handbook of Pharmaceutical Excipients(药用辅料手册)[M].4th Ed(第四版).Bei jing(北京):Chemical Industry Press(化学工业出版社),2004.584