高效液相色谱法测定人血液和尿中帕尼培南-倍他米隆的浓度

目的 :建立高效液相色谱法测定人血浆及尿样品中帕尼培南 (panipenem ,PAPM)、倍他米隆(betamipron,BP)浓度。方法 :以 1mol·L-1MOPS(pH 7.0 )为稳定剂 ,PAPM浓度测定采用外标法 ,BP则采用内标法 ,内标物为N 苯甲酰 DL 丙氨酸。PAPM测定的色谱条件 :色谱柱为C18Diamosil(TM ) (2 5 0× 4.6mm ,5 μm) ;血浆中流动相为CH3 OH∶0 .0 4mol·L-1CH3 COONH4(pH 4.0 ) / 5∶95 ,尿中流动相为CH3 OH∶ 0 .0 4mol·L-1CH3 COONH4(pH 4.0 ) / 2∶98;检测波长为 2 99.3nm ;流速为 1 .6mL·min-1。BP测定色谱条件 :色谱柱同PAPM ;血浆中流动相为CH3 CN∶CH3 COOH∶H2 O/ 2 0∶1∶79,尿中流动相为CH3 CN∶CH3 COOH∶H2 O/ 1 2∶1∶87;检测波长为 2 40nm ;血浆中流速为1 .4mL·min-1,尿中流速为 1 .6mL·min-1。结果 :血浆中PAPM在 0 .5～ 5 0 .0mg·L-1范围、BP在0 .2～ 5 0mg·L-1范围内具良好线性关系 (r =0 .9999) ;尿中PAPM和BP在 2～ 2 0 0mg·L-1浓度范围内具良好线性关系 (r =0 .9999)。PAPM和BP平均方法回收率为 1 0 1 .69% ;日内、日间精密度RSD均 <5 %。结论 :本方法简便、快捷、灵敏、准确 ,适用于临床药动学及药效学的研究

Determination of panipenem/betamip-ron concentration in blood and urine by high performance liquid chromatography

AIM: To use HPLC to determine panipenem(PAPM) and betamipron(BP) concentration in human plasma and urine. METHODS: The stabilizer of plasma and urine was a mixture of 1 mol·L -1 3 (N mopholine) propanesulfonic acid (pH 7.0). BP was examined by HPLC with internal standard of N benzoyl DL alanine, while PAPM was examined with external standard. HPLC conditions for PAPM were as follows: a chromatogram column of C 18 Diamosil (TM) was 250×4.6 mm, 5 μm. Mobile phases were CH 3OH∶CH 3COONH 4 (0.04 mol·L -1 , pH 4.0)/5∶95 in plasma and CH 3OH∶CH 3COONH 4 (0.04 mol·L -1 , pH 4.0)/2∶98 in urine. And the flow rate was 1.6 mL·min -1 . They were determined at UV wavelength of 299.3 nm. The conditions for BP: the column was the same as described above. Mobile phases were CH 3CN∶CH 3COOH∶H 2O/20∶1∶79 and CH 3CN∶CH 3COOH∶H 2O/ 12∶1∶87, the determination wavelength was 240 nm and the flow rates were 1.4 mL ·min -1 and 1.6 mL·min -1 in plasma and urine, respectively. RESULTS: The linear range was 0.5 50.0 mg·L -1 of plasma PAPM concentration( r = 0.9999) , while 0.2～50.0 mg·L -1 for BP; The linear range was 2～200 mg·L -1 of urine PAPM and BP concentrations( r = 0.9999 ). The average recovery of PAPM and BP from plasma and urine was 101.69%, The intra day( n =6) and inter day( n =3) precision rates were less than 5 %. CONCLUSION:This method is simple, rapid, accurate, and suitable for study of pharmacokinetics and pharmacodynamics of panipenem/betamipron in clinic.