LC-MS/MS法测定人体血浆中左旋舒必利的浓度及药物动力学应用

Determination of levosulpiride in human plasma and the application in pharmacokinetics by LC- MS / MS

目的建立测定人血中左旋舒必利浓度的高效液相色谱串联质谱电喷雾检测法（LC-MS/MS）。方法血浆样品用乙酸乙酯-二氯甲烷（V∶V=4∶1）提取,色谱柱为Agilient TC-C18柱,流动相为甲醇-乙腈-5 mmol·L-1醋酸铵（含体积分数为2%的甲酸）（V∶V∶V=85∶10∶5）,流速为0.8 m L·min-1;质谱：采用多反应检测模式,API2000电喷雾离子源。结果左旋舒必利线性为1~600μg·L-1;定量下限为1μg·L-1;日间日内精密度（RSD）均不大于14.8%。准确度（RE）：-6.8%~9.5%。健康受试者单剂量口服左旋舒必利25、50和100 mg及多剂量口服50 mg后,受试制剂的主要药物动力学参数为：tmax分别为（3.67±0.65）、（3.75±1.14）、（3.33±0.49）和（2.75±1.14）h,ρmax分别为（40.28±11.13）、（74.75±16.21）、（179.08±35.78）和（131.83±43.18）μg·L-1,t1/2分别为（13.19±3.69）、（13.21±4.71）、（11.74±2.45）、（18.59±10.81）h,AUC0-t分别为（473.6±125.0）、（841.20±203.67）、（1687.62±373.98）和（1848.17±610.65）μg·h·L-1。单剂量口服左旋舒必利后,在25~100 mg剂量内,AUC0-t、ρmax呈线性动力学特征趋势,多剂量给药7 d,左旋舒必利在体内有一定蓄积。各剂量组的药物动力学参数在男女性别间无显著性差异。结论该法适合于左旋舒必利的人体药物动力学研究。

Objective To develop an LC-MS/MS method for the study of the pharmacokinetics of levo- sulpiride in volunteers. Methods The plasma samples were extracted with ethyl acetate-methylene chloride （ V： V = 4： 1 ） and the separation was performed on a reversed-phase Agilitent TC-C18 column. The mobile phase contained methanol-acetonitrile-5 mmol·L- 1 ammonium acetate with 2% （φ）formic acid （ V： V： V = 85： 10：5 ）. Detection was carried out by multiple reactions monitoring with an API 2 000 triple quadrupole mass spectrometer equipped with an electrospray ionization source. Results The linear calibration curve was obtained in the range of 1- 600 ktg· L - 1 for levosulpiride. The lower limit of quantitation was 1 μg. L - 1. The intra-day and inter-day precision RSD was less than 14.8%, and the accuracy （relative error）was within 6. 8%-9.5%. The main pharmacokinetic parameters of levosulpiride were as follows： tmax were（3.67 ± 0. 65 ）, （3.75 ± 1.14 ）, （ 3.33 ± 0. 49 ） and （ 2. 75 ± 1.14 ） h, respectively; Pmax were （ 40. 28 ± 11.13 ）, （74. 75 ± 16. 21 ）, （ 179. 08 ± 35.78 ） and （ 131.83 ± 43.18 ） μg· L - 1, respectively; t1/2 were （ 13.19 ± 3.69 ）, （ 13.21 ± 4.71 ）, （ 11.74 ± 2.45 ） and （ 18.59 ± 10. 81 ） h, respectively; AUC0-t were （ 473.6 ± 125.0 ）, （841.20 ± 203.67 ）, （ 1687.62 ± 373.98 ） and （ 1848. 17 ± 610. 65 ） μg. h. L- 1, respectively. Oral adminis- trate single dose of 25 mg ,50 mg, 100 mg ,AUC0-t and Pmax presented liner pharmacokinetics. And after oral administrating multiple doses of 50 mg, the parameters showed slight drug accumulation. The pharmacokinet- ic parameters of levosulpirde among different dose groups were found no significant difference between male and female. Conclusions The method is suitable for pharmacokinetics study of levosupiride.