HPLC-MS/MS 法测定白血病患者血浆甲氨蝶呤浓度
王磊,张阳,刘红星,赵建超,陶蓓蓓
河北燕达医院陆道培血液肿瘤中心,河北 三河 065201
Determination of methotrexate in leukemia patients plasma by HPLC-MS/MS
WANG Lei, ZHANG Yang, LIU Hong-xing, ZHAO Jian-chao, TAO Bei-bei
Hematology and Oncology Center,Yanda Hospital of Hebei Province, sanhe 065201, China
摘要 目的:建立测定白血病患者使用大剂量甲氨蝶呤化疗后血浆中甲氨蝶呤浓度的HPLC-MS/MS 法,并应用于临床治疗药物监测,为临床合理用药提供可靠依据。方法:采用多索茶碱作内标,血浆样品经含内标甲醇沉淀蛋白处理。色谱柱为Ultimate XB-C18,柱温40 ℃,流速0.75 mL·min-1,流动相为含0.1% 甲酸和2 mmol·L-1 乙酸铵的水溶液和含0.1% 甲酸的甲醇溶液,梯度洗脱。质谱检测方式为电喷雾离子阱正离子模式,MRM 扫描,监测甲氨蝶呤m/z 455.200 → 308.200,多索茶碱m/z 267.100 → 181.100。结果:甲氨蝶呤浓度在0.022~2.200μmol·L-1 内线性关系良好,Y= 4.56x + 0.081(r = 0.996 0)。结论:该方法简便、准确、快速,检测的特异性及灵敏度高,适用于甲氨蝶呤的血药浓度测定。
关键词 :
甲氨蝶呤 ,
高效液相质谱联用 ,
治疗药物监测
Abstract :Objective : To establish a HPLC-MS/MS method to determine methotrexate in Leukemia patients plasma. Methods :The method employs a direct precipitation with methanol containing doxofylline as the internal standard and a reversed phase separation in an Ultimate XB-C18 column. Mobile phase was a mixture of water (containing 2mmol?L-1 ammonium acetate and 0.1% formic acid) and methanol (containing 0.1% formic acid). The flow rate was 0.75 mL?min-1 and the column temperature was 40℃ . Mass spectrometric analysis was performed in positive ion mode and the multiple reaction monitoring transitions of m/z 455.200 → 308.200 and m/z 267.100 →181.100 were used to quantify methotrexate and doxofylline (internal standard), respectively. Results : The calibration curve of methotrexate was linear within range of 0.022-2.200 μmol?L-1, Y=4.56X+0.081, r=0.996 0. conclusion : This method is convenient,accurate and rapid and it is suitable for detecting methotrexat e in leukemia patients plasma.
Key words :
methotrexate
HPLC-MS/MS
TDM
收稿日期: 2014-11-28
[1] 刘桂兰, 李红丽, 殷惠君, 等. 儿童髓外白血病中氨甲蝶呤防治剂量的研究( 附13 例报告)[J]. 临床儿科杂志, 1993, 11(3):17-19.
[2] 李照民, 陆婉琴, 孔令琪, 等. 甲氨蝶呤的临床药理- 血药浓度与毒性相关的初步观察[J]. 上海第一医学院学报, 1980, 7(6):406-408.
[3] 曾东向, 向芳. 反相高效液相色谱法监测患者血浆中甲氨蝶呤浓度[J]. 医药导报, 2004, 23(3):194-196.
[4] Kremer J M,Lee J g. The safety and efficacy of the use of methotrexate in long-term therapy for rheumatoid arthritis.[J].ArthritisRheum, 1986, 29(10):822-831.
[5] Ramamn A V.Whitworth P,Baildam E M.Use of methotrexate in juvenile idiopathic arthritis.[J].Arch Dis child, 2003, 88(5):197-200.
[6] Tanaka E.Taniguchi A,Urano W,et al.Pharmacogenetics of Diseasemodifying anti-rheumatic drugs.[J].Best Pract Res Clin Bheumatol,2004, 18(15):233-247.
[7] Assadullahi T P,Dagli E,Warner J O.High-performance liquid chromatography method for serum methotrexate levels in children with severe steroid-dependent asthma.[J].J Chromatogr, 1991,565(32):349-356.
[8] Rule G.Chapple M,Henion J.A 384-well solid-phase extraction for LC/MS/MS determination of methotrexate and its 7-hydroxy metabolite in human urine and plasma.[J].Anal Chem, 2001,73(25):439-443.
[9] Guo P,Wang X,Liu L,et al.Determination of methotrexate and its major metaholite 7-hydroxymethotrexate in mouse plasma and brain tissue by liquid chromatography-tandem mass spectrometry.[J].J Pharm Biomed Anal, 2007, 43(15):1789-1795.
[10] 李海菊, 王峰, 李焕德.HPLC 监测甲氮蝶呤虹药浓度方法的改进与优化[J]. 中国药物应用与监测, 2004, l(1):51-53.
[11] 张善堂, 杨林海, 孙言才, 等. HPLC 法测定急性淋巴细胞白血病患儿血清中甲氨蝶呤浓度[J]. 中国药房, 2008, 19(11):825-827.
[12] 王璐璐, 朱林, 倪坤仪, 等. HPLC 研究甲氨蝶呤磁靶向脂质体在小鼠体内的组织分布[J]. 中国药学杂志, 2006, 41(20):1572-1575.
[13] 张强, 徐丽婷, 谢景文, 等. HPLC 法测定人血清中氨甲蝶呤血药浓度[J]. 解放军药学学报, 2000, 16(1):40-42.
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[9]
李慧博;卢晟晔;赵荣生. 来氟米特与甲氨蝶呤合用所致的间质性肺炎 [J]. , 2013, 11(6): 52-55.