右兰索拉唑缓释胶囊在Beagle犬体内的生物等效性研究

Bioequivalence Study of Dexlansoprazole Sustained-release Capsules in Beagle Dogs

目的:建立测定Beagle犬血浆中右兰索拉唑浓度的方法,评价两种右兰索拉唑缓释胶囊的生物等效性。方法:6只Beagle犬随机分为参比制剂组和受试制剂组,每组3只,采用双周期交叉随机试验(清洗期为1周)设计,分别空腹单次口服右兰索拉唑缓释胶囊参比制剂和受试制剂各60 mg,并于给药前及给药后0.5、1、1.5、2、2.5、3、3.5、4、4.5、5、6、7、8、10、12 h自前肢头静脉取血2mL,采用液相色谱-串联质谱法(LC-MS/MS)测定Beagle犬血浆中右兰索拉唑的浓度。以奥美拉唑为内标,色谱柱为Inertsil ODS,流动相为甲醇-0.1%甲酸溶液(90∶10,V/V);采用电喷雾离子源,以多反应监测模式进行正离子扫描,用于定量分析的离子对分别为m/z 369.9→251.7(右兰索拉唑)、m/z 345.7→197.9(内标)。采用DAS 3.1.6软件计算药动学参数,以双侧t检验及(1-2α)%置信区间(CI)法、Wilcoxon检验考察两种制剂的生物等效性。结果:右兰索拉唑血药浓度的线性范围为10~4 000 ng/mL,定量下限为10 ng/mL;批内、批间RSD<10%,准确度为94.40%~105.20%。空腹单次口服参比制剂或受试制剂的AUC_(0-t)分别为(8 892.48±1 399.67)、(8 683.71±1 167.88)ng·h/mL,AUC_(0-∞)分别为(8 925.73±1 399.64)、(9 053.08±1 553.46)ng·h/mL,t_(max)分别为(3.50±0.71)、(3.75±1.21)h,c_(max)分别为(2 980.00±487.40)、(2 863.33±331.34)ng/mL。受试制剂AUC_(0-t)、AUC_(0-∞)、c_(max)的90%CI均在参比制剂相应参数的80%~125%范围内,两制剂t_(max)间的差异无统计学意义(P>0.05)。结论:本研究建立的LC-MS/MS法适用于Beagle犬血浆中右兰索拉唑浓度的测定,且两种制剂在Beagle犬体内具有生物等效性。

OBJECTIVE：To establish a method for the determination of dexlansoprazole concentration in plasma of Beagle dogs,and to evaluate bioequivalence of 2 kinds of Dexlansoprazole sustained-release capsules. METHODS：A total of 6 Beagle dogs were randomly divided into reference preparation group and test preparation group,with 3 dogs in each group. In double cycle cross randomized trial（cleaning period lasting for one week）design,they were given reference preparation and test preparation of Dexlansoprazole sustained-release capsules 60 mg once. The blood sample 2 mL of forelimb vein were collected before medication,0.5,1,1.5,2,2.5,3,3.5,4,4.5,5,6,7,8,10,12 h after medication,respectively. The concentration of dexlansoprazole in plasma of Beagle dogs was determined by LC-MS/MS. Using omeprazole as internal standard,the determination was performed on Inertsil ODS with mobile phase consisted of methanol-0.1% formic acid solution（90 ∶ 10,V/V）. ESI was used for positive ion scanning by multiple reaction monitoring mode. The ion pair for quantitative analysis were m/z 369.9→251.7（dexlansoprazole）and m/z 345.7→197.9（internal standard）,respectively. The pharmacokinetic parameters were calculated by using DAS 3.1.6 software.The bioequivalence of two preparations were investigated by bilateral t test,（1-2α）confidence interval（CI）method and Wilcoxon test. RESULTS：The linear range of dexlansoprazole concentration was 10-4 000 ng/mL. The lower limit of quantitation was 10 ng/mL;RSDs of inter-batch and intra-batch were lower than 10%. The accuracy ranged 94.40%-105.20%. The pharmacokinetic parameters of reference preparation or test preparation of single fasting dose were as follows：AUC0-twere（8 892.48±1 399.67）,（8 683.71±1 167.88）ng·h/mL,AUC0-∞were（8 925.73±1 399.64）,（9 053.08±1 553.46）ng·h/mL,tmaxwere（3.50±0.71）,（3.75±1.21）h,cmax were（2 980.00±487.40）,（2 863.33±331.34）ng/mL,respectively. 90%CI of AUC0-t,AUC0-∞and cmaxfor test preparation were80%-125% of corresponding parameters of reference preparation;there was no statistical significance in tmaxbetween 2 preparations（P〈0.05）. CONCLUSIONS：LC-MS/MS is suitable for the determination of dexlansoprazole concentration in Beagle dogs plasma.Moreover,the two preparations are bioequivalent in Beagle dogs.