高效液相色谱-荧光法与高效液相色谱-二极管阵列法测定食品中维生素B1含量的比较研究

Comparison of the determination of vitamin B_1 content in the food by high performance liquid chromatography-fluorescence method and high performance liquid chromatography–diode array method

目的比较研究GB 5009.84-2016中高效液相色谱-荧光法(high performance liquid chromatography-fluorescence method,HPLC-FLD)和GB/T 5009.197-2003中高效液相色谱-二极管阵列法(high performance liquid chromatography–diode array method,HPLC-DAD)检测食品中维生素B_1含量的适用性。方法以实验室常见样品奶粉(基质相对复杂)和维生素原料粉(基质简单)为研究对象,分别采用HPLC-FLD法与HPLC-DAD法对其维生素B_1含量进行测定。HPLC-FLD法中样品经过水解、酶解过夜、萃取过滤后经HPLC分析;HPLC-DAD法中样品经溶解、离心、过滤后经HPLC分析。结果 2种方法在各自线性范围内,线性关系均良好(r=0.9995、0.9994),回收率范围分别为92.8%~101.6%和106.3%~109.9%,均能满足检测要求。对于基质相对简单、维生素B_1含量较高的食品,HPLC-DAD法的检测结果更为准确,重现性好且更接近样品声称值;对于基质较为复杂的食品,HPLC-DAD法的检测结果与HPLC-FLD法相差不大。结论 HPLC-DAD法操作简便、准确度高、精密度好,是测定食品中维生素B_1含量的可行方法,能够缩短检验时间,提高检验效率,具有较好的实用价值。

Objective To explore and compare the applicability of the 2 national standard methods GB 5009.84-2016 with high performance liquid chromatography-fluorescence method （HPLC-FLD） and GB/T 5009.197-2003 with high performance liquid chromatography–diode array method （HPLC-DAD） for determination the content of vitamin B1 in food. Methods The common samples in laboratory were chosen as research objects, such as milk powder with relatively complex matrix and vitamin raw material powder with relatively simple matrix, and the content of vitamin B1 was determined by HPLC-FLD and HPLC-DAD, respectively. The samples were analyzed by HPLC after hydrolysis, enzymolysis overnight, extraction, and filtration by HPLC-FLD, while the samples were analyzed by HPLC after dissolution, centrifugation, and filtration by HPLC-DAD. Results Both methods showed good linear relationships （r=0.9995, 0.9994）, the recoveries were 92.8%~101.6% and 106.3%~109.9%, respectively, which could satisfy the detection requirements. The determination results of HPLC-DAD were more accurate and reproducible and closer to the claims of samples which were relatively simple and had high content of vitamin B1. The determination results were similar by HPLC-FLD and HPLC-DAD for those samples which were relatively complex. Conclusion HPLC–DAD is simple, accurate and precise. It is a feasible method to determine vitamin B1 in food and can save test time, improve efficiency and has good practical value.