心脏移植患者磺胺甲噁唑血药浓度快速检测分析方法的构建及临床应用

Construction and clinical application of An analytical method for rapidly detecting sulfamethoxazole plasma concentration in heart transplant patients

关键词:  磺胺甲噁唑  二维液相色谱  大体积  血药浓度检测  合理用药

Key words:  sulfamethoxazole  two-dimensional liquid chromatography  large volume  blood concentration monitoring  rational administration of drug

摘要:

目的 探索建立心脏移植患者磺胺甲噁唑血药浓度分析方法,以指导临床合理用药。方法 采用柱切换技术的新型二维液相色谱仪(2D-LC-UV),使用一维色谱柱Aston SC2(3.5 mm×25 mm,5 μm)对血浆中磺胺甲噁唑进行在线固相萃取,然后经中间柱Aston SBR(3.5 mm×10 mm,5 μm)截取保留,随后转移至第二维色谱柱Aston SNX4(4.6 mm×130 mm,5 μm)对目标分析物进行完全分离检测。色谱条件:一维流动相为乙腈-甲醇-水(10:10:70),流速0.8 mL·min^-1;二维流动相比例为BPI-1碱性移动相-API-3酸性移动相-甲醇(20:40:40),流速为1.2 mL·min^-1;紫外检测波长为240 nm。结果 磺胺甲噁唑浓度在9.96~200.04 μg·mL^-1内与峰面积呈良好的线性关系(相关系数R²=0.999 6);低、中、高3个浓度的日内和日间精密度RSD均<15%、相对回收率为85%~115%。对在院56例患者血药浓度进行测定,仅有30例(53.57%)患者血样磺胺甲噁唑浓度为100~150 μg·mL^-1,其余均不满足浓度治疗窗;泌尿系统/血液系统/肝脏等发生III~IV级不良反应为0例。结论 本方法前处理简单,自动化程度高,可大体积进样,准确度、灵敏度高,能够满足临床应用,研究结果可为临床治疗药物监测提供方法学参考。

Abstract:

OBJECTIVE To explore and establish an analysis method for sulfamethoxazole blood concentration in patients with heart transplantation to guide clinical rational drug use. METHODS A new two-dimensional liquid chromatograph (2D-LC-UV) based on column switching technology was used. The on-line solid-phase extraction of sulfamethoxazole in serum was performed using a one-dimensional column Aston SC2(3.5 mm×25 mm, 5 μm), then it was intercepted and retained through the middle column Aston SBR(3.5 mm×10 mm, 5 μm), and the target analytes were completely separated and detected by transfer to the second-dimension column Aston SNX4(4.6 mm×130 mm, 5 μm). The chromatographic conditions were obtained by optimization. The one-dimensional mobile phase was acetonitrile-methanol-H₂O(10:10:70), the flow rate was 0.8 mL·min^-1; the two-dimensional mobile phase ratio was BPI-1 basic mobile phase-API-3 acidic mobile phase-methanol(20:40:40) the flow rate was 1.2 mL·min^-1. The UV detection wavelength was 240 nm. RESULTS The correlation coefficient between the concentration of sulfamethoxazole and peak area was in the range of 9.96‒200.04 μg·mL^-1 with R²=0.999 6, it showed a good linear relationship. The intra-day and inter-day precisions(RSD) at low, medium and high concentrations were <15%, and the relative recoveries were between 85%‒115%. The blood concentration of 56 patients in the hospital was measured, and only 30 patients(53.57%) had the blood sample sulfamethoxazole concentration of 100‒150 μg·mL^-1, and the rest were not comfortable about the concentration treatment window; there were 0 cases of grade III-IV adverse reactions in the urinary system/blood system/liver, etc. CONCLUSION This method has a simple pre-treatment, high automation, can be sampled in large volumes, and has high accuracy and sensitivity. It can meet the requirements of clinical applications. The research results can provide a methodological reference for clinical therapeutic drug monitoring..