H-酸高效液相色谱分析新方法的建立及评价

Set up and Evaluation on a New Analysis Method of H-Acid by HPLC

经典的HPLC方法分析H-酸,采用低有机相、低pH值、高盐流动相,色谱柱使用寿命短、分析成本高。本文采用反相离子对分离模式,建立了一种H-酸HPLC分析的新方法。以C18柱(4.6mm×150mm,5μm)作分析柱,甲醇:30mM四丁基溴化铵(pH=2.7)=50:50(V/V)作流动相,流速1.0mL/min,检测波长234nm。此色谱条件下H-酸在100～500μg/mL范围内,进样浓度与峰面积值具有良好的线性关系(γ>0.9997),其对应H-酸的检测限为0.11μg/mL。另外还考察了此方法的稳定性及耐用性。该方法不仅能基线分离H-酸及其中的杂质,而且稳定性及耐用性均可达到应用要求。

Low organic phase,low pH and high salt mobile phase were adopted according to the classical analysis method of H-acid by HPLC.The service life of used columns was shortened and analysis cost was increased.A new analysis method of H-Acid by HPLC with the ion-pair reversed-phase separation mode was established.The conditions were：the column was C18 column（4.6mm × 150mm,5μm）,the mobile phase was methanol：30mM tetrabutylammonium bromide（pH = 2.7）= 50∶50（V/V）,the flow rate was 1.0mL/min and detection wavelength was 234nm.The concentration of H-acid had a good linear relationship（γ0.9997）with peak area value between 100 ~ 500μg/mL,the corresponding detection limit of H-acid was 0.11μg/mL.Moreover,the stability and durability of the method were also studied.Not only impurities could be separated from H-acid by this method,but also stability and durability could meet application requirements.