Protein hydrolysates in cosmetics production, part II

COSME TICS HYDR OLYSED PR OTEINS

M. Ścibisz, J. Arct*, K. Pytkowska

Protein hydrolysates in cosmetics production, part II Key words: skin care, hair care, cell growth stimulation, undesired effect, modified hydrolysed proteins, formulations

Cosmetic properties of proteins

and their hydrolysates

Proteins have been used in cosmetics from time immemorial, mainly in form of plant or animal extracts. Nevertheless, just 40 years ago first hydrolysates of isolated proteins were introduced. Nowadays, they belong to the most often used group of active ingredients, primarily for hair care and top quality cleansing products. Typology of available hydrolysates is various, with regard to source of their origin and diversity of production processes. However, there is a strong dependence between their cosmetic activity and physicochemical properties, such as average molecular weight, charge or hydrophobicity. Substantivity Many conducted studies concerning cosmetic activity of proteins and their hydrolysates revealed, that the most important feature determining their effectiveness is substantivity to the surface of the skin and hair. The outermost layer of skin and hair is built of fewer or more compact keratin structures. As a result of non-covalent interactions, during treatment with proteins and their hydrolysates water solutions, layer of adsorbed polypeptides is formed on the surface. Its properties and strength of the force which keeps it connected with the surface strictly depends 12

on proteins or their hydrolysates chemical and physicochemical properties and in some cases (i.e. during damaged hair treatment) also on the state of the adsorbing surface. In the event of proteins, term “substantivity” first of all concerns ability to form weak or strong bonds with keratin of the stratum corneum or hair cuticle. In case of protein molecules with molecular weights lower than 3000 Da barely few studies confirmed their ability to penetrate deeper areas of skin (dermis) and hair (cortex). Experiment which revealed, that peptides can easily penetrate cuticle and tends to accumulate in cortex were carried out in the 70ties. In this study quantity of peptides was determined by detection technique based on fragmentary ninhidrine staining of considered hair. It should be noticed that this assay is characterized by very low precision. Hence, credibility of obtained results seems to be questionable. Human hair (Fig. 1) is composed of two main layers: outer shield containing high quantities of sulfur amino acids and cortex (60 – 90% of the fiber) responsible for mechanical properties of the hair. Empty core (medulla) is not always present - it usually appears in thick hairs. Hair ending and hair root are regiments where medulla is not observed. Substances used for hair treatments (oxidants, alkalies), environmental factors (UV radiation, alkaline pollutions), and friction or bending create cuticle and

cortex damages. Long-term and frequent exposition to damaging factors and inappropriate hair care entails cuticle structure alterations manifested by worsen hair appearance. Studies concerning factors, which influence the proteins substantivity, such as time of contact, concentration, hair condition (normal and bleached hair) were carried out. It was stated, that in case of both healthy and bleached hair, sorption of peptides on the surface occurs mainly during the first 15 minutes of treatment by water solution. After this period increase in quantity of adsorbed peptides is much slower. Also quantity of bounded proteins and their hydrolysates clearly increased as hair condition worsened. Surprisingly, this dependence was limited – hair bleached twice adsorbed much more polypeptide molecules than hair after single bleaching. Further damage caused by repeated application of bleaching agent didn’t affect the adsorption level. Sorption of polypeptides wasn’t also influenced by their concentration (in range between 5 and 20%). Similar results were obtained during studies concerning substantivity of collagen hydrolysates with various molecular weights. In case of these experiments, method based on hydroxyproline (amino acid characteristic for collagen) assay was used. It was revealed, that absorption of peptides increases simultaneously with hair damage. At the SÖFW-Journal Wydanie Polskie | 1 | 4-2008


