Zou Lianpeng, Zhang Xuesong, Huang Baifen, Yang Jingming, Xu Weisheng, Wang Zhu
National Institute for Nutrition and Health, Chinese Center for Disease Control and Prevention, Beijing 100050, China.
Zhejiang Provincial Center for Disease Control and Prevention, Hangzhou 130050, China.
Wei Sheng Yan Jiu. 2019 May;48(3):468-473.
To establish a high performance liquid chromatography method with fluorescence detection for a variety of vitamin K and to assess the content of vitamin K in animal foods.
Animal foods were hydrolyzed by 0.2 g lipase and 0.1 g protease in pH 8 for 4 hours, extracted with isooctane followed by rotary evaporation and reconstitution. The mobile phase was 900 mL methanol and 100 mL tetrahydrofuran which contained 5 mmol glacial acetic acid, 11 mmol zinc chloride and 6 mmol anhydrous sodium acetate. The content of vitamin K_1, menaquinone-4(MK-4), and menaquinone-7(MK-7)were separated on Atlantis T3 column(4.6 mm×250 mm, 5 μm)by high-performance liquid chromatography. The fluorescence detector was set at a wavelength of 320 nm for excitation and 410.3 nm for emission.
The linear range of the method was 0.01-0.40 μg/mL, and coefficient of determination was > 0.999. The spiked recoveries were 84.4%-124.2% with relative standard deviation was <6%(n=6). MK-4 was the main form of vitamin K in pork and chicken. The highest content of vitamin K_1 was found in beef, and MK-7 could be detected in aquatic products.
The proposed method is successfully applied for the determination of vitamin K in animal foods. A variety of vitamin K are distributed differently in distinct animals.
建立一种用于多种维生素K的高效液相色谱荧光检测法，并评估动物性食品中维生素K的含量。
将动物性食品在pH 8条件下用0.2 g脂肪酶和0.1 g蛋白酶水解4小时，用异辛烷萃取，然后旋转蒸发并复溶。流动相为900 mL甲醇和100 mL四氢呋喃，其中含有5 mmol冰醋酸、11 mmol氯化锌和6 mmol无水醋酸钠。维生素K1、甲萘醌-4（MK-4）和甲萘醌-7（MK-7）的含量在Atlantis T3柱（4.6 mm×250 mm，5 μm）上通过高效液相色谱分离。荧光检测器设定的激发波长为320 nm，发射波长为410.3 nm。
该方法的线性范围为0.01 - 0.40 μg/mL，测定系数>0.999。加标回收率为84.4% - 124.2%，相对标准偏差<6%（n = 6）。MK-4是猪肉和鸡肉中维生素K的主要形式。牛肉中维生素K1的含量最高，水产品中可检测到MK-7。
所提出的方法成功应用于动物性食品中维生素K的测定。多种维生素K在不同动物中的分布存在差异。
