Innovative Folate

With Quatrefolic® Gnosis by Lesaffre has managed to develop an innovative product with significant advantages over previous folate generations while maintaining the same biological properties of 5-methyltetrahydrofolate. This remarkable step forward is attributed to the development of the glucosamine salt of Quatrefolic®, through two main steps:

  • Improving solubility in water of 5-methyltetrahydrofolate, Quatrefolic® is defined as water soluble, a significant opportunity for improving bioavailability, whereas the calcium salt version is defined as sparingly soluble.

  • Guaranteeing a safety and high-quality stable profile by choosing glucosamine as natural compound naturally present in the body.


What is glucosamine salt
made from?

It is a natural compound endogenously formed throughout the human body by cellular glucose metabolism and found in almost all human tissues. Glucosamine is made up of glucose and amino acid glutamine and it is a key building block for several structures. In the body it is needed to produce glycosaminoglycans, molecules used in the formation and reparation of cartilage and other body tissues.

Glucosamine has an extremely safe profile and it has been recognized as GRAS for its use as a nutritional supplement to promote joint health. It has been widely used as a human dietary supplement for years, both alone and in combination with other products.


As a result of our clinical research, Quatrefolic® can claim key relevant benefits allowing the product to be suitable for dietary supplements as well as food and beverage uses, specifically for combinations of different vitamins and minerals.

Quatrefolic® is manufactured under controlled conditions according to cGMP, and since the glucosamine is animal- and shellfish-free, Quatrefolic® is an excellent option for vegetarians.

The maximum flexibility and versatility for the best quality finished products

Long-lasting stability

Quatrefolic® shows a long-lasting chemical stability at both room and refrigerated conditions, guaranteeing purity and ensuring easy handling and storage. Stability at room temperature might be a useful indicator to predict shelf life especially if there is potential for combining the Quatrefolic® with other components such as vitamins and minerals.

Stability studies of Quatrefolic® have been performed according to ICH (International Conference on Harmonisation of Technical Requirements for Registration of Pharmaceuticals for Human Use) guidelines. Entering into its third decade of activity, the Global Quality Expert Working Group of ICH undertakes to rationalise and harmonise regulation in the world (the USA, Europe and Japan) to ensure that safe, effective, and high quality medicines are developed and registered in the most resource-efficient manner.

Gnosis by Lesaffre has deliberately chosen to carry out stability tests of Quatrefolic® according to the most strict guidelines, specifically required for pharmaceutical ingredients guaranteeing undependable and reliable criteria to claim the long-lasting chemical stability to its clients.

Improved Bioavailability

Based on the superior water solubility of Quatrefolic®, Gnosis by Lesaffre has undertaken several pre-clinical and clinical studies in order to evaluate and assess the improved bioavailability of Quatrefolic® in comparison to the (6S)-5-methyltetrahydrofolate calcium salt. Several protocols have been carried out both on animal and human models. The first test was a direct comparison between Quatrefolic®, (6S)-5-methyltetrahydrofolate calcium salt and folic acid in rats that has allowed to know the difference of bioavailability in-vivo between the three folate generations. Once the animal studies were completed, a human clinical trial was performed, confirming Quatrefolic® owns a superior bioavailability profile over the (6S)-5-methyltetrahydrofolate calcium salt. It was a single dose, balanced, two sequences, two periods, two treatments randomised crossover study.

(6S)-5-methyltetrahydrofolate haematic peak produced by an equivalent dose of the indicated folate source.

Higher water solubility

In addition to the high chemical stability, Quatrefolic® demonstrates a surprisingly high solubility in water, compared with the slight solubility of the reference compound (6S)-5-methyltetrahydrofolate calcium salt. Solubility in water is extremely important for an active ingredient considering the human body is composed of more than 65% of water and that the absorption of the compound may happen after dispersion and solution in gastrointestinal fluid. High water solubility means the product may be better absorbed by mucosal cells which may facilitate access to the blood and circulation with the potential for improved bioavailability.

Comparison of solubility in water has been performed evaluating the quantity of solvent needed to solubilise the same amount of each folate derivative, at standard temperature. Quatrefolic® is solubilised by a quantity of water 100 times lower than calcium salt. The improved solubility of Quatrefolic® over the (6S)-5-methyltetrahydrofolate calcium salt represents another advantage achieved by Gnosis by Lesaffre in creating a product for applications in the food and beverage industries.

Established Safety

The safety of 5-methyltetrahydrofolate has been extensively investigated in a number of published and unpublished experimental animal studies and human clinical trials. Furthermore glucosamine itself has GRAS status. Quatrefolic®, as glucosamine salt of (6S)-5-methyltetrahydrofolate has been the subject of an extensive and relevant number of biological and toxicological studies in order to prove the safety and tolerability of this new revolutionary folate. Gnosis by Lesaffre has performed several in vitro and in-vivo studies such as mutagenicity, genotoxicity and acute toxicity on Quatrefolic® and the product has met all the safety requirements.

Toxicological studies


  • Bacterial mutation in S.typhimurium and E.coli

    The bacterial mutation assay was performed in order to assess the compound’s ability to induce gene mutations in S.typhimurium and E.coli. The reverse mutation assay was run in bacterial strains already mutant at a locus whose phenotypic effects are easily detected, and, since many chemicals can demonstrate mutagenic activity only after metabolism to some reactive forms, the test was performed in the presence and in absence of a rat liver metabolic system (S9 microsomal fraction).

  • Mutation in L5178YTK+/- mouse lymphoma cells

    The essay was done in order to confirm the inability of Quatrefolic® to induce mutations in L5178YTK+/- mouse lymphoma cells cultured after in vitro treatment, and in absence or presence of a rat liver microsomal system.

  • Chromosome aberrations in Chinese hamster ovary cells (CHO) in vitro

    The essay was made in order to demonstrate the inability of Quatrefolic® to induce any chromosomal aberration in presence or absence of a S9 liver microsomal fraction.


  • Single dose oral toxicity

    The acute toxicity of Quatrefolic® was assessed in rats of both sexes, dosing the product by gavage at 500 mg/kg level. No mortality occurred at this dose and during the observation time, and no clinical significant signs were observed in any animal. Changes in body weight were not relevant, and no anomalies were recorded at the autopsy performed after the observation time. The lack of mortality points out that the maximum tolerated dose is a dose some thousand times the one suggested for human use.

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