Peat is a complex combination of partially degraded plant-derived biomass, varying in composition due to differences in peat-forming plants and stage of decomposition. Peat contains a variety of compounds soluble in organic solvents, known as the lipid or bitumen fraction, consisting of compounds usually categorised as waxes and resins, and further sterols, fatty acids, hydroxy acids, alkanes, alcohols, phenolic acids and terpenoids. Also water-soluble humic substances, such as polyphenolic compounds, and aromatic compounds probably originating from lignine, fulvic substances and carbohydrates are found in peat. The exact composition of peat and its variation as well as the rationale for differences in peat compositions are basically unknown.

Peat has traditionally been used as such in peat baths and peat masks, which have been used to soothe, nourish and invigorate skin. Difficulties have been encountered in handling of dark coloured, tarnishing peat for skin care.

Both solvent extraction and super-critical fluid extraction methods for peat extraction have been used or proposed. Many methods use destructive steps, e. g. acid hydrolysis or extraction with hot liquids, which lead to both polymerisation and decomposition of organic compounds. Furthermore, most of the methods use organic solvents resulting in solvent residues in the final extracts.

In RU 2050853 is described an extraction of peat using carbon dioxide. The used extraction temperature was 25-29 °C, pressure 6.0-7. 0 MPa and extraction time 2 - 3 hours.

The document SU 1754106 discloses the use of a peat wax extract including fat- soluble components in cosmetic-oral hygiene composition, that ensures better cleaning of mouth cavity, prevents inflammation and combats caries.

Fagernas (1986) has extracted dried peat by using supercritical carbon dioxide. The temperature used was 40-80°C and pressure 20-50 MPa. The peat samples originated from Karanginneva (an aapa mire located in Central Finland), Piipsanneva (an aapa mire located in Central Finland) and Kakkeriasuo (a raised bog in southwest Finland). This document does not give any use for the product obtained.

An extraction of peat with an aliphatic solvent with at least 5 carbon atoms under conditions of elevated pressure and temperature for isolation of asphaltic materials was described in GB 2 001 670. There the preferred solvent was n-pentane with preferred temperatures starting from 40 % of the critical temperature of the solvent, which is for pentane about 80 °C.

Peat extracts have been proposed as ingredients in cosmetic formulations and medicinal products. Majority of peat extracts used have been defined as product-by- use or product-by-process, as it has not been possible to define the compounds in the extract. Peat extracts has been used in skin care based on the tradition without thorough knowledge of the bioactive components and effecting mechanism of peat.

It is known that peat contains modest levels of lipophilic substances, of which plant sterols, long-chain alcohols and long-chain fatty acid esters are of particular interest with regard to their physiological effects especially on skin.

In WO 9216216 a peat-derived combination of water-soluble salts of cosmetically and pharmaceutically active compounds is proposed. DE 3402223 proposes a slightly coloured lipoid extract suitable for use in cosmetic products due to its visual properties, which extract was obtained from peat by extraction with myristinic acid isopropylester and/or by paraffin oil. US 4,272, 527 proposes an ethanol extract of peat wax resin containing sterols and estrogenic compounds to be effective in treating of skin diseases. In WO 9858655 an extract containing humic acid is proposed to be used as a prophylactic to haemopoietic system injuries, the extract being produced with aqueous extraction of a specific peat type. In JP 63-222105 (A) an"extracted essence"consisting higher fatty acid esters and amino acids and isolated from low-to high-grade peat with supercritical C02 has been used in skin cosmetic compositions as a water-soluble component.

Sterols and steryl esters are important constituents of cell membranes and epidermal and sebaceous lipids. Incorporation of certain plant sterols into human keratinocyte plasma membrane have resulted in modulation of membrane fluidity as described by Mora et al. , 1999. Topically applied phytosterols have been shown to be effective in improving dry skin by regulating and normalizing the turnover speed of corneocytes in the stratum corneum as described by Kashibuchi et al. , 1993. Plant sterols have also shown anti-inflammatory and skin conditioning properties without significant hormone-like effects. This makes these compounds possible alternatives for partially replacing cortisone and corticosteroids in skin treatment. Phytosterols may also play a role in regulating immune functions of skin.

