HPLC法测定人血浆中伏立康唑及其代谢物的浓度
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摘要
目的建立一种快速、灵敏、准确、稳定的测定人血浆中伏立康唑及其代谢物浓度的方法,用于伏立康唑临床治疗药物监测。方法采用高效液相色谱(HPLC)-紫外(UV)检测法,以罂粟碱为内标,乙腈蛋白沉淀法处理血浆样品。色谱柱为ACE5C_(18)-AR (150 mm×4.6 mm),流动相为0.025 mol/L磷酸二氢钠(含三乙胺400μl/L,用0.25 mol/L的氢氧化钠调至pH=7.0)-乙腈(67∶33),流速为l ml/min,柱温为40℃,检测波长为255、276 nm(双波长检测),进样量为20μl。结果伏立康唑氮氧化物、伏立康唑和罂粟碱的保留时间分别为4.5、11.3、13.7 min;血浆中伏立康唑、伏立康唑氮氧化物线性范围均为0.5～20.0μg/ml(r=0.999 5),定量下限均为0.5μg/ml,批内、批间RSD均<11%,定量下限、低、中、高梯度浓度提取回收率在90.3%～109.9%之间。结论该方法操作简便,结果准确,适用于伏立康唑的临床治疗药物监测和对其主要代谢物的测定。
Objective To establish a HPLC method for the determination of the voriconazole concentration in human plasma,which can be used for voriconazole monitoring clinically.Methods High performance liquid chromatography(HPLC-UV) detection method was used with papaverine as internal standard.Acetonitrile protein precipitation method was employed to treat plasma samples.The chromatographic column was ACE5 C_(18)-AR150 mm×4.6 mm with the mobile phase 0.025 mol/L sodium dihydrogen phosphate(containing triethylamine 400 μl/L,pH=7.0 adjusted by 0.25 mol/L sodium hydroxide)-acetonitrile(67∶33).The flow rate was l ml/min,the column temperature at 40 ℃,the detection wavelength at 255,276 nm(double wavelength detection) and sampling amount 20 μl.Results The retention time of voriconazole nitrogen oxide,voriconazole and papaverine were 4.5,11.3 and 13.7 min respectively.The linear ranges of voriconazole and voriconazole nitrogen oxides in plasma were 0.5-20.0 μg/ml(r=0.999 5).The quantitative lower limits were 0.5 μg/ml with RSD< 11%.The recovery rates of extraction were between 90.3% and 109.9%.Conclusion This simple and accurate method is suitable for clinical monitoring of voriconazole.
Objective To establish a HPLC method for the determination of the voriconazole concentration in human plasma,which can be used for voriconazole monitoring clinically.Methods High performance liquid chromatography(HPLC-UV) detection method was used with papaverine as internal standard.Acetonitrile protein precipitation method was employed to treat plasma samples.The chromatographic column was ACE5 C_(18)-AR150 mm×4.6 mm with the mobile phase 0.025 mol/L sodium dihydrogen phosphate(containing triethylamine 400 μl/L,pH=7.0 adjusted by 0.25 mol/L sodium hydroxide)-acetonitrile(67∶33).The flow rate was l ml/min,the column temperature at 40 ℃,the detection wavelength at 255,276 nm(double wavelength detection) and sampling amount 20 μl.Results The retention time of voriconazole nitrogen oxide,voriconazole and papaverine were 4.5,11.3 and 13.7 min respectively.The linear ranges of voriconazole and voriconazole nitrogen oxides in plasma were 0.5-20.0 μg/ml(r=0.999 5).The quantitative lower limits were 0.5 μg/ml with RSD< 11%.The recovery rates of extraction were between 90.3% and 109.9%.Conclusion This simple and accurate method is suitable for clinical monitoring of voriconazole.
引文
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[6] QI F,ZHU L,LI N,et al.Influence of different proton pump inhibitors on the pharmacokinetics of voriconazole[J].Int J Antimicrob Agents,2017,49(4):403-409.
