建立了用ＨＰＬＣ同时测定人血清中硫唑嘌呤（ＡＺＰ）和６－巯基嘌呤（６－ＭＰ）浓度的方法。血清样品经乙腈除蛋白后在常压和３７℃下用氮气吹干，残余物用１００μＬ洗脱液溶解，离心后上清液直接进样。以ＳｐｈｅｒｉｓｏｒｂＣ１８固定相，采用梯度洗脱，从Ｖ（乙腈）∶Ｖ（０．０１ｍｏｌ／Ｌ磷酸二氢钾）＝３∶９７经５ｍｉｎ时变为１８∶８２，１５ｍｉｎ后再变为５０∶５０，检测波长开始为３２５ｎｍ，１０ｍｉｎ后为２７８ｎｍ，甲硝唑（ＭＮＺ）为内标。ＡＺＰ和６－ＭＰ平均回收率分别为（１００．６±４．２）％和（１０２．４±４．５）％，最低检出质量浓度均为５μｇ／Ｌ。所建方法简便、灵敏、准确。 A method for the simultaneous determination of azathioprine (AZP) and 6-mercaptopurine (6-MP) in human serum by HPLC was established. Serum samples were deproteinized with acetonitrile and dried under nitrogen at 37 ° C under atmospheric pressure. The residue was dissolved in 100 μL of the eluate and the supernatant was injected directly after centrifugation. With SpherisorbC18 ​​stationary phase, the gradient was used to change from V (acetonitrile): V (0.01mol / L potassium dihydrogen phosphate) = 3:97 to 18:82 after 5min and then to 50:50 with gradient elution. The detection wavelength starts at 325 nm, 278 nm after 10 min, and metronidazole (MNZ) is the internal standard. The average recoveries of AZP and 6-MP were (100.6 ± 4.2)% and (102.4 ± 4.5)%, respectively, with the lowest detectable concentrations of 5 μg / L. The method is simple, sensitive and accurate.