二维液相色谱测定伏立康唑血药浓度的研究

目的：采用全自动二维液相色谱（2D-LC-UV）建立快速测定伏立康唑血药浓度的方法，并应用于临床。方法：样品去除蛋白后直接进样，待测物在一维柱Aston SC2（4.6 mm×25 mm，5μm）上初步分离，通过中间柱Aston SH（3.0 mm×10 mm，5μm）截取保留，转移到二维色谱柱Aston SCB（4.6 mm×100 mm，5μm）上进一步分离。一维流动相为VCV-1D移动相，流速0.7 mL·min^-1；二维流动相为OPI-1有机移动相-BPI-1碱性移动相-MPI-1移动相（30：48：22），流速1.0 mL·min^-1；柱温40℃；紫外检测波长262 nm。结果：伏立康唑与各杂质分离良好，在0.38~12μg·mL^-1范围内呈良好的线性关系（r=0.9999），可在10 min内完整出峰，方法回收率高于95%；日内、日间的精密度RSD均小于5%；稳定性试验RSD小于5%。所建立方法对79名肾移植住院患者进行了伏立康血药浓度检测共280例次，常规剂量未达到治疗浓度的占55%，超过安全浓度的占10.4%，在治疗浓度内的占34.6%。结论：本方法简便快速，结果准确，稳定性良好，自动化程度高，不依赖于专门的技术人员，适用于临床伏立康唑快速检测。  

Objective: To establish a method for rapid determination of voriconazole blood concentration by automated two-dimensional liquid chromatography (2D-LC-UV)and apply it to clinical practice. Methods: The sample was directly injected for determination after being deproteinized. The sample was preliminarily separated on one-dimensional Aston SC2 column, then was trapped on intermediate Aston SH column (3.0 mm×10 mm, 5μm) and separated by two-dimensional Aston SCB column (4.6 mm×100 mm, 5 μm). The mobile phase of the onedimensional and two-dimensional liquid chromatography were VCV-1D (flow rate was 0.7mL·min^-1)and mixed mobile phase[VOPI-1 organic-VBPI-1 basic -VMPI-1 acidic (30:48:22)] (flow rate of 1.0 mL·min^-1), respectively. The column temperature was maintained at 40℃ and the UV adsorption wavelength was set at 262nm. The established method was applied to the patients with kidney transplantation to monitor the voriconazole blood concentration and the preliminary results were evaluated. Results: Under the established chromatographic conditions, voriconazole was separated well from other impurities and linear range was 0.38-12 μg·mL^-1 (r=0.999 9). Voriconazole had a complete chromatographic peak within 10 minutes. The recovery of the method was higher than 95%. Intraday and interday precisions were less than 5% and the RSD of stability test was less than 5%. A tolal of 280 voriconazole blood concentrations were measured in 79 patients addmitted to the kidney transplant department. In the routine dose of voriconazole, 55% of the patients did not reach the therapeutic comcentratin, and 10.4% exceeded the safe concentration, accountcd for 34.6%. Conclusion: The eatablished method is simple, rapid, accurate, stable, highly automated, independent of specialized technicians, and is suitable for rapid detection of voriconazole in hospitals. 

