摘要
以2，3，4，6-四-0-乙酰基-β-D-葡萄糖异硫氰酸酯（GITC）为柱前手性衍生化试剂，建立了柱前衍生化反相高效液相色谱（RP—HPLC）法测定盐酸托莫西汀中S．异构体杂质的方法。考察了碱化与衍生化试剂的浓度及反应时间对衍生化产率的影响，以及流动相组成对柱效、分离度、保留时间的影响，并讨论了结构因素对手性衍生和分离的影响。采用AgilentZorbaxSB—C，8（250mm×4．6mm，5μm）色谱柱，以乙腈-四氢呋喃-水（体积比为41：14：45）为流动相，流速1．0mL／min；检测波长254nm，柱温35℃。在优化实验条件下，盐酸托莫西汀与S-异构体的分离度R〉1．5；S-异构体的检出限为0．15μg／L，在0．5～8mg／L范围内线性良好；精密度RSD为1．8％；按标准加入法计算，加标回收率为92％-99％；按外标法计算，加标回收率为94％～102％。该方法简便、可靠，可作为盐酸托莫西汀原料药中S-异构体杂质限量的质控方法。
A pre-column derivatization high performance liquid chromatographic （HPLC） method for the determination of S-isomer impurity of atomoxetine hydrochloride was developed by using 2,3,4,6- tetra-O-acetyl-β-D-glucopyranosyl isothiocyanate （GITC）as derivative reagent. Effects of GITC concentration and reaction time on derivative reaction and influences of types and concentrations of organic modifier on chiral separation were investigated. The effects of structural factors on ehiral derivatization and separation efficiency were discussed. The separation of two isomers was performed on a Zorbax SB-C18 （250 mm ×4. 6 mm, 5 μm） column using acetonitrile - tetrahydrofuran - water （ 41 ： 14 ： 45, by volume） as mobile phase with a flow rate of 1.0 mL/min. The column temperature was maintained with 35 ℃ and the detection wavelength was set at 254 nm. Under the optimal conditions, the resolution of atomoxetine hydrochloride and its S-isomer was greater than 1.5. The linear range of S-isomer was 0. 5 - 8 mg/L with a limit of detection （ LOD ） of 0. 15 mg/L. The relative standard deviation（RSD） was 1.8% . The spiked recoveries of the S-isomer obtained from the general external standard method and the standard addition method were in the range of 94% -102% and 92%- 99% , respectively. The results showed that the established HPLC method was reliable and simple, and could be used for the quality control of active pharmaceutical ingredient.
出处
《分析测试学报》
CAS
CSCD
北大核心
2010年第8期817-821,共5页
Journal of Instrumental Analysis
