尼美舒利分散片在健康人体内的药代动力学及生物等效性的LC-MS/MS研究

目的: 建立人血浆中尼美舒利浓度的LC-MS/MS测定法,并用于尼美舒利分散片的药代动力学和生物等效性研究。方法: 采用自身双交叉试验设计,20名男性受试者随机分成2组,分别单剂量口服100 mg受试制剂或参比制剂,0～24 h间隔采集血样。以LC-MS/MS内标法测定尼美舒利血药浓度,使用Agilent TC-C₁₈色谱柱(250 mm×4.6 mm,5 μm),流动相为甲醇-0.1%甲酸溶液(82:18,v/v);多反应监测[M+H]⁺离子通道分别为m/z 309.1→m/z 153.9(尼美舒利)和m/z 237.1→m/z 194.0(内标卡马西平),DAS 2.1计算药动学参数。结果: 建立的LC-MS/MS法在0.075～12 μg·mL^-1范围内色谱响应与浓度相关性良好,最低定量限为0.075 μg·mL^-1,批内及批间精密度RSD均小于15%。受试制剂与参比制剂的T_max分别为(3.0±0.7)h和(3.5±1.0)h,C_max分别为(4.863±1.194)μg·mL^-1和(4.657±1.038)μg·mL^-1,t_1/2分别为(3.2±1.0)h和(3.3±1.1)h,AUC_{0-24 h}分别为(32.35±12.50)h·μg·mL^-1和(32.32±11.69)h·μg·mL^-1,相对生物利用度F为(105.2%±35.0)%。结论: 建立的LC-MS/MS法准确、灵敏,结果可靠,测得尼美舒利受试制剂和参比制剂生物等效。

Objective: To establish an LC-MS/MS method for the determination of nimesulide in human plasma, used for the study of the clinical pharmocokinetics and bioequivalence of nimesulide dispersible tablets. Methods: In a randomized two-way self-crossover study, 20 healthy male volunteers were divided into two groups, and were administered respectively either with a single oral dose of test or reference preparations containing 100 mg of nimesulide.The blood samples were collected at predetermined time intervals within 24 hours.The plasma concentration of nimesulide was determined by LC-MS/MS.The chromatographic separation was performed on an Agilent TC-C₁₈ (250 mm ×4.6 mm, 5 μm)column with a mobile phase consisting of methanol and water solvent with 0.1% formic acid (82: 18, v/v).The analytes were detected by multiple reaction monitoring of the [M+H]⁺ions with transitions of m/z 309.1→m/z 153.9 and m/z 237.1→m/z 194.0 for nimesulide and carbamazepine, respectively.The pharmacokinetic parameters were estimated by DAS 2.1. Results: The peak areas and concentrations showed good correlation in the range of 0.075-12 μg·mL^-1 with a LOQ of 0.075 μg·mL^-1 for quantitation of nimesulide in human plasma.The intra- and inter- batch precision RSDs were less than 15%.The pharmacokinetic parameters of the test and reference tablets were as follows:T_max (3.0±0.7) and (3.5±1.0)h, C_max (4.863±1.194) and (4.657±1.038)μg·mL^-1, t_1/2 (3.2±1.0) and (3.3±1.1)h, AUC_{0-24 h}(32.35±12.50) and (32.32±11.69)h·μg·mL^-1, respectively.The relative bioavailability F was(105.2%±35.0)%. Conclusion: The method is accurate, sensitive, reliable and suitable for the pharmacokinetic study of nimesulide.The two preparations are bioequivalent.