Long-chain alcohols and long-chain fatty acids are building materials of ceramides, a group of compounds essential for the lipid barrier structure of the stratum corneum of epidermis. Wax esters are important components of the lipid barrier structure.

Aliphatic long-chain alcohols have been reported to be effective in promoting tissue regeneration and wound healing by Mironov et al, 1989.

It has not been possible to control the extract composition obtained from peat by extraction. This far, extracts obtained from peat with different extracting methods have been used as such without clear understanding of the compositions and without possibility to influence the proportions of the different fractions in the extract. It has not been possible to produce a peat extract with clearly defined composition, where the amount of important bioactive ingredients could be tailored according to the demands of the end use. Peat extracts have contained bioactive ingredients in same proportions, in which they exist originally in peat. No method of modification of these proportions has been proposed.

The object of the present invention is to solve or minimise the problems and disadvantages existing in the prior art.

One object of the present invention is to control the extract composition obtained by carbon dioxide peat extraction.

Another object of the present invention is an extract with clearly defined composition.

In order to achieve the above-mentioned objects the present invention is characterised in what is defined in the characterising parts of the independent claims presented hereafter.

In a typical method of extract composition control in extraction of peat by carbon dioxide the extraction temperature is 5-120 °C and pressure 70-1000 bar and the extraction is conducted in presence of a modifier.

A typical peat extract according to the present invention comprises 1-60 weight-%, preferably 1-20 weight-% of sterols.

Now it has been surprisingly found out that by using carefully chosen conditions during carbon dioxide extraction of peat composition of the obtained extract can be controlled and modified according to the specific needs of the end use. Important parameters with which the extract composition can be controlled and modified are the extraction pressure and temperature as well as the type and the concentration of modifier that is used in the extraction together with carbon dioxide.

The present invention can be applied to extraction of peat with carbon dioxide, either at sub-or supercritical conditions, preferably supercritical condition. Using carbon dioxide instead of organic solvents, the process has clear advantages of solvent free extract, safety, low cost and environmental friendliness. Supercritical carbon dioxide extraction is carried out in a system substantially free of oxygen or under low oxygen pressure. As the extraction conditions are generally mild the bioactive components and ingredients can be extracted in their native forms and the extract obtained free of harmful organic solvent residues.

The term"peat"when used in the context of the present invention relates to a material consisting largely of organic residues accumulated as a result of incomplete decomposition of dead plant constituents under conditions of excessive moisture with the aid of a wide spectrum of microbes. The abundance of microflora is a special feature of peat. The formation of peat requires special bog environment and an especially long time for complex changes by microbes. On the average, it takes 3000 to 4000 years to accumulate a meter of peat. Peat for commercial use is generally from layers of 3-12 meters, which has been formed during time periods of up to 10000 years. Thus peat is different from the plant sediments found in normal forests and deserted lands.

The term"skincare product"when used in the context of the present invention relates to moisturising and anti-wrinkle face creams, aftershave creams, moisturising and anti-wrinkle eye creams, moisturising and softening hand and foot creams, face and body creams for anti-inflammation caused e. g. by UV-irradiation, face and body creams for atopic diseases, face and body creams for preventing skin cancer induced e. g. by UV-irradiation and to sun creams for protecting skin from UV damage.

The term"keratin fibre care product"when used in the context of the present invention relates to shampoos, conditioners and balsam products for hair, hair styling products, such as mousses, waxes and gels, hair colours and toners, hair oils and hairsprays, shaving creams, beard and moustache colours and toners, as well as beard and moustache waxes. The term"keratin fibre"comprises both hairs of human and animal origin. The peat extract according to present invention can also be applied to products meant for animals, for example for pets, such as dogs or cats.

