CA2286094A1 - Use in cosmetics of a protein fraction extracted from okra seeds - Google Patents

Use in cosmetics of a protein fraction extracted from okra seeds Download PDF

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CA2286094A1
CA2286094A1 CA002286094A CA2286094A CA2286094A1 CA 2286094 A1 CA2286094 A1 CA 2286094A1 CA 002286094 A CA002286094 A CA 002286094A CA 2286094 A CA2286094 A CA 2286094A CA 2286094 A1 CA2286094 A1 CA 2286094A1
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cosmetic composition
composition according
seeds
protein fraction
protein
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Gilles Pauly
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BASF Health and Care Products France SAS
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    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61QSPECIFIC USE OF COSMETICS OR SIMILAR TOILETRY PREPARATIONS
    • A61Q17/00Barrier preparations; Preparations brought into direct contact with the skin for affording protection against external influences, e.g. sunlight, X-rays or other harmful rays, corrosive materials, bacteria or insect stings
    • A61Q17/04Topical preparations for affording protection against sunlight or other radiation; Topical sun tanning preparations
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61KPREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K8/00Cosmetics or similar toiletry preparations
    • A61K8/02Cosmetics or similar toiletry preparations characterised by special physical form
    • A61K8/04Dispersions; Emulsions
    • A61K8/06Emulsions
    • A61K8/068Microemulsions
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61KPREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K8/00Cosmetics or similar toiletry preparations
    • A61K8/18Cosmetics or similar toiletry preparations characterised by the composition
    • A61K8/30Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds
    • A61K8/64Proteins; Peptides; Derivatives or degradation products thereof
    • A61K8/645Proteins of vegetable origin; Derivatives or degradation products thereof
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61KPREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K8/00Cosmetics or similar toiletry preparations
    • A61K8/18Cosmetics or similar toiletry preparations characterised by the composition
    • A61K8/96Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution
    • A61K8/97Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution from algae, fungi, lichens or plants; from derivatives thereof
    • A61K8/9783Angiosperms [Magnoliophyta]
    • A61K8/9789Magnoliopsida [dicotyledons]
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61QSPECIFIC USE OF COSMETICS OR SIMILAR TOILETRY PREPARATIONS
    • A61Q17/00Barrier preparations; Preparations brought into direct contact with the skin for affording protection against external influences, e.g. sunlight, X-rays or other harmful rays, corrosive materials, bacteria or insect stings
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61QSPECIFIC USE OF COSMETICS OR SIMILAR TOILETRY PREPARATIONS
    • A61Q19/00Preparations for care of the skin
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61QSPECIFIC USE OF COSMETICS OR SIMILAR TOILETRY PREPARATIONS
    • A61Q19/00Preparations for care of the skin
    • A61Q19/08Anti-ageing preparations
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61QSPECIFIC USE OF COSMETICS OR SIMILAR TOILETRY PREPARATIONS
    • A61Q5/00Preparations for care of the hair
    • A61Q5/06Preparations for styling the hair, e.g. by temporary shaping or colouring
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61QSPECIFIC USE OF COSMETICS OR SIMILAR TOILETRY PREPARATIONS
    • A61Q5/00Preparations for care of the hair
    • A61Q5/12Preparations containing hair conditioners
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61KPREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K2800/00Properties of cosmetic compositions or active ingredients thereof or formulation aids used therein and process related aspects
    • A61K2800/74Biological properties of particular ingredients
    • A61K2800/75Anti-irritant
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61KPREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K2800/00Properties of cosmetic compositions or active ingredients thereof or formulation aids used therein and process related aspects
    • A61K2800/80Process related aspects concerning the preparation of the cosmetic composition or the storage or application thereof
    • A61K2800/805Corresponding aspects not provided for by any of codes A61K2800/81 - A61K2800/95

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  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Veterinary Medicine (AREA)
  • Public Health (AREA)
  • General Health & Medical Sciences (AREA)
  • Animal Behavior & Ethology (AREA)
  • Epidemiology (AREA)
  • Birds (AREA)
  • Dermatology (AREA)
  • Mycology (AREA)
  • Microbiology (AREA)
  • Botany (AREA)
  • Biotechnology (AREA)
  • Engineering & Computer Science (AREA)
  • Gerontology & Geriatric Medicine (AREA)
  • Chemical & Material Sciences (AREA)
  • Dispersion Chemistry (AREA)
  • Cosmetics (AREA)
  • Peptides Or Proteins (AREA)

Abstract

The invention concerns the use of at least one protein fraction extracted from okra seeds and a cosmetic composition containing same. It concerns the use of a soluble protein fraction extracted from Hibiscus esculentus or Okra seeds as a substitute for casein in a cosmetic composition or product, said composition comprising between 0.01 % and 50.00 % of said fraction.

