摘要：
Objective To develop an ion exchange chromatography method for determination of metformin in human plasma,and further apply the method in pharmacokineties study.Methods Human plasma was precipitated with acetonitrile containing internal standard (chlorpheniramine maleate),after mixing and centrifugation,the supernatant requried nitrogen drying at 40 C of water bath,and then the residue was redissolved by mobile phase,after mixing and centrifugation,then injected the supernatant to analyzed on column.Using Capcell PAK SCX UG80 (4.6 mm × 150 mm,5 μm) column.The mobile phase,consisting of acetonitrile and 0.15 mol · L-1 ammonium dihydrogen phosphate (adjusted to pH 3 with 10％ phosphoric acid solution) (52∶ 48,v/v).The samples were eluted at a flow rate of 1 mL · min-1,35 °C of column temperature and assessed by a UV detector set at 233 nm.Urine samples were diluted by ultra pure water,then analyzed by HPLC.Results The calibration curve in plasma and urine samples exhibited a linear dynamic range of 0.05-5 μg · mL-1 (r =0.999 3) and 0.20-200 μg · mL-1 (r =0.999 1) respectively.The limit of quantification were 0.05,0.20μg · mL-1 respectively.The extraction recoveries were 88.65％-95.42％in serum and 91.22％-108.18％ in urine.The inter-day and intra-day RSDs were less than 15％.And the plasma and urine samples were all in good stability.Conclusion This method is widely applicable,strong specificity,good reproducibility and high sensitivity,it is suitable for the pharmacokinetic study of metformin hydrochloride tablets.%目的 建立离子交换色谱法人血浆和尿液中盐酸二甲双胍浓度的测定方法,并应用于人体药代动力学研究.方法 血浆样本采用加入含有内标(马来酸氯苯那敏)的乙腈溶液沉淀蛋白,离心后的上清液置40°C水浴下氮气挥干,残渣用流动相复溶,涡旋混合,高速离心后吸取上清液进样分析.色谱柱:Capcell PAK SCX UG80(4.6 mm×150 mm,5μm);流动相:乙腈-0.15 mol·L-1磷酸二氢铵(用10％磷酸调至:pH 3)=52∶48;流速:l mL·min-1;柱温:35°C;检测波长:233 nm.尿液样本用超纯水稀释后直接进HPLC分析.结果 二甲双胍的血浆和尿液样本的线性范围分别为0.05～5 μg· mL-1(r=0.999 3)和0.20 ～ 200 μg·mL-1(r=0.999 1);最低定量下限分别为0.05,0.20 μg· mL-1;回收率分别为88.65％～95.42％和91.22％ ～ 108.18％;日内和日间RSD均＜15％;血浆和尿液质控样本在稳定性考察范围内均良好.结论 该方法适用性广、准确性好、专属性强、重现性好且快速灵敏,适用于盐酸二甲双胍片的药代动力学研究.
