HPLC法测定他克莫司的含量及有关物质  

HPLC determination of tacrolimus and its related substances

作　　者：陈悦[1] 

机构地区：[1]浙江省药品检验所,杭州310004

出　　处：《药物分析杂志》2008年第2期309-313,共5页Chinese Journal of Pharmaceutical Analysis

摘　　要：目的:建立一种适用于他克莫司有关物质及含量测定的 HPLC 分析方法。方法:色谱柱为 Agilent 公司 Eclipse XDB—C_(18)(5 μm,4.6 mm×150 mm);以含0.2%聚氧乙烯月桂醚(Brij35)的0.05 mol·L^(-1)磷酸二氢钾溶液(用10%磷酸溶液调节 pH至3.0±0.1)-乙腈(50:50)为流动相;柱温为50℃;流速为1.5 mL·min^(-1);检测波长为210 nm;进样体积为50μL。选用乙腈-水(1:1)混合溶液为溶解介质制备浓度为0.25 mg·mL^(-1)的试验溶液,室温放置3 h 使其立体异构体相互转换趋于平衡后,取样测定;以他克莫司3个异构体峰面积之和进行定量计算。结果:他克莫司3个异构体的色谱峰之间及与各自相邻杂质峰之间均能良好分离;理论板数按他克莫司主峰 FK506峰计算不低于2000。方法的线性范围按他克莫司3个异构体总和计为5.15～412.2μg·mL^(-1),r=0.9999;重复性试验测得他克莫司的平均含量(n=9)为97.4%,RSD 为0.3%;加样回收率(n=3)分别为99.7%,101.1%,100.7%;RSD 分别为1.2%,1.5%,0.68%;以主成分 FK506计,他克莫司的检测限为1μg·mL^(-1),定量限为4.5μg·mL^(-1)。结论:本法专属、简便、实用,适合于他克莫司的有关物质检测与含量测定,能有效控制产品质量。Objective：To establish an HPLC method for the quality control analysis of tacrolimus. Methods：The HPLC assay was carried out using an Agilent Eclipse XDB -Cs column （5μm,4. 6 mm × 150 mm） where the col- umn temperature was 50 ℃ and detection wavelength was 210 nm, and the mobile phase was 50% acetonitrile and 50% monobasic potassium phosphate buffer （0. 05 mol · L^- 1 ） , which contained 0. 2% polyoxyethylene lauryl ether （Brij35） with pH adjusted to 3.0 ＋0. l with phosphoric acid. The flow rate was 1.5 mL per minute. The test solution of tacrolimus was prepared with a mixture of water and acetonitrile （ 1 ： 1 ）, incubated for 3 hours at ambient temperature to allow the dynamic equilibrium of three stereo - isomers of tacrolimus. The total content of the three stereo - isomers of tacrolimus was calculated based on the sum of their respective peak areas. Results： Clear separation was achieved among the stereo - isomers of tacrolimus and its related substances by this method. The column efficiency was not less than 2000 theoretical plates indicated by the major peak （FK506）. The standard curves for tacrolimus were linear over the range of S. 15 -412. 2 μg · mL^-1. The limit of detection （LOD） was 1 μg · mL^-1 indicated by the major peak and the limit of quantification （LOQ） was 4. 5μg ·mL ^- 1. The average content of tacrolimus obtained from the precision test was 97.4% with RSD of 0. 3% （n = 9）. The average recoveries of this method for assay were 99.7 %, 101.1% and 100. 7 % with RSD of 1.2%, 1.5% and 0. 68% （ n = 3 ）. Conclusion： This assay is proved to be specific ,reproducible and practical. It can be effectively applied in the field of the quality control analysis of tacrolimus.

关 键 词：他克莫司 立体异构体 含量 有关物质 高效液相色谱法 

分 类 号：R917[医药卫生—药物分析学]