Description

a t CA 02286094 1999-10-14 r3!~':Z.~

Use of at least one protein fraction extracted from Hibiscus esculentus seeds and cosmetic composition containing such a fraction The present invention concerns the field of cosmetology, in particular cutaneous and capillary applications, and relates to the use of at least one protein fraction extracted from Hibiscus esculentus and to a composition containing at least one such extract.
The Hibiscus esculentus (Abelmoshus esculentus or okra from the Malvaceae family) is a plant of African origin introduced into the United States and East Indies under the Spanish name of gumbo. It is one of the botanical species which has been cultivated for its pods for more than 2,000 years.
okra grows in numerous regions of the world such as India, Malaysia, the Philippines, America (Mid-West), Mediterranean regions, Africa and, more generally, in tropical regions.
The fruits (pods) which are eaten young as vegetables are long, green and tapering; they have a delicate flavour and a mucilaginous internal texture.
Apart from the pods, which are of interest on account of the gum which they contain, research has also been carried out on the seeds of Hibiscus esculentus in order to study their potential as a new source of proteins.
To this end, the chemical composition of the whole seed of different varieties of okra (outer skin plus endosperm) has been determined.
Similarly, the properties (protein solubility, amino acid composition, emulsification capacity, foaming capacity, nutritional value) of various products obtained from the seeds (whole flour prepared from skinned seeds, delipidated flour, concentrate and protein isolate) have been studied for food purposes (see, for example, Bryant LA, Montecalvo J, Morey KS, Loy B: "Processing, functional and nutritional properties of okra seed products", Journal of food science, vol. 53, No. 3, 818-816).
okra seed mainly contains the following substances in o by weight, relative to the dry material:
17.7 to 21.80 of proteins - 14.7 to 20.060 of lipids - 4.33 to 4.620 of ash - 6.84 to 7.920 of water - 0.0032a of gossypol, and, in particular, traces of the following substances:
- Calcium: 282.26 mg/100 g - Iron: 10.26 mg/100 g - Thiamine: 0.69 mg/100 g - Riboflavin: 0.14 mg/100 g - Niacin: 4.01 mg/100 g - a-tocopherol: 30.4 mg/100 g.

