高效液相色谱法测定调制乳粉中维生素K2的含量
Determination of Vitamin K2 Content in Formulated Milk Powder by High Performance Liquid Chromatography
陈昭 1李春阳 2张小丽 3孙锦 2程倩1
作者信息
- 1. 农业微生物资源发掘与利用全国重点实验室,湖北宜昌 443003;宜昌市营养健康食品工程技术研究中心,湖北宜昌 443003
- 2. 宜昌市营养健康食品工程技术研究中心,湖北宜昌 443003;酵母功能湖北省重点实验室,湖北宜昌 443003
- 3. 农业微生物资源发掘与利用全国重点实验室,湖北宜昌 443003;酵母功能湖北省重点实验室,湖北宜昌 443003
- 折叠
摘要
建立一种快速的高效液相色谱法用于分析检测调制乳粉中维生素K2的含量.取调制乳粉样品,加入脂肪酶充分酶解,以一定比例的无水乙醇/正丁醇(10∶10,V∶V)提取,异丙醇充分稀释,定容,0.2 μm滤膜过滤过滤.采用高效液相色谱法进行测定,色谱柱为Welch Ultimate® AQ-C18(4.6 mm×150 mm,5 μm),流动相为 100％甲醇,流速 1.0 mL/min,进样体积20 μL,柱温35℃,检测波长254 nm,以外标法定量.维生素K2在0.1～5.0 μg/mL浓度范围内呈良好的线性关系,标准曲线的相关系数R2为1.000.加标回收率在103.00％～106.68％之间,精密度试验中维生素K2含量的相对标准偏差为1.7％,检出限为0.0508 μg/mL.此方法操作简单、准确性好、精密度高,适用于调制乳粉中维生素K2含量测定.
Abstract
A rapid high performance liquid chromatography(HPLC)method for analyzing the content of vitamin K2 in milk powder was established.Fortified milk powder samples were enzymatically hydrolyzed with lipase,extracted with a mixture of anhydrous ethanol/normal butanol(10∶10,V∶V)in a certain proportion,fully diluted with isopropanol,and filtered through a 0.2 μm membrane filter.Using HPLC for analysis,the chromatographic column used was the Welch Ultimate® AQ-C18(4.6 mm×150 mm,5 μm),the mobile phase was 100％methanol,the flow rate was 1.0 mL/min,and the injection was 20 μL,the column temperature was 35 ℃,and a detection wavelength of 254 nm.The standard curve of vitamin K2 was linear in the range of 0.1-5.0 μg/mL(R2=1.000).The standard recovery was 103.00％-106.68％,the relative standard deviation(RSD)of vitamin K2 in precision experiment was 1.7％,the limit of detection was 0.0508 μg/mL.The method is simple,accurate and accurate,making it suitable for the determining the content of vitamin K2 in milk powder.
关键词
高效液相色谱法/维生素K2/脂肪酶/调制乳粉/检测Key words
high performance liquid chromatography/vitamin K2/lipase/milk powder/detection引用本文复制引用
基金项目
国家重点研发计划(2023YFF1104405-2)
出版年
2024
NETL
NSTL
万方数据