Inside the Science of Vitamin B12 Testing
What are the challenges of testing for Vitamin B12?
There are several challenges to testing Vitamin B12 (cobalamins) that create issues for analytical laboratories to accurately measure these compounds. The challenges include the relative low levels generally found in various product matrices, instability of certain cobalamins, complexity of product matrices, and the lack of standardized industry test methods to name a few. Eurofins Supplement Testing in Madison, WI understands these challenges and has consistently strived to develop updated methodologies utilizing state-of-the-art technology to improve overall method performance.
How has Eurofins addressed these challenges?
Eurofins has recognized that Vitamin B12 methods over the years have performed inadequately in certain complex matrices, especially for forms like methylcobalamin. With the breadth of experience and understanding the need to develop a more robust, specific, and accurate way to test Vitamin B12 forms was made a priority. Where other laboratories may be limited to more traditional test methods for Vitamin B12 like microbiological assays, spectrophotometry, or chromatographic techniques, Eurofins has recently added a more precise and specific methodology to their toolbox. This more advanced technique utilizes High Performance Liquid Chromatography coupled with Inductively Coupled Plasma Tandem Mass Spectrometry (HPLC-ICP-MSMS) to leverage cobalt as the inherent atom in the structure of cobalamins. By doing so, it greatly reduces the background noise and matrix interferences that plague other less advanced methods and allows for more accurate quantitation.
Why is Methylcobalamin so difficult to test for?
Methylycobalamin is a difficult form of Vitamin B12 to measure accurately, primarily due to its instability under certain exposures to light, heat, and pH extremes. This instability creates a pathway for methylcobalaimn to convert to other forms of Vitamin B12 (hydroxocobalamin, adenosylcobalamin) during manufacturing, production, and even during analysis of the test method. The utmost care must be taken when handling the product in the laboratory to prevent these unwanted conversions taking place. Methylcobalamin will often convert to hydroxocobalamin, making it difficult to determine the original amount of methylcobalamin from its degradation products. Eurofins has multiple methods of analysis for Vitamin B12 and can use numerous techniques depending on the forms, levels, and varying matrices that are available in the market.
How can I better understand the stability of Vitamin B12 in my product?
Vitamin B12 is found in multiple active forms and can have varying degrees of instability. Cyanocobalamin is the most stable form of Vitamin B12 and does not pose the high risk of degradation that other forms, like methylcobalamin, may exhibit. To assess the stability (or instability) of cobalamins in your product may be best suited in a stability study specifically designed to assess the conversions to other breakdown components. Eurofins can design a scientifically sound formalized plan to help assess any changes to your cobalamin levels over time. This will allow you to make proper formulation decisions and produce high quality products.
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