(See Karakoltsidis PA, Constantidines SM: "okra seed: a new protein source", J. Agric Food Chem., 1975, 23 No. 6, 1204-1207/Wandawi AL: "Chemical composition of seeds of two okra cultivars Abelmoschus esculentus", Journal of agricultural and food chemistry, 1983, 31 No. 6, 1355-1358).
The amino acid composition of concentrated Hibiscus esculentus seed proteins or protein isolates resembles that of Soya proteins and is close to that of casein (see table below).
Amino Hibiscus HibiscusProtein Protein Soya Casein acid (seeds) (seeds) concentrateisolate (seed) Karakot-g/16 KarakoltsidisBryant Bryant Bryant Karakot-sidis g et al et al LA LA et sidis et of Mandawi AL et al al et al nitrogen al Asp 11.82 to 15.4711.57 10.89 12.12 17.00 7.11 Thr 3.02 to 4.38 2.86 3.37 2.90 5.47 4.65 Ser 5.25 to 6.71 5.07 5.23 4.97 7.42 6.02 Glu 20.48 to 22.0815.91 19.15 17.35 21.05 21.19 Pro 3.83 to 6.06 3.79 4.88 4.98 7.71 11.54 Gly 5.79 to 6.66 4.78 7.77 4.16 4.32 1.97 Ala 5.89 to 6.66 4.83 4.53 4.59 6.13 3.07 Val 4.0 to 6.4 4.24 4.42 4.33 5.26 6.72 Cys 1.54 to 2.53 3.63 1.89 1.90 1.61 0.36 Met 1.29 to 1.85 1.83 2.18 2.21 1.25 2.78 Ile 3.15 to 4.65 2.96 3.06 3.13 4.46 5.40 Leu 6.68 to 8.47 6.21 6.96 6.97 9.35 9.49 Tyr 3.6 to 3.83 3.46 5.15 4.03 3.72 5.81 Phe 3.93 to 4.7 4.41 4.80 4.85 5.26 5.23 Lys 7.24 to 8.9 6.22 6.19 6.47 8.00 8.80 His 1.78 to 2.99 2.34 3.61 3.83 2.67 2.91 Arg 11.04 to 12.4611.17 10.02 9.98 10.07 3.74 Trp 0.85 to 0.96 2.02 2.57 2.03 nd nd With regard to the casein, close contents of threonine, serine, glutamic acid, valine, isoleucine, leucine, phenylalanine, lysine and histidine are noted in particular.
The various aforementioned studies therefore demonstrate the value of Hibiscus esculentus seeds as potential sources of proteins from a food point of view.
Furthermore, the use for dermatological purposes of a viscous liquid product obtained by hot extraction and/or extraction under pressure from the fruits of Hibiscus esculentus (FR-A-2 679 443) as well as the use of a powdered material composed of polysaccharide extracted from immature Hibiscus esculentus seeds in a cosmetic product (JP-A-57/199969) are also known.
Now the inventors have found that it was possible to use extracts of Hibiscus esculentus seeds directly in cosmetics and that the use of at least one preferably soluble protein fraction extracted from Hibiscus esculentus or okra seeds, in particular as a substitute for casein, in a cosmetic composition or product yielded a composition or product having surprising and advantageous specific properties.
A strong cellular nutritive power, a smoothing and biofilm-forming effect, conditioning, restructuring and repairing effects as well as anti-irritant, light-protecting, soothing and cutaneous anti-ageing effects have thus been found.
The aforementioned extracts can be used not only for skin care and hygiene applications (products for the face or for the body, day or night products, solar products, anti-wrinkle hygiene products, slimming products), but also in the field of hair care and hygiene (lotions or shampoos; creams; mousses;
protective products, repairing products, softeners, film-forming agents and light protectors; perming and colouring products).
Proteins can be prepared by conventional methods of extraction of vegetable proteins and preparation of protein concentrates or isolates known to a person skilled in the art and described, in particular, in the aforementioned article by Bryant LA, Montecalvo J, Morey KS and Loy B.
The raw material consists of Hibiscus esculentus seeds or seed flour (protein content - 21.6o relative to the dry material).
The flour thus obtained may be delipidated by extraction in hexane at 45°C (advantageously 3 successive extractions).

The oil yield is 16.2 to 23.90 by weight, depending on whether the starting material is a whole seed flour or a flour from seeds partially freed of the outer skins or shells by sifting.
The delipidated flours partially freed of the seed shells contain between 37 and 44% by weight of proteins (N x 6.25).
Various processes for obtaining and preparing extracts of Hibiscus esculentus or okra seeds will be described hereinafter by way of illustrative, non-limiting examples.
Example 1 100 g of non-delipidated enriched flour (partially freed of the shell waste) in one litre of the following media:
- distilled water, - distilled water containing 1 g/1 of NaCl, - distilled water containing 5 g/1 of NaCl, - distilled water containing 10 g/1 of NaCl are added.
After stirring for 15 minutes, the pH of the solution is adjusted to 9 with NaOH 4 N.
Extraction is carried out for one and a half hours at ambient temperature while keeping the pH at 9.
After centrifugation, the upper lipidic layer is removed and the aqueous supernatant liquid is collected.
The golden yellow supernatant liquid is adjusted to pH - 7.5 then. filtered to 0.22 ~.m; it is noted that the higher the salt content of the solvent, the easier filtration is.

The protein content of the filtrate is determined by the biuret method. The supernatant liquids remain opalescent.
The following results are obtained:
Extraction solvent Proteins in the biuret filtered extract (g/1) distilled water 16.9 NaCl 1 g/1 13.3 NaCl 5 g/1 9,5 NaCl 10 g/1 10.2 Example 2 25 g of enriched delipidated flour (freed from shell debris) are added in 250 ml of distilled water containing 5 g/1 of sodium chloride.
After stirring for 15 minutes, the pH of the solution is adjusted to the following pH according to the test: 6-6.5-7-7.5.
Extraction is carried out for one hour at ambient temperature.
After centrifugation, the supernatant liquid is recovered, then filtered over 5 Vim.
The following results are obtained:
pH Biuret proteins Proteins (N x 6.25) (g/1) (g/1) 6 12.5 13.8 6.5 13.3 14.8 7 14.6 15.1 7.5 15.5 . 16.1 Example 3 25 g of enriched delipidated flour are added in 250 ml of distilled water containing 5 g/1 of sodium chloride.
The pH of the solution is adjusted to 8 after stirring for 15 minutes.
Extraction is carried out for 6 hours at ambient temperature.
After centrifugation, the supernatant liquid is recovered and the pH adjusted to 7.5, then the solution is filtered over 5 ~.m .
The following results are obtained:
Duration (hours) Biuret proteins (g/1) 2 18.45 4 20.3 19.7 Example 4 25 g of enriched delipidated flour are added in 250 ml of distilled water containing 5 g/1 of sodium chloride.
After stirring for 15 minutes, the pH of the solution is adjusted to pH 7.5.
Extraction is carried out for 6 hours at 45°C.
After centrifugation, the supernatant liquid is recovered, its pH is adjusted to 7.5 then the solution is filtered over 5 Vim.

