目的：建立反相高效液相测定人血清中来氟米特（leflunomide，Lef）活性代谢物A771726浓度的方法。方法：采用VP-ODS C18柱，0.01 mmol·L-1醋酸盐缓冲液(pH4.6)-乙腈(67∶33)为流动相，紫外检测波长为288 nm，流速1.0 mL·min-1，血清样品用乙酸乙酯提取。结果：A771726在血清中得到很好的分离，没有明显的干扰峰，出峰时间为6.4 min。线性范围0.39～50 mg·L-1（r=0.9996），最低检测限为0.098 mg·L-1，血清中日内和日间RSD均小于7%，回收率为89.4%～95.3%。结论：本法简便灵敏，符合生物制品分析要求。
Abstract
OBJECTIVE To establish a method for determination of A771726, leflunomide's active metabolite, in human serum by reversed-phase high performance liquid chromatograph.METHODS The chromatographic column was VP-ODS C18 column(5 μm，150 mm×4.6 mm).The mobile phase was 0.01 mmol·L-1 acetate buffer(pH4.6)-acetonitrile(67∶33). The flow rate was 1.0 mL·min-1.The ultraviolet wavelength for measurement was 288 nm. The serum samples were extracted with ethyl acetate. RESULTS Under the condition of test, A771726 was well separated in serum. The retention time was 6.4min. The linear ranges was 0.39～50 mg·L-1（r=0.9996）.The lowest concentration of detection was 0.98 mg·L-1. Recovery of extraction was 89.4%～95.3%, and RSD of intra-day and inter-day were lower than 7%.CONCLUSIONS This method was simple, rapid and sensitive for determination of A771726 in human serum.
关键词
活性代谢物 /
高效液相色谱法 /
血清浓度 /
来氟米特
{{custom_keyword}} /
Key words
active metabolite（A771726） /
RP-HPLC /
serum concentration /
leflunomide
{{custom_keyword}} /
{{custom_sec.title}}
{{custom_sec.title}}
{{custom_sec.content}}
参考文献
[1］ Bruneau JM, Yea CM, Spinella-Jaegle S, et al. Purification of human dihydro-orotate dehydrogenase and its inhibition by A 771726 , the active metabolite of leflunomide［J］. J Biochem，1998, 336 (2):299.
[2］ Xu X, Blinder L, Shen J, et al. In vivo mechanism by which leflunomide controls lymphoproliferative and autoimmune disease in MRL/MpJ-Ipr/Ipr mice［J］. J Immunol，1997，159(1):167.
[3］ Elder RT, Xu X, Wlliams JW, et al. The immunosuppressive metabolite of leflunomide, A 771726 , affects murine T cells through two biochemical mechanisms［J］. J Immunol，1997，159(1):22.
[4］ Dias VC, Lucien J, Legatt DF, et al. Measurement of the active leflunomide metabolite (A 771726 ) by reverse-phase high-performance liquid chromatography［J］. Ther Drug Monit，1995, 17(1):84.
[5］ Chong AS, Huang WY, Liu W, et al. In vivo activity of leflunomide: Pharmacokinetic analyses and mechanism of immunosuppression［J］. Transplantation,1999, 68(1): 100.
{{custom_fnGroup.title_cn}}
脚注
{{custom_fn.content}}
