固相萃取HPLC检测生物样品中甲苯磺丁脲和代谢产物及其人体药代动力学研究

目的:建立固相萃取高效液相色谱测定人血浆和尿液中的甲苯磺丁脲和代谢产物浓度的方法,并用于研究甲苯磺丁脲人体代谢过程。方法:固相萃取净化和富集样品,建立高效液相色谱法测定人血清和尿中甲苯磺丁脲和代谢产物的浓度。色谱条件:色谱柱为Waters Spherisorb 5 μm Phenyl色谱柱(4.6 mm×250 mm),流动相为甲醇-0.02 mmol·L-1pH 3.3乙酸钠缓冲液(28∶72),流速1 ml·min-1,检测波长:230 nm。用该方法测定10名健康志愿者单次口服500 mg甲苯磺丁脲后血清和尿液中药物及其代谢产物浓度,应用3p97计算其的药动学参数。结果:血浆甲苯磺丁脲线性范围为2~100μmol·L-1(r=0.999),回收率为105.1％~103.9％。尿液羧基甲苯磺丁脲、4-羟基甲苯磺丁脲和甲苯磺丁脲的线性范围为2~50μmol·L-1(r=0.999)、1~50μmol·L-1(r=0.999)和1~50μmol·L-1(r=0.999),回收率为98.8％~100.1％、95.4％~103.5％和97.7％~106.6％。各样品的日内、日间精密度均≤15％。健康志愿者单次口服500 mg甲苯磺丁脲的AUC0-∞为 2644.6 ±472.8 μmol·h-1·L-1,Tmax是1.4±0.6 h,Cmax是235.8±47.3 μmol·L-1,T1/2为6.9±2.1 h,MR0-24=277.5±125.6。结论:甲苯磺丁脲及其代谢产物的固相萃取高效液相色谱法灵敏、准确,重复性好,可用于甲苯磺丁

HPLC assay for tolbutamide and its metabolites in biological sample by solid phase extraction and study of its pharmacokinetics in human

AIM: To establish a HPLC method by solid-phase extraction which was used for determining tolbutamide and its metabolites in human plasma and urine,and to study its pharmacokinetics. METHODS: The concentrations of tolbutamide and its metabolites in human plasma and urine by solid-phase extraction were determined by HPLC. Waters Spherisorb 5 μm Phenyl column (4.6 mm × 250 mm) was used as the analytical column and the mobile phase was consisted by methanol - 0.02 nm. The concentrations of tolbutamide and its metabolites in plasma and urine in healthy volunteers were determined by HPLC and the pharmacokinetic parameters were analyzed by 3p97. RESULTS: The linear range was 2 - 100 by solid phase extraction. The recovery was 105.1% -0.999) for carboxytolbutamide in human urine. The recovery was 98.8% - 100.1%. The linear range was 1 human urine. The recovery was 95.4% - 103.5%. The tamide in human urine. The recovery was 97.7% -106.6%. The within-day and between-day derivations of all samples were less than 15%. A single oral dose of 500 mg tolbutamide was administrated to the volunteers,the main pharmacokinetic parameters AUC0-∞, T1/2,Tmax, Cmax and MR0-24 were 2644.6 ± 472.8 CONCLUSION: The method is simple and accurate,and it can be used determining tolbutamide and its metabolites in biological samples. Tolbutamide is mainly excreted through urine in form of carboxytolbutamide and hydroxytolbutamide.