人血浆中甲氨蝶呤的高效液相色谱法测定及其代谢物的质谱定性分析

王漪璇; 毋丹; 曹军宁; 钱隽

doi:10.3969/j.issn.1006-0111.2015.02.013

留言板

尊敬的读者、作者、审稿人, 关于本刊的投稿、审稿、编辑和出版的任何问题, 您可以本页添加留言。我们将尽快给您答复。谢谢您的支持!

姓名
邮箱
手机号码
标题
留言内容
验证码

x 关闭永久关闭

应中央军委要求，2022年9月起，《药学实践杂志》将更名为《药学实践与服务》，双月刊，正文96页；2023年1月起，拟出版月刊，正文64页，数据库收录情况与原《药学实践杂志》相同。欢迎作者踊跃投稿！

人血浆中甲氨蝶呤的高效液相色谱法测定及其代谢物的质谱定性分析

王漪璇, 毋丹, 曹军宁, 钱隽

文章导航 > 药学实践与服务 > 2015 > 33(2): 143-146,158

王漪璇, 毋丹, 曹军宁, 钱隽. 人血浆中甲氨蝶呤的高效液相色谱法测定及其代谢物的质谱定性分析[J]. 药学实践与服务, 2015, 33(2): 143-146,158. doi: 10.3969/j.issn.1006-0111.2015.02.013

引用本文:

WANG Yixuan, WU Dan, CAO Junning, QIAN Jun. Determination of methotrexate in human plasma by HPLC and qualitative analysis of its metabolite by mass spectrometry[J]. Journal of Pharmaceutical Practice and Service, 2015, 33(2): 143-146,158. doi: 10.3969/j.issn.1006-0111.2015.02.013

Citation:

WANG Yixuan, WU Dan, CAO Junning, QIAN Jun. Determination of methotrexate in human plasma by HPLC and qualitative analysis of its metabolite by mass spectrometry[J]. Journal of Pharmaceutical Practice and Service, 2015, 33(2): 143-146,158. doi: 10.3969/j.issn.1006-0111.2015.02.013

人血浆中甲氨蝶呤的高效液相色谱法测定及其代谢物的质谱定性分析

doi: 10.3969/j.issn.1006-0111.2015.02.013

复旦大学附属肿瘤医院肿瘤内科, 复旦大学上海医学院肿瘤学系, 上海 200032

Determination of methotrexate in human plasma by HPLC and qualitative analysis of its metabolite by mass spectrometry

Department of Medical Oncology, Cancer Hospital Affilliated to Fudan University; Department of Oncology, Shanghai Medical College, Fudan University, Shanghai 200032, China

摘要: 目的建立固相萃取-高效液相色谱法测定人血浆中的甲氨蝶呤浓度,用于临床治疗的监测,并通过质谱对甲氨蝶呤的人体代谢物进行定性分析。方法血浆样本经C₁₈固相萃取小柱净化洗脱后直接进样,以Diamonsil C₁₈分析柱(150 mm×4.6 mm, 5 m)分离,流动相为甲醇和0.5%乙酸溶液(含0.3%三乙胺),采用梯度洗脱;流速为1.0 ml/min;紫外检测波长为306 nm。通过质谱全扫描(QLMS)和多反应监测-信息关联采集-增强子离子扫描(MRM-IDA-EPI)获得甲氨蝶呤代谢物的质谱信息。结果甲氨蝶呤在0.05~100 μmol/L范围内线性关系良好(r=0.999 9),提取回收率>95%,准确度在97%~105%之间,日内与日间精密度(RSD)均<5%。分析甲氨蝶呤代谢物的质谱信息,推断该代谢物为7-羟基甲氨蝶呤。结论本方法灵敏度高、重现性好、操作简便,适用于甲氨蝶呤的血药浓度监测。
- 甲氨蝶呤 /
- 高效液相色谱法 /
- 固相萃取 /
- 代谢物
Abstract: Objective To develop an SPE-HPLC method for the determination of methotrexate (MTX) in human plasma to monitor the clinical drug use of MTX, and to identify the human metabolite of MTX by mass spectrometry. Methods MTX was extracted from human plasma using C₁₈ SPE column and analyzed directly after elution. The separation of MTX was performed on Diamonsil C₁₈ column (150 mm×4.6 mm, 5 m) with a gradient mobile phase of methanol and 0.5% acetic acid solution (containing 0.3% triethylamine) at a flow rate of 1.0 ml/min. The detection wavelength was 306 nm. QLMS and MRM-IDA-EPI scan modes were used to obtain the mass spectrum of MTX metabolite. Results The calibration curve of MTX in plasma was linear over the range of 0.05-100 μmol/L(r=0.999 9). The extraction recovery was above 95% and accuracy was between 97% and 105%. Both intra-and inter-day precision were less than 5%. 7-OH MTX was identified to be the metabolite through the analysis of the mass spectrum. Conclusion The method is sensitive, reproducible, easy to operate and suits for monitoring the concentration of MTX in clinical practice.
- methotrexate /
- HPLC /
- solid phase extraction /
- metabolite

