HPLC法测定自发性腹膜炎患者腹腔积液中安妥沙星的方法学及其药动学研究  

Methodological and pharmacokinetic study on the determination of antofloxacin in peritoneal effusion of patients with spontaneous peritonitis by HPLC

作　　者：胡浩然 孙学强[3] 邢亚群[1,2] 韩卓越 林翔 HU Haoran;SUN Xueqiang;XING Yaqun;HAN Zhuoyue;LIN Xiang(Department of Pharmacy,Second Affiliated Hospital of Bengbu Medical College,Bengbu 233040,China;School of Pharmacy,Bengbu Medical College;Department of Infection Division,Second Affiliated Hospital of Bengbu Medical College)

机构地区：[1]蚌埠医学院第二附属医院药剂科,蚌埠233040 [2]蚌埠医学院药学院 [3]蚌埠医学院第二附属医院感染科

出　　处：《山西医科大学学报》2023年第2期249-254,共6页Journal of Shanxi Medical University

基　　金：蚌埠医学院自然科学重点项目(2021byzd188)。

摘　　要：目的建立一种高效液相色谱法检测肝硬化自发性腹膜炎患者腹腔积液安妥沙星的浓度,同时研究其药动学特征。方法收集6名肝硬化自发性腹膜炎患者服用盐酸安妥沙星片400 mg后0,0.5,1,1.5,2,2.5,3,4,6,8,12,24,36,48,72 h的腹腔积液,加入内标氧氟沙星,甲醇沉淀蛋白,采用内标法测定安妥沙星含量。色谱柱条件:Phenomenex C18柱(150 mm×4.6 mm,5μm);流动相为0.05 mol/L KH 2PO 4缓冲液-乙腈85∶15(0.05 mol/L KH 2PO 4缓冲液含0.005 mol/L乙烷磺酸钠,0.007%乙二胺四乙酸二钠,磷酸调pH=2.4);柱温30℃,检测波长297 nm,流速1.0 ml/min,进样量20μl。结果腹腔积液中安妥沙星在0.125~20μg/ml范围内与安妥沙星和内标物的峰面积之比呈良好的线性关系(R 2=0.9996),定量下限0.125μg/ml,低、中、高浓度质控样品的日内精密度RSD 0.96%~2.09%(n=6)、日间精密度RSD 2.52%~3.54%(n=6),准确度97.68%~101.60%(n=6),加样回收率97.76%~106.22%(n=6),稳定性试验RSD 0.93%~7.78%(n=6)。实验条件下自发性腹膜炎患者腹腔积液中安妥沙星的主要药动学参数C max,T max,AUC 0-72,AUC 0-∞,T 1/2分别为(1.53±0.17)μg/ml,(7.67±0.82)h,(49.35±3.01)μg·h/ml,(57.16±3.88)μg·h/ml,(23.43±1.08)h。结论本实验中建立的方法灵敏、可靠、准确度高、重现性好,样品处理简单方便,出峰速度快,分析成本较低,适用于肝硬化自发性腹膜炎患者腹腔积液安妥沙星浓度测定及相关药动学研究。Objective To establish a high performance liquid chromatography(HPLC)method for determining antofloxacin in the peritoneal effusion of cirrhotic patients with spontaneous peritonitis and investigate its pharmacokinetic characteristics.Methods The peritoneal effusion of 6 patients with spontaneous peritonitis was respectively collected at 0,0.5,1,1.5,2,2.5,3,4,6,8,12,24,36,48,72 h after taking antofloxacin hydrochloride tablets(400 mg).Antoloxacin content was determined by internal standard method with ofloxacin as internal standard and methanol for protein precipitation.The study was performed on Phenomenex C18 column(150 mm×4.6 mm,5μm),with 0.05 mol/L KH 2PO 4 buffer-acetonitrile 85∶15 as mobile phase.The 0.05 mol/L KH 2PO 4 buffer contained 0.005 mol/L sodium ethane sulfonate and 0.007%ethylenediamine tetraacetic acid disodium,and adjusted to pH=2.4 with phosphoric acid.The column temperature was 30℃,the detection wavelength was 297 nm,the flow rate was 1.0 ml/min,and the injection volume was 20μl.Results There was a good linear relationship between antofloxacin in abdominal fluid and the peak area ratio of antloxacin and internal standard in the range of 0.125-20μg/ml(R 2=0.9996).The lower limit of quantification was 0.125μg/ml.The intra-day precision RSD of low,medium and high concentration quality control samples,the inter-day precision RSD,the RSDs for the accuracy,the recovery and the stability tests were 0.96%-2.09%,2.52%-3.54%,97.68%-101.60%,97.76%-106.22%,and 0.93%-7.78%(n=6),respectively.The main pharmacokinetic parameters C max,T max,AUC 0-72,AUC 0-∞and T 1/2 of antofloxacin in peritoneal effusion of patients with spontaneous peritonitis were(1.53±0.17)μg/ml,(7.67±0.82)h,(49.35±3.01)μg·h/ml,(57.16±3.88)μg·h/ml,and(23.43±1.08)h,respectively.Conclusion The method established in this expe-riment was of high sensitivity,high accuracy and good reproducibility,and has the advantages of simple sample processing,fast peak output speed,and low cost.It is suitable for the concentration dete

关 键 词：高效液相色谱法 安妥沙星 自发性腹膜炎 腹腔积液 药代动力学 

分 类 号：R969[医药卫生—药理学]