反相高效液相色谱法测定2型糖尿病人群血浆中盐酸二甲双胍浓度

Determination of plasma concentration of metformin hydrochloride in type 2 diabetic patients by RP-HPLC

目的:建立盐酸二甲双胍血药浓度测定的反相高效液相色谱法,并测定2型糖尿病人群血浆中盐酸二甲双胍药物浓度。方法:建立反相高效液相色谱法测定盐酸二甲双胍血药浓度,以阿替洛尔为内标,并考察方法学的专属性、线性、定量下限、精密度、准确度、回收率和稳定性。12名2型糖尿病患者连续服药3天以上,利用反相高效液相色谱法测定血浆中盐酸二甲双胍的稳态谷浓度和稳态峰浓度。结果:盐酸二甲双胍在血浆中浓度线性范围是0.2～5μg·mL^(-1),标准方程为Y=1.153 X+0.036 6(R2=0.999 9),定量下限为0.2μg·mL^(-1)。准确度(RE)在±5.58%以内,批内、批间精密度(RSD)≤9.96%,提取回收率在65%～76.9%之间。治疗药物监测显示,12名2型糖尿病患者稳态谷浓度平均值为0.87μg·mL^(-1)(0.4～1.39μg·mL^(-1)),稳态峰浓度平均值为2.47μg·mL^(-1)(0.7～3.41μg·mL^(-1))。结论:该法专属性强,灵敏度高,适用于2型糖尿病人群二甲双胍治疗药物监测。

OBJECTIVE To develop a reversed phase high performance liquid chromatographic method for the determination of the plasma concentration of metformin hydrochloride in type 2 diabetes mellitus patients.METHODS The concentrations of metformin hydrochloride in human plasma were determined by RP-HPLC.Atenolol was used as the internal standard.The methodology was examined for specificity,linear range,lower limit of quantitation,precision,accuracy,recovery and stability.Twelve patients with type 2 diabetes received metformin hydrochloride orally for at least 3 days to achieve the pharmacokinetic steady state.The trough and peak concentrations of metformin hydrochloride in plasma were determined by reversedphase high performance liquid chromatography.RESULTS The linear range of the calibration curve was 0.2-5μg·mL-1,with the equation of Y=1.153 X＋0.036 6（R2=0.999 9）and the lower limit of quantitation of 0.2μg·mL-1.The inter-and intra-day precisions were less than 9.96% and accuracy was within±5.58%.The extraction recoveries were 65%-76.9%.Therapeutic drug monitoring showed that a total of 12 metformin hydrochloride steady-state trough concentrations were available with a mean value of 0.87μg·mL-1（range 0.44μg·mL-1 to 1.39μg·mL-1）,and the steady-state peak concentrations were available with a mean value of 2.47μg·mL-1（range 0.7μg·mL-1 to 3.41μg·mL-1）.CONCLUSION This method has specificity and high sensitivity,is suitable for the monitoring of concentration of metformin hydrochloride in plasma of type 2 diabetic patients.