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Development of Pretreatment Method for Analysis of Vitamin B12 in Cereal Infant Formula using Immunoaffinity
Jung Min Park1, Jong Ho Koh2, Jin Man Kim1
1Department of Food Marketing and Safety, Konkuk University, Seoul 05029, Korea.
Insights
A new analytical method accurately detects vitamin B12 in powdered milk using immunoaffinity-HPLC. This rapid, labor-saving technique ensures reliable vitamin B12 quantitation, crucial for infant and adult health.
Area of Science:
- Analytical Chemistry
- Food Science
- Biochemistry
Background:
- Vitamin B12 deficiency poses significant health risks.
- Accurate quantitation of vitamin B12 in food matrices like powdered milk is essential for public health.
- Existing methods may be time-consuming or labor-intensive.
Purpose of the Study:
- To develop a rapid, reliable, and labor-efficient analytical method for vitamin B12 detection in powdered milk.
- To validate the method's accuracy, precision, and recovery rates.
- To establish a technique suitable for constructing a vitamin B12 database for various powdered milk compositions.
Main Methods:
- Sample pretreatment involving enzymatic treatment and cyanide addition under acidic conditions.
- Separation using an immunoaffinity column.
- Quantitation via high-performance liquid chromatography (HPLC) with ultraviolet detection.
Main Results:
- The method achieved detection and quantitation limits of 2.71 μg/L and 8.21 μg/L, respectively.
- Intra-day and inter-day precisions showed low relative standard deviations (0.98%-5.31% and 2.16%-3.90%).
- Recovery rates ranged from 83.41% to 106.57%, indicating acceptable accuracy. Analysis of SRM 1849a yielded a vitamin B12 content of 54.10 μg/kg with 112.24% recovery.
Conclusions:
- The developed immunoaffinity-HPLC method is a valid and reliable technique for vitamin B12 analysis in powdered milk.
- The method is efficient, saving time and labor, making it suitable for laboratory-scale applications.
- This technique facilitates the creation of a dependable vitamin B12 content database, particularly for complex powdered milk formulations.
Abstract:
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 (HPLC) 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-HPLC/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.
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