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
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摘要
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 C_(18)-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~(-2)μg·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~(-2)μg·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 3C_(18)-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~(-2)μg·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~(-2)μg·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.
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 3C_(18)-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~(-2)μg·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~(-2)μg·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.
引文
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[9]梁键谋,王知坚,陈贵斌,等.液相色谱-质谱测定注射用头孢哌酮舒巴坦钠有关物质[J].医药导报,2013,32(3):378-380.
[10]叶显撑,范国荣,崔晓如,等.液相色谱-串联质谱法快速测定人血浆中头孢哌酮浓度及药动学[J].中国新药与临床杂志,2010,29(5):345-349.
[2]中华医学会神经外科学分会中国神经外科重症管理协作组.中国神经外科重症患者感染诊治专家共识(2017)[J].中华医学杂志,2017,97(21):1607-1614.
[3]胡付品,郭燕,朱德妹,等. 2017年CHINET中国细菌耐药监测[J].中国感染与化疗杂志,2018,18(3):241-251.
[4]卫生部合理用药专家委员会.中国医师药师临床用药指南[M].重庆:重庆出版社,2009:103-104.
[5] ZHOU Y,ZHANG J,GUO B,et al. Liquid chromatography/tandem mass spectrometry assay for the simultaneous determination of cefoperazone and sulbactam in plasma and its application to a pharmacokinetic study[J]. J Chromatogr B Analyt Technol Biomed Life Sci,2010,878(30):3119-3124.
[6]张宇一,郁继诚,林东昉,等.人血浆及脑脊液中美罗培南浓度的HPLC测定方法研究[C].中华医学会第一届感染与抗微生物治疗论坛,第八届全国感染性疾病及抗微生物化疗学术会议论文集.复旦大学附属华山医院,2009:203-206.
[7] Ch. P(2015)VolⅡ,中国药典2015版.二部[S]. 2015:1425-1426.
[8]黄琦,季晓娟. HPLC法测定头孢哌酮舒巴坦钠(2∶1)的含量[J].浙江科技学院学报,2012,24(1):44-47.
[9]梁键谋,王知坚,陈贵斌,等.液相色谱-质谱测定注射用头孢哌酮舒巴坦钠有关物质[J].医药导报,2013,32(3):378-380.
[10]叶显撑,范国荣,崔晓如,等.液相色谱-串联质谱法快速测定人血浆中头孢哌酮浓度及药动学[J].中国新药与临床杂志,2010,29(5):345-349.