高效液相色谱柱后衍生法测定食品中的V(_K_1)

应用高效液相色谱柱后衍生法测定食品中的维生素K1,样品经过均质、酶解后 ,用正己烷萃取 ,再经水洗涤、浓缩、过滤 ,以 10 0mL二氯甲烷 +90 0mL甲醇 (包含 10mmol氯化锌 ,5mmol冰醋酸 ,5mmol无水乙酸钠 )做流动相 ,利用锌粉柱进行柱后衍生 ,在Ex =2 43nm ,Em =430nm下进行荧光检测 ,外标法定量。该方法比国家标准中紫外检测灵敏度提高了 10 0倍 ,在 0 1～ 10 μg/mL的浓度范围内 ,线性相关系数达到 0 999,相对标准偏差为 4 1% ,回收率为10 0 2 %。

Determination of Vitamin K_1 in Food by HPLC with Post-column Derivation

Vitamin K 1(Phylloquinone)in food was determined with post-column derivated HPLC(high-performance liquid chromatography). The sample was operated following homogenization,enzymatic digestion,extraction with haxance, washing with water,evaporation,filtering and a single-stage HPLC technique utilizing post-column reduction with zinc and fluorescence detection at Ex=243nm, Em=430nm. The mobile phase:10% dichloromethane in methanol containing 10?mmol zinc chloride and 5?mmol acetic acid-sodium acetate,On-column RF monitoring and quantification by external standard method.The sensitivity of the method is 100 times higher than that of national standard with UV detective method. In the concentration range of 0.1～10?μg/mL have a regression of 0.999. The RSD were within 4.1%.The percentage of recovery was 100.2%.