[1]	Green MR,Chowdhary S,Lombardi KM,et al.Clinical utility and pharmacology of high-dose MTX in the treatment of primary CNS lymphoma[J].Expert Rev Neurother,2006,6(5):635-652.
[2]	Lennard L.Therapeutic drug monitoring of antimetabolic cytotoxic drugs[J].Br J Clin Pharmacol,1999,47:131-143.
[3]	彭波.高效液相色谱法测定甲氨蝶呤的血药浓度[J].中国医院药学杂志,2011, 31 (11):951-952.
[4]	张春燕,顾健.反相高效液相色谱法同时测定人血浆中亚叶酸、5-甲基四氢叶酸及甲氨蝶呤的浓度及临床应用[J].中国药学杂志,2010,45(7):543-547.
[5]	许耘川,袁向亮,沈立松.HPLC法测定血清甲氨蝶呤浓度的方法建立及其与荧光偏振法的比较[J].检验医学,2010,25(9):701-704.
[6]	谢军平,冯玲玲,匡霞.HPLC 测定大剂量甲氨蝶呤患者血药浓度[J].中国实验方剂学杂志,2010,16(11):69-72.
[7]	霍晶,袁盛华,方芸,等.反相高效液相色谱法测定人血浆中甲氨蝶呤的浓度[J].药学与临床研究,2007,15(5):390-392.
[8]	Turci R,Micoli G,Minoia C.Determination of methotrexate in environmental samples by solid phase extraction and high performance liquid chromatography: ultraviolet or tandem mass spectrometry detection?[J].Rapid Commun Mass Spectr,2000,14:685-691.
[9]	周卿,潘雪君.固相萃取-高效液相色谱法测定人乳中甲氨蝶呤的含量[J].药物分析杂志,2011,31(12):2305-2307.
[10]	吴雪梅, 丘宏强, 方令平, 等.固相萃取-高效液相色谱法测定甲氨蝶呤的血药浓度[J].中国医院药学杂志,2008,28(18):1617-1619.
[JP2]张华年,文玲莉,张少文, 等.固相萃取高效液相色谱法检测生物样本中甲氨喋呤[J].药物分析杂志,2000,20(6):401-404.
[12]	曹一辰,孟祥乐,朱金辉,等.LC-MS/MS法测定人血清中甲氨蝶呤及其代谢产物7-羟基甲氨蝶呤的浓度[J].中国药师,2012,15(2):191-194.
[13]	Guo PI, Wang XM, Liu LS, et al.Determination of methotrexate and its major metabolite 7-hydroxymethotrexate in mouse plasma and brain tissue by liquid chromatography-tandem mass spectrometry[J].J Pharm Biomed Anal,2007,43:1789-1795.