The following results are obtained:
Duration (hours) Biuret proteins (g/1) 18.0 4 18.1 Example 5 300 g of enriched delipidated flour are added in 3 1 of distilled water containing 5 g/1 of sodium chloride.
After stirring for 15 minutes, the pH of the solution is adjusted to pH 7.5.
Extraction is carried out for 2 hours at 50°C.
After centrifugation, the supernatant is recovered then filtered over 5 ~.m.
The following results are obtained:
Biuret proteins: 13.7 g/1 Kjeldahl proteins: 14.6 g/1 One litre of supernatant liquid is removed and its pH is adjusted to 4.5 using 4N sulphuric acid .
After 30 minutes of stirring, the solution is centrifuged and the precipitate is collected, then washed with water at pH
4.5. It is then freeze-dried: a powder of which the protein content is 85.30 by weight is obtained.
Example 6 600 g of enriched delipidated flour in 6 litres of distilled water containing 5 g/1 of NaCl are added while stirring in a thermostatically controlled reactor. The pH measured is adjusted to 7.5.
The solution is pumped into an ultrasonic tube with an integral passage by means of a peristaltic pump (ultrasonic power 600 4~1 - frequency 20,000 Hz), the flow rate being 60 1/h.
Two discontinuous passages by successive charges are produced (the extract is collected after passage in the ultrasonic tube in a second reactor).
After centrifugation, then filtration over 5 ~.m, the following results are obtained:
- proteins from the extract after the first passage: 14.2 g/1 - proteins from the extract after the second passage: 15.2 g/1 Example 7 300 g of enriched delipidated flour in 6 litres of distilled water containing 5 g/1 of NaCl are added in a thermostatically controlled reactor while stirring. The measured pH is adjusted continuously to 7.5.
The solution is introduced continuously for 1 hour in a closed circuit in the integral passage ultrasonic tube from example 6 (ultrasonic power: 700 W - frequency: 20,000 Hz), the flow rate being 60 1/h.
A circulation of cold water allows the temperature of the solution in the vessel to be kept at about 33°C.
Samples are taken after 15 min, 30 min, and 45 min of extraction.

After centrifugation, then filtration of the extracts over 5 ~,m, the following results are obtained:
Duration of extraction (min) Proteins in the biuret filtered extract (g/1) 8.23 30 8.56 45 8.89 60 9.89 Example 8 25.0 kg of distilled water are introduced into a reactor and the following operations are carried out in succession:
- dispersing with stirring 2.5 kg of whole flour obtained by crushing whole hibiscus seeds, - adjusting the pH to 9 with NaOH 4N after 15 minutes of dispersion, - extracting with stirring for 2 hours at ambient temperature while keeping the pH at 9 by addition of NaOH 4N, - centrifuging for 10 minutes at 5,000 g, - recovering the cloudy beige supernatant liquid, - adjusting the pH to 7.5 by addition of H2S04 4N, - centrifuging, - clarifying by renewed centrifugation, - recovering the opalescent supernatant liquid, - filtering to 0.5 ~tm, - recovering the supernatant liquid, - spraying.
A clear beige powder can therefore be recovered with a spray yield of 62.70 by weight.

