同位素稀释-超高效液相色谱-串联质谱法测定人血浆中氟康唑的浓度
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摘要
目的:建立测定人血浆中氟康唑浓度的方法。方法:血浆样品经乙腈沉淀蛋白后,以同位素氟康唑-d4为内标,采用超高效液相色谱-串联质谱法测定。色谱柱为ACQUITY UPLC BEH C_(18),流动相为0.1%甲酸溶液-乙腈(梯度洗脱),流速为0.3 mL/min,柱温为40℃,进样量为3μL;采用电喷雾离子源,以多反应监测模式进行正离子扫描,用于定量分析的离子对分别为m/z 307.1→220.0(氟康唑)和m/z 311.1→223.0(内标)。结果:氟康唑血药浓度检测的线性范围为10～5 000 ng/mL(r=0.998 1),定量下限为10 ng/mL;日内、日间RSD均低于8%,准确度为95.8%～106.7%;提取回收率为97.3%～107.3%(RSD<5.0%,n=6),基质效应、稀释效应及残留效应均不影响待测物的定量分析。结论:该方法简便、快速、专属性强、准确度高,可用于氟康唑治疗药物监测及药动学研究。
OBJECTIVE:To establish a method for determination of fluconazole concentration in human plasma. METHODS:UPLC-MS/MS method was adopted to determine plasma after precipitated with acetonitrile. Using isotope fluconazole-d_4 as internal standard,the determination was performed on ACQUITY UPLC BEH C_(18) column with mobile phase consisted of 0.1% formic acid-acetonitrile(gradient elution)at the flow rate of 0.3 mL/min. The column temperature was 40 ℃,and the sample size was 3 μL.ESI was used for positive ion scanning by multiple reaction monitoring mode. The ion pairs for quantitative analysis were m/z307.1→220.0(fluconazole)and m/z 311.1→223.0(internal standard). RESULTS:The linear range of fluconazole was 10-5 000 ng/mL(r=0.998 1). The limits of quantitation was 10 ng/mL. RSDs of intra-day and inter-day were less than 8%;accuracy ranged 95.8%-106.7%. The extraction recovery ranged 97.3%-107.3%(RSD<5.0%,n=6),and matrix effect,dilution effect and residual effect didn't influence quantitative analysis of the substance to be measured. CONCLUSIONS:The method is simple,rapid,specific and accurate,which can be used for therapeutic drug monitoring and pharmacokinetic study of fluconazole.
OBJECTIVE:To establish a method for determination of fluconazole concentration in human plasma. METHODS:UPLC-MS/MS method was adopted to determine plasma after precipitated with acetonitrile. Using isotope fluconazole-d_4 as internal standard,the determination was performed on ACQUITY UPLC BEH C_(18) column with mobile phase consisted of 0.1% formic acid-acetonitrile(gradient elution)at the flow rate of 0.3 mL/min. The column temperature was 40 ℃,and the sample size was 3 μL.ESI was used for positive ion scanning by multiple reaction monitoring mode. The ion pairs for quantitative analysis were m/z307.1→220.0(fluconazole)and m/z 311.1→223.0(internal standard). RESULTS:The linear range of fluconazole was 10-5 000 ng/mL(r=0.998 1). The limits of quantitation was 10 ng/mL. RSDs of intra-day and inter-day were less than 8%;accuracy ranged 95.8%-106.7%. The extraction recovery ranged 97.3%-107.3%(RSD<5.0%,n=6),and matrix effect,dilution effect and residual effect didn't influence quantitative analysis of the substance to be measured. CONCLUSIONS:The method is simple,rapid,specific and accurate,which can be used for therapeutic drug monitoring and pharmacokinetic study of fluconazole.
引文
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[16]王伟,丁仁奎,李清艳,等.UPLC-MS/MS法同时测定人血浆中卡马西平、文拉法辛、罗格列酮和硝苯地平的浓度[J].中国药房,2018,29(2):194-198.
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[19]马欢,葛庆华.特殊基质样品及内源性物质生物分析方法的验证[J].中国医药工业杂志,2017,48(10):1424-1432.
[2]元子青云,陈安九,沈怡雯,等.三唑类抗真菌药物临床应用研究进展[J].药学与临床研究,2018,26(2):125-129.
[3]王剑斐.氟康唑与其他药物的相互作用[J].临床合理用药杂志,2010,3(21):58-59.
[4]COKER RJ,TOMLINSON DR,PARKIN J,et al.Interaction between fluconazole and rifampicin[J].BMJ,1990,301(6755):818.
[5]胡钟,张华,朱文莉,等.氟康唑致不良反应文献分析[J].中国药业,2018,27(7):79-82.
[6]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):1038-1045.
[7]宋青,张华峰,戴博,等.HPLC法测定人血浆中氟康唑的浓度[J].解放军药学学报,2015,31(2):142-144.
[8]盛晓燕,熊歆,杨文领,等.RP-HPLC法测定人血清氟康唑浓度及其临床应用[J].中国新药杂志,2010,19(9):772-775.
[9]杜旭召,赵曦,张彦玲,等.人血浆中氟康唑的LC-MS/MS法测定[J].中国医药工业杂志,2010,41(9):683-685.
[10]任进民,孙倩,张国旗,等.人血浆中氟康唑的LC-MS/MS法测定及药动学[J].中国医药工业杂志,2010,41(12):929-931.
[11]李晓冰,石富国,宋沁馨,等.药物分析研究进展[J].药学进展,2013,37(8):360-367.
[12]周亚飞,王月婷,于嘉屏.HPLC-MS/MS同位素稀释法测定人体血清中类固醇激素的研究[J].检验医学,2015,30(5):427-432.
[13]国家药典委员会.中华人民共和国药典:四部[S].2015年版.北京:中国医药科技出版社,2015:363-368.
[14]时欣欣,徐红蓉,储楠楠,等.LC-MS/MS法测定人血浆中卡马西平的浓度[J].中国临床药学杂志,2015,24(3):158-161.
[15]黄成坷,周伶俐,孙未,等.UPLC-MS/MS联用技术测定人血浆中的利奈唑胺[J].中国药学杂志,2015,50(11):974-977.
[16]王伟,丁仁奎,李清艳,等.UPLC-MS/MS法同时测定人血浆中卡马西平、文拉法辛、罗格列酮和硝苯地平的浓度[J].中国药房,2018,29(2):194-198.
[17]HUGHES NC,BAJAJ N,FAN J,et al.Assessing the matrix effects of hemolyzed samples in bioanalysis[J].Bioanalysis,2009,1(6):1057-1066.
[18]HUANG Y,SHI R,GEE W,et al.Matrix effect and recovery terminology issues in regulated drug bioanalysis[J].Bioanalysis,2012,4(3):271-279.
[19]马欢,葛庆华.特殊基质样品及内源性物质生物分析方法的验证[J].中国医药工业杂志,2017,48(10):1424-1432.