[1] THEURETZBACHER U,IHLE F,DERENDORF H. Pharmacokinetic/pharmacodynamic profile of voriconazole[J]. Clin Pharmacokinet, 2006, 45(7):649
[2] PATTERSON TF, Ⅲ GRT, DENNING DW, et al. Practice guidelines for the diagnosis and management of aspergillosis:2016 update by the infectious diseases society of America[J]. Clin Infect Dis Off Pub Infect Dis Soc Am,2016, 63(4):e1
[3] PURKINS L,WOOD N,GHAHRAMANI P,et al. Pharmacokinetics and safety of voriconazole following intravenous-to oral-dose escalation regimens[J]. Antimicr Agents Chemother,2002,46(8):2546
[4] DOLTON MJ, MCLACHLAN AJ. Clinical importance of the CYP2C19*17 variant allele for voriconazole[J]. Br J Clin Pharmacol,2011, 71(1):137
[5] 王晶晶,王茜,何瑾,等. 伏立康唑血药浓度监测及其影响因素分析[J]. 中国医院药学杂志, 2018, 38(23):2402 WANG JJ, WANG Q, HE J, et al. Retrospectiveof voriconazole therapeutic drug monitoring[J]. Chin J Hosp Pharm, 2018, 38(23):2402
[6] 易文燕,刘滔滔,严汝庆,等. 基于基因多态性的儿童患者伏立康唑药代动力学研究进展[J]. 中南药学,2018, 16(6):799. YI WY, LIU TT,YAN RQ,et al. Advance in pharmacokinetics and pharmacodynamics of voriconazole in child patients based on genetic polymorphisms[J]. Cent South Pharm,2018, 16(6):799
[7] 中华人民共和国药典2015年版. 四部[S]. 2015:363 ChP 2015. Vol IV[S]. 2015:363
[8] 中华医学会器官移植学分会. 中国实体器官移植受者侵袭性真菌病临床诊治指南(2016年版)[J]. 中华器官移植杂志,2016, 37(5):300 The Transplantation Society of Chinese Medical Association, The Transplantation Society of Chinese Medical Doctor Association. The Clinical Guideline of Invasive Fungal Disease in the Solid Organ Transplantation Recipients in China(2016 Edition)[J]. Chin J Organ Transplant,2016, 37(5):300
[9] 中华医学会器官移植学分会. 中国实体器官移植受者侵袭性真菌病临床诊治指南(2016年版)(续)[J]. 中华器官移植杂志, 2016, 37(6):368 The Transplantation Society of Chinese Medical Association, The Transplantation Society of Chinese Medical Doctor Association. The Clinical Guideline of Invasive Fungal Disease in the solid organ Transplantation Recipients in China(2016)[J]. Chin J Organ Transplant,2016, 37(6):368
[10] 张寒娟,陈永妍,王高彪,等. 新型二维液相色谱快速测定丙戊酸血药浓度的研究[J]. 中国医院药学杂志, 2018, 38(13):1394 ZHANG HJ,CHEN YY,WANG GB,et al. Measurement of valproic acid in blood by a novel two-dimensional liquid chromatographic system[J]. Chin J Hosp Pharm,2018, 38(13):1394
[11] 萧伟斌,蹇阳,李桦. 二维液相色谱在药物和毒物分析中的应用进展[J]. 分析化学, 2014, 42(12):1851 XIAO WB, SAI Y, LI H. Application of two-dimensional liquid chromatography analysis of drugs and toxicants[J]. Chin J Anal Chem,2014, 42(12):1851.
[12] 高辉,温学森,马小军,等. 二维液相色谱技术在药物分析中的应用[J]. 药物分析杂志,2007, 27(4):616 GAO H,WEN XS,MA XJ,et al. The technique of two-dimensional liquid chromatography (2D-LC)and its applications in drug analysis[J]. Chin J Pharm Anal,2007, 27(4):616
[13] 丁坤,吴大朋,关亚风. 二维液相色谱接口技术[J]. 色谱,2010, 28(12):1117 DING K, WU DP, GUAN YF. Interface of two dimensional liquid chromatography[J]. Chin J Chromatogr, 2010,28(12):1117
[14] 张素洁,孙贺伟,郭君君,等. 液相色谱-串联质谱法同时测定血清中5个三唑类抗真菌药物浓度及其治疗药物监测应用[J]. 药物分析杂志, 2017, 37(6):1038 ZHANG SJ, SUN HW, GUO JJ, et al. Quantitative determination of five azole antifungals in human serum by LC-MS/MS and its application to therapeutic drug monitoring[J]. Chin J Pharm Anal, 2017, 37(6):1038
[15] 王峰,朱运贵,罗雪梅,等. 新型二维液相色谱测定百草枯尿液浓度的研究[J]. 中国药学杂志,2017, 52(10):871 WANG F, ZHU YG, LUO XM, et al. Measurement of paraquat in urine by a novel two-dimensional liquid chromatographic system[J]. Chin Pharm J,2017, 52(10):871