Example 9 In a first variation of example 8, the following operations can also be carried out:
- adjusting the pH of a proportion of the extract prepared according to example 8 to 4.1 by addition of H2S04 4N, - leaving to rest at +4°C (formation of a precipitate - about 500 of the total volume), - centrifuging to 5,000 g for 15 min, - washing the caps with distilled water at pH = 4.1, - centrifuging to 5,000 g for 15 min, - collecting the precipitate, - reconstituting the precipitate in distilled water (l00 of the volume prior to precipitation), - adjusting the pH to 7.5 by adding NaOH 4N while stirring, - homogenising to assist solubilisation and homogenisation, - centrifuging for 10 min at 5,000 g to eliminate the insoluble material, - spraying the protein concentrate (spray yield based on the dry extract: 82.90a by weight).
Example 10 In a second variation of example 8, the following operations can be carried out:
- adjusting the pH of a proportion of the extract prepared in example 8 to pH 5 by addition of H2S04 4N, - leaving to rest for one hour at ambient temperature, - centrifuging, - collecting the precipitate, - reconstituting the precipitate in l00 of the volume prior to precipitation without carrying out the washing stage, - adjusting the pH to 7.5 by adding NaOH 4N while stirring, - homogenising to assist solubilisation and homogenisation, - centrifuging for 10 min to 5,000 g to eliminate the insoluble material, - atomising the protein concentrate.
Gel permeation analysis over a column of the superose 12 HR
type of the fraction extracted by the various methods described hereinbefore allows at least 6 different fractions to be characterised as shown in the diagrams in Fig. 1 to 6 of the accompanying drawings, in which:
Fig. 1 shows the molecular weight distribution of the proteins of an extract of whole seed flour of Hibiscus esculentus (extraction: 2 hours at ambient temperature in water, pH = 9), Fig. 2 shows the molecular weight distribution of the proteins of an extract of dehusked seed flour of Hibiscus esculentus (extraction: 1.5 hour at ambient temperature in water, pH -9) , Fig. 3 shows the molecular weight distribution of the proteins of an extract of dehusked seed flour of Hibiscus esculentus (extraction: 1.5 hour at ambient temperature in NaCl 10 g/l, pH = 9), Fig. 4 shows the molecular weight distribution of the proteins of an extract of dehusked seed flour of Hibiscus esculentus (extraction in an ultrasonic tube, 1 hour in NaCl 5 g/l, pH -7.5), Fig. 5 shows the molecular weight distribution of the proteins of a protein concentrate obtained by precipitation at pH = 4.1 of an extract of whole seed flour of Hibiscus esculentus, and Fig. 6 shows the molecular weight distribution of the proteins of a protein concentrate obtained by precipitation at pH - S
of an extract of whole seed flour of Hibiscus esculentus.
The following summary table (in two parts) shows the distribution of the various protein fractions extracted.
Extraction pH = pH = 9 pH = pH = pH = pH =
9 9 9 6.5 8 conditions water NaCl 1 NaCl NaCl NaCl NaCl g/1 g/1 g/1 g/1 g/1 4 h PM > 500,000 13.7 9.8 4.4 3.7 2.9 2,7 Da PM between 100,000 and 500,000 Da 12.2 11.2 10.7 14.4 9.7 17.3 PM between 30,000 and 100,000 Da 4 3.3 7.6 7.7 14.7 7.9 PM between 5,000 and 30,000 Da 41.2 44.7 46.3 51.2 49.6 45.9 PM lower than or equal to 5,000 28.9 31 31 23 23.1 26.2 Da Extraction pH = pH = pH = pH Protein Protein 7.5 7.5 7.5 =

conditions NaCl NaCl NaCl Hz0 concentrateconcentrate g/1 g/1 g/1 whola 6 h 15 min 60 min flour (pH 4.1) (pH 5) US US