The term"modifier"when used in the context of the present invention relates to different solvents, such as different oils, water, acetone, different alcohols or mixtures of these. Examples of suitable oils are different vegetable oils like rapeseed or sunflower seed oils or the like and examples of suitable alcohols are methanol or ethanol. Mixtures of these modifiers may also be used. The term "modifier"is used in the context of the present application interchangeably with the term"co-solvent".

According to an embodiment of the present invention, the modifier is water and it is used in an amount of 1-10 % of carbon dioxide as solvent used for extraction.

According to another embodiment of the present invention, the modifier is ethanol and it is used in an amount of 5-20 % of carbon dioxide as solvent used for extraction.

It is desirable to develop a control method for the extraction process of peat, so that important bioactive ingredients such as plant sterols, steryl esters, long-chain alcohols and long-chain fatty acid esters, can be selectively isolated from peat and/or enriched into obtained extract. These selectively isolated and/or enriched bioactive extracts can be used as ingredients for skin and keratin fibre care.

The present invention relates to a method of control of an extract composition, especially method of control of sterol content, in extracts obtained from non- processed and processed peat by means of carbon dioxide extraction with co- solvents and/or modifiers. The present invention relates especially to a method of control of an extract composition in extracts obtained from non-processed and processed peat by means of supercritical carbon dioxide extraction with co-solvents and/or modifiers. The extracts according to present invention comprise carefully and selectively isolated groups of lipophilic compounds, whose amounts can be controlled to desired levels.

An advantage of the present method over for instance that disclosed in Fagernas (1986) is an increased yield due to the presence of a modifier.

Usually the obtained extracts according to one aspect of the invention are especially enriched with sterols, steryl esters or other sterol derivatives, long-chain alcohols and/or long-chain fatty acids. The long-chain alcohols and fatty acids have preferably main chain length of 16 to 40 carbon atoms. Typically the peat extract comprises 10-60 weight-%, preferably 20-40 weight-% of longs chain alcohols and 10-60 weight-%, preferably 10-40 weight-% long-chain fatty acids.

According to one embodiment of the invention the peat extract can comprise up to 70 weight-% long chain alcohols. The major alcohols are usually octadecanol, eicosanol, docosanol, tetracosanol, hexacosanol, and octacosanol.

In addition, wax esters, acylglycerols and their derivatives as well as some special fatty acids such as hydroxyl fatty acids can be present in the extracts. The dominating fatty acids are usually palmitic, stearic, eicosanoic, docosanoic, tetracosanoic, pentacosanoic, hexacosanoic, and octacosanoic acids. Typically the peat extract comprises 30-50% wax esters comprising esters of alcohols and fatty acids with chain-length of typically 20-30 carbon atoms. The long-chain alcohols and long-chain fatty acids are normally in free form or in the form of wax esters.

It is believed that the waxes, long chain alcohols and sterols are important compounds for the skin health, for example. These components are either essential constituents of skin lipids or bioactive compounds with beneficial effects on skin.

By using the present invention the total sterol content can be increased up to 60 weight-% of the extract. Typically the total sterol content can be increased up to 20 weight-% of the extract. According to one embodiment of the invention the sterol content in peat extract can be increased up to 10 weight-%. The major sterols found in the extract are sitosterol, campesterol, sitostanol, stigmasterol, cycloartenol and 24-methylenecycloartanol. The structures of sitosterol (A), campesterol (B), sitostanol (C), stigmasterol (D), cycloartenol (E) and 24-methylenecycloartanol (F) are shown in Figure 1. The sterols are normally in form of free sterols, steryl esters, or acylated steryl glycosides. In comparison it can be stated that the corresponding sterol content in extracts obtained with conventional uncontrolled processes is normally lower than 1 %.

The nature of the extraction process and of the control method according to the present invention is non-destructive, which makes it possible to isolate the compounds in peat without causing any changes in their chemical structure during the extraction. Use of the method according to the present invention enables the

production of peat extracts, which differ from extracts produced with conventional methods in both chemical composition and physical properties. The extracts obtained are usually light-coloured, free-flowing powders having a high content of sterols, long chain alcohols and/or fatty acids.

