Vitamin B12 in BERTRAND

What exactly is the challenge?

Vegan nutrition has grown in popularity in recent years—for ethical, environmental, and health reasons. While a plant-based diet offers many benefits, it also presents challenges, particularly when it comes to obtaining vitamin B12. This essential vitamin, which is indispensable for the formation of red blood cells, the functioning of the nervous system, and energy metabolism, occurs naturally almost exclusively in animal-based foods.

For people following a vegan diet, obtaining vitamin B12 is often difficult. Although some plant-based foods, such as algae or fermented products, contain small amounts of B12, these are often inactive analogues that the body cannot use. These analogues may even block the absorption of active B12. That is why many people seek reliable, bioavailable, vegan sources of this essential vitamin—and this is exactly where our innovative approach comes in.

Where does the vitamin B12 in vegan BERTRAND come from?

A detailed look: extraction, purification, and detection of active vitamin B12 from shiitake

In the vegan varieties of BERTRAND, a very small amount of B12 comes from the microalga Chlorella.  However, the effective portion comes from the shiitake mushroom. These contain a mixture of bioactive forms of vitamin B12 (cobalamin) and inactive analogues. Active B12 (such as methylcobalamin, adenosylcobalamin, and hydroxycobalamin) can be used directly by the human body, while inactive analogues have a similar structure but no vitamin activity. To ensure safe and effective use, it must be guaranteed that only the active forms are present in the final product. Obtaining bioactive vitamin B12 from shiitake mushrooms involves several highly specialized steps:

Hydrolysis of the mushroom material

After harvesting, the fruiting bodies of the shiitake mushrooms are hydrolyzed mechanically and chemically. This means that the cell walls are broken down to release the nutrients contained within them, particularly the cobalamins. Water and special enzymes are used to support this process by breaking down the mushroom’s complex carbohydrates and proteins. This makes the vitamin B12 available.

Flocculation

In this step, the cobalamins are isolated from the mushroom material. Substances such as zinc and lime are used to help separate the cobalamins from other components such as proteins, polysaccharides, and possible inactive B12 analogues. The resulting solution is filtered to obtain a clear suspension of cobalamins.

Specialized purification and stabilization

The main focus is on the elimination of inactive B12 analogues. These analogues are removed through a combination of: chromatographic methods (e.g., ion-exchange chromatography): This method separates molecules based on their charge. Since inactive B12 analogues often have a slightly different chemical structure, they can be separated from the active forms.

Enzymatic purification: Specific enzymes are used here that act only on inactive B12 analogues, breaking them down or binding to them so that they can be removed.

After purification, a highly concentrated solution containing active forms of vitamin B12 remains.

What is pseudo-B12?

The difference between active and inactive forms of vitamin B12

Active forms of vitamin B12, such as methylcobalamin and adenosylcobalamin, are the two biologically active variants that the human body can use directly. Hydroxycobalamin, on the other hand, is a precursor that the body can readily convert into these active forms. These forms are essential for numerous vital functions, including energy metabolism, cell division, DNA synthesis, and nervous system function.

Inactive B12 analogues, by contrast, have a similar structure to active B12 but cannot be incorporated into the metabolism. One well-known example is pseudovitamin B12, which is commonly found in algae such as spirulina. These inactive analogues can even be problematic because they compete with active B12 for absorption by binding to transport proteins in the intestine, thereby reducing the availability of active B12.

To ensure that our product uses active forms of vitamin B12 that the body can use, the standardized test method AOAC 2011.09 was applied in the laboratory to determine the B12 content of our shiitake extract.

Laboratory verification

Many vegan products advertise sometimes very high vitamin B12 levels. However, as described above, these are often pseudo-forms of B12. To ensure that only active forms of vitamin B12 are present, standardized testing methods are used. One of the most important methods is AOAC 2011.09, a microbiological method specifically designed to detect and quantify bioactive vitamin B12.

In this method, a microorganism, Lactobacillus delbrueckii subsp. lactis, is cultured in a nutrient medium that is deliberately kept free of vitamin B12. The microorganism depends entirely on active vitamin B12 for its growth.

The growth, which can be measured through the turbidity or cell count of the medium, therefore provides a direct indication of the bioactive B12 content in the tested sample. The stronger the microorganism’s growth, the higher the proportion of active B12.

AOAC 2011.09 is highly specific, because only biologically active forms such as methylcobalamin, adenosylcobalamin, and hydroxycobalamin can promote growth. Inactive forms or analogues of vitamin B12 that are structurally similar but cannot be utilized do not contribute to the microorganism’s growth and can therefore be reliably excluded. This precise method ensures that the products contain exclusively bioavailable and effective forms of the vitamin.