In 2018, we commissioned the Muséum d’Histoire Naturelle of Paris and the University of Normandy in 2019 for a series of scientific analyses in chromatography and mass spectrometry. This researches aim to demonstrate the presence of proteins and peptides in the snail slime that we manually harvest and that we process by low pressure cold extraction.
Chromatographic and mass spectrometric analyses of our snail filtrate extract
Harvest and Extraction
Step 1 : Breeding
The entire cycle of organic farming of our snails (Hélix Aspersa Maxima - Gros Gris) takes place in Le Creux de la Tine, in an exceptional region between Drôme and Ardèche. We do not outsource any of our work. Patrice, our snail breeder, was the first in France to be certified in Organic Agriculture.
Step 2 : Respect of the snail’s life cycle
Snail slime is a antic secret. Ancient Greece recognized its benefits very early on. And at this time, it was identified as an effective solution with properties: healing, regenerating, highly moisturising or even exfoliating. These are the main curative and cosmetic powers attributed to it as by today. After carrying out our own research with the Muséum d’Histoire Naturelle in Paris and the University of Rouen, we were able to confirm all these benefits known for more than 2,500 years by the presence of very interesting proteins for the skin (see our page “scientific research”). For all these reasons, snail slime is the main ingredient of our organic skin care products.
Step 3 : Manual harvest
This is the most delicate step. It requires great dexterity because if the animal is stressed it will produce a foamy and liquid slime without viscosity. Only this manual harvesting process can guarantee the welfare of the animal. The manual- collected spawn mucus (an average 3 gr per snail) is very thick and viscous with a lot of protein bonds. (see page 100% manual harvest).
Step 4 : Mucus stabilization
The low pressure cold extraction process for mucus allows for filtration without causing an emulsion to form and the product to become oxidized. This extraction system helps preserve the water and lipid-soluble parts of the mucus. This is a key point in the manufacturing process, which lowers the number of bacteria naturally present in the filtrate without damaging the nutritional qualities and organoleptic characteristics of the mucus.
MUSÉUM D’HISTOIRE NATURELLE, PARIS
Since Antiquity, snails have been considered as a natural source of many therapeutic and biological properties. Today, snail filtrate extract is revolutionizing the world of cosmetics and skincare. The regenerative and antioxidant properties of Helix Aspersa Maxima (Gros Gris) secretion have been identified by several studies (Brieva et al 2008 - Iglesias de la Cruz et al 2012). The topical application of a cream with a snail filtrate base to cutaneous burns shows an improvement in the healing process of open wounds by accelerating skin regeneration activity and decreasing pain (Tsoutsos et al 2009).
Our scientific analyses aim at highlighting the wealth of proteins in the snail filtrate used in Mademoiselle Agathe cosmetics, as well as validating, through proteomics, the manual harvesting system and low pressure cold extraction method that are specific to the Mademoiselle Agathe brand.
Like all successful markets, the cosmetic sector using snail filtrate formulas is increasingly buoyant in Europe and worldwide. It is therefore important for a fast-growing brand such as Mademoiselle Agathe to provide product traceability (breeding location, harvesting conditions, extraction method, etc.), while adopting a solid scientific approach.
The database research shows that the precipitation protocol is effective. Hemocyanin and actin are the two proteins that are primarily present. Approximately one hundred proteins were detected with the identification of 23 proteins. The research carried out in bibliographies on the role of the identified proteins show that they all have a link to the skin, its immune system or its collagens.
PROTEINS LINKED TO THE SKIN
Proteins are highly diverse molecules, and our body produces approximately 100,000 different ones. Each of these proteins has a three-dimensional shape and specific chemical properties. They play many roles in human cells, performing a vast array of functions, including: Structure: protein fibers, Movement, Transportation of substances in the blood, Transportation of substances through cell membranes Hormones, Cell identification, Defense: antibodies, Enzymes.
PROTEINS LINKED TO THE SKIN’S IMMUNE SYSTEM
These proteins play a role in the body’s immune system and consist of a coordinated set of recognition and defense elements. An immune response is activated when our body is confronted with a stress factor or disorder, leading to immune resilience. This concept covers all the body’s effective defense mechanisms in the face of a pathogenic agent. These proteins are part of natural, nonspecific defense mechanisms. The skin is the first and most important element in our defense system, as it prevents most foreign particles from penetrating the body.