end of the trial healthy hair contained about 0.02% of the peptides, while bleached hair 0.2% and hairs additionally after permanent waving contained nearly 3% of polypeptides. Hydrolysates with low molecular weight showed maximum substantivity. Other experiment was conducted on the human skin. Surface of the skin was treated for 10 minutes with 9% solution of tested hydrolysate (m.w. 2000 Da). Next, using hydroxyproline assay content of the collagen hydrolysate in each layer of stripped skin was estimated. Experiment revealed, that collagen peptides were present in the first 6 layers (21 verified) of stratum corneum. In layers 1 to 3 content of collagen amounted to 2.78µg/cm2, while in layers 4 to 6 only 0,33µg/cm2. In comparison, control probe in layers 1-3 contained 0.15µg/cm2 of collagen peptides. Influence of the pH on the collagen hydrolysate substantivity to hair after bleaching and permanent waving (hydrogen peroxide, ammonium thioglycolate, sodium bromate) was also investigated. It was observed, that absorption is the highest in neutral pH (7.0). Maximum substantivity in alkaline pH was observed only in the event of hair pretreated with ammonium thioglycolate. It was also found, that in case of hair pretreated with hydrogen peroxide and ammonium thioglycolate, adsorption of peptides significantly increases by the first 5-10 minutes in the course of action. Conducted studies concerning substantivity of peptide fractions with various molecular weights, which were isolated from hydrolysates available on market proved, that fractions with m.w. about 1000 Da showed the strongest adsorption. Additionally, higher substantivity was observed for hair after bleaching and permanent waving than for only bleached hair. Again, increasing absorption with increasing hair damage was confirmed. In order to investigate substantivity of collagen hydrolysates to the surface of the skin and hair with respect to applied concentration, type of cosmetic vehicle, hair condition and time of product application experiments were carried out. Peptide quantity determination was based on their labeling with 125I isotope. 14

Fig. 1 Cross-section of the hair shaft

As a result following conclusions were drawn: •

•

•

•

•

increase of the polypeptides concentration causes practically rectilinear growth of adsorbed substances quantity, this dependence is also true for concentration higher than 5%, what is clearly not in conformity with previous results, type of cosmetic vehicle (water, shampoos with various formulations) and conditions of its usage have influence on the peptides substantivity to hair, adsorption is higher for damaged than for healthy hair. Dependence is true only in case of low concentration of peptides and short time of application, peptides with molecular weights in range 7500 – 15000 Da demonstrate the highest substantivity, hydrolysates with the lowest value of isoelectric point show the highest substantivity.

Part of these conclusions confirms results obtained in previous studies. However, there are some differences especially in matter of substantivity and molecular weight of hydrolysates dependence. In order to evaluate relationship between peptide structure and its ability to bind with hair keratin, 20 hydrolysates with various molecular weights were ex-

amined. Tested hydrolysates were obtained from both animal and plant sources; hydrolysates from following materials were used: collagen, keratin, elastin, wheat and corn gluten. Peptides obtained from keratin demonstrated high substantivity, higher in compassion to hydrolysates fro other animal and plant sources. As a result of conducted studies such order of hydrolysed proteins with respect to decreasing substantivity has been proposed: keratin > wheat gluten > collagen In case of elastin hydrolysates obtained results were ambiguous, whereas small quantity of assayed samples for corn hydrolysates did not allow placing it in presented line. For compared hydrolysates received from the same protein and characterized by the same isoelectric point (pI) value, substantivity increased in conjunction with decreasing molecular weight. Additionally, absorption of peptides especially from LMW hydolysate fractions, was preferred both for damaged and healthy hair. Thus results confirmed that substantivity depends on hydrophobicity. The higher hydrophobicity of the peptide the smaller its effective size in the water solutions. It is caused by forming such shape of the structure, thanks to which peptide reduces to minimum its contact with water molecules. As a result, small peptide molecules much easier apSÖFW-Journal Wydanie Polskie | 1 | 4-2008


