Vitamin B12 is a water-soluble vitamin belonging to the family of cobalamins having a cobalt ion in the center of the corrin ring. It exists in several forms including cyano-, methyl-, hydroxyl-, and adenosyl-cobalamins. There is lack of national database on vitamin B12 in food consumed in Korea. Th...
Vitamin B12 is a water-soluble vitamin belonging to the family of cobalamins having a cobalt ion in the center of the corrin ring. It exists in several forms including cyano-, methyl-, hydroxyl-, and adenosyl-cobalamins. There is lack of national database on vitamin B12 in food consumed in Korea. This study was performed to validate the immunoaffinity-HPLC/DAD method and apply for analysis of vitamin B12 in noodles, cereals, wheat products, dairy products, and meat products. Vitamin B12 was extracted by enzyme treatment(α-amylase and pepsin) and heating in the presence of cyanide at acidic condition. The vitamin B12 extract was cleaned up and concentrated on an immunoaffinity column(IAC) and then analyzed by HPLC-DAD. The analytical method was validated by the determination of analytical performance parameters. Accuracy and precision were good with high recovery and low %RSD, which meets the acceptable guidelines by AOAC. Limits of detection(LOD) and quantification(LOQ) were 0.01 and 0.03 μg/100 g, respectively. The QC chart obtained by running infant formula with food sample once a week showed that all assay was under the control during the entire of study. The z-score of the IAC-HPLC/DAD assay for vitamin B12 in FAPAS Proficiency Test was less than 2, assuring reliability of analytical performance. Vitamin B12 content in noodles, cereals, wheat products, daily products, and meat products ranged by 0.03-2.67, 0.03-2.67, 0.02-0.25, 0.05-1.11 and 0.20-2.79 μg/100 g, respectively. Method validation and analytical quality control data proved that analytical vitamin B12 data produced using the IAC-HPLC/DAD is reliable and could be used for the construction of national vitamin B12 database for the selected Korean foods.
Vitamin B12 is a water-soluble vitamin belonging to the family of cobalamins having a cobalt ion in the center of the corrin ring. It exists in several forms including cyano-, methyl-, hydroxyl-, and adenosyl-cobalamins. There is lack of national database on vitamin B12 in food consumed in Korea. This study was performed to validate the immunoaffinity-HPLC/DAD method and apply for analysis of vitamin B12 in noodles, cereals, wheat products, dairy products, and meat products. Vitamin B12 was extracted by enzyme treatment(α-amylase and pepsin) and heating in the presence of cyanide at acidic condition. The vitamin B12 extract was cleaned up and concentrated on an immunoaffinity column(IAC) and then analyzed by HPLC-DAD. The analytical method was validated by the determination of analytical performance parameters. Accuracy and precision were good with high recovery and low %RSD, which meets the acceptable guidelines by AOAC. Limits of detection(LOD) and quantification(LOQ) were 0.01 and 0.03 μg/100 g, respectively. The QC chart obtained by running infant formula with food sample once a week showed that all assay was under the control during the entire of study. The z-score of the IAC-HPLC/DAD assay for vitamin B12 in FAPAS Proficiency Test was less than 2, assuring reliability of analytical performance. Vitamin B12 content in noodles, cereals, wheat products, daily products, and meat products ranged by 0.03-2.67, 0.03-2.67, 0.02-0.25, 0.05-1.11 and 0.20-2.79 μg/100 g, respectively. Method validation and analytical quality control data proved that analytical vitamin B12 data produced using the IAC-HPLC/DAD is reliable and could be used for the construction of national vitamin B12 database for the selected Korean foods.
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