Huang S, Caldwell M J, Simpson K L
Department of Food Science and Nutrition, University of Rhode Island, Kingston 02881.
Int J Vitam Nutr Res. 1993;63(3):217-22.
Reverse phase open column chromatography was used to determine urinary riboflavin. An aliquot of a urine specimen was added directly onto the reverse phase open column. The riboflavin was collected and measured as the compound was eluted. The mobile phase (MeOH/H2O/HOAc 37/63/0.1 v/v/v) and stationary phase (C18, 50 microns) were used. The method shows good reproducibility. Recovery of added Vit.B2 was 98.89-100%. The coefficient variation of reproducibility of within day was 1.53%. The coefficients of variation of reproducibility of day-to-day were 0.39% and 1.74% for low and high concentration samples, respectively. The reverse phase open column chromatography was compared to the traditional fluorometric method and HPLC method for riboflavin. The coefficient of linear regression of these comparisons are Y = 0.331 + 1.010X (n = 37, r = 0.984) and Y' = -0.004 + 1.036X' (n = 15, r = 0.999) for RP open column vs. traditional method and RP open vs. HPLC, respectively, where X or X' is the reverse phase open column chromatography method and Y or Y' is the traditional fluorometric method or HPLC, respectively. Concentrations of riboflavin as low as 0.05 micrograms/ml were measured with a linear relation of response to concentration to at least 10 micrograms/ml.
采用反相开放柱色谱法测定尿中核黄素。将一份尿液标本的等分试样直接加到反相开放柱上。当化合物被洗脱时，收集并测定核黄素。使用的流动相为（甲醇/水/醋酸 37/63/0.1 v/v/v），固定相为（C18，50 微米）。该方法具有良好的重现性。添加的维生素 B2 的回收率为 98.89 - 100%。日内重现性的变异系数为 1.53%。低浓度和高浓度样品日间重现性的变异系数分别为 0.39%和 1.74%。将反相开放柱色谱法与传统荧光法和高效液相色谱法用于核黄素的测定进行了比较。这些比较的线性回归系数分别为反相开放柱与传统方法时 Y = 0.331 + 1.010X（n = 37，r = 0.984），反相开放柱与高效液相色谱法时 Y' = -0.004 + 1.036X'（n = 15，r = 0.999），其中 X 或 X' 分别为反相开放柱色谱法，Y 或 Y' 分别为传统荧光法或高效液相色谱法。核黄素浓度低至 0.05 微克/毫升时即可测定，其响应与浓度的线性关系至少可达 10 微克/毫升。