proach to skin and hair keratins’binding centres and penetrate keratin fibres. Weak, but meaningful substantivity increase was observed, when isoelectric point value of particular hydrolysate was higher. This dependence was clear for damaged hair (after bleaching or permanent waving). It should be noticed, that two types of wheat hydrolysates with different pI values (pI1= 3.9 and pI2=5.6) were compared. Experiment was carried out in pH = 6.0, hence both hydrolysates were negatively charged, same as surface of the hair. Probably, this could significantly reduce their substantivity. Reduction of surfactants’ irritant potential Frequent and accumulative skin exposition to the surfactants, especially to anionic, can lead to skin barrier damage and provoke such adverse effects as: dryness, roughness, and even irritation. This phenomenon is caused by surfactant penetration and interaction with liquid crystal structures of stratum corneum, as well as cell membranes in viable epidermis. On the other hand, hair damages caused by activity of surfactants are result of cuticle structure relaxing and weakness of its mechanical properties. When critical micelle concentration (CMC) is reached, which is characteristic for particular surfactant and temperature, large aggregates (micelles) are formed. Remaining surfactant monomers present in solution are able to penetrate keratin structures of skin and hair. Anionic surfactants may affect the skin by following three postulated mechanisms: • hydrocarbon chain of the surfactant penetrates through non-polar regions of keratin where it disturbs hydrophobic bonds stabilizing proteins conformation, • negative charge of surfactant generates forces causing attraction or repulsion, which interact with charged fragments of keratin and cause its structure alteration, • presence of hydrophilic ionic structures leads to increase of osmotic SÖFW-Journal Wydanie Polskie | 1 | 4-2008

pressure, which increases permeability of the stratum corneum and hairs cuticle. Addition of proteins is one of the most examined and effective method of epidermis protection from negative effects caused by surfactants. Activity of proteins and their derivatives may be due to: •

•

lowering the CMC value by surfactant molecules complexing (ionic bonds, hydrogen, hydrophobic and mixed micelle formation), which reduces quantity of free, unbounded monomers in the solution; surface activity remain unchanged. Sorption of surfactant molecules on the surface of proteins is shown in Fig. 2. binding the protein to skin keratin by weak, but numerous bonds, ipso facto causing formation of protective colloidal layer which is able to bind aggressive chemical substances.

Protective properties of proteins and their hydrolysates, preventing skin irritation and hair damaging, were evaluated in in vitro and in vivo tests carried out in human and in animals. Initially, derivatives of proteins were used – fatty acids condensates, utilized as a mild surfactants. Dependence of their cleansing

properties and irritation potential with respect to the chain length of the polypeptide was investigated. It was stated, that molecules containing in their structure 750 Da chain have five times lower irritancy potential than those containing 500 Da chain. Molecular weight of proteins’chain higher than 750 Da insignificantly lowered irritation potential of surfactant. On the other hand, molecules containing short chains had the best cleansing properties. Presence of chain with higher molecular weight than 750 Da significantly reduced cleansing properties. It was also observed, that combination of anionic surfactant (Sodium Lauryl Sulfate, SLS) and protein derivative (20% of the mixture) reduced irritation potential by one third in comparison to solution containing only SLS. Other trial revealed, that pure Sodium Laureth Sulfate completely deactivated enzyme for saccharose, while its combination with protein derivative in ratio 35 : 65 practically didn’t affect this enzyme. Many studies concerning influence of proteins and their derivatives on reduction of surfactant irritant potential were conducted. Effectiveness of their ability to diminish negative activity of surfactants was evaluated among others by usage of two hydrolysates combination, characterized by different molecular weights, 2200 Da (pI=5.1) and 800 Da (pI=5.1). In experiment, surfactants with

Fig. 2 Interaction of surfactant molecules with protein

15


soothing properties, available on the market, were used. They were: Cocamidopropyl Betaine and CocamidopropylAmine Betaine. Similar to proteins, activity of these compounds is based on interactions with surfactant molecules and mixed micelle formation. Evaluation of such parameters as skin hydration and transepidermal water loss allowed estimating ability of protein additives to reduce undesirable effects caused by surfactants. Effectiveness was estimated after single and multiple skin exposition to tested substances. Results of conducted experiments allowed drawing following conclusions: •

•

•

•

influence on TEWL of protein hydrolysates is similar to Cocamidopropyl Betaine and CocamidopropylAmine Betaine after single skin treatment with surfactant solution, hydrolysates with high molecular weight demonstrate higher protective properties in comparison with l.m.w. hydrolysed proteins, there is a slight, but noticeable increase of the skin tolerance in conjunction with protein concentration increase, wheat proteins demonstrate synergy with soothing surfactants.