Peat or pre-treated peat, from which is used as raw material in the extraction process, can be of various compositions. Differences in the composition relate to the differences in peat forming plants and to differences in the processes, which have turned them into peat. The composition of fractions isolated can also be influenced by the choice of peat and/or pre-treated peat from which the fractions are isolated.

According to one preferred embodiment of the invention the peat, which is used as a raw material in the extraction, is highly decomposed peat, such as grade 7-8 von Post humification scale, from e. g. Spagnum and Carex plants.

According to the present invention by adjusting of the extraction temperature, pressure as well as the types and concentrations of the modifiers used, the solvating property of carbon dioxide, preferably supercritical carbon dioxide, can be modulated considerably, especially in the region close to the critical temperature or pressure. This will yield extracts, wherein sterol content can be adjusted according to the needs. The selectivity of the method can be controlled by extraction pressure and temperature as well as by application of co-solvents and/or modifiers. These parameters govern the solvating power of carbon dioxide, preferably supercritical carbon dioxide, for different groups of compounds.

The extraction pressure varies according to the present invention from 70 to 1000 bar, preferably from 480 to 900 bars. The extraction temperature varies from 5 to 120 °C, preferably from 40 to 80 °C.

According to the present invention a modifier is added to influence the solubility of specific compounds during extraction. Most of the modifiers are used for increasing the polarity of C02 and thus solubility of certain slightly more polar components.

The concentration of modifiers should be adjusted according to the polarity of the modifier and target solutes to be extracted.

According the present invention the sterol content in the obtained extract can be adjusted to a level ranging from 1 to 60 weight-%, typically from 1 to 20 weight-%, by modifying the above-mentioned process parameters, i. e. temperature, pressure, type or concentration of a modifier. For example, when increasing extraction temperature from 20 °C to 60 °C and extraction pressure from 100 bars to 500 bars, the yield of total sterols and long chain alcohols are significantly increased. Use of water as modifier, in amounts ranging from 0.1-5 weight-%, typically 0. 5-1 weight-%, will increase the yield of components with slightly higher polarity such as free sterols. According to one embodiment of the invention the amount of modifier varies between 1-3 weight-%.

The present invention also relates to a peat extract obtainable by the present method, provided that the peat originates from a mire that is not selected from the group consisting of Karanginneva, Piipsanneva and Kakkeriasuo. These mires are the ones mentioned in Fagernas (1986). It is suspected that the extract obtained depends on the mire the peat comes from.

Since peat is a complex of biomass, the extraction process according to present invention also reduces the content of microbes in the extract to a minimum level. A high content of microbes would limit the application of the extract, therefore the extraction procedure reduces the amount of microbes in the extracts to a minimum level, by adjusting the density of supercritical C02 and the particle size of the peat raw material. For example, under extraction conditions around 60 °C and 500 bars

the microbes will be partially inactivated and remain mainly with the extraction residues.

The method according to invention can also include a pre-treatment of peat and/or post-extraction separation. According to one embodiment of the invention the peat is processed by drying, heating, milling, pelletising before the extraction. A preferred pre-treatment is pelletising, which often improves the economics of the process significantly due to the improved penetration and flow of carbon dioxide through the raw material.

The present invention can be carried out in a single autoclave or in two or several autoclaves connected in various configurations to each other e. g. in parallel and/or in series. Two, three or several autoclaves may, for example, be connected to each other to enhance the capacity and profitability of the process. Extraction can be carried out at any scale from laboratory scale and pilot-scale to various full-scale industrial processes; and it can be conducted as a batch-wise, semi-continuous or continuous process. It is possible to intensify the extraction process using e. g. supporting materials, such as diatomaceous earth, which can be mixed with raw materials to enhance the extraction efficiency. The extraction process according to the invention is also faster than common solvent extraction used previously due to e. g. a low number of unit processes. Process steps can also include feeding of peat or pre-treated peat using any feeding system, e. g. screws, belts or conveyors.

