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

Clinical pharmacokinetics and bioequivalence of nimesulide dispersible tablets by LC-MS/MS method

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,5μm),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0.075～12μg·mL-1范围内色谱响应与浓度相关性良好,最低定量限为0.075μg·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Tmax分别为(3.0±0.7)h和(3.5±1.0)h,Cmax分别为(4.863±1.194)μg·mL-1和(4.657±1.038)μg·mL-1,t1/2分别为(3.2±1.0)h和(3.3±1.1)h,AUC0-24 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 nime- sulide. 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 transi- tions 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 pharmacokinetie parameters of the test and reference tablets were as follows ： T_max （ 3.0 ± 0.7 ） and （ 3.5 ± 1.0 ） h, C （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, AUC0-24 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 ac-curate, sensitive, reliable and suitable for the pharmacokinetic study of nimesulide. The two preparations are bio- equivalent.