Modified EMR-lipid method combined with HPLC-MS/MS to determine folates in egg yolks from laying hens supplemented with different amounts of folic acid

Abstract Egg yolks are a good source of folates. However, the method for analyzing the naturally occurring folates in egg yolks is complicated and time-consuming. In this study, a simplified pre-treatment method followed by validated HPLC–MS/MS was developed to determine native folates in eggs from laying hens treated with different amounts of folic acid. The modified enhanced matrix removal –lipid method to purify samples showed good performance in lipid elimination, reduction of steps and time savings. According to experimental analysis, yolks contained total folate amounts ranging from 147 to 760 μg/100 g when laying hens’ diet was supplemented with folic acid from 0 to 10 mg/kg. Four folate vitamers were detected in egg yolks: 5-methyltetrahydrofolate accounted for 91–98% of total folates, whereas folic acid, 5-formyltetrahydrofolate and 10-formylfolic acid together accounted for 2–9%. Therefore, laying hens efficiently converted folic acid in feed into 5-methyltetrahydrofolate in eggs with little folic acid deposition.