The obtained peat extract can be isolated from the extraction process by e. g. transferring the extract from the extraction system (s) with fluids or gases into separators or depressurising devices. A separator may be a dual separator in which the extract can be fractionated at different temperatures and pressures to achieve additional fractionation of the product. Specific fractions in the obtained extract can be isolated by purifying or fractionating the original extract, removing the more

soluble compounds in the original extract with fluid or gas. This can be done in either a dynamic or static process in an autoclave or autoclaves or in fractionation or separation columns with e. g. a counter-current extraction process.

Extracts obtained from peat by the method according to the invention are preferably odourless solid powders, which are free of organic solvent and microbes and have a light, yellow-white colour. These powders are typically grainy and free-flowing when dry, which makes their handling and use easy. Thus they are especially useful as medicinal and/or cosmetic products or as ingredients in such products.

The extract according to the invention comprises plant sterols, plant steryl esters, long-chain alcohols, long-chain fatty acids and wax esters. These compounds are either direct constituents or molecular units of the lipids of skin barrier structure controlling the permeability of water, lipids as well as water-and lipid-soluble substances through skin. For example long-chain alcohol and long-chain fatty acids are molecular units of ceramides, an important component of skin barrier lipids. A preferable composition of the lipids in the skin barrier system is advantageous for the health and for the normal functions of skin, such as water-retaining capability and sensitivity of skin. When applied topically, the extract according to the present invention can provide skin with nutrients essential for building the barrier system and also have a series of beneficial physiological effects on skin. The uses and products of the present application are not limited to peat extracts obtained by the present method, but also to peat extracts obtained by carbon dioxide extraction without using a modifier.

As some examples, it may be said that by extracting at low temperature of 20-40 °C and pressure e. g. at 100-200 bars, and without the use of modifiers, the extract obtained will consist of 80-90 % waxes with low contents of free fatty acid (1-5 %), free alcohols (1-5 %) and sterols (0.1-1 %).

On the other hand, by extracting at low temperature and pressure e. g. at 300-600 bars, 40-60 °C and without the use of modifiers, extracts can be obtained that consist of 40-70 % waxes with content of free fatty acid (10-20 %), free alcohols (10-20 %) and sterols (1-5 %).

By extracting at low temperature and pressure e. g. at 400-900 bars, 40-100 °C with the use 5-20 % ethanol or 1-10 % water as a modifier, the extracts can be obtained, consisting of 5-20 % waxes with high content of free fatty acid (10-40 %), free alcohols (10-40 %) and sterols (10-50 %).

Typically an extract according the present invention comprises 40-70 weight-% sitosterol, 10-20 weight-% campesterol, 2-5 weight-% cycloartenol and 2-5 weight-% 24-methylenecycloartanol, all calculated as proportion of the total sterols.

The method according to the present invention can be made specific compared to e. g. organic solvent extraction processes.

Peat extract according to the present invention can be used in skin care or keratin fibre care products, general hygiene products and/or cosmetic products. The amount of the peat extract in the different products is 0.1-99. 9 weight-%, typically 1-90 weight-%, more typically 5-40 weight-%.

Thus, according to one preferred embodiment of the present invention the sterol- enriched peat extract, is used as bioactive ingredient in cosmetics, skincare products for improving skin moisture, decreasing skin sensitivity, retarding skin aging, alleviating skin inflammation, reducing the risk of skin cancers, and improving the general appearance of skin surface. The peat extract can also be used as an ingredient or component in keratin fibre care products to improve the health and outlook of keratin fibres, such as human or animal hair.

Based on the skin-beneficial effects plant sterols and long-chain alcohols described in this application, the extracts according to present invention can furthermore be applied as bioactive ingredients in skincare and keratin fibre care products with functional targets of skin-moisturising, anti-inflammation, anti-skin cancer, decreasing skin sensitivity, regulating immune function, anti-UV products, promoting tissue regeneration as well as improving the general health condition of skin.

