在线衍生高效液相色谱荧光光谱法测定食品中维生素B_1

Determination of vitamin B_1 in food by high performance liquid chromatography-fluorescence spectrometry coupled with on-line derivatization

目的建立以过硫酸钾为衍生试剂的在线衍生-高效液相色谱(high performance liquid chromatography,HPLC)荧光光谱测定食品中维生素B_1的方法。方法分别研究了流动相pH值、荧光检测波长、衍生反应温度和衍生管线规格参数对测试方法的影响,确立各个参数的数值,建立测试方法 ;对测试方法的标曲稳定性、线性范围、相关系数、精密度和准确度进行考察,并同时采用该方法与GB 5009.84-2016的第一法对常见样品基质进行测试。结果测试方法的参数如下:流动相pH值为6.0,荧光激发波长为350 nm,荧光发射波长为480 nm,衍生反应温度为60℃,衍生管线规格为长10 m,内径0.25 mm;该测试方法在0.1～10.0μg/mL线性范围内的相关系数r>0.999,方法检出限为0.033 mg/100 g,定量限为0.092 mg/100 g,精密度在5%以内;与GB 5009.84-2016第一法测试结果偏差在10%以内。结论本研究建立的方法操作简单,准确度高,重复性好,适合于大批量样品中维生素B_1的测试,可显著提高测试效率。

Objective To establish a method for the determination of vitamin B1 in food by high performance liquid chromatography（HPLC）-fluorescence spectrometry couple with on-line derivatization. Methods The influence of pH value of mobile phase, fluorescence detection wavelength, derivative reaction temperature and derivative line specifications on the test method were studied, respectively, the each parameter value were confirmed and the test method was established. The calibration stability, linear range, correlation coefficient, precision, and accuracy were investigated. The vitamin B1 content in common samples was determined by this method and it was compared with GB 5009.84-2016 the first method. Results The parameters of the test method were as follow： mobile phase pH value 6.0, fluorescence excitation wavelength 350 nm, fluorescence emission wavelength 480 nm, derivative reaction temperature 60 ℃, derivative line of 10 m length, and inner diameter 0.25 mm. In the linear range of 0.1-10.0 μg/mL, the method had good linear relationship with correlation coefficient r greater than 0.999. The limit of detection was 0.033 mg/100 g and the limit of quantitation was 0.092 mg/100 g. The precision was less than 5%, and the relative standard deviations of testing results obtained from this method and GB 5009.84-2016 the first method were less than 10%. Conclusion The established method is simple, accurate and reproducible, which is suitable for detecting vitamin B1 in large batches of samples. This method can improve the efficiency of testing significantly.