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医药导报  2019, Vol. 38 Issue (6): 786-791    DOI: 10.3870/j.issn.1004-0781.2019.06.023
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液相色谱-串联质谱法快速同时测定人血浆中卡培他滨及其活性代谢物
熊璐琪1(),赵博欣2()
1.广东省惠州市中心人民医院药学部,惠州 516001
2.南方医科大学南方医院药学部,广州 510515
A Rapid and Simultaneous Determination of Capecitabine and Its Activated Metabolites in Human Plasma by LC-MS/MS
Luqi XIONG1(),Boxin ZHAO2()
1.Department of Pharmacy, the Central Hospital of Huizhou City, Guangdong Province, Huizhou 516001,China
2.Department of Pharmacy, Southern Hospital of Southern Medical University, Guangzhou 510515,China
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摘要 

目的 建立一种简单、快速、同时检测人血浆中卡培他滨及其活性代谢产物氟尿嘧啶(5-FU)浓度的液相色谱-串联质谱(LC-MS/MS)的方法,并用于临床实践。方法 取血浆样本 100 μL,以乙腈沉淀蛋白后,高速离心后取上清液直接进样分析,以乙腈-25 mmol·L-1乙酸铵溶液(含0.1%甲酸)(95:5)为流动相,通过Hilic硅胶柱进行液相分离,采用 ESI 负离子源以及多离子反应监测(MRM)对血浆中卡培他滨及5-FU进行定量分析,卡培他滨及5-FU 离子对分别为m/z 358.1→154.1,m/z 129→42.1。接受卡培他滨化疗患者20例,监测血浆中卡培他滨及5-FU稳态谷浓度,并结合卡培他滨及5-FU单点浓度的相关性评价该检测方法的临床应用性。结果 卡培他滨及5-FU的线性范围均为 39.062 5~10 000 ng·mL-1(R2=0.999),最低定量限为 39.062 5 ng·mL-1。卡培他滨在低(78.125 ng·mL-1)、中(625 ng·mL-1)、 高(5000 ng·mL-1)3种质控浓度的提取回收率分别为(78.22±3.15)%,(75.37±2.99)%及(81.05±2.78)%,5-FU低、中、高3种浓度的提取回收率分别为(70.45±2.19)%,(72.56±1.68)%及(72.02±1.72)%。 日间和日内精密度差异均小于 5%,各浓度间基质效应无显著差异。20例临床患者标本检测结果表明卡培他滨及其活性产物5-FU的血药浓度存在个体差异,且卡培他滨与其对应活性产物血药浓度间相关性较差。结论 该实验建立测定卡培他滨及5-FU的方法灵敏度高,专属性强,样品处理过程简单快捷,适用于同时检测血浆中卡培他滨及5-FU的浓度,应用于临床以提高卡培他滨相关化疗方案的有效性和安全性。

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关键词 卡培他滨氟尿嘧啶液相色谱-串联质谱法    
Abstract

Objective To develop a simple and rapid method for simultaneous quantification of capecitabine and its active metabolite(5-fluorouracil )in human plasma by liquid chromatography—tandem mass spectrometry (LC-MS/MS) and apply it to clinical practice. Methods After a protein precipitation using acetonitrile, the supernatant taken from the centrifugal plasma samples was analyzed directly. With a mobile phase comprising acetonitrile/0.1% aqueous formicacid and 25 mmol·L-1ammonium acetate (95:5) and an electrospray ion (ESI) source, samples contained capecitabine and 5-fluorouracil were quantitative analyzed. The MS/MS detection was performed using multiple reaction monitoring (MRM), in positive mode. The mass transitions of the protonated precursor/product ion pairs were used to record the selected ion mass chromatograms of capecitabine and 5-fluorouracil were m/z 358.1→154.1 and m/z 129→42.1, respectively. 20 patients were receiving chemotherapy with capecitabine were then the steady state trough concentration of capecitabine and 5-fluorouracil in the patients’ plasma were monitored. The clinical application of the detection method was evaluated through the correlation of single point concentration of capecitabine and 5-fluorouracil. Results The linear calibration curve was obtained in the concentration range from 39.062 5-10 000 ng·mL-1(R2=0.999)for both capecitabine and 5-fluorouracil with the limit of quantification