目的 建立一种同时测定人血浆中亚叶酸、5-甲基四氢叶酸及甲氨蝶呤（MTX）浓度的反相高效液相色谱法。方法 乙腈沉淀后，上清液用氮气吹干，残留物流动相复溶后进样。色谱条件：分析柱为Spherisob C18反相柱，流动相为0.05 mol·L-1醋酸钠 + 0.07 mmol·L-1 EDTA缓冲液（pH 4.4）：甲醇梯度洗脱，流速：1.0 mL·min-1，紫外及荧光同时检测。结果 亚叶酸、5-甲基四氢叶酸及MTX的保留时间分别为11.6，16.0，27.0 min； 检测限分别为0.5，0.05，0.05 mg·L-1；线性关系良好，r分别为0.998 0、0.999 2、0.998 9 (n=6)；低、中、高浓度的回收率、日内及日间精密度均符合方法学要求。结论 本方法结果稳定，重现性好，可以用于临床大剂量MTX化疗体内药物浓度监测。
Abstract
OBJECTIVE To develop a HPLC method using a reversed-phase C18 column for the determination of folinic acid, 5-methylenetetrahydrofolate and methotrexate for plasma drug monitoring. METHODS The samples were precipitated with CH3CN, dried with N2 ,then dissolved with mobile phase and injected into HPLC. The separation was performed on a Spherisob C18 column with gradient elution. The mobile phase constitute of 0.05 mol·L-1sodium acetate, 0.07 mmol·L-1 EDTA buffer (pH 4.4) and methanol with the flow rate of 1.0 mL·min-1. The samples were determined by ultraviolet and fluorescence detection at the same time. RESULTS The retention time of folinic acid, 5-methyltetrahydrofolate and methotrexate were 11.6, 16.0 and 27.0 min. The detection limits were 0.5, 0.05 and 0.05 mg·L-1. The linearity was obtained in the quantitative range. r were 0.998 0, 0.999 2 and 0.998 9 (n=6), respectively. CONCLUSION It is a repeatable and stable method for simultaneous determination of folinic acid, 5-methylenetetrahydrofolate and methotrexate in plasma.
关键词
反相高效液相色谱法 /
亚叶酸 /
5-甲基四氢叶酸 /
甲氨蝶呤 /
血浆药物浓度
{{custom_keyword}} /
Key words
RP-HPLC /
5-methyltetrahydrofolate /
methotrexate /
plasma drug monitoring
{{custom_keyword}} /
{{custom_sec.title}}
{{custom_sec.title}}
{{custom_sec.content}}
参考文献
［1］ YE H, GU L J. Research on high-dose methotrexate in the treatment of acute lymphoblastic leukemia ［J］. Chin J Hematol(中华血液学杂志),1999,20 (2):110-112.
［2］ LU X T, LI Y Z, HUA Y. Study on leucovorin rescue after intravenous infusion of high-dose methotrexate［J］. Chin J Pediatric Hematol(中国小儿血液)，1999，4（2）：59-61.
［3］ GUO Y, ZHU X F. Research on molecular pharmacological mechanism of high-dose methotrexate in the treatment of acute lymphoblastic leukemia［J］. Int J Blood Transfus Hematol(国际输血及血液学杂志), 2006，29（4）：350-352.
［4］ YE H, GU L J, CHEN J,et al. Exploration of leucovorin rescue program after intravenous infusion of high-dose methotrexate［J］. Chin J Hematol(中华血液学杂志),2000, 21(2):92-93.
［5］ LI S, HE Y J, LIAO H. High dose methotrexate treatment for malignant tumor and the serum concentration monitoring［J］. Chin Hosp Pharm J(中国医院药学杂),2001, 21(4):201-203.
［6］ ZHANG S T, YANG L H, SUN Y C, et al. Determination of serum concentration of methotrexate in children with acute LM hoblastic leukemia by HPLC［J］. J China Pharm(中国药房),2008,11(19):825-827.
［7］ XIONG F Y, WU H W, LI J G. Detection of blood methotrexate concentration with HPLC［J］. Chin J Clin Pharm(中国临床药学杂志),2006，15(4):243-244.
［8］ CHENG B, LIN N M, FANG L. RP-HPLC Determination of methotrexate in human plasma［J］. China J Pharm Anal(药物分析杂志)，2006，26（5）：598-601.
［9］ SCHLEYER E, REINHARDT J, UNTERHALT M, et al. Highly sensitive coupled-column high-performance liquid chromatographic method for the separation and quantitation of the diastereomers of leucovorin and 5-methyltetrahydrofolate in serum and urine［J］. J Chromatogr B, 1995，669:319-330.
［10］ THOMAS O, VINCENT T R, GERHARD H, et al. Analysis of 5-methyltetrahydrofolate in serum of healthy children［J］. Mol Genet Metab, 2006，87（1）：61-65.
［11］ ORMAZABAL A, GARCIA-CAZORLA A, PEREZ-DUENAS B, et al. Determination of 5-methyltetrahydrofolate in cerebrospinal fluid of paediatric patients:Reference values for a paediatric population［J］. Clin Chim Acta, 2006，371（1-2）:159-162.
［12］ JAMES A S, DANIELE S, WILLIAM T W. Pharmacokinetics of the diastereoisomers of leucovorin after intravenous and oral administration to normal subjects［J］. Cancer Res, 1984，44（2）:3114-3119.
［13］ EDWARD M N, JAMES A S, JAMES H D. Pharmacokinetics of the diastereoisomers of (6R-s)-folic acid(leucovorin) in human during constant high-dose intravenous infusion ［J］. Cancer Res, 1989, 49（10）:5755-5760.
［14］ BELZ S, FRICKEL C, WOLFROM C, et al. High-performance liquid chromatographic determination of methotrexate, 7-hydroxymethotrexate, 5-methyltetrahydrofolic acid and folinic acid in serum and cerebrospinal fluid［J］. J Chromatogr B, 1994，661:109-118.
{{custom_fnGroup.title_cn}}
脚注
{{custom_fn.content}}
PDF(948 KB)