In case of multiple tests, proteins were more effective than soothing surfactants. It confirms the fact, that beyond direct interactions with SLS, protein molecules bind with skin keratin forming continuous barrier which protects the stratum corneum. Using similar method, effects of collagen, elastin, keratin and wheat gluten hydrolysates with various molecular weights were investigated. All tested hydrolysates showed ability to reduce irritation potential of SLS. TEWL measurements allowed creating following dependence (decreasing ability to reduce irritant potential of the surfactant) with respect to hydrolysates origin: elastin and gluten > collagen >> keratin Results of conducted studies showed better protection ability of hydrolysates 16

with higher hydrophobicity and molecular weight. Other studies concerned ability of hydrolysates to interact with Sodium Lauryl Sulfate (SLS). It was demonstrated, that proteins which are short in sulphurcontaining amino acids show tendency to bind over 50% more SLS than proteins with ability to form disulfide bonds (i.e. keratin). CMC values of complexes formed by protein and tested surfactant were similar in case of all proteins regardless of their molecular weight. Hair and skin care In skin care, proteins and hydrolysed proteins are used mainly as moisturizing agents. Polypeptides adsorbed on the skin surface supplement outer hydrophilic skin coat and due to ability of hydrogen bonds formation they bind water molecules. Many amino acids may act as a donors (ie. arginine and tryptophan) and acceptors (asparagine, glutamine, serine and threonine) in connection with presence of special groups localized in the side chains. On the other hand, depending on the pH value, acidic (glutamic and aspartic acid) and alkaline (lysine, tyrosine, histidine) amino acids play role as hydrogen acceptors or donors. Highly hydrated layer, which consist of proteins bounded to the skin surface hampers diffusion and prevents water evaporation. Hydrolysates with low molecular weight may form strongly adsorbed monomolecular layer, which is difficult to remove. Large molecules tend to form weakly anchored film, but with greater ability to bound water molecules. Hence, HMW polypeptides are more often used as moisturizing film formers. LMW peptides, on the other hand constitute ingredients of products, which require high substanivity, i.e. designed for hair regeneration, where peptides after building in hair keratin restore its proper structure. Conducted studies confirmed beneficial influence of proteins on the skin hydration. In vivo comparative test using emulsion containing 5% of the water soluble collagen and placebo, was carried out. After 10 days of application,

evaluation of each skin layer revealed significant difference of their thickness strictly related to the hydration level. Studies concerning moisturizing properties of proteins and their hydrolysates delivered from various physicochemical forms of cosmetics, such as o/w emulsions, gels, water solutions and form which contained surfactants, were conducted. HMW proteins showed excellent moisturizing properties when used in o/w emulsions and cleansing products, whereas low molecular acted as a substances which astringe and tighten the skin, even the surface and have anti-wrinkle properties. Results of studies allowed to define advantages of proteins and hydrolysed proteins as cosmetic ingredients: • they increase elasticity and hydration level of the skin after application of leave on (creams) as well as rinse–off cosmetics (soaps, shampoos), • they improve skin tightness, • they improve skin ability to respond to the deformation, thus temporary even wrinkles. Proteinaceous ingredients are used in cosmetics for skin care mainly at concentration of 0.1 to 2% (Formulations 1-2). Hydrolysed proteins are often used in combination with other moisturizing agents, such as glycerin, sodium lactate, or free amino acids. Polar peptides, first of all constitute ingredients of emulsion water phase. Their hydrophilic nature permit to suppose that they stabilize o/w emulsions, due to increasing viscosity of the external phase. On the other hand hydrophobic proteins, characterized by low solubility might stabilize emulsions, thanks to water-oil interface affinity. Proteins and hydrolysed proteins are widely applied in hair care cosmetics as conditioning agents – improving softness, elasticity, and gloss and hair resilience. They are most often used as main ingredients of shampoos and both - leave on and rinse off conditioners for damaged and dry hair. In this kind of formulations their concentration is usually not higher than 1-2% (Formulations 34). Proteins play crucial role in bleaching, SÖFW-Journal Wydanie Polskie | 1 | 4-2008


permanent waving and hair straightening products.They have unique ability to limit undesirable effects of oxidizing and reducing agents on the hair structure. Permanent waving is based on disulfide bonds reduction and formation of a new bonds determining geometry of fibrils. Addition of HMW protein hydrolysates, which are rich in sulfur-containing amino acids causes formation of covalent bonds between keratin and delivered in cosmetic polypeptides. As a result, durable connections are formed, which reduce damages occurring during treatment. Additionally, hydrolysates which are easily accessible for oxidizing and reducing agents constitute substrates in side reactions, which normally occur between mentioned agent and hair keratin. As keratins plant equivalent used in these kind of treatments wheat proteins are mainly used.

