HPLC检测三苄糖苷中的有关物质

Determination of related substances of Tribenoside

目的 检查三苄糖苷中的有关物质。方法 采用HPLC法,Hypersil C18色谱柱(250 mm×4.6 mm,10 μm),甲醇-水(75∶25)为流动相,流速1.0 ml·min-1,在波长224 nm处采用不加校正因子的主成分自身对照法对有关物质进行检查。结果 杂质与主成分能完全分离,有关物质的限量控制在2.0%以下。三苄糖苷的α体在1.28～25.60μg·ml-1范围内,峰面积与其浓度的线性关系良好(r=0.9998),最低检测限为10.40ng(S/N=3);β体在3.76-75.20μg·ml-1范围内,峰面积与其浓度的线性关系良好(r=0.9998),最低检测限为4.16 ng(S/N=3)。RSD=1.17%(n=6)。结论 通过对流动相强度、检测波长以及进样浓度的考察,建立了检测三苄糖苷中有关物质的最佳条件。

OBJECTIVE To establish an HPLC method for determination of related sudstances of Tribenoside( TBS) . METHODS The related substances of TBS were determined on the HPLC instrument fixed a Hypersil C18 column (250 mm×4.6 mm, 10 μm) with UV detection at 224 nm. The mobile phase consisted of Methyl alcohol and water( 75 :25). The flow rate was 1.0 ml· min-1. RESULTS The related substances of Tribenoside could be determined with no correction factors. TBS could be separated from all related substances completely. The contents of all related substances was lower than 2.0% .The calibration curves were linear (r = 0.9998) within the range of 1.28 - 25.60 μg· ml-1 for body a and linear ( r = 0.9998) within the range of 3.76 - 75.20 μg· ml -1 for body β. The precision RSD of this analytical method was 1.17%(n = 6).The limits of detection were 10.40 ng (S/N = 3) for body a and 4.16 ng (S/N = 3)for body β. CONCLUSION This article studied the mobile phase, the wavelength and the concentration, then established the best conditions for determination of related sudstances of TBS.