was 39.062 5 ng·mL-1. The extraction recoveries of capecitabine at three concentration levels(78.125,625 and 5000 ng·mL-1) and that of the 5-fluorouracil were (78.22±3.15)%,(75.37±2.99)%,(81.05±2.78)% and (70.45±2.19)%,(72.56±1.68)%,(72.02±1.72)%,respectively. The intra-and inter-run precisions,expressed as the relative standard deviation(RSD)were less than 5% for both of capecitabine and 5-fluorouracil. The detection result acquired from the plasma samples indicated that there was differences for the plasma concentration of capecitabine and 5-fluorouracil between individuals and that there was a poor correlation of plasma concentration between the capecitabine and its active metabolite. Conclusion The method is sensitive,specific and validate rapid for simultaneous quantification of capecitabine and 5-fluorouracil in human plasma by liquid chromatography—tandem mass spectrometry and will be successfully applied for clinical practice to improve the effectiveness and security of chemotherapy regimens with capecitabine.

Key wordsCapecitabine    5-Fluorouraci    Liquid chromatography-tandem mass spectrometry
收稿日期: 2017-10-30      出版日期: 2019-06-11
引用本文:   
熊璐琪,赵博欣. 液相色谱-串联质谱法快速同时测定人血浆中卡培他滨及其活性代谢物[J]. 医药导报, 2019, 38(6): 786-791.
Luqi XIONG,Boxin ZHAO. A Rapid and Simultaneous Determination of Capecitabine and Its Activated Metabolites in Human Plasma by LC-MS/MS. Herald of Medicine, 2019, 38(6): 786-791.
链接本文:  
http://www.yydbzz.com/CN/10.3870/j.issn.1004-0781.2019.06.023      或      http://www.yydbzz.com/CN/Y2019/V38/I6/786
图1  卡培他滨及5-FU离子扫描质谱图
A.卡培他滨;B.5-FU
图2  卡培他滨和5-FU多离子监测图
A.空白样品;B.卡培他滨;C.5-FU
图3  空白血浆样品多离子监测图
A.TIC;B.卡培他滨;C.5-FU
浓度/
(ng·mL-1)
卡培他滨 5-FU
日间 日内 日间 日内
第1天 第2天 第3天 第1天 第2天 第3天
78.125 74.378±2.230 74.013±2.370 76.156±3.240 74.849±1.147 75.374±1.960 78.573±2.310 79.354±3.030 77.767±2.109
625 617.394±11.480 624.782±9.620 627.573±9.670 623.250±5.260 624.421±13.240 621.853±12.740 628.844±13.970 625.039±3.536
5000 4936.234±45.230 4899.118±51.920 4968.352±49.520 4934.568±34.647 4897.823±39.750 5012.834±45.820 4978.752±42.940 4963.136±59.074
表1  3种质量控制浓度的日内及日间精密度
药品与浓度 新鲜配制 室温放置1 h 室温放置24 h 反复冻融1 h 反复冻融24 h
测定 RE/% 测定 RE/% 测定 RE/% 测定 RE/%
卡培他滨
低浓度 74.842±1.974 76.183±2.045 1.792 75.748±2.874 1.211 74.893±1.939 0.068 74.084±2.62 -1.015
中浓度 618.428±11.736 620.312±14.159 0.305 618.074±11.620 -0.057 621.858±12.518 0.555 623.717±13.290 0.855
高浓度 5001.980±58.210 4987.295±46.749 -0.294 4975.390±46.765 -0.532 5005.237±45.374 0.065 5041.741±52.566 0.795
5-FU
低浓度 77.916±1.841 76.018±1.991 -2.44 76.386±1.746 -1.964 74.532±2.369 -4.343 77.090±2.062 -1.060
中浓度 625.931±12.375 623.641±14.159 -0.366 624.856±11.6e0 -0.172 618.243±11.354 -1.228 617.322±12.929 -1.375
高浓度 5012.364±46.253 4997.452±52.443 -0.298 4985.563±46.737 -0.535 5011.756±47.560 -0.012 4997.432±52.467 -0.298
表2  卡培他滨及5-FU在人血浆中的稳定性
图4  临床患者卡培他滨与其活性代谢产物5-FU血药浓度相关性
[1] TOWNSEND D M,TEW K D.The role of glutathione-S-transferase in antrcancer® drug resistance[J].Oncogene,2003,22:7369-7375.