PM > 500,000 4 13 20.9 30.5 35.5 46.6 Da PM between 100,000 and 500,000 Da 13.1 14.2 3.8 15 12.2 17.5 PM between 30,000 and 100,000 Da 4.3 6.1 15.1 5.9 8.1 7,3 PM between 5,000 and 30,000 Da 50.2 46 35.3 26.4 35.4 22.6 PM lower than or equal to 5,000 28.4 20.7 24.9 22.2 8.8 6 Da The aforementioned 6 protein fractions consist of:
- a fraction (designated F1) having a very high molecular weight (between 1,000,000 and 1,500,000 Da according to column calibration), - a fraction (designated F2) having a molecular weight between 250,000 and 350,000 Da visible mainly on the extracts of whole seed flour, - a fraction (designated F3) having a molecular weight between 130,000 and 180,000 Da, - a fraction (designated F4) corresponding to a molecular weight between 17,000 and 22,000 Da, - two molecular weight fractions of about 3,300 Da (F5) and about 2,300 to 2,600 Da (F6) respectively.
According to the extraction conditions, the fraction F1 constitutes between 2.7 and 50.OOo of the proteins and the fraction F4 constitutes between 16 and 520 (average 400) of the total proteins.
It is noted that, the higher the salt content of the extraction solvent at the outset, the smaller the fraction F1.
The extracts obtained by means of the examples of the described processes can be used directly in liquid form or after drying by conventional dehydration techniques (spraying, freeze-drying, etc.).
The protein fractions extracted according to the aforementioned examples have the advantage, relative to the natural state in which they occur in the seeds, of having an identical native chemical structure, of being able to be partially or completely purified (that is freed of other seed components such as lipids, fibres, saccharoses or the like) and of having a composition which can be varied as a function of the extraction and/or purification process employed (total proteins of the crude extract, protein concentrate or purified protein fraction (s) .
In relation to lactic casein, improved solubility is noted in addition to the desired renewable vegetable origin in the cosmetics field, while maintaining an amino acid composition close to casein, of which the nutritional, moisturising and film-forming properties for skin and the superficial body growths are known.
The protein fractions can be used either in their native form without modification of the initial structures or in the form of natural associations of two or of all the extracted fractions corresponding to the various peaks of the chromatograms shown in the accompanying drawings or again in isolated form, that is the use of one or more fractions corresponding to one or more peaks of the aforementioned chromatograms.
The protein fractions can also be used in the form modified or functionalised by any one of the following treatments:
- polymerisation, - chemical hydrolysis of the hibiscus proteins - enzymatic hydrolysis of the hibiscus proteins. To this end, proteases originating from extracts of animal, vegetable, microbial or fungal origin can be used to modify the hibiscus proteins: pepsin, trypsin, chymotrypsin/papain, pronase, bromelain/endoproteinase, thermitase, proteases of Bacillus subtilis, Aspergillus Niger and Aspergillus Oryzae/subtilisine, alcalase, neutrase.
- microbiological transformation with use of hibiscus proteins as substrate for fermentation by various microorganisms such as yeasts (Saccharomyces), mould fungus (of the Aspergillus type), bacteria such as Bacillus or the like.
- chemical or enzymatic functionalisation by processes such as desamidation, succinylation or phosphorylation.
- quaternisation.
- grafting of saccharidic or lipidic molecules.
The invention also relates to a cosmetic composition, in particular for topical application, for the skin and/or the superficial body growths, characterised in that it contains at least one preferably soluble protein fraction extracted from Hibiscus esculentus seeds as substitute for casein.
According to a characteristic of the invention, the protein fractions) is (are) extracted from non-delipidated or delipidated flour of whole or dehusked Hibiscus esculentus seeds by water or saline solutions at various pH.
According to a preferred embodiment of the invention, the protein fracbon(s) is (are) extracted in aqueous solution under the influence of ultrasound and this fracb on(s) is (are) purified by a purification process selected from the group formed by precipitation, absorption, ion or affinity exchange chromatography and ultrafiltration.
The total or native protein fraction is selected from the group of total and native protein fractions extracted from Hibiscus esculentus seeds and having apparent molecular weights when filtered over gel of 1,000,000 to 1,500,000 Da, 250,000 to 350,000 Da, 130,000 to 180,000 Da, 17,000 to 22,000 Da, 3,300 Da and 2,300 to 2,600 Da.
The aforementioned protein fracbon(s) can consist of~ a chemical or enzymatic hydrolysate prepared from native proteins, can be obtained by polymerisation or depolymerisation of native proteins or can be chemically modified by grafting.
According to a first embodiment of the invention, the cosmetic composition contains at least two protein fractions of different apparent molecular weights.
According to a second embodiment of the invention, the cosmetic composition contains an extract of Hibiscus esculentus seeds consisting of all the soluble protein fractions naturally present in these seeds.
Said cosmetic composition according to one of claims 1 to 11, characterised in that it preferably contains between O.Olo and 50.OOo by weight, preferably between 1 and 25a by weight, of protein fractions) extracted from Hibiscus esculentus seeds as obtained in any one of the examples of processes described hereinbefore.
As non-limiting embodiments of the invention, various cosmetic products or compositions containing at least one soluble protein fraction extracted from Hibiscus esculentus or okra seeds will be described hereinafter.
Example 1 A cosmetic product according to the invention, in the form of a moisturising smoothing day cream for the face and body can, for example, have a composition by weight made up of the following fractions A, B, C and D, as indicated hereinafter.
Fraction A:
- Cutina MD 14.OOo - Eutanol G 6.OOo - Cetiol B 6.OOo - Eumulgin B1 1.500 - Eumulgin B2 1.500 Fraction B:
- Extracts of total native proteins from hibiscus according to the aforementioned example 1 8.OOo Fraction C:
- Allantoin 0.200 - Methylparaben 0.200 - Germall 115 0.300 - Distilled water 62.OOo Fraction D:
- Fragrance 0.300 The process for the preparation and production of the aforementioned day cream essentially involves heating fraction A to 75°C, preparing fraction C at 75°C, pouring fraction A
into fraction B with turbine stirring, adding fraction C at about 50°C then continuing planetary stirring to ambient temperature and finally adding fraction D.
Example 2 A cosmetic product according to the invention in the form of a restructuring, anti-wrinkle nourishing repairing night cream could, for example, have a composition by weight consisting of the following fractions A, B, C and D, as mentioned hereinafter.
Fraction A:
- Miglyol 810 6.OOo - Myrj 51 3.OOo - Arlatone 983 S 2.OOo - Stearic acid TP 4.00%

