摘要
目的:建立他克莫司软膏体外释放评价方法,比较国内仿制制剂和参比制剂的体外释放行为一致性。方法:采用A型浸没池建立他克莫司软膏的体外释放方法,选择无水乙醇-生理盐水(30∶70)为释放介质,孔径0.45μm的聚四氟乙烯膜为人工膜,转速200 r·min^(-1)。以液相色谱-串联质谱法测定释放量,采用Agilent Zorbax SB C_(18)(150 mm×2.1 mm,3.5μm)色谱柱,以0.1%甲酸溶液(含5 mmol·L^(-1)甲酸铵)(A)-甲醇溶液(B)为流动相,梯度洗脱,柱温55℃,进样体积10μL。测定他克莫司仿制制剂与参比制剂的释放量,并计算释放速率。采用基于Wilcoxon的秩检验和Mann-Whitney U检验的统计学方法计算仿制制剂与参比制剂的释放速率比值的90%标准置信区间,评价仿制与参比他克莫司软膏体外释放一致性。结果:所建检测方法检测限达到0.026 ng·mL^(-1),回收率在92.6%~99.3%。2家国产他克莫司软膏仿制制剂中,仿制制剂1与参比制剂的释放速率比值的90%置信区间为97.92%~122.19%,在75.00%~133.33%范围内,显示体外释放速率与参比制剂一致,初、复试中仿制制剂2与参比制剂供试品释放速率比值的90%置信区间均超出75.00%~133.33%的范围,表明仿制制剂2的体外释放速率与参比制剂不一致。浸没池方法的研究结果与扩散池方法的研究结果一致。结论:本文所建方法符合他克莫司软膏体外释放评价要求且结果可靠,浸没池体外释放法可用于他克莫司软膏的体外释放行为评价。
Objective:To establish an in vitro release evaluation method for tacrolimus ointment,and compare the consistency of in vitro release behavior between domestic generic products and the reference product.Methods:An in vitro release method for tacrolimus ointment was established using immersion cell-Model A.The mixture of anhydrous ethanol and physiological saline(30∶70)was selected as the release medium,and 0.45μm polytrafluoroethylene(PTFE)membrane was used as the artificial membrane,the rotation speed was set at 200 r·min^(-1).High-performance liquid chromatography with tandem mass spectrometry(LC-MS/MS)was adopted to determine the release amount and release rate of tacrolimus,using Agilent Zorbax SB C_(18)(150 mm×2.1 mm,3.5μm)column and gradient elution.0.1%formic acid solution(containing 5 mmol·L^(-1) ammonium formate)(A)and methanol(B)were used as the mobile phase,with column temperature of 55℃,and an injection volumn of 10μL.The release amounts of the generic and reference products of tacrolimus ointment were measured,and their release rates were calculated.Wilcoxon and Mann-Whitney U statistical methods were employed to calculate the 90%standard confidence interval of the ratio of release rates between the generic product and the reference product,evaluating the consistency of in vitro release.Results:The LOD of the established method was 0.026 ng·mL^(-1),and the recovery of released tacrolimus in the receptor solution(mass balance)was within 92.6%-99.3%.Among the two domestic generic tacrolimus ointments,the 90%standard confidence interval of the ratio of the release rate between generic product 1 and reference product was 97.92%-122.19%which fell within the acceptable range of 75.00%-133.33%,indicating consistency in in vitro release rate with the reference product.The 90%confidence intervals of the ratio of the release rate between the generic product 2 and the reference product were all out of range of 75.00%-133.33%in both the initial and repeat tests,suggesting inconsistency in in vitro release rate compared to the reference product.The results obtained using the immersion cell method were consistent with those obtained using the Franz diffusion cell method.Conclusion:The developed immersion cell method meets the requirements for evaluating the in vitro release of tacrolimus ointment and the results are reliable.The immersion cell in vitro release method can be used to evaluate the in vitro release behavior of tacrolimus ointment.
作者
宋诗颖
崔潆心
凌霞
叶晓霞
乐健
许旭
SONG Shi-ying;CUI Ying-xin;LING Xia;YE Xiao-xia;LE Jian;XU Xu(Shanghai Institute of Technology,Shanghai 201418,China;Shanghai Institute for Food and Drug Control,NMPA Key Laboratory for Quality Analysis of Chemical Drug Preparations,Shanghai 201203,China)
出处
《药物分析杂志》
北大核心
2025年第4期671-678,共8页
Chinese Journal of Pharmaceutical Analysis
