高效液相色谱法测定大鼠和人血浆中坦西莫司的蛋白结合率

Determination of Plasma Protein Binding Rate of Temsirolimus in Rat Plasma and Human Plasma by HPLC

目的建立高效液相色谱法测定坦西莫司在大鼠和人血浆中的含量,并结合超滤法测定坦西莫司的血浆蛋白结合率。方法色谱柱为Inertsil ODS-SP C_(18)柱(250 mm×4.6 mm,5μm),流动相为乙腈-0.5%冰醋酸(65∶35,V/V),检测波长为277 nm,柱温为40℃,流速为1.0 mL·min^(-1)。结果坦西莫司在0.26~21.58 mg·L^(-1)浓度范围内与峰面积线性关系良好(R>0.999 5),定量下限为0.086 mg·L^(-1),坦西莫司大鼠血浆样品的平均提取回收率为(96.70±0.57)%(RSD为0.59%),坦西莫司人血浆样品的平均提取回收率为(96.33±1.54)%(RSD为1.60%)。坦西莫司在2.57,5.15,10.29 mg·L^(-1)浓度水平与大鼠的平均蛋白结合率为(87.48±0.61)%,与人血浆的的平均蛋白结合率为(85.31±0.63)%。结论该法操作简便、快速灵敏,能满足坦西莫司血药浓度的测定。在试验研究浓度范围内,血浆蛋白结合率与药物浓度无关,坦西莫司与大鼠和健康人血浆均有较高强度结合,坦西莫司与大鼠的血浆蛋白结合率高于人血浆(P<0.05)。

OBJECTIVE To establish HPLC method combined with ultrafiltration for the determination of temsirolimus and its plasma protein binding rate in rat plasma and human plasma.METHODS The HPLC analysis of temsirolimus was achieved on Inertsil ODS-SP C18 column with mobile phase consisted of 0.05%acetic acid-acetonitrile(65∶35,V/V).The flow rate was 1.0 mL·min-1,the detection wavelength was 277 nm,and column temperature was set at 40℃.RESULTS The calibration curve for temsirolimus was linear in the range of 0.26 to 21.58 mg·L-1(R>0.9995),and the lower limit of quantification was 0.086 mg·L-1.The average recovery rate of temsirolimus in rat plasma sample was(96.70±0.57)%(RSD=0.59%)and that of human plasma sample was(96.33±1.54)%(RSD=1.60%).At 2.57,5.15,10.29 mg·L-1,the average protein binding rate to rat plasma was(87.48±0.61)%,and to human plasma was(85.31±0.63)%.CONCLUSION This method is simple,rapid and sensitive,and can be used for the determination of temsirolimus in plasma.Plasma protein binding characteristics of temsirolimus show high intensity of binding in rat plasma and human plasma,while the protein binding rates of temsirolimus in rat plasma is higher than that in human plasma(P<0.05),which is independent of its plasma concentration.