According to one embodiment of the invention skin care product comprises peat extract obtained by a method according to the present invention at levels ranging from 0.1-99 weight-%, preferably 5-30 weight-%. The extract can be used as single bioactive ingredient or in combination with other ingredients such as natural and/or synthetic antioxidants and vitamins such as vitamin C, tocopherols and tocotrienols. Also other natural extracts or synthetic ingredients containing skin- beneficial compounds can be added. Preferably the extract according to the present invention is used in combination with other bioactive ingredients such as vitamins, antioxidants, proteins, and/or amino acids.

According to one embodiment of the invention the peat extract can be used in general hygiene products, such as antiperspirants, talcum powders, baby powders, facemasks, skin exfoliating products or washing agents, such as soaps and shower gels. According to one embodiment of the invention the amount of extract in the final product can be 0.1-99 weight-%, preferably 5-20 weight-%. The extracts can be used either as sole bioactive compounds or preferably in combination with other ingredients beneficial to skin.

According to another embodiment of the invention the peat extract can be used in other cosmetic products, such as lipsticks, lip-glosses, lip creams, mascaras, eye-

brow pencils, kohl pencils, lip liners, cover sticks, blushers, foundation creams, face powders, eye shadows, cuticle creams, nail oils or colours. According to one embodiment of invention the amount of peat extract is usually 0.1-99. 9 weight-%, preferably 20-40 weight-%, either as sole bioactive ingredient or preferably in combination with other beneficial ingredients such as fatty acids, vitamins C and E, and ceramides.

The present invention also relates to a skin care formulation that comprises 0.1-99, 9 % of a peat extract according to the present invention. This kind of skin care formulation may be used for example as a facial cream, a body lotion or a hand cream. The amount of the peat extract is selected according to the target and the effect wished to be obtained. It may be for example 1-10 %, 5-25 %, 1-50 %, 10-55 %, 20-80 %, 50-90 %, 70-99 % or 60-87 %.

The present invention further relates to lipstick formulation that comprises 0.1-99, 9 % of a peat extract according to the present invention. This kind of lipstick formulation may have moisturising and anti-wrinkle properties. The amount of the peat extract is selected according to the target and the effect wished to be obtained.

It may be for example 1-10 %, 5-25 %, 1-50 %, 10-55 %, 20-80 %, 50-90 %, 70-99 % or 60-87 %.

The present invention still relates to a hair care formulation that comprises 0.1-99, 9 % of a peat extract according the present invention. This kind of hair care formulation may have moisturising properties. The amount of the peat extract is selected according to the target and the effect wished to be obtained. It may be for example 1-10 %, 5-25 %, 1-50 %, 10-55 %, 20-80 %, 50-90 %, 70-99 % or 60-87.

Without wishing to be bound by a theory, it is believed that peat extracts (0.1-99 % in the final formulation) smooth the skin and reduce wrinkles. It is also believed that

peat extracts increase moisturisation and improve elasticity of skin. Peat extracts also promote the turnover (renewal process) of stratun corneum of skin. This will reduce the hyperpigmentation and photoageing of skin after sun bath. Thus the extract can be used as skin lightening ingredients. Furthermore, peat extracts can be used as effective anti-ageing ingredients in skin formulations. Also, peat extracts rich in long chain alcohols extraction are skin regenerative ingredients for skin care.

On the other hand, peat extracts rich in sterols are effective in protecting skin from UV irradiation and suitable for suncare formulations.

EXPERIMENTAL PART Example 1 The invention was carried out as follows. Peat was dried and milled in a hammer mill. A volume of 100 1 of milled peat was placed in an extraction basket of 300 1, transferred in an autoclave and extracted by supercritical carbon dioxide at a pressure on 480 bar and at a temperature of 70 °C. The flow rate of the circulating fluid was 30 1/min. Extract was separated during the fluid circulation at lowered pressure in a separator at a pressure of 50 bars and temperature of 30 °C. The carbon dioxide was be re-circulated by pumps and heat exchangers back to the autoclave.

Water was used as a modifier at a level of 1 % of the C02.

