UPLC法测定大鼠血浆中那格列奈的浓度及药动学研究

Concentration Determination of Nateglinide in Rats' Plasma by UPLC and Study on Its Pharmacokinetics

目的:建立测定大鼠血浆中那格列奈浓度的方法,并研究其在大鼠体内的药动学特征。方法:采用超高效液相色谱法(UPLC)。色谱柱为Acquity UPLC~? BEH C_(18),流动相为乙腈-10 mmol/L磷酸二氢钾缓冲盐溶液(41:59,V/V),流速为0.38 mL/min,柱温为35℃,检测波长为210 nm,进样量为2μL。18只Wistar大鼠分别ig那格列奈16 mg/kg,分别于给药前及给药后10、20、30、45、60、90、120、180、240、360、480 min于眼内眦取血0.4 mL,测定血浆中那格列奈的浓度;并采用DAS 2.1.1软件计算那格列奈药动学参数。结果:那格列奈质量浓度在0.05~6.4μg/mL范围内线性关系良好(r=0.999 3),定量下限为0.05μg/mL;日内(n=5)、日间(n=3)精密度和稳定性(n=3)试验的RSD均小于10%;提取回收率和方法回收率分别为78.71%~80.56%、91.78%~100.42%(RSD<10%,n=5)。大鼠ig那格列奈后,AUC_(0-8h)为(5.87±2.32)μg·h/mL,AUC_(0-∞)为(6.11±2.48)μg·h/mL,t_(1/2)为(1.72±0.55)h,t_(max)为(0.67±0.29)h,c_(max)为(3.34±1.23)μg/mL。结论:该方法准确快速、专属性强,可用于大鼠体内那格列奈浓度的测定;那格列奈在大鼠体内吸收迅速、代谢较快。

OBJECTIVE：To establish a method for the concentration determination of nateglinide in rats＇ plasma and study its pharmacokinetic characteristics in rats in vivo. METHODS：UPLC was performed on the column of Acquity UPLC~？ BEH C（18） with mobile phase of acetonitrile-10 mmol/L potassium dihydrogen phosphate buffer（41：59,V/V）at flow rate of 0.38 mL/min,with column temperature of 35 ℃,detection wavelength of 210 nm and volume of 2 μL. 18 Wister rats were intragastrically administrated nateglinide 16 mg/kg. Blood sample 0.4 mL was taken from medial canthus before administration and after 10,20,30,45,60,90,120,180,240,360,480 min of administration. The concentration of nateglinide in rats＇ plasma was determined;then DAS2.1.1 software was used to calculate its pharmacokinetic parameters. RESULTS：Nateglinide showed good linear relationship in 0.05-6.4 μg/mL（r=0.999 3）,lower limit of quantification was 0.05 μg/mL;RSDs of inter-day（n=5）,intra-day（n=3）and stability（n=3） tests were lower than 10%;extraction recovery rate and method recovery rate were 78.71%-80.56%,91.78%-100.42%（RSD10%,n=5）,respectively. After rats were intragastrically administrated nateglinide,AUC（0-8 h） was（5.87 ± 2.32） μg·h/mL,AUC（0-∞） was（6.11 ± 2.48）μg·h/mL,t（1/2） was（1.72 ± 0.55）h,t（max） was（0.67 ± 0.29）h and c（max） was（3.34 ± 1.23）μg/mL.CONCLUSIONS：The method is accurate,rapid with strong specificity,and can be used for the concentration determination of nateglinide in rats in vivo;nateglinide is absorbed and metabolized quickly in rats in vivo.