两种高效液相色谱法测定维生素B1含量的比较

Comparison of two high performance liquid chromatography methods for the content determination of Vitamin B1

目的:比较高效液相-紫外检测器与高效液相-荧光检测器测定维生素B1含量的优劣,为维生素B1的含量测定提供指导意见,供实验者参考。方法:采用安捷伦1260型全自动高效液相色谱仪,中汇达C 18色谱柱(250 mm×4.6 mm,5μm),柱温:30.0℃,分别采用紫外检测器与荧光检测器及不同的流动相测定维生素B1的含量。结果:紫外检测器维生素B1在2.175~217.534μg·mL^-1的浓度范围内线性关系良好(r=0.1,n=7),平均回收率100.0%,RSD=0.58%(n=9)。荧光检测器在0.1088~108.7669μg·mL^-1的浓度范围内线性关系良好(r=0.9997,n=10),平均回收率96.2%,RSD=1.1%(n=9)。结论:两种方法均可以准确测定维生素B1的含量,紫外检测器法准确度与精密度都优于荧光检测器法,可作为维生素B1含量测定的首选方法,荧光检测器法专属性更好,可作为维生素B1含量测定的备用检测方法。

Objective:To compare the advantages and disadvantages of HPLC-ultraviolet detector and HPLC-fluorescence detector for the content determination of vitamin B1 content,and to provide the reference for the content determination of vitamin B1.Methods:The test was performed in Zhonghuida C 18 column(250 mm×4.6 mm,5μm)with Agilent 1260 full-automatic high-performance liquid chromatograph.The column temperature was 30.0℃.Vitamin B1 was determined with UV detector and fluorescence detector under the different mobile phases.Results:The UV detector method showed that the linear range of vitamin B1 fell into 2.175-217.534μg·mL^-1(r=0.9999,n=7),the average recovery was 100.0%with RSD=0.58%(n=9).The fluorescence detector method showed that the linear range of vitamin B1 fell into 0.1088μg·mL^-1 to 108.7669μg·mL^-1(r=0.9997,n=10),and the average recovery was 96.2%with RSD=1.1%(n=9).Conclusion:The established two mcethods can accurately determine the content of vitamin B1.The accuracy and precision of the UV detector method are better than the fluorescence detector method.The UV detector method is recommended as the preferred method for the content determination of vitamin B1.The fluorescence detector method has better specificity,and can be used as a backup method for the content determination of vitamin B1.