高效液相色谱法同时测定预混合饲料中维生素C、维生素B_2及其磷酸盐

Simululanrous determinution of vitamin C,vitamin B_2 and their phosphate in additive premix by high performance liquid chromatography

本研究建立了高效液相色谱（HPLC）同时测定预混合饲料中维生素c、维生素B：及其磷酸盐形式维生素C-2-磷酸酯（VCP）、维生素琰-5-磷酸钠（VB，P）的方法。预混合饲料用1．5mmol／L四丁基硫酸氢铵缓冲溶液提取后，进行HPLC检测。采用ZorbaXEclipsePlusC18色谱柱（250rTlfnX4．6mm，5μm），以1．5mmol／L四丁基硫酸氢铵缓冲溶液（pH=3．0）与甲醇作为流动相，梯度淋洗。在此色谱条件下，维生素c、维生素B。VCP和VB2P的线性关系良好，检出浓度分别为3、1、1、3mg／kg，定量限为10、5、5、10mg／kg。应用该法测定了预混合饲料中维生素c、维生素B2、VCP和VB2P，平均回收率为81．5％～106％，相对标准偏差（RSD）均小于10％。

A high performance liquid chromatographic （HPLC） method with ultraviolet detection was established to determine vitamin C,vitamin B2 and its phosphate （vitamin C-2- phosphate （VCP）and vitamin B2-5- phosphate （VB2P））in additive premix simultaneously.Additive premix were extracted by 1.5 mmol/L tetrabutylammonium hydroxide,then detected by HPLC.A Zorbax Eclipse Plus-C18 column （250 minx 4,6 ram,5μm）was used in the analysis.The separation was ear- ried out using the mobile phase consisting of 1.5 mmol/L tetrabutylammonium hydroxide （pH 3.0）-methanol at a flow rate of 1.0 mL/min.The eluates were monitored by the ultraviolet detection setting at 255 nm.The linearities were from 1 mg/L to 500 mg/L,for vitamin C ,vitamin B2,VCP,VB2P.The detection limit was 3,1,1,3 mg/kg,and 10,5,5,10 mg/kg for vita- min C,vitamin B2,VCP,VB2P.The mean recoveries were from 81.5% to 106% ,precision were less than 10% ,respectively. The method was very reliable for the simultaneous determination of vitamin C, vitamin B2, VCP,VB2P in additive premix.