[2] DAHL O,FLUGE O,CARLSEN E,et al.Final results of a randomized phase Ⅲ study on adjuvant chemotherapy with 5-FU and levamisol in colon and rectum cancer stage Ⅱand Ⅲ by the Norwegian Gastrointestinal Cancer Group[J].Acta Oncol,2009,48(3):368-376.
doi: 10.1080/02841860902755244
[3] ZACHARIAE R,PAULSEN K,MEHLSEN M,et al.Chemot herapy-in-duced nausea,vomiting,and fatigue-the role of individual differences related to sensory perception and autonomic reactivity[J].Psychother Psychosom,2007,76(6):376-384.
doi: 10.1159/000107566
[4] GAMELIN E,DELVA R,JACOB J,et al.Individual fluorou-racil dose adjustment based on pharmacokinetic follow-up compared with conventional dosage:results of a multicenter randomized trial of patients with metastatic colorectal cancer[J].J Clin Oncol,2008,26(13):2099-2105.
doi: 10.1200/JCO.2007.13.3934
[5] DHANANJEYAN M R,LIU J,BYKOWSKI C,et al.Rapid and simultaneous determination of capecitabine and its metabolites in mouse plasma,mouse serum,and in rabbit bile by high-performance liquid chromatography[J].J Chromatography A,2007,1138(1/2):101-108.
doi: 10.1016/j.chroma.2006.10.038
[6] YUKO T,YUKIO K,MASAKO U,et al.Pharmacokinetic analysis of capecitabine,a triple prodrug of 5-FU in humans:the mechanism for tumor selective accumulation of 5-FU[J].Pharm Res,2001,18(8):1190-1201.
doi: 10.1023/A:1010939329562
[7] 叶敏,付强,朱珠.高效液相色谱法测定人血浆中卡培他滨浓度[J].中国医院药学杂志,2004,24(11):678-680.
[8] 左家信,廖声华,严拯宇.HPLC法测定卡培他滨的含量及有关物质[J].海峡药学,2013,25(1):48-51.
[9] SALVADOR A,MILLERIOUX L,RENOU A.Simultaneous LC-MS-MS analysis of capecitabine and its metabolites after off-line SPE from human plasma[J].Chromatographia,2006,63:609-615.
doi: 10.1365/s10337-006-0799-5
[10] HERMES L P,SHERRY W,CHEESTER B.Development of a sensitive and selective LC-MS/MS method for the determination of α-fluoro-β-alanine,5-fluorouracil and capecitabine in human plasma[J].J Chromatogr B,2009,877:1040-1046.
doi: 10.1016/j.jchromb.2009.02.025
[11] MONTANGE D,BDRARD M,DEMARCHI M,et al.An APCI LC-MS/MS method for routine determination of capecitabine and its metabolites in human plasma[J].J Mass Spectrom,2010,45(6):670-677.
[12] TSUME Y,PROVODA C J,AMIDON G L.The achievement of mass balance by simultaneous quantification of floxuridine prodrug,floxuridine,5-fluorouracil,5-dihydrouracil,α-fluoro-β-ureidopropionate,α-fluoro-β-alanine using LC-MS[J].J Chromatogr B,2011,879(13/14):915-920.
doi: 10.1016/j.jchromb.2011.02.045
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