Analytical Determination of Vitamin B₁₂Content in Infant and Toddler Milk Formulas by Liquid Chromatography Tandem Mass Spectrometry (LC-MS/MS)

Abstract

The development of a sample preparation method and optimization of the analytical instrumentation conditions were performed for the determination of the vitamin B12 content in emulsified baby foods sold on the Korea market. After removal of the milk protein and fats by chloroform extraction and centrifugation, the vitamin B12 was water extracted from the sample. Following filtration of the solution through a nylon filter, the water-soluble extract was purified by solid-phase extraction using a Liquid Chromatography Tandem Mass Spectrometry (LC-MS/MS). The solution eluted from the cartridge was dried under a stream of nitrogen gas and reconstituted with 1 mL of water. The sample solution was injected into an LC-MS/MS system after optimizing the mobile phase for vitamin B12 detection. The calibration curve showed good linearity with the coefficient of correlation (r2) value of 0.9999. The limit of detection was 0.03 µg/L and the limit of quantitation was 0.1 µg/L. The method of detection limit was 0.02 µg/kg. The vitamin B12 recovery from a spiking test was 99.62% for infant formula and 99.46% for cereal-based baby food. The sample preparation method developed in this study would be appropriate for the rapid determination of the vitamin B12 content in infant formula and baby foods with emulsified milk characteristics. The ability to obtain stable results more quickly and efficiently would also allow governments to exercise a more extensive quality control inspection and monitoring of products expected to contain vitamin B12. This method could be implemented in laboratories that require time and labor saving.

Development of Pretreatment Method for Analysis of Vitamin B12 in Cereal Infant Formula using Immunoaffinity Chromatography and High-Performance Liquid Chromatography

“…In addition, it is difficult to detect vitamin B 12 in some food samples using only one pretreatment method ( http://foodsafetykorea.go.kr/foodcode/01_03.jsp?idx=324 ). Since vitamin B 12 exists in different forms at very low concentrations in powdered milk containing cereal, the sample preparation methodology is extremely crucial ( Lee et al, 2015 ). In this study, individual pretreatment methods were developed by modifying the Association of Official Analytical Chemists ( AOAC, 2002 ) method to detect vitamin B 12 in powdered milk containing starch in high amounts.…”

“…Especially, powdered milk add starch as well as protein and/or fat to improve its nutrition value ( Seo et al, 2018 ). The presence of these additional components renders the analysis of vitamin B 12 extremely difficult ( Bito et al, 2016 ; Lee et al, 2015 ). Hence, the analysis of vitamin B 12 in powder milk must include a pretreatment step.…”

Influence of food groups on plasma total homocysteine for specific MTHFR C677T genotypes in Chinese population

“…Serum folate concentration was measured using a dual count Solid Phase Boil Radioassay (Diagnostic Products, Los Angeles, CA). Serum Vit B12 was detected by liquid chromatography–tandem mass spectrometry (HPLC‐MS/MS, Agilent Model 6410) . The blood glucose levels, including FPG(fasting plasma glucose) and G2h (postprandial 2 hours blood glucose) were measured using the hexokinase method; the total cholesterol (TC), high‐density lipoprotein (HDL), low‐density lipoprotein (LDH), triglyceride (TG), hemoglobin (HGB), blood uric acid (BUA), alanine aminotransferase (ALT), aspartate transaminase (AST), and blood creatinine (Cr) were measured with an autoanalyzer (Cobas c 501 autoanalyzer, Roche Diagnostics, Germany).…”

Determination of Vitamin B12 in Milk and Dairy Products by Isotope-Dilution Liquid Chromatography Tandem Mass Spectrometry

“…e absolute recovery of immunoaffinity SPE cartridge for vitamin B 12 is not very well; meanwhile, quality differences between batches would influence the accuracy of determination. Luo [24] developed an analytical method for the determined of vitamin B 12 in food products and multivitamin multimineral tablets by HPLC-ESI-MS, and ginsenoside Re was used as an internal standard (I.S.). Ginsenoside Re is not a perfect internal standard of vitamin B 12 for the obvious contrast between the molecular structure of ginsenoside Re and vitamin B 12 .…”

“…e existing reported methods for determination of VB 12 in infant formula mainly include microbiological assays [9,10], high-performance liquid chromatography (HPLC) [11][12][13][14][15][16][17][18][19], liquid chromatography-tandem mass spectrometry (LC-MS/MS) [20][21][22][23][24][25][26], and liquid chromatography and inductively coupled plasma-mass spectrometry (LC-ICP-MS) [27][28][29]. e microbiological assays usually have a long measurement time and complicated operation.…”