- Cetyl alcohol 3.00%

Fraction B:
- Propylene glycol 3.OOo - Elestab LS 388 (Laboratoires Serobiologiques) 2.500 - Distilled water 61.200 Fraction C:
- Sprayed extracts of native hibiscus proteins according to the aforementioned example 9 2.OOo - Distilled water 13.OOo Fraction D:
- Fragrance 0.30%
The process for preparing and producing the aforementioned night cream essentially involves preparing and heating fractions A and B to 75°C, pouring fraction A into fraction B
with turbine stirring, preparing (separately and extemporaneously) the solute of fraction C by stirring of the atomisate in distilled water, adding fraction C to emulsion A
+ B while cooling to about 50°C, then continuing planetary stirring from 45°C and to ambient temperature while adding the fragrance at about 40°C.
Example 3 A cosmetic product according to the invention in the form of a protective conditioning covering light-protective microemulsion for hair could, for example, have a composition by weight made up of the following fractions A, B, C and D, as mentioned hereinafter.
Fraction A:
- Brij 96 11.800 - Arlatone G 10.000 - Paraffin oil 13.OOo Fraction B:
- Distilled water 53.700 - Methylparaben 0.20a - Elestab 4112 (Laboratoires Serobiologiques) 0.300 Fraction C:
- Native hibiscus proteins in the form of a protein concentrate according to the aforementioned example 6 5.OOo - Distilled water S.OOo Fraction D:
- Fragrance 0.20%
- Tween 20 0.80%
The process for preparing and producing the aforementioned microemulsion essentially involves preparing and heating fraction A and B to 75°C, pouring fraction A into fraction B
with turbine stirring, carrying out progressive cooling and, at about 50°C, adding the fraction C then the fraction D and continuing stirring until cooling to ambient temperature and perfect homogenisation.
The invention is obviously not limited to the embodiments described and illustrated in the accompanying drawings.
Modifications are possible, in particular with regard to the composition of the various elements or by substitution of technical equivalents, without departing from the scope of protection of the invention.

Claims (12)

Claims
1. Use of at least one protein fraction from Hibiscus esculentus or okra seeds in a cosmetic composition or product.
2. Cosmetic composition, characterised in that it contains at least one protein fraction extracted from Hibiscus esculentus seeds as a substitute for casein.
3. Cosmetic composition according to claim 2, characterised in that the protein fraction(s) is (are) extracted from non-delipidated or delipidated flour of whole or dehusked Hibiscus esculentus seeds by water or saline solutions at various pH.
4. Cosmetic composition according to any one of claims 2 and 3, characterised in that the protein fractions) is (are) extracted in aqueous solution under the influence of ultrasound.
5. Cosmetic composition according to any one of claims 2 to 4, characterised in that the protein fraction(s) is (are) purified by a purification process selected from the group formed by precipitation, adsorption, ion or affinity exchange chromatography and ultrafiltration.
6. Cosmetic composition according to one of claims 2 to 5, characterised in that the total or native protein fraction is selected from the group of total and native protein fractions extracted from Hibiscus esculentus seeds and having apparent molecular weights when filtered over gel of 1,000,000 to 1,500,000 Da, 250,000 to 350,000 Da, 130,000 to 180,000 Da, 17,000 to 22,000 Da, 3,300 Da and 2,300 to 2,600 Da.
7. Cosmetic composition according to any one of claims 2 to 6, characterised in that the protein fraction(s) consist(s) of a chemical or enzymatic hydrolyzate prepared from native proteins.
8. Cosmetic composition according to any one of claims 2 to 6, characterised in that the protein fraction(s) is (are) obtained by polymerisation of native proteins.
9. Cosmetic composition according to any one of claims 1 to 7, characterised in that the protein fraction(s) is (are) chemically modified by grafting.
10. Cosmetic composition according to any one of claims 2 to 9, characterised in that it contains at least two protein fractions having different apparent molecular weights.
11. Cosmetic composition according to any one of claims 2 to 9, characterised in that it contains an extract of Hibiscus esculentus seeds made up of all the soluble protein fractions naturally present in these seeds.
12. Cosmetic composition according to one of claims 2 to 11, characterised in that it contains between 0.01% and 50.00% by weight of protein fraction(s) extracted from Hibiscus esculentus seeds.
CA002286094A 1997-04-16 1998-04-08 Use in cosmetics of a protein fraction extracted from okra seeds Abandoned CA2286094A1 (en)

