[关键词]
[摘要]
目的 建立高效液相色谱-荧光检测法测定α-生育酚的方法。方法 建立高效液相色谱-荧光检测法,采用Merck LiChrosorb Si60(4×250 mm,5 μm)硅胶柱、荧光检测器,以0.75%异丙醇的正己烷溶液为流动相,考察方法专属性、线性关系、检测限与定量限、精密度、加样回收率、稳定性,并与高效液相色谱-紫外检测法进行灵敏度比较;检测ω-3羧酸胶囊、维生素A棕榈酸酯和维生素E中α-生育酚含量,与高效液相色谱-紫外检测法检测结果进行比较。结果 建立的高效液相色谱-荧光检测法中α、β、γ和δ-生育酚分离良好,在3~30 μg/mL呈良好的线性关系(r=0.999 8);信噪比约为1:10,定量限为0.15 μg/mL;信噪比约为1:3,检测限为0.05 μg/mL;添加ω-3羧酸、维生素A棕榈酸酯、维生素E,加样回收率分别为(98.7±1.4)%、(97.6±0.8)%、(99.5±2.3)%(n=9);精密度、稳定性均符合要求;荧光检测器较紫外检测器灵敏度高3~5倍,且受其它主药成分干扰小;应用两种方法检测的3种产品中α-生育酚含量相近。结论 该方法灵敏、准确、简便,专属性好,可用于测定多种脂溶性药物中α-生育酚的含量。
[Key word]
[Abstract]
Objective To establish a method to determine the content of α-tocopherol in fat-soluble drugs by high performance liquid chromatography (HPLC) with fluorescence detection. Method Merck LiChrosorb Si60 silica gel column (4 mm × 250 mm, 5 μm) and fluorescence detector were adopted to establish HPLC-fluorescence detection. The mobile phase was 0.75% isopropyl alcohol in n-hexane. The specific properties, linear relations, detection limits and quantitative limits, precision, rate of recovery, and stability of the established methods were investigated, and its sensitivity was compared with that of HPLC-UV detection. The contents of α-tocopherol in Omega -3 carboxylic acid capsules, vitamin A palmitate and vitamin E were detected and compared with those of HPLC with UV detection. Results From the method established in this study, good separation of α, β, γ, and δ-tocopherol could be achieved, and the standard curve has a good linear relation in the range of 3~30 μg/mL (r= 0.999 8). The signal-to-noise ratio was 1:10, and the limit of quantification was 0.15 μg/mL; Signal-to-noise ratio was 1:3, and the detection limit was 0.05 μg/mL. After adding Omega -3 carboxylic acid, vitamin A palmitate and vitamin E, the recoveries were (98.7 + 1.4)%, (97.6 + 0.8)% and (99.5 + 2.3)%, respectively (n= 9). Precision and stability are in line with the requirements of a UV detector. The sensitivity of fluorescence detector is 3~5 times higher than that of UV detector, and is less disturbed by other main components. The contents of α-tocopherol in three products detected by two methods were similar. Conclusion This method is sensitive, accurate, simple, and good specificity, and will be used for the determination of the content of α-tocopherol in various fat-soluble drugs.
[中图分类号]
[基金项目]