目的: 研究复方克霉唑乳膏中抑菌剂降解产物的分子结构和产生路径,为药品处方改进提供依据。方法: 针对复方克霉唑乳膏中的抑菌剂及其降解产物、以及尼泊金类抑菌剂与辅料氢氧化钾的相容性,采用HPLC和UPLC-HRMS方法进行分析研究。HPLC采用Phenomenex Prodigy ODS3 C18色谱柱(150 mm×4.6 mm, 5 μm),以0.02 mol·L-1醋酸铵溶液(A)-甲醇(B)为流动相梯度洗脱,检测波长为257 nm,柱温为35 ℃;UPLC-HRMS采用Q-Orbitrap质谱,YMC C18色谱柱(250 mm×4.6 mm, 5 μm),以0.1%甲酸水(A)-0.1%甲酸乙腈(B)为流动相梯度洗脱,HESI负离子化检测。结果: 通过乳膏相容性试验和质谱分析,阐明了辅料氢氧化钾促进了尼泊金类抑菌剂较为严重的降解,二者的不相容是造成多批次抑菌剂不符合规定(<处方量的80%)的根本原因。结论: 个别生产企业复方克霉唑乳膏处方中选择氢氧化钾作辅料存在抑菌剂含量不合格的风险,建议企业优化制剂处方,降低因辅料-辅料不相容带来的安全性风险。
Objective: To research the molecular structure and formation route of the degrade products of bacteriostatic agents in compound clotrimazole cream, and to provide evidence for improvement of preparation formulation. Methods: This work, which included the determination of bacteriostatic agents and their degrade products and the compatibility study between paraben bacteriostatic agent and excipient potassium hydroxide, was carried out by using HPLC to measure the content of bacteriostatic agents and their degrade products and by using UPLC-HRMS to characterize the structure of the degrade impurity. HPLC was applied with a Phenomenex Prodigy ODS3 C18 column (150 mm×4.6 mm, 5 μm), and gradient elution with 0.02 mol·L-1 ammonium acetate solution (A)-methanol(B) under the condition of detection wavelength 257 nm and column temperature 35 ℃. UPLC-Q-Orbitrap HRMS was applied with a YMC C18 column (250 mm×4.6 mm, 5 μm), and gradient elution with 0.1% formic acid in water(A)-0.1% formic acid in acetonitrile(B) under the MS condition of HESI source and negative ion mode. Results: Through compatibility test simulating cream process conditions and HRMS analysis, it was clarified that excipient potassium hydroxide promoted the more serious degradation of paraben antibacterial agents, and the incompatibility of the above two components was the cause of multiple batches of antibacterial agents that did not meet the requirement(<80% of the prescription amount). Conclusion: The selection of potassium hydroxide as an excipient in the prescription of compound clotrimazole cream by individual company has the risk of unqualified antibacterial content. It is recommended that the related company optimizes the cream prescriptions and strives to reduce the safety risks caused by the excipient-excipient incompatibility.
[1] SMITH RJ, WEBB ML. Analysis of Drug Impurities[M].Oxford: Blackwell Publishing, 2007: 27
[2] 李敏. 药物降解的有机化学[M].北京: 化学工业出版社, 2019:136
LI M. Organic Chemistry of Durg Degradation[M].Beijing: Chemical Industry Press, 2019:136
[3] SONG Y, SCHOWEN RL, BORCHARDT RT, et al. Formaldehyde production by Tris buffer in peptide formulations at elevated temperature[J].J Pharm Sci, 2001, 90(8):1198
[4] 程杰, 李显庆, 王静静, 等. 亚硫酸氢钠甲萘醌注射液中4个降解杂质的UPLC-Q-Orbitrap HRMS结构鉴定[J].药物分析杂志, 2021, 41(9):1538
CHENG J, LI XQ, WANG JJ, et al. Qualitative analysis of 4 degradation impurities in menadione sodium bisulfite injection by UPLC-Q-Orbitrap HRMS[J].Chin J Pharm Anal, 2021, 41(9):1538
[5] WANG G, FISKE JD, JENNINGS SP, et al. Identification and control of a degradation product in AvaproTM film-coated tablet: low dose formulation[J].Pharm Devel Technol, 2008, 13(5):393
[6] ZHANG F, NUNES MJ. Structure and generation mechanism of a novel degradation product formed by oxidatively induced coupling of miconazole nitrate with butylated hydroxytoluene in a topical ointment studied by HPLC-ESI-MS and organic synthesis[J].J Pharm Sci, 2004, 93(2):300
[7] 张恩娟, 曹健, 刘同华, 等. 复方克霉唑乳膏的制备与质量控制[J].中国药业, 2004, 13(7):46
ZHANG EJ, CAO J, LIU TH, et al. Preparation and determination of compound clortimazole cream[J].China Pharm, 2004, 13(7):46
[8] 曹筱琛, 贾飞, 陶巧凤. 药物与辅料相容性研究进展[J].中国现代应用药学, 2013, 30(2):223
CAO XC, JIA F, TAO QF. Progress in the study of drug-excipient compatibility in dosage forms[J].Chin J Mod Appl Pharm, 2013, 30(2):223
[9] 杨晓梅, 梁勇坤, 余良钟, 等. 克霉唑乳膏中有关物质的定性与定量分析[J].药学学报, 2018, 53(12):2093
YANG XM, LIANG YK, YU LZ, et al. Qualitative and quantitative analysis of related substances in clotrimazole cream[J].Acta Pharm Sin, 2018, 53(12):2093
[10] 中华人民共和国药典2020年版. 四部[S].2020: 966
ChP 2020. Vol Ⅳ[S].2020: 966
[11] 吴春敏, 朱精英, 池文杰, 等. HPLC法快速检测苦参素注射液中8个抑菌剂及苯甲醛[J].药物分析杂志, 2012, 32(7):1200
WU CM, ZHU JY, CHI WJ, et al. HPLC rapid screening and simultaneous determination of eight bacteriostatic agents and benzaldehyde in oxymatrine injection[J].Chin J Pharm Anal, 2012, 32(7):1200
[12] 王静静, 堵伟锋, 卫星红, 等. 替硝唑葡萄糖注射液杂质的色谱-质谱结构鉴定[J].药学学报, 2019, 54(9):1655
WANG JJ, DU WF, WEI XH, et al. Identification of impurity profile in tinidazole glucose injection by UPLC-MS[J].Acta Pharm Sin, 2019, 54(9):1655
[13] 胡昌勤. 药物杂质谱分析[M].北京: 化学工业出版社, 2015: 9
HU CQ. Impurity Profiling of Pharmacetuticals[M].Beijing: Chemical Industry Press, 2015: 9
[14] 杨园, 李苗, 王珊珊, 等. 盐酸西替利嗪片有关物质分析[J].药物分析杂志, 2019, 39(12):2136
YANG Y, LI M, WANG SS, et al. Analysis of related substances in cetirizine hydrochloride tablets[J].Chin J Pharm Anal, 2019, 39(12):2136
[15] FARSA O, SUBERT J, MAREKOVÁ M. Hydrolysis and transesterification of parabens in an aqueous solution in the presence of glycerol and boric acid[J].J Excip Food Chem, 2011, 2(2):41
[16] KOSOVÁ M, HRÁDKOVÁ I, MÁTLOVÁ V. Antimicrobial effect of 4-hydroxybenzoic acid ester with glycerol[J].J Clin Pharm Ther, 2015, 40(4):436