Determination of Vitamin K2 Content in Formulated Milk Powder by High Performance Liquid Chromatography
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.
high performance liquid chromatographyvitamin K2lipasemilk powderdetection
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