Cells growth stimulation Many publications concerning proteins and their hydrolysates pay special attention to their stimuli effect on cells divisions. It was mainly observed in treated with wheat proteins fibroblasts. Level of cells divisions increased simultaneously with extending concentration of the hydrolysate (range between 0.01 and 0.2%). In other study, influence of collagen, gelatin and hydrolysed collagen (m.w. < 15000Da) on mouse keratinocytes division was determined. Proliferation was observed during 8 to 20h of the trial. Significant stimulation was noticed just after 8h, but only in case of collagen. Results obtained for hydrolysates and gelatin, were comparable with results for control probes. Effects of conducted studies allow deducing that proteins show activity in deeper parts of skin, as they stimulate fibroblast proliferation. Thus they can delay aging process and improve skin elasticity and firmness. However, advantageous effects on hairs and skin condition are due to mentioned before interactions between proteins and peptides with their surfaces. Studies concerning permeability didn’t confirm SÖFW-Journal Wydanie Polskie | 1 | 4-2008

their penetration ability into deeper parts of skin. That’s why their activity in these areas seems to be questionable. On the other hand, in case of low molecular weight peptides penetration through stratum corneum is possible, and is supported by alkyl substituents. Undesired activity Protein based ingredients are of natural origin. The same INCI name may used for substances obtained in different technological processes. That implicates risk of irritant activity of these substances. Safety reports published by the American College of Toxicology classified collagen hydrolysates as non-toxic, substances with minimal irritant effect (evaluated in Draize test), without sensitization in guinea-pig test. LD50 value for rats and mouses was at level of 10-20 grams of pure protein per kg of the body mass. Despite of low toxicity of proteins, one can find studies indicating undesired activity of various proteins and their derivatives used in hair care cosmetic products. Such ingredients are obtained from collagen, elastin, keratin, milk proteins, wheat, silk and almonds. Hydrolysates of these proteins showed irritant potential in 0.5% of examined patients. In studied groups of hydrolysates the larger group of undesired reaction in patients, especially with atopic dermatitis, were observed for collagen derivative (i.e. Hydroxypropyl Trimonium Hydrolyzed Collagen). Such reaction was associated with contact urticaria symptom. There are as well studies confirmed irritant activities of wheat and bovine hydrolysed proteins, used in skin and hair care products.

Foam boosting effect Softness and smoothness to skin and hair, together with anti-irritating effect are the main factors determining use of such ingredients in cosmetic products. Proteins may also be applied as “technical function” ingredients (buffering properties, viscosity control). One of such function is also foam boosting effect. Water soluble proteins, as other hydrophilic polymers stabilise foams. Hydrolysed proteins of different origin (i.e. collagen, elastin, keratin, corn gluten, and wheat proteins) were studied regarding their foam boosting activity. The best effect was obtained for hydrolysed wheat proteins (foam height at approx 100-140mm, with high stability) with exception of hydrolysate obtained by means of enzymatic hydrolysis. Hydrolysed keratin formed foam with significant height, yet unstable. In the 5-8 pH range foam boosting effect was similar for various hydrolysed proteins. That effect was considerable worse in the pH below 5. Foam boosting behaviour relied also on molecular weight, especially in the case of hydrolysed proteins with low hydrophobicity. Such tendency was not observed for hydrolysed wheat protein. In the case of hydrolysed collagen foam boosting effect decreased with molecular weight reduction. Modified hydrolysed protein The significance of proteins and their hydrolysates in cosmetics is also reflected by use of theirs modified derivatives. Properties of such compound are strictly associated with protein’ part in the

