Vitamin B12 quantification in human milk – Beyond current limitations using liquid chromatography and inductively coupled plasma – Mass spectrometry

Dubascoux, Stéphane; Payot, Janique Richoz; Sylvain, Paul; Nicolas, Marine; Giménez, Esther

doi:10.1016/j.foodchem.2021.130197

Cited by 14 publications

(10 citation statements)

References 32 publications

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“…Samples were hydrolysed in hydrochloric acid (100 °C, 35 min), neutralized, and filtered (0.22 µm, Millipore). The subsequent analysis was based on method LI-00.610 [ 78 ] using UHPLC-MS/MS with positive electrospray ionization and quantification via the peak area ratio of each analyte and an internal standard (Nestle Quality Assurance Center, Dublin, Republic of Ireland).…”

Section: Methodsmentioning

confidence: 99%

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Co-cultures of Propionibacterium freudenreichii and Bacillus amyloliquefaciens cooperatively upgrade sunflower seed milk to high levels of vitamin B12 and multiple co-benefits

et al. 2022

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Background Sunflower seeds (Helianthus annuus) display an attractive source for the rapidly increasing market of plant-based human nutrition. Of particular interest are press cakes of the seeds, cheap residuals from sunflower oil manufacturing that offer attractive sustainability and economic benefits. Admittedly, sunflower seed milk, derived therefrom, suffers from limited nutritional value, undesired flavor, and the presence of indigestible sugars. Of specific relevance is the absence of vitamin B12. This vitamin is required for development and function of the central nervous system, healthy red blood cell formation, and DNA synthesis, and displays the most important micronutrient for vegans to be aware of. Here we evaluated the power of microbes to enrich sunflower seed milk nutritionally as well as in flavor. Results Propionibacterium freudenreichii NCC 1177 showed highest vitamin B12 production in sunflower seed milk out of a range of food-grade propionibacteria. Its growth and B12 production capacity, however, were limited by a lack of accessible carbon sources and stimulants of B12 biosynthesis in the plant milk. This was overcome by co-cultivation with Bacillus amyloliquefaciens NCC 156, which supplied lactate, amino acids, and vitamin B7 for growth of NCC 1177 plus vitamins B2 and B3, potentially supporting vitamin B12 production by the Propionibacterium. After several rounds of optimization, co-fermentation of ultra-high-temperature pre-treated sunflower seed milk by the two microbes, enabled the production of 17 µg (100 g)−1 vitamin B12 within four days without any further supplementation. The fermented milk further revealed significantly enriched levels of l-lysine, the most limiting essential amino acid, vitamin B3, vitamin B6, improved protein quality and flavor, and largely eliminated indigestible sugars. Conclusion The fermented sunflower seed milk, obtained by using two food-grade microbes without further supplementation, displays an attractive, clean-label product with a high level of vitamin B12 and multiple co-benefits. The secret of the successfully upgraded plant milk lies in the multifunctional cooperation of the two microbes, which were combined, based on their genetic potential and metabolic signatures found in mono-culture fermentations. This design by knowledge approach appears valuable for future development of plant-based milk products.

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Section: Methodsmentioning

confidence: 99%

“…The level of cobalt was quantified after sample work up by a high pressure asher (HPA-S High Pressure Asher, Anton Paar, IGZ Instruments, Zurich, Switzerland) using ICP-MS [ 78 , 80 ].…”

Section: Methodsmentioning

confidence: 99%

Co-cultures of Propionibacterium freudenreichii and Bacillus amyloliquefaciens cooperatively upgrade sunflower seed milk to high levels of vitamin B12 and multiple co-benefits

et al. 2022

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“…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.…”

Section: Introductionmentioning

confidence: 99%

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

Huang

Zhang

Cai

2022

Journal of Food Quality

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An isotope-dilution liquid chromatography tandem mass spectrometry method was established for the determination of vitamin B12 in milk and dairy products. The samples were spiked with stable isotope-labeled vitamin B12 and digested by pepsin and amylase. The various forms of cobalamin were transformed to cyanocobalamin by potassium cyanide after they were released from the enzymatically digested samples. Cyanocobalamin was extracted and purified by an immunoaffinity SPE cartridge and then measured in multiple reaction monitoring mode (MRM). The linear correlation coefficient (R2) of this method was greater than 0.999 in the range of 2–100 ng/mL. The detection limit and the quantification limit were 0.5 μg/kg and 1.0 μg/kg, respectively. The spiked recoveries ranged from 92.0% to 99.4% at the three spiked levels with the relative standard deviation (RSD) between 1.89% and 4.51%. The measured results of NIST SRM1849a and NIST SRM1869a by the current method are all within the reference value range. The Z value was 0.8 during participating in the FAPAS proficiency test using the developed method in 2021. The method is simple, rapid, accurate, and sensitive, and it is suitable for the determination of vitamin B12 in different types of milk and dairy products such as whey powder, whole milk powder, pure milk, fermented milk, infant formula, and prescription food for special medical purposes.

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“…Current analysis of cobalamins in foods are carried out using various techniques, among which the microbiological assay (MBA) and high-performance liquid chromatography (HPLC)-UV − being the most common ones. Recently, a new method using liquid chromatography (LC) and inductively coupled plasma (ICP)-MS has been developed for analyzing B12 in human milk . These approaches, however, suffer from different drawbacks.…”

Section: Introductionmentioning

confidence: 99%

“…Recently, a new method using liquid chromatography (LC) and inductively coupled plasma (ICP)-MS has been developed for analyzing B12 in human milk. 18 These approaches, however, suffer from different drawbacks. The MBA is based on the growth of bacteria, such as L. leichmannii, which reveals a reasonable sensitivity but lacks the specificity and selectivity.…”

Section: Introductionmentioning

confidence: 99%

Development of Stable Isotope Dilution Assays for the Analysis of Natural Forms of Vitamin B12 in Meat

Wang

Asam

Chen

et al. 2021

J. Agric. Food Chem.

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The first multiple stable isotope dilution assay method was developed for the simultaneous determination of four cobalamins, namely, hydroxocobalamin (OHCbl), adenosylcobalamin (AdoCbl), methylcobalamin (MeCbl), and cyanocobalamin (CNCbl), in their native forms. The sample preparation was optimized with enzyme treatment and immunoaffinity purification. The analysis was performed by LC-MS/MS using respective 15 N-labeled cobalamins as internal standards. Method validation resulted in limits of detection ranging from 0.19 to 0.58 ng/g and limits of quantification ranging from 0.68 to 1.73 ng/g. Recoveries at three levels were between 82 and 121%. Intra-day and inter-day precisions were below 6% and 11% RSD, respectively. The analysis of a reference material resulted in a variance of <1% from the certified value. The newly developed method demonstrated excellent sensitivity, recovery, accuracy, and reproducibility and was further applied to quantitate the four cobalamins in various meats.

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Vitamin B12 quantification in human milk – Beyond current limitations using liquid chromatography and inductively coupled plasma – Mass spectrometry

Cited by 14 publications

References 32 publications

Co-cultures of Propionibacterium freudenreichii and Bacillus amyloliquefaciens cooperatively upgrade sunflower seed milk to high levels of vitamin B12 and multiple co-benefits

Co-cultures of Propionibacterium freudenreichii and Bacillus amyloliquefaciens cooperatively upgrade sunflower seed milk to high levels of vitamin B12 and multiple co-benefits

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

Development of Stable Isotope Dilution Assays for the Analysis of Natural Forms of Vitamin B12 in Meat

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