LC-MS/MS法测定Beagle犬血浆中布洛芬对映体的浓度  

Vailidation of a LC- MS/MS method for quantification of ibuprofen enantiomers in Beagle dog plasma

作　　者：赵秀红[1,2] 夏媛媛[2] 黄玉荣[2] 张爱杰[2] 魏广力[2] 司端运[2] 

机构地区：[1]天津医科大学研究生院,天津300070 [2]天津药物研究院释药技术与药代动力学国家重点实验室,天津300193

出　　处：《中国药理学通报》2015年第4期570-575,共6页Chinese Pharmacological Bulletin

基　　金：国家科技部"重大新药创制"科技重大专项资助项目(No2012ZX09304002);天津市自然科学基金资助项目(No10SYSYJC28600)

摘　　要：目的 建立简便、快速、灵敏的LC-MS/MS法测定Bea-gle犬血浆中手性药物布洛芬浓度的分析方法,进行系统的方法学验证,并用于布洛芬在Beagle犬体内的药代动力学研究。方法 以酮洛芬为内标,血浆样品经正己烷（含5%异丙醇）液萃取后,以甲醇：1mmol·L^-1甲酸铵（含0.2%甲酸,10%甲醇）（82∶18,V/V）为流动相,采用Lux5uCellu-lose-3（250mm.4.6mm,5μm）柱分离,流速为0.8mL·min^-1。通过电喷雾电离源以多反应监测（MRM）方式进行负离子检测,用于定量分析的离子对分别为m/z205.2/161.2（布洛芬）和m/z253.1/209.2（内标,酮洛芬）。方法验证包括基质效应、提取回收率、线性、定量下限、批内与批间精密度、稳定性稀释效应等。结果 犬血浆中左/右旋布洛芬的线性范围为0.2～50mg·L^-1,最低定量限为0.2mg·L^-1,批内、批间精密度（RSD）介于1.01%～13.1%之间。Beagle犬单次口服给予右旋布洛芬后的主要药动学参数Cmax、T1/2、AUC（0-t）分别为：（82.98±14.83）mg·L-1、（3.217±0.7298）h、（362.0±58.67）h·mg·L^-1。单次口服给予消旋布洛芬后,其左/右旋布洛芬的主要药动学参数Cmax、T1/2、AUC（0-t）分别为：（70.62±14.81）/（74.48±20.08）mg·L^-1、（1.520±0.9286）/（5.432±1.234）h、（177.8±34.18）/（649.6±200.2）h·mg·L^-1。结论 该方法简便、快速、灵敏度高、重复性好,能够同时测量血浆中左/右旋布洛芬的药物浓度,可用于布洛芬在Beagle犬体内的药代动力学研究。Aim To develop a simple,rapid and accurate analysis method for determination of chiral ibuprofen in Beagle dog plasma. Method The plasma sample was submitted to liquid – liquid extraction using hexane / isopropanol（ 95 ∶ 5,V / V）,with ketoprofen as the internal standard（ IS）. The separation was accomplished in a Lux 5u Cellulose-3（ 250 mm·4. 6mm,5 μm） column,and the mobile phase consisted of methanol and a mixture of 1 mmol·L- 1ammonium acetate-methanol-formic acid（ 90 ∶ 10 ∶ 0. 2,V / V / V）with the volume ratio of 82 ∶ 18 at a flow rate of 0. 8m L · min- 1. The mass spectrometer consisted of an ESI interface operating at negative ionization mode and the detection was performed using multiple reaction monitoring at the transitions of m / z 205. 2 /161. 2 for ibuprofen and m / z 253. 1 /209. 2 for ketoprofen（ IS）.Method validation included the evaluation of the matrix effect, extraction recovery, linearity, lower LOQ,within-run and between-run precision,stability and dilution effect. Results The calibration curve was linear across the concentration range of 0. 2 - 50 mg·L^- 1for each ibuprofen enantiomer with a lower LOQ of 0. 2mg·L^-1. The within-run and between-run precision（ RSD%） was in the range 1. 01% - 13. 1% for each ibuprofen. The pharmacokinetic parameters for orally single dose of（ S ＋） and racemic ibuprofen in Beagle dogs were as follows： Cmax, T1 /2, AUC（ 0-t）were（ 82. 98 ± 14. 83） mg·L^- 1,（ 3. 217 ± 0. 7298） h,（ 362. 0 ± 58. 67） h·mg·L^- 1for（ S ＋） ibuprofen and 70. 62 /74. 48 mg·L^-1,1. 520 /5. 432 h,177. 8 /649. 6 h·mg·L^- 1for（ R-） /（ S ＋） ibuprofen,respectively. Conclusions A simple,rapid,accurate,high sensitivity and repeatability method has been successfully developed,which can analyze the concentrations of（ R-） /（ S ＋） ibuprofen in Beagle dog plasma simultaneously. The method could be applied for the investigation of pharmacokinetics of ibuprofen enantiomers in Beagle dogs.

关 键 词：布洛芬 LC-MS/MS 血药浓度 BEAGLE犬 药代动力学 对映异构体 

分 类 号：R332[医药卫生—人体生理学] R969.1[医药卫生—基础医学]