RP-HPLC法测定大鼠血浆中3-正丁基苯酞的含量及其药动学

Determination of 3-n-butylphthalide in rat plasma by RP-HPLC and its pharmacokinetics

目的建立大鼠口服及静脉注射3-正丁基苯酞(NBP)的药动学研究方法。方法以雄性Wistar大鼠为实验对象,分别采取静脉注射和口服给药2种方式,于给药后不同时间点取血,采用高效液相反相色谱检测法(RP-HPLC)测定大鼠血浆中NBP的含量,选用Hypersil ODS色谱柱(250 mm×4.6 mm,5μm),以0.05 mol·L^(-1)醋酸钠溶液-乙睛(50:50,V/V)为流动相,流速为1.0 mL·min^(-1),检测波长为228 nm。应用3P97药动学软件处理血药浓度数据,获得药动学参数。结果 NBP在0.050 38～21.23μg·mL^(-1)(r=0.999 7)范围内线性关系良好,其中定量下限为0.050 38μg·mL^(-1);日内和日间精密度均小于4.8%,低、中、高浓度的NBP提取回收率均在80%以上。NBP药动学过程符合双隔室开放模型。大鼠静脉注射NBP 5、10、20 mg·kg^(-1)的主要药动学参数分别为t_(1/2)(2.67±0.72)h、(2.23±0.92)h、(2.27±0.52)h;AUCo_(0-t)(8.15±1.46)mg·h·L^(-1)、(7.23±1.55)mg·h·L^(-1)、(14.05±1.67)mg·h·L^(-1)。口服60、240和360 mg·kg^(-1)NBP的主要药动学参数依次为P_(max)(2.90±0.94)μg·mL^(-1)、(5.46±0.56)μg·mL^(-1)、(9.46±1.61)μg·mL^(-1);AUC_(0-t)(7.11±2.90)mg·h·L^(-1)、(14.00±0.82)mg·h·L^(-1)、(24.33±2.86)mg·h·L^(-1)。结论本法灵敏度高、专属性强、易操作,结果准确可靠,可用于NBP的药动学研究。

AIM To study the pharmacokinetics of 3-n-butylphthalide （NBP） in rats. METHODS Male Wistar rats were given NBP by intravenous and oral administration at different doses. The serial blood samples were collected at different time following administration. A highperformance liquid chromatographic （HPLC） method with a reversedphase Hypersil ODS column （250 mm x 4.6 mm, 5 μm） was used for the determination of NBP in rats. The mobile phase was consisted of 0.05 moL. L-1 sodium acetate-acetonitrile （50 ：50, V/V）, and adjusted to pH 4.5 with acetic acid. The flow rate was 1.0 mL .rain and the detection wavelength was 228 nm. 3P97 software was used to deal with data of the concentration in plasma and calculate pharmacokinetic parameters for NBP in rats. RESULTS The method demonstrated that good linearity ranged from 0.050 38 to 21.23 μg.mL-1 （r = 0.999 7） . The lower limit of quantification for NBP was 0.050 38 μg-mL-1. The intra-and inter-day precision values were below 4.8% and the reeovery of NBP at different eoneentrations was all higher than 80%. The absorption of NBP in rats of two administrations was consistent with twocompartment model. The main pharmacokinetic parameters of NBP in rat plasma after intravenous administration at the dose of 5, 10 and 20 mg＇kg-1 were tin （2.67 ± 0.72） h, （2.23 ± 0.92） h, （2.27 _± 0.52） h; AUC0 （8.15 ± 1.46） mg.h.L-1, （7.23± 1.55） mg.h.L-1, （14.05 ± 1.67） mg.h.L-1. The main pharmacokinetic parameters of NBP in rat plasma after oral administration at the dose of 60, 240 and 360 mg. kg-1 were pr （2.90 ± 0.94） μg-mL-1, （5.46 ± 0.56） μg.mL-1, （9.46 ± 1.61） μg-mL-l; AUCo （7.11 ± 2.90） mg.h.L-1, （14.00 ± 0.82） mg.h.L-1, （24.33 ± 2.86） mg.h.L-L CONCLUSION The method is simple, sensitive, stability and operated easily. It can be used to study the pharmacokinetics of NBP in rats.