Optimization of Method for Folate Content Determination in Eggs and Study on Its Enrichment Rule

BAI Yan; WANG Rui; YANG Yu; LI Ruirui; WU Xiaotian

doi:10.3969/j.issn.1006-267x.2021.10.043

Chinese Journal of Animal Nutrition >

2021 , Vol. 33 >Issue 10: 5838 - 5848

DOI: https://doi.org/10.3969/j.issn.1006-267x.2021.10.043

EXPERIMENTAL METHOD AND ANIMAL

Optimization of Method for Folate Content Determination in Eggs and Study on Its Enrichment Rule

BAI Yan ,
WANG Rui ,
YANG Yu ,
LI Ruirui ,
WU Xiaotian

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1. College of Animal Science, Shanxi Agricultural University, Taigu 030801, China;
2. Department of Life Sciences, Lyuliang University, Lyuliang 033000, China

Received date: 2021-03-18

Online published: 2021-10-16

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Abstract

This experiment aimed to develop a simple, accurate and relatively fast determination method for folate in eggs, to study the effect of dietary folate supplementation on the folate content in eggs, and to study the enrichment rules of folate in eggs during the period of egg-laying. Experimental methods:1) The optimal detection method was determined by comparing the performance of fluorescence detector and ultraviolet detector of high-performance liquid chromatography, and comparing the detection value of folate in yolks extracted by sonicate, heat treat (water-bath at 90℃), enzymatic hydrolysis and their combination. 2) The suitable dietary folic acid supplemental level for folate-enriched eggs production was screened under the feeding experiment lasted for 35 days. Six hundred and forty-eight Jinghong laying hens (30 weeks of age) were randomly divided into 6 groups with 6 replicates in each group and 18 hens in each replicate. Laying hens in one group (control group) was fed a basal diet, while those in the other five groups were fed the basal diet supplemented with 1, 3, 6, 12 and 24 mg/kg folic acid, respectively. 3) Given these results of screening test, a long-term experiment was conducted by feeding 16 000 laying hens (20 to 60 weeks of age) with 7.5 mg/kg folic acid supplementation in the diet, and detecting the content of folate in 10 egg monitoring points at 20, 21, 22, 23, 24, 25, 27, 29, 33, 37, 41, 46, 50, 55 and 60 weeks of age. The results showed as follows:1) the limit of detection (LOD) and the limit of quantitation (LOQ) of fluorescence detector were lower, and the peak area and peak height of fluorescence detector were extremely significantly higher than those of ultraviolet (UV) detector (P<0.01). The detected value of folate content in yolk extracted by heat treatment and sonicate for 5 min+heat treatment was significantly higher than that extracted by sonicate for 5 min+enzymolysis and heat treat treatment+enzymolysis (P<0.05). The best condition for extracting folate from yolk was 5 min of sonicate followed by 5 min of 90℃ water bath, and the fluorescence detector had good detection performance. The relative standard deviation (RSD) of precision and stability were 2.15% and 0.57%, respectively; the spiked recoveries averaged 98.01%. 2) The folate content in eggs of 6 and 12 mg/kg groups were extremely significantly higher than those of control group and 1 mg/kg group (P<0.01), and there was no significant difference with 3 and 24 mg/kg groups (P>0.05). Regression analysis showed that there was an extremely significant cubic linear relationship between dietary folic acid supplemental level and whole egg folate content (determinate coefficient=0.679; P<0.001). 3) Adding 7.5 mg/kg folic acid to laying hen diet from 20 to 60 weeks of age, the content of folate in yolk tended to be saturated at 23 weeks of age, and the content of folate in whole egg gradually increased from 36.9 μg/egg at 20 weeks of age to 87.7 μg/egg at 33 weeks of age, then remained stable. In conclusion:1) the egg folate detection method determined in this experiment is simple to operate, high accuracy accurate and rapid. 2) Due to add 7.5 mg/kg folic acid to diet from 20 weeks of age, the folate content in yolk remains at more than 5 μg/g from 23 weeks of age, and the folate content in whole egg remains at more than 85 μg/egg from 33 weeks of age.

Key words： yolk; 5-methyltetrahydrofolate; high performance liquid chromatography; folate enrichment

Cite this article

BAI Yan , WANG Rui , YANG Yu , LI Ruirui , WU Xiaotian . Optimization of Method for Folate Content Determination in Eggs and Study on Its Enrichment Rule[J]. Chinese Journal of Animal Nutrition, 2021 , 33(10) : 5838 -5848 . DOI: 10.3969/j.issn.1006-267x.2021.10.043

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