反相高效液相色谱法同时测定维生素B6、烟酰胺和泛酸钙

Simultaneous Determination of Vitamin B6,Nicotinamide and Calcium Pentothenate by RP-HPLC

应用反相高效液相色谱法同时测定了雏生素营养品中维生素B6、烟酰胺和泛酸钙。样品用由磷酸盐缓冲溶液（pH3．0）和甲醇组成的提取液超声提取5min后，再置于65-70℃的水浴中加热10min，所得提取液供高效液相色谱法分析用。色谱分析中用Inertsil pH-3苯基色谱柱作固定相，用由0．05mol·L^-1磷酸二氢钾、甲醇及乙腈以90比6比4的体积比混合的溶液为流动相，并在210nm波长处进行紫外检测。以测得的峰面积值对上述3种化合物的浓度制作标准曲线，其线性关系通过回归分析予以检验，3个回归方程的相关系数均大于0．9999。以实样为基体，用标准加入法对所提出的方法进行了回收试验，测得上述3种化合物的回收率依次为97．4％，102．4%及100．3%。

The method of RP-HPLC was applied to simultaneous determination of vitamin B6, nicotinamide and calcium pentothenate in some vitamin nutriments. The sample was extracted ultrasonically with an extractant containing phosphate buffer solution （pH 3.0） and methanol for 5 min and then heated at 65-70 ℃ in a water bath for 10 min. The extract obtained was used for the HPLC analysis. The Inertsil pH-3 Phenyl column was used as stationary phase, and a mixed solution of 0. 05 mol · L^-1 KH2PO4, methanol and acetonitrile mixed in the ratio of 90 to 6 to 4 by volume, was used as mobile phase. UV detection was made at the wavelength of 210 nm Linearity of the standard curves between the values of peak area and concentration of the 3 compounds was checked by regression analysis, giving values of correlation coefficients greater than 0. 999 9. Test for recovery was made by standard addition method with a substantial sample as the matrix, the results of recovery were found to be 97. 4% for vitamin B6, 100. 3% for calcium pentothenate and 102.4% for nicotinamide.