[1]	凯丽比努尔·奥布力艾散, 李倩, 谢志, 贾文彦, 尹东锋. 星点设计-效应面法优化仑伐替尼混合胶束的制备工艺 . 药学实践与服务, 2024, 42(11): 1-8. doi: 10.12206/j.issn.2097-2024.202403019
[2]	姚小静, 计佩影, 陆峰, 施国荣, 傅翔. 表面增强拉曼光谱法快速测定尿液中曲马多的研究 . 药学实践与服务, 2024, 42(): 1-5. doi: 10.12206/j.issn.2097-2024.202401072
[3]	景凯, 杨慈荣, 张圳, 臧艺蓓, 刘霞. 黄芪甲苷衍生物治疗慢性心力衰竭小鼠的药效评价及作用机制研究 . 药学实践与服务, 2024, 42(5): 190-197. doi: 10.12206/j.issn.2097-2024.202310004
[4]	冯志惠, 邓仪卿, 叶冰, 安培, 张宏, 张海军. 雀梅藤石油醚提取物诱导三阴性乳腺癌细胞凋亡的实验研究 . 药学实践与服务, 2024, 42(6): 253-259. doi: 10.12206/j.issn.2097-2024.202311055
[5]	孙丹倪, 黄勇, 张嘉宝, 王培. 代谢相关脂肪性肝病的无创诊断与药物治疗 . 药学实践与服务, 2024, 42(10): 411-418. doi: 10.12206/j.issn.2097-2024.202403049
[6]	毛智毅, 王筱燕, 陈晓颖, 汤逸斐. 度拉糖肽联合二甲双胍对肥胖型2型糖尿病患者机体代谢、体脂成分及血清脂肪因子的影响 . 药学实践与服务, 2024, 42(7): 305-309. doi: 10.12206/j.issn.2097-2024.202305032
[7]	张艺昕, 关欣怡, 王博宁, 闻俊, 洪战英. 二氢吡啶类钙离子拮抗药物手性分析及其立体选择性药动学研究进展 . 药学实践与服务, 2024, 42(8): 319-324. doi: 10.12206/j.issn.2097-2024.202308062
[8]	赖立勇, 夏天爽, 徐圣焱, 蒋益萍, 岳小强, 辛海量. 中药青蒿抗氧化活性的谱效关系研究 . 药学实践与服务, 2024, 42(5): 203-210, 216. doi: 10.12206/j.issn.2097-2024.202211012

点击查看大图

计量

文章访问数: 2767
HTML全文浏览量: 281
PDF下载量: 581
被引次数: 0

人血浆中甲氨蝶呤的高效液相色谱法测定及其代谢物的质谱定性分析

doi: 10.3969/j.issn.1006-0111.2015.02.013

复旦大学附属肿瘤医院肿瘤内科, 复旦大学上海医学院肿瘤学系, 上海 200032

收稿日期: 2014-04-03
修回日期: 2014-06-26

关键词:

摘要: 目的建立固相萃取-高效液相色谱法测定人血浆中的甲氨蝶呤浓度,用于临床治疗的监测,并通过质谱对甲氨蝶呤的人体代谢物进行定性分析。方法血浆样本经C₁₈固相萃取小柱净化洗脱后直接进样,以Diamonsil C₁₈分析柱(150 mm×4.6 mm, 5 m)分离,流动相为甲醇和0.5%乙酸溶液(含0.3%三乙胺),采用梯度洗脱;流速为1.0 ml/min;紫外检测波长为306 nm。通过质谱全扫描(QLMS)和多反应监测-信息关联采集-增强子离子扫描(MRM-IDA-EPI)获得甲氨蝶呤代谢物的质谱信息。结果甲氨蝶呤在0.05~100 μmol/L范围内线性关系良好(r=0.999 9),提取回收率>95%,准确度在97%~105%之间,日内与日间精密度(RSD)均<5%。分析甲氨蝶呤代谢物的质谱信息,推断该代谢物为7-羟基甲氨蝶呤。结论本方法灵敏度高、重现性好、操作简便,适用于甲氨蝶呤的血药浓度监测。