Short Communication

Development of pretreatment method for analysis of vitamin B12 in cereal infant formula using immunoaffinity chromatography and high-performance liquid chromatography

jung min Park1, Jong Ho Koh2, Jin Man Kim1,*
Author Information & Copyright
1Konkuk University, Seoul 05029, Korea.
2Korea Polytechnic University, Nosan 32940, Korea.
*Corresponding Author: Jin Man Kim, Konkuk University, Seoul 05029, Korea, E-mail:

© Copyright 2020 Korean Society for Food Science of Animal Resources. This is an Open-Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License ( which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.

Received: Sep 03, 2020 ; Revised: Nov 05, 2020 ; Accepted: Nov 16, 2020

Published Online: Nov 23, 2020


Vitamin B12 deficiency may lead to serious health issues in both infants and adults. A simple analytical method involving sample pretreatment with enzyme, followed by cyanide addition under acidic conditions; separation on an immunoaffinity column; and high-performance liquid chromatography was developed for the rapid detection and quantitation of vitamin B12 in powdered milk. Detection limit and powdered milk recovery were determined by quantitative analysis. The limits of detection and quantitation were 2.71 and 8.21 μg/L, respectively. Relative standard deviations of the intra-day and inter-day precisions varied in the ranges of 0.98–5.31% and 2.16–3.90%, respectively. Recovery of the analysis varied in the range of 83.41–106.57%, suggesting that the values were acceptable. Additionally, vitamin B12 content and recovery in SRM 1849a were 54.10 μg/kg and 112.24%, respectively. Our results suggested that the analytical method, including the sample pretreatment step, was valid. This analytical method can be implemented in many laboratory-scale experiments that seek to save time and labor. Therefore, this study shows that immunoaffinity–high-performance liquid chromatography/ultraviolet is an acceptable technique for constructing a reliable database on vitamin B12 in powdered milk containing starch as well as protein and/or fat in high amounts.

Keywords: vitamin B12; powdered milk; HPLC; analytical method

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The name of the journal has been changed from Korean Journal for Food Science of Animal Resources to Food Science of Animal Resources from January 2019.

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