Kosovec Juliann E, Egorin Merrill J, Gjurich Susanna, Beumer Jan H
Molecular Therapeutics/Drug Discovery Program, University of Pittsburgh Cancer Institute, Pittsburgh, PA 15213, USA.
Rapid Commun Mass Spectrom. 2008;22(2):224-30. doi: 10.1002/rcm.3362.
5-Fluorouracil (5-FU) has long had a place in the treatment of many malignancies. 5-FU plasma concentrations have been correlated with toxicity and efficacy, and therapeutic drug monitoring has been reported to result in an improved response/toxicity balance. We report validation, according to FDA guidelines, of a hydrophilic interaction chromatography (HILIC) liquid chromatography/tandem mass spectrometry (LC/MS/MS) assay for the sensitive, accurate and precise quantitation of 5-FU in human plasma. The assay employed an isotopically labeled 5-FU internal standard and ethyl acetate extraction. Separation was achieved with an amino column and an isocratic mobile phase of 0.1% formic acid in acetonitrile/water (97:3, v/v), followed by a wash. Detection consisted of electrospray, negative-mode ionization tandem mass spectrometry in the multiple reaction monitoring (MRM) mode. The accuracy was 96.0-102.2%, and precision was 2.1-7.5% in the concentration range of 10-10 000 ng/mL. Recovery from plasma was 46.0-72.6%, and ion suppression was 9.8-25.7%. Plasma freeze/thaw stability was 87.5-104.3%, and stability for 4 h at room temperature was 98.7-100.0%. This assay is currently being used to quantitate 5-FU in human plasma samples.
5-氟尿嘧啶(5-FU)长期以来在多种恶性肿瘤的治疗中占有一席之地。5-FU的血浆浓度与毒性和疗效相关,据报道治疗药物监测可改善反应/毒性平衡。我们根据美国食品药品监督管理局(FDA)的指导原则,报告了一种亲水相互作用色谱(HILIC)液相色谱/串联质谱(LC/MS/MS)法的验证情况,该方法用于灵敏、准确和精确地定量测定人血浆中的5-FU。该测定法采用同位素标记的5-FU内标和乙酸乙酯萃取。使用氨基柱和乙腈/水(97:3,v/v)中含0.1%甲酸的等度流动相进行分离,随后进行冲洗。检测采用电喷雾、负模式电离串联质谱的多反应监测(MRM)模式。在10 - 10000 ng/mL的浓度范围内,准确度为96.0 - 102.2%,精密度为2.1 - 7.5%。血浆回收率为46.0 - 72.6%,离子抑制率为9.8 - 25.7%。血浆冻融稳定性为87.5 - 104.3%,在室温下4小时的稳定性为98.7 - 100.0%。该测定法目前正用于定量测定人血浆样本中的5-FU。