PROTEINS LINKED TO COLLAGEN
These proteins are part of the structural protein family, most often found in fibrillar form. They are involved in cell migration, as well as play a role in adhesive junctions by participating in the cohesion of epithelial cells along their basal sides. These proteins and collagen form a network, which is the backbone of the basal membrane in order to maintain the structure and function of tissues. Cytoskeletons. The majority of cells have an internal framework of protein fibers. This cytoskeleton is responsible for the particular shape of each cell and its level of resistance to observed tensions and movements.
COLLAGEN PROTEINS Collagen is a fibrous protein whose main function is to provide tissue with mechanical resistance to extension. It is the most abundant protein in the animal kingdom, accounting for approximately one quarter of our total protein mass. The collagen family includes 23 types of collagen, composed of 38 distinct chains. This is why we should speak about collagens rather than collagen in singular form. Each type of collagen has its own structure and is found in specific organs. However, in snail filtrate, collagens I, V and VI are linked to connective tissue, such as skin.
ROUEN UNIVERSITY (Normandy) – November 2019
STUDY OF THE ANTIBACTERIAL ACTIVITY OF THE SNAIL SLIME
A bacteriostatic effect of the purifying gel for acne-prone skin in agar medium against S. epidermidis has been demonstrated. The bacterial species tested is Staphylococcus epidermidis or white staphylococcus, it is a bacterium of the genus Staphylococcus. It is a bacteria typical of the skin flora in humans. This microorganism is linked to certain skin infections of the skin such as acne. We also carried out on our purifying gel another clinical test under dermatological control with 22 volunteers (20 women and 2 men) of average age of 27 years (between 18 and 35 years) and with an acne-prone skin. This test demonstrated that the majority of volunteers (95%) appreciated its sebum-regulating properties and its effectiveness on traces of skin imperfections after 21 days. Result: rebalanced, purified and mattified skin
SEPARATION AND IDENTIFICATION OF THE PREDOMINANT PROTEINS
3 proteomic analysis methods using 1-D and 2-D electrophoresis to assess extraction quality and proteomic concentration and to identify the observed proteins. And shotgun analysis, which is a more sensitive method that allows for the identification of minority proteins. Hemocyanin and actin are identified as the predominant proteins in our snail filtrate. Hemocyanin is used in cosmetics to activate cellular respiration (Source: Patent Application FR 280432 A1). It is an essential element in the skin regeneration process and in preserving the skin’s extracellular proteins (Source: Abdulghani – A.A et al,1988 – Gadi Borkow, 2004). Actin plays an important role in healing skin cells and regulating the production of collagen, which helps maintain skin firmness (Source: Zhaoping Qui et al, 2018-PIMB: 29888864).
NON-TARGETED APPROACH TO SECONDARY
A non-targeted approach for non-polar compounds, preparation of protein precipitation with blank protocol, LC/UV/MS/MS coupling - RSLC U3000 chromatographic system (Thermo) coupled to a QTOF mass spectrometer (Synapt G2, Waters). Apolar compounds using a 7890A gas chromatograph coupled to an AccuTOF time-of-flight mass spectrometer (JEOL). The presence of small proteins such as lipids, nucleic bases, alkaloids, and flavonoids was observed. Identification of oxygenated compounds such as vitamin D (Vitamin D3, D4, Ergosterol), which play a significant role in the skin renewal process, protect the skin from free radicals, are essential for healing wounds and support the skin’s immune function.
Our studies show that hemocyadin and actin are predominant proteins, both known for their roles in the skin regeneration and healing processes, through in vitro research and clinical studies. 2,500 years after Hippocrates, science has confirmed the observations made during Antiquity. It is important that the mucus be extracted in extract form, which is essential to its high proteomic quality and hence its properties. In this work, we have shown that the snail filtrate extract used by Mademoiselle Agathe cosmetics contains numerous proteins and that all these proteins have a more or less direct link with the skin. Interestingly, the oxygenated compounds found, such as vitamin D, which play an essential role in the skin’s health, had never before been documented. This work will provide a basis for a future study, which could show the connection between the hydrolipidic film of human skin and a distant ancestor of snail filtrate. Additional research is needed to identify the other proteins present.
SYNERGY OF SNAIL SLIME AND PLANT
We are looking for the perfect synergy with your skin. Indeed, for a cosmetic cream to be well assimilated by the skin, it is important to produce an emulsion (oily phase and aqueous phase) which will be penetrating and will not leave any greasy and unpleasant film on the skin. And as the “miracle” active ingredient does not exist, it is important to specify that it is the perfect combination between our snail slime extract and plant extracts, oils and vegetable wax elaborated and worked in the respect of the nature which gives all the effectiveness of our care. Therefore, the excellent interaction between the plant world and the animal world is in our opinion the quintessence of organic and ethical cosmetics.