单次和多次口服氯乙酰左卡尼汀片在健康人体的药动学研究

Pharmacokinetics Study of A Single and Multiple Oral Chloroacetyl Acetyl-L-carnitine Tablet in Healthy Volunteers

关键词:  乙酰左卡尼汀  高效液相色谱-质谱联用  药动学  血浆

Key words:  acetyl-L-carnitine  LC-MS/MS  pharmacokinetics  plasma

摘要:

目的 采用LC-MS/MS测定人血浆中乙酰左卡尼汀的浓度并用于氯乙酰左卡尼汀片在健康受试者体内的药动学研究。方法 健康受试者单次给药0.5，1.0，1.5 g和多次给予0.5 g后，0~24 h采集血样。通过测定单次和多次给药后血浆中乙酰左卡尼汀的绝对浓度，计算其药动学参数。米屈肼为内标，经甲醇沉淀蛋白后进行LC-MS/MS分析。ESI离子源正离子模式监测，检测离子m/z 204.3→145.2（乙酰左卡尼汀），m/z 147.2→58.2（米屈肼）；色谱柱为EC 250/4.6 NUCLEOSIL100-5CN，流动相为甲醇-10 mmol·L^-1乙酸铵溶液（含0.1%甲酸）（85：15）。结果 乙酰左卡尼汀在20~3 000 μg·L^-1内线性良好（r=0.999 1），最低定量限为20 μg·L^-1。批内、批间精密度及基质效应RSD均<15%。单次给药3个剂量组（0.5，1.0，1.5 g）的主要药动学参数为：AUC_0-t为（4 181.77±2 473.24）μg·h·L^-1、（6 099.54±1 939.41）μg·h·L^-1和（8 064.71±3 575.99）μg·h·L^-1，C_max为（611.42±270.76）μg·L^-1，（830.92±233.19）μg·L^-1和（1 004.67±414.95）μg·L^-1，t_1/2z为（4.50±2.93）h、（6.25±3.65）h和（5.76±3.94）h；多次给药后主要的药动学参数：AUC_0-t为（13 728.82±6 493.04）μg·h·L^-1，C_max为（1 129.00±374.05）μg·L^-1，t_1/2z为（8.57±4.42）h。结论 本方法准确、灵敏、专属性强，适用于人体内乙酰左卡尼汀药动学研究。单次和多次给予氯乙酰左卡尼汀片后药动学参数有明显差异，性别间无差异，健康受试者对药物的耐受性良好。

Abstract:

OBJECTIVE To develop a LC-MS/MS method for the content determination of acetyl-L-carnitine in human plasma, and to study the pharmacokinetics of chloroacetyl acetyl-L-carnitine tablet in healthy volunteers. METHODS Chloroacetyl acetyl-L-carnitine tablet at the single administration of 0.5, 1.0, 1.5 g and multiple administration of 0.5 g were given to healthy volunteers. Blood samples were collected at 0-24 h. The pharmacokinetic parameters were calculated by measuring the absolute concentration of acetyl-L-carnitine in plasma of healthy volunteers after single and multiple administration. Mildronate was used as the internal standard(IS), the plasma samples were treated with methanol to precipitate protein and analyzed by LC-MS/MS. The detection was accomplished with ESI source operated in positive ionization mode, and ion reaction pairs were selected to be m/z 204.3→145.2(acetyl-L-carnitine), and m/z 147.2→58.2(mildronate). Chromatographic separation had been achieved on EC 250/4.6 NUCLEOSIL 100-5CN with methanol-10 mmol·L^-1 ammonium acetate(with 0.1% formic acid)(85:15) as the mobile phase. RESULTS The linear calibration curves were obtained in the entire range (20-3 000 μg·L^-1) for acetyl-L-carnitine(r=0.999 1), inter-and intra-batch RSDs and matrix effect were <15%. The pharmacokinetic parameters of single dose(0.5, 1.0, 1.5 g) were:AUC_0-t(4 181.77±2 473.24)μg·h·L^-1, (6 099.54±1 939.41)μg·h·L^-1 and (8 064.71±3 575.99)μg·h·L^-1; C_max (611.42±270.76)μg·L^-1, (830.92±233.19)μg·L^-1 and (1 004.67±414.95)μg·L^-1; t_1/2z (4.50±2.93)h, (6.25±3.65)h and (5.76±3.94)h. The main pharmacokinetic parameters of multiple doses were:AUC_0-t (13 728.82±6 493.04)μg·h·L^-1; C_max (1 129.00±374.05)μg·L^-1; t_1/2z (8.57±4.42)h. CONCLUSION The method is proved to be accurate, sensitive and specific to study on the pharmacokinetics of acetyl-L-carnitine in healthy volunteers. There is a significant difference in the pharmacokinetic parameters after single and multiple doses of chloroacetyl acetyl-L-carnitine tablets, and there is no difference between the sexes, healthy subjects are well tolerated.