Applications Claiming Priority (3)

Application Number Priority Date Filing Date Title
FR9704853A FR2762211B1 (en) 1997-04-16 1997-04-16 USE OF AT LEAST ONE PROTEIN FRACTION EXTRACTED FROM HIBISCUS ESCULENTUS SEEDS AND COSMETIC COMPOSITION COMPRISING SUCH A FRACTION
FR97/04853 1997-04-16
PCT/FR1998/000715 WO1998046205A1 (en) 1997-04-16 1998-04-08 Use in cosmetics of a protein fraction extracted from okra seeds

Publications (1)

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CA2286094A1 true CA2286094A1 (en) 1998-10-22

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KR (1) KR20010006358A (en)
AT (1) ATE214266T1 (en)
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CA (1) CA2286094A1 (en)
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WO (1) WO1998046205A1 (en)

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EP1179339B1 (en) * 2000-08-10 2007-03-21 Henkel Kommanditgesellschaft auf Aktien Cosmetic agent containing extracts of malvaceae seeds
FR2814063B1 (en) * 2000-09-15 2002-11-29 Clarins Lab COSMETIC COMPOSITION FOR LIPS CARE
EP1245222A3 (en) * 2001-03-23 2004-03-17 L'oreal Topical composition containing fibres and enzymes
DE10120175A1 (en) * 2001-04-24 2002-10-31 Cognis Deutschland Gmbh Use of milk protein hydrolyzates
FR2829694B1 (en) * 2001-09-14 2003-12-05 Jacques Andhrel NOVEL COSMETIC COMPOSITIONS COMPRISING PLANT EXTRACTS AND THEIR USE AS ANTIRADICAL AGENT
FR2834718B1 (en) * 2002-01-15 2004-12-24 Cognis France Sa COSMETIC AND / OR PHARMACEUTICAL ACTIVE SUBSTANCES
FR2832062B1 (en) * 2002-04-18 2004-02-27 Oreal COMPOSITION IN THE FORM OF AN OIL IN WATER EMULSION AND ITS USES, IN PARTICULAR COSMETICS
EP1398019A1 (en) * 2002-09-13 2004-03-17 Cognis France S.A. Method for protecting and modulating the dermal-epidermal junction
EP2223608A4 (en) * 2007-10-22 2010-12-08 Beijing Dongsheng Agricultural Abelmoschus manihot (linn.) medius kernel product for natural nutritional edible and pharmaceutical raw material
ES2485490T3 (en) * 2012-01-30 2014-08-13 Thd S.P.A. A composition and use of it in the treatment of anal fissures
JP2016033137A (en) * 2015-11-06 2016-03-10 ティアッカディ ソチエタ ペル アツィオニ Composition in treatment of anal fissure and use thereof
JP6807535B2 (en) * 2017-01-20 2021-01-06 ビーエイチエヌ株式会社 Fibroblast growth factor
CN107510038B (en) * 2017-07-24 2021-03-23 华中农业大学 Preparation method of water chestnut gel food
KR102299846B1 (en) * 2021-03-04 2021-09-09 농업회사법인 (주)로가 Cosmetic composition comprising collagen peptides derived from Hibiscus
FR3141066A1 (en) 2022-10-24 2024-04-26 Basf Beauty Care Solutions France Sas New use of a peptide to improve the extracellular matrix of the dermis of the skin and/or mucous membranes

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JPS5888305A (en) * 1981-11-19 1983-05-26 Nikko Kemikaruzu Kk Cosmetic
FR2679443A1 (en) * 1991-07-11 1993-01-29 Faure Jean Paul Vegetable extract and animal extract having dermatological properties

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KR20010006358A (en) 2001-01-26
DE69804203T2 (en) 2002-10-24
WO1998046205A1 (en) 1998-10-22
ATE214266T1 (en) 2002-03-15
DE69804203D1 (en) 2002-04-18
JP2001518910A (en) 2001-10-16
AU7340098A (en) 1998-11-11
EP0975322B1 (en) 2002-03-13

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