R=-CH3, n-alkyl group

Fig. 3 Chlorhydrine with quarterbary nitrogen atom 17


molecule, but through other, non-protein part they may demonstrate better solubility, substantivity, or better foam boosting effect. Production of protein derivatives takes quite big part of cosmetic ingredients and raw materials segment. One of the most popular, used for 30 years, group of modified hydrolysed proteins are condensates of proteins with fatty acids. Protein or hydrolysed proteins, most often obtained by means of enzymatic hydrolysis, are N-acetylated with long chain fatty acids chlorides in neutral or weak alkaline pH (7-9). Products of that reaction are described as ones of the mildest surface active agents. Since theirs irritant potential is lower than observed one for surfactants form betaines group, they are use in shampoos, skin, face cleansing products and shower gels (Formulations 5-6). They also indicate a good skin and eyes tolerance, have very well cleansing and foam boosting properties, even in hard water. They are as well far soluble in organic solvents like ethanol. Comparing to condensates of proteins with fatty acids, more important from the commercial point of view are quaternary protein derivatives. Such compounds are obtained in alkylation reaction of hydrolysed protein with chlorhydrine, compound with quaternary nitro-

gen atom (Fig. 3), in alkaline environment. To increase conditioning effect of such modified protein, linear chains (12 and more carbon atoms) with lipophilic properties, are introduced as R alkyl group. Isoelectric point of hydrolysed proteins lies in pH range of 4 to 7. After reaction yielding quaternary derivative obtaining, isoelectric point value increases to pH 9-12. Furthermore, quaternary hydrolysed proteins indicate positive charge in whole pH range, while unmodified hydrolysed proteins bear it only in pH below the isoelectric point. Increased cationic character of protein derivatives increases substantivity of these compound to both skin and hair keratin, which in physiological pH indicate total negative charge. Such properties may be useful in washable products. Quaternary derivatives are used as conditioning and film-forming agents, especially in hair care products. They are compatible with ionic and non-ionic surfactants; thereby they are easily introduced into shampoo formulation. One can find studies that showed cationic derivative of wheat protein soothing properties of irritant activity of surfactants. They also decreased irritant potential formulation in which skin compatible ingredient such as betaines were introduced. Furthermore skin

Formulation 1 Skin tone cream (Sinerga)

Potassium Palmitoyl Hydrolyzed Wheat Protein

(and) Glyceryl Stearate (and) Cetearyl Alcohol Ethylhexyl Ethylhexanoate Dimethicone Aqua Phenethyl Alcohol (and) Methylparaben (and) Propylparaben (and) Glycerin Squalane Potassium Caproyl Tyrosine Algae Hydrolyzed Vegetable Protein Sodium Carbomer Parfum Carbomer Aminomethyl Propanol Disodium EDTA Tocopherol (and) Lecithin (and) Citric Acid (and) Ascorbyl Palmitate

18

10,00 5,00 0,50 Do 100 1,00 10,00 5,00 2,50 1,50 0,10 0,70 0,40 0,05 0,10 0,05

compatibility of quaternary protein derivatives increased with molecular weight increasing of protein part of molecule. Recently such protein derivatives as silicone copolymers and phosphorylated derivatives are applied as cosmetic ingredients.

Literature (1)E.D. Goddard, J.V. Gruber; Principles of polymer science and technology in cosmetics and personal care; Marcel Dekker, New York, 1999 (2)V.L. Johnsen; Proteins in cosmetics and toiletries; Drug&Cosmetic Industry, 6, p.36, 1980 (3)V.L. Johnsen; Innovation in protein products and technology; Cosmetics&Toiletries, 92, 12,p.29, 1977 (4)A. Teglia, G. Mazzola, G. Secchi; Chemical characteristics and cosmetic properties of protein hydrolysates; Cosmetics&Toiletries, 108, 11, p.56, 1993 (5)J.Arct; Proteiny; Wiadomości PTK, 2, 1, p.17, 1999

