液相色谱-串联质谱法同时测定人血浆及脑脊液中头孢哌酮与舒巴坦浓度

Simultaneous determination of cefoperazone and sulbactam concentrations in human plasma and cerebrospinal fluid by HPLC-MS/MS

目的建立液相色谱-串联质谱(HPLC-MS/MS)检测法同时测定人血浆及脑脊液样品中头孢哌酮和舒巴坦的浓度。方法用HPLC-MS/MS法,以倒千里光碱、葛根素为内标,色谱柱ACE Excel 3 C18-PFP(150 mm×4. 6 mm,1. 7μm)进行分离,血浆样品流动相A为0. 1%甲酸-水溶液,B为0. 1%甲酸-乙腈溶液,脑脊液样品流动相A为0. 1%甲酸-水溶液,B为0. 1%甲酸-乙腈溶液。用于定量分析的离子对分别为m/z 646. 2→m/z 143. 2(头孢哌酮)和m/z352. 1→m/z 290. 0(倒千里光碱);m/z 232. 2→m/z 140. 1(舒巴坦)和m/z 415. 1→m/z 266. 9(葛根素)。结果脑脊液样品中头孢哌酮、舒巴坦保留时间分别为6. 47,5. 56 min,在0. 4×10-^2~2μg·mL^-1线性关系良好(r=0. 997 8,0. 9992),最低检测浓度为0. 4×10^-2μg·mL^-1;血浆样品中头孢哌酮、舒巴坦保留时间分别为6. 52,5. 56 min,在0. 4×10^-2~2μg·mL^-1线性关系良好(r=0. 9968,0. 998 4),最低检测浓度为0. 4×10^-2μg·mL^-1;脑脊液及血浆中头孢哌酮日内、日间精密度RSD均<12%;方法回收率> 95%,提取回收率> 93%;舒巴坦日内、日间精密度RSD均<10%;方法回收率> 96%,提取回收率> 98%。结论该方法快速、简便、准确、灵敏,适用于人血浆和脑脊液中头孢哌酮和舒巴坦的含量测定。

Objective To establish a LC-MS/MS method for the simultaneously determination of cefoperazone and sulbactam in human plasma and cerebrospinal fluid samples. Methods Taking retrorsine and puerarin as internal standard,separate,the plasma sample mobile phase A:0. 1% formic acid to aqueous solution,B: 0. 1% formic acid to acetonitrile solution and the cerebral fluid sample mobile phase A: 0. 1% formic acid to aqueous solution,B: 0. 1% formic acid to acetonitrile solution was separated with chromatographic column ACE Excel 3C18-PFP( 150 mm × 4. 6 mm,1. 7 μm) by HPLC-MS/MS method. Using the ion pairs of m/z 646. 2→m/z 143. 2( cefoperazone) and m/z 352. 1→m/z 290. 0( retrorsine),m/z 232. 2→m/z 140. 1( sulbactam) and m/z 415. 1→m/z266. 9( puerarin) were taken quantitative analysis respectively. Results The retention time of cefoperazone and sulbactam in cerebrospinal fluid were6. 47 and 5. 56 min,respectively. The range of 0. 4 × 10^-2-2 μg·mL^-1 as the best linear correlation with the lowest limit at 0. 4 × 10^-2μg·mL^-1;the retention time of cefoperazone and sulbactam in plasma were 6. 52 and 5. 56 min,respectively. The range of 0. 4 × 10^-2-2 μg·mL^-1 as the best linear correlation with the lowest limit at 0. 4 × 10^-2μg·mL^-1. Precision( RSD) of inter-day and intra-day of cefoperazone in plasma and cerebrospinal fluid were lower than 12%. Method recovery was higher than 95%,and extraction recovery was higher than 93%. Precision( RSD) of inter-day and intra-day of sulbactam in plasma and cerebrospinal fluid were lower than 10%. Method recovery was higher than 96%,and extraction recovery was higher than 98%. Conclusion This method is rapid,simple,accurate and sensitive. It is suitable for the determination of cefoperazone sulbactam in human plasma and cerebrospinal fluid.