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[8] BRESSáN I G,MENDEZ M L,GIMENEZ M I.Validation of a reversed-phase ultra-high-performance liquid chromatographic method with photodiode array detection for the determination of voriconazole in human serum and its application to therapeutic drug monitoring[J].Ther Drug Monit,2018,40(2):276-283.
[9] LIN S C,LIN S W ,CHEN J M,et al.Using sweeping-micellar electrokinetic chromatography to sweeping-micellar electrokinetic[J].Talanta,2010,82(2):653-659.
[10] PEREA S,PENNICK G J, MODAK A,et al.Comparison of high-performance liquid chromatographic and microbiological methods for determination of voriconazole levels in plasma[J].Antimicrob Agents Chemother,2000,44(5):1209-1213.
[11] 刘学松,何为群,陈文英,等.肺部真菌感染患者伏立康唑血药浓度监测的临床应用[J].国际呼吸杂志,2016,36(20):1521-1526.
[12] GROLL A H,TOWNSEND R,DESAI A,et al.Drug- drug interactions between triazole antifungal agents used to treat invasive aspergillosis and immunosuppressants metabolized by cytochrome P450 3A4[J].Transpl Infect Dis,2017,19(5):e12751.
[2] CHUWONGWATTANA S,JANTARAROUNGTONG T,CHITASOMBAT M N,et al.A prospective observational study of CYP2C19 polymorphisms and voriconazole plasma level in adult Thai patients with invasive aspergillosis[J].Drug Metab Pharmacokinet,2016,31(2):117-122.
[3] XU G Q,ZHU L Q,GE T Y,et al.Pharmacokinetic/pharmacodynamic analysis of voriconazole against Candida spp.and Aspergillus spp.in children,adolescents and adults by Monte Carlo simulation[J].Int J Antimicrob Agents,2016,47(6):439-445.
[4] CABRAL-GALEANO E,RUIZ-CAMPS I,LEN-ABAD O,et al.Clinical usefulness of therapeutic drug monitoring of voriconazole in a university hospital[J].Enferm Infecc Microbiol Clin,2015,33(5):298-302.
[5] LI Z W,PENG F H, YAN M,et al.Impact of CYP2C19 genotype and liver function on voriconazole pharmacokinetics in renal transplant recipients[J].Ther Drug Monit,2017,39(4):422-428.
[6] QI F,ZHU L,LI N,et al.Influence of different proton pump inhibitors on the pharmacokinetics of voriconazole[J].Int J Antimicrob Agents,2017,49(4):403-409.
[7] WANG Y,WANG T,XIE J,et al. Risk factors for voriconazole-associated hepatotoxicity in patients in the intensive care unit[J].Pharmacotherapy,2016,36(7):757-765.
[8] BRESSáN I G,MENDEZ M L,GIMENEZ M I.Validation of a reversed-phase ultra-high-performance liquid chromatographic method with photodiode array detection for the determination of voriconazole in human serum and its application to therapeutic drug monitoring[J].Ther Drug Monit,2018,40(2):276-283.
[9] LIN S C,LIN S W ,CHEN J M,et al.Using sweeping-micellar electrokinetic chromatography to sweeping-micellar electrokinetic[J].Talanta,2010,82(2):653-659.
[10] PEREA S,PENNICK G J, MODAK A,et al.Comparison of high-performance liquid chromatographic and microbiological methods for determination of voriconazole levels in plasma[J].Antimicrob Agents Chemother,2000,44(5):1209-1213.
[11] 刘学松,何为群,陈文英,等.肺部真菌感染患者伏立康唑血药浓度监测的临床应用[J].国际呼吸杂志,2016,36(20):1521-1526.
[12] GROLL A H,TOWNSEND R,DESAI A,et al.Drug- drug interactions between triazole antifungal agents used to treat invasive aspergillosis and immunosuppressants metabolized by cytochrome P450 3A4[J].Transpl Infect Dis,2017,19(5):e12751.