(6)U. Griesbach, M. Klingels, V. Homer; Pro-

Formulation 2 Anti hyperpigmentation skin cream (Sinerga) Potassium Palmitoyl Hydrolyzed Wheat Protein (and) Glyceryl Stearate (and) Cetearyl Alcohol Squalane Ethylhexyl Ethylhexanoate Cocoglycerides Dimethicone Aqua Potassium Azeloyl Diglycinate Prunus Amigdalus var. Dulcis (and) Hydrolyzed Sweet Almond Protein (and) Potassium Palmitoyl Hydrolyzed Wheat Protein Avena Sativa (and) Hydrolyzed Oat Protein (and) Potassium Palmitoyl Hydrolyzed Oat Protein

10,00 12,00 5,00 2,00 0,50 Do 100 7,00

2,50 2,50

SÖFW-Journal Wydanie Polskie | 1 | 4-2008


teins: classic additives and actives for skin and hair care; Cosmetics&Toiletries, 113, 11, p.69, 1998 (7)G.Y. Li, S. Fukunaga, K. Takenouchi, F. Nakamura; Comparative study of the physiological properties of collagen, gelatin and collagen hydrolysate as cosmetic materials; International Journal of Cosmetic Science, 27, p. 101, 2005 (8)UIImann’s Encyclopaedia of Industrial Chemistry, Fifth Edition, A22 (9)E.S. Stern. V.L. Johnsen; Studies on the molecular weight distribution of cosmetic protein hydrolysates; Journal of the Societies of Cosmetic Chemists, 28, 8, p.447, 1977 (10) B.W. Gesslein, R.T. Jones; Kerasol, a new keratin protein; Cosmetics&Toiletries, 102, 6, p.52, 1987 (11) F. Pasche-Koo, M. Claeys, C. Hauser; Contact urticaria with systemic symptoms caused by bovine collagen in a hair conditioner; American Journal of Contact Dermatitis, 1, 1, p.56. 1996 (12) US Patent 4,279,996 (13) 24. E.S. Cooperman, V.L. Johnsen; Penetration of protein hydrolysates into human hair strands; Cosmetic&Perfumery, 88, p.19, 1973 (14) S.A. Karjala, RJ. Bouthilet, J.E. Williamson; Some factors affecting the substantivity of proteins to hair; Proceedings of Scientific Section the Toilet Goods Association, 45, 5, p.6. 1966 (15) M.D. Ranganayaki, T.S. Ranganathan, K.S. Jayaraman; A novel technique for the preparation of collagen derivatives for cosmetics. Part 2. Preparation and characterisation of collagen derivative; Research and Industry, 34, p.183, 1989 (16) M.D. Ranganayaki, T.S. Ranganathan, K.S. Jayaraman; A novel technique for the preparation of collage derivatives for cosmetics. Part 3. Functional properties of collagen derivatives; Research and Industry, 35, p.l, 1990 (17) S.A. Karjala A. Karier, J.E. Williamson; The effect of pH on the sorption of collagen – derived peptides by hair; Journal of the Societies of Cosmetic Chemists, 18, 10, p.599, 1967


Formulation 3 Hair conditioner (Croda) Quaternium-91 (and) cetrimonium methosulfate (and) cetearyl alcohol Stearyl alcohol Cetyl alcohol C10-C30 cholesterol/lanosterol esters PPG-3 benzyl ether myristate PG-hydroxyethyl cellulose steardimonium chloride Hydrolyzed silk Hydrolyzed wheat protein Hydrolyzed vegetable protein PG-propyl silantriol Glycerin (and) butylene glycol (and) water (and) camellia sinensis leaf extract Glycerin (and) butylene glycol (and) water (and) chamomilla recutita (Matricaria) extract Glycerin (and) butylene glycol (and) water (and) hippophae rhamnoides fruit extract Phenoxyethanol (and) methyl paraben (and) ethyl paraben (and) butyl paraben (and) propyl paraben (and) isobutyl paraben Aqua

2,23 2,00 0,50 1,00 4,50 0,20 1,00 1,00 1,00 0,10 0,10 0,10 1,00 85,27

Formulation 4 Regenerating shampoo (Cognis) Sodium coco-sulfate Coco-glucoside Lauryl glucoside (and) stearyl citrate Coco-glucoside (and) glyceryl oleate Hydrolyzed wheat protein Sodium benzoate Perfume Citric acid (50%) Sodium chloride Aqua

