CAJ | 학술논문

目的：建立比格犬血浆中TBI-166的LC-MS/MS测定方法，研究比格犬口服TBI-166的血浆药代动力学特征和生物利用度。方法比格犬血浆样品中加入TBI-166和内标普萘洛尔后经乙腈沉淀蛋白。 HPLC分离采用Symmetry C8（2.1 mm×50 mm，3.5μm）柱，流动相为含0.1%甲酸的乙腈-水，梯度洗脱，流速为0.2 ml/min质谱检测采用电喷雾离子源，正离子选择反应监测（SRM）模式检测m/z 590→478（TBI-166）和m/z 260→183（普萘洛尔，内标）。应用上述LC-MS/MS方法测定比格犬口服TBI-166（3 mg/kg）和静脉注射（0.5 mg/kg）后的血浆药物浓度，用WinNonlin软件计算药代动力学参数和绝对生物利用度。结果比格犬血浆中TBI-166在2～1000 ng/ml浓度范围内线性关系良好，日内和日间精密度<10%、回收率>98.6%、无明显基质效应且血浆样品稳定性好。雌雄比格犬口服TBI-166（3mg/kg）后血药浓度分别于（4.4±3.5）和（1.4±0.5）h达峰，血药峰浓度分别为（122.0±34.6）和（65.4±2.3）ng/ml，AUC（0-t）为（2615.1±1524.4）和（897.2±318.6）h·μg/L。雌雄比格犬静注TBI-166（0.5mg/kg）后t1/2z为（112.9±25.3）和（69.6±35.3）h、CL为（0.3±0.1）和（0.2±0.1）L/（h·kg）、AUC（0-t）为（3222.4±1656.2）和（1798.0±729.2）h·μg/L。雌雄比格犬口服TBI-166生物利用度分别为13.5%和8.3%。结论本研究建立了比格犬血浆中TBI-166的LC-MS/MS测定方法，该方法准确、简便、灵敏。比格犬口服TBI-166后体内消除较慢，具有一定性别差异，生物利用度为8.3%～13.5%。
목적：건립비격견혈장중TBI-166적LC-MS/MS측정방법，연구비격견구복TBI-166적혈장약대동역학특정화생물이용도。방법비격견혈장양품중가입TBI-166화내표보내락이후경을정침정단백。 HPLC분리채용Symmetry C8（2.1 mm×50 mm，3.5μm）주，류동상위함0.1%갑산적을정-수，제도세탈，류속위0.2 ml/min질보검측채용전분무리자원，정리자선택반응감측（SRM）모식검측m/z 590→478（TBI-166）화m/z 260→183（보내락이，내표）。응용상술LC-MS/MS방법측정비격견구복TBI-166（3 mg/kg）화정맥주사（0.5 mg/kg）후적혈장약물농도，용WinNonlin연건계산약대동역학삼수화절대생물이용도。결과비격견혈장중TBI-166재2～1000 ng/ml농도범위내선성관계량호，일내화일간정밀도<10%、회수솔>98.6%、무명현기질효응차혈장양품은정성호。자웅비격견구복TBI-166（3mg/kg）후혈약농도분별우（4.4±3.5）화（1.4±0.5）h체봉，혈약봉농도분별위（122.0±34.6）화（65.4±2.3）ng/ml，AUC（0-t）위（2615.1±1524.4）화（897.2±318.6）h·μg/L。자웅비격견정주TBI-166（0.5mg/kg）후t1/2z위（112.9±25.3）화（69.6±35.3）h、CL위（0.3±0.1）화（0.2±0.1）L/（h·kg）、AUC（0-t）위（3222.4±1656.2）화（1798.0±729.2）h·μg/L。자웅비격견구복TBI-166생물이용도분별위13.5%화8.3%。결론본연구건립료비격견혈장중TBI-166적LC-MS/MS측정방법，해방법준학、간편、령민。비격견구복TBI-166후체내소제교만，구유일정성별차이，생물이용도위8.3%～13.5%。
Objective To develop a LC-MS/MS method for the quantification of TBI-166, a novel antituberculotic, in beagle dog plasma, and apply it to the pharmacokinetic and bioavailability study. Methods The preparation of plasma sample was a simple deproteinization by the addition of acetonitrile followed by centrifugation. The separation was performed on a Symmetry C 8 column (2.1 mm × 50 mm,3.5μm) with mobile phase of acetonitrile/water containing 0.1%formic acid(V/V) using a gradient elution mode at a flow rate of 0.2 ml/min. The detection was performed in positive selected reaction monitoring (SRM) mode with an electrospray ionization source. The analytes were quantified at m/z 590→478 for TBI-166 and m/z 260→183 for propranolol (internal standard, IS). Results Linear detection responses were obtained for TBI-166 in dog plasma ranging from 2 to 1000 ng/ml. The intra- and inter-day precisions (RSD%) were no more than 10%. The average recovery was greater than 98.6%, and there was good stability and no obvious matrix effect for the quantification. The method was successfully applied in the pharmacokinetic study of TBI-166 in beagle dogs. The AUC(0-t), Cmax and Tmax of TBI-166 in male and female dogs were(897.2±318.6) and(2615.1±1524.4) h·μg/L,(65.4±2.3) and(122.0±34.6) ng/ml and(1.4±0.5) and(4.4±3.5) h after oral administration at 3 mg/kg. The t1/2z, AUC(0-t), and CL of TBI-166 in male and female dogs were (69.6±35.3) and (112.9±25.3) h, (1798.0±729.2) and (3222.4±1656.2) h·μg/L,(0.2±0.1) and (0.3 ±0.1) L/(h·kg) after intravenous administration at 0.5 mg/kg. Conclusion An accurate, simple and sensitive LC-MS/MS method for the determination of TBI-166 in beagle dog plasma was developed and validated. This method was applied to the pharmacokinetic study of TBI-166 in beagle dogs. There was a gender difference on the pharmacokinetic profiles of TBI-166 in dogs. TBI-166 was eliminated slowly and the bioavailability of TBI-166 was 8.3-13.5% in male and female dogs.