Formulation 5 Shower gel (Sinerga)

Sodium Laureth Sulfate Lauramidopropyl Betaine Disodium Cocoamphodiacetate (and) Sodium Laureth Sulfate Disodium Laureth Sulfosuccinate (and) Sodium Lauryl Sulfoacetate Potassium Cocoyl Hydrolyzed Soy Protein Caprylyl/Capryl Glucoside Linoleamidopropyl Dimethylamine Lactate Hydrolyzed Vegetable Protein Yucca Schidigera Extract Parfum Preservative Disodium EDTA Sodium Chloride Aqua

10,81 19,24 2,00 2,00 1,00 0,50 q.s. q.s. 0,50 63,95

25,00 6,00 10,00 5,00 5,00 2,50 2,50 2,00 1,00 1,00 q.s. 0,10 0,50 Do 100

19


Formulation 6 Shower emulsion(Sinerga)

Sodium Laureth Sulfate Sodium Laureth Sulfate (and) Cocamide MEA Disodium Laureth Sulfosuccinate (and) Sodium Lauryl Sulfoacetate Potassium Cocoyl Hydrolyzed Soy Protein Caprylyl/Capryl Glucoside Potassium Cocoyl Hydrolyzed Oat Protein (and) Glyceryl Stearate Cocodimonium Hydroxypropyl Hydrolyzed Wheat Protein Hydrolyzed Vegetable Protein Almond (Prunus amigdalus var, dulcis) Oil (and) Hydrolyzed Almond Protein (and) Potassium Palmitoyl Hydrolyzed Wheat Protein Aqua Preservatives Disodium EDTA Lauramidopropyl Betaine Parfum

30,00 12,50 5,00 5,00 2,50 10,00 1,00 1,50 2,50 Do 100 q.s. 0,10 1,50 q.s.

(22)A. Niinimaki:, M. Niinimaki, S. MakinenKiljunen, M. Hannuksela; Contact urticaria from protein hydrolysates in hair conditioners; Allergy, 53, p.1078, 1998 (23)E.Yarjonen, L. Petman, S. MakinenKiljunen; Immediate contact allergy from hydrolyzed wheat in a cosmetic cream; Allergy, 55, p.294, 2000 (24)F. Sanchez-Perez, T. Sanz, A. Garcia-Diez; Allergic contact dermatitis from hydrolyzed wheat protein in cosmetic cream; Contact Dermatitis, 42, p.360, 2000

Formulation 7 Mild, hair care shampoo (Cognis) Hydroxypropyl guar hydroxypropyl- trimonium chloride Xanthan gum Decyl glucoside Lauryl glucoside Coco-glucoside (and) glyceryl oleate Dicaprylyl ether (and) lauryl alcohol Laurdimonium hydroxypropyl hydrolyzed wheat protein Sodium benzoate Citric acid (50%) Aqua (18) A. Turowski, B.C. Adelmann-Grill; Substantivity to hair and skin of 125I-labelled collagen hydrolysates under application simultaning conditions; International Journal of Cosmetic Science, 7, p7I, 1985 (19) A. Teglia, G. Secchi; Minimizing the cutaneous effects of anionic detergents; Cosmetics&Toiłetńes, 111, 8, p.61, 1996

20

0,1 1,0 30,0 6,0 3,0 0,5 2,0 0,5 q.s. 56,9

(20) R. Pitt-Rivers, F.S.A. Imptombato; The bindin; of Sodium Dodecyl Sulphate to various proteins; Biochemical Journal, 109, p.825, 1968 (21)N.I. Challoner, S.P. Chahal, R.T. Jones; Cosmetic proteins for skin care; Cosmetics&Toiletries, 112, 12, p.51, 1997

Authors’ address: Marta Ścibisz Academy of Cosmetics and Health Care Jacek Arct, PhD Academy of Cosmetics and Health Care, Podwale St. 13, 00-252 Warsaw, Poland Warsaw University fo Technology, Faculty of Chemistry * Email: [email protected] Katarzyna Pytkowska Academy of Cosmetics and Health Care