Using the extraction pressure and temperature as described in Example 1, and using a vegetable oil as modifier, a homogenous gel form extract can be obtained. This extract contains 30-80 % oil and 20-70 % peat extract. The extract is easy to be incorporated in various cosmetic products.

Example 2 Highly decomposed peat of grade 7-8 von Post humification scale was extracted according to method of Example 1. The obtained product, i. e. extract contained up

to 10-20 weight-% sterols and up to 60-70 weight-% long-chain alcohols and fatty acids. The extract had light yellowish colour and was devoid or nearly devoid of any smell and free of microbes.

Example 3 Peat material was obtained from natural bogs and mires of selected locations in Finland. After being air-dried, the peat was milled to a particle size 0.2-0. 9 mm and pelletised before extraction. The supercritical extraction was carried out under conditions of temperature 40-60 °C and pressure 300-500 bars. The extraction time was 1-2 hours. Water was used as modifier. The extract was dried at 40 °C to water content less than 10 %.

After saponification of the peat extract, the unsaponifiables were separated, and the long-chain alcohols, acids and sterols were analysed with GC-FID and GC-MS. A capillary column DB-1701 (30 m x 0.25 mm, df 0.20 Om) was used for GC analysis.

In Figure 2 a typical GC-FID chromatogram of peat extract is shown. The column temperature program was 230 °C hold 1 min, followed by an increase to 275 °C at a rate of 1 °C/min, and hold at 275 °C for 30 min. The injector temperature was 280 °C, and detector temperature 300 °C.

Example 4 One embodiment of the present invention is shown in Figure 3, which shows a flow diagram for supercritical fluid extraction utilising carbon dioxide and using milled peat as raw material. In Figure 1 the milled peat is fed batch wise into autoclaves, i. e. extractors 1,2, 3. Carbon dioxide is fed from the carbon dioxide storage tank 4 through a sub-cooler 5 and pumped with pumps 6,6'through a pre-heater 7 to the extractors 1,2, 3, which are set up in series. Extract is fractioned in two successive decompression stages and the extracted fractions are collected from separators 8, 9.

The carbon dioxide can be returned to the storage tank 4.

Example 5 The effect of a single application of a peat extract on cutaneous moisturisation was studied.

Methods Twenty (20) female volunteers of different skin types at ages between 18 and 60 were selected. The volunteers stayed in rest for 30 minutes, in an acclimatized room with temperature 20 2 °C and relative humidity 50 5 %. Symetrical areas of 25 cm2 of the skin on the anterior area of the legs or arms were chosen at random and demarcated: an area for the application of the test product, an area for base cream, an area to be kept untreated as negative control.

The test product consisted of a base cream wherein 5 % of a peat extract was added.

The base cream contained the following incredients: aqua, hydrogenated polydecene, isopropyl miristate, steareth-2, cetearyl alcohol, steareth-21, propylene glycol, imidazolidinyl urea, methylparaben and propylparaben. The base cream was used as a placebo.

The peat extract used contained mainly waxes (60 %), free (C18-C30, 10 %) straight chain alcohols and free fatty acids (C16-C28 saturated and monounsaturated, 10 %).

The total sterol content in the extract is 8 %, of which 60 % is sitosterol, 5 % sitostanol, 5 % isofucosterol, 10 % campesterol.

Before the application of the product or placebo, the measurement of skin hydration was carried out with corneometry, determining the electric capacity of each area.

The average value of each area was obtained from the arithmetic mean of twenty measures.

0,05 g of the test product or 0,05 g of the placebo were applied once on their corresponding previously demarcated areas on each volunteer. The electric capacity of the areas was measured after 60, 120, 180 and 360 minutes, while the volunteers were staying in rest in an acclimatized room. The final value for each area was obtained from the arithmetic mean of twenty measures.

Exploratory data analysis was performed (means, standard deviations, minimum, maximum, median, boxplots, line plots and bar plots) using the software SAS System for Windows version 8.0. The comparative analysis was carried out using Student t Test.

Results A single application of the test product significantly increased the skin moisture compared to the same area before treatment (p < 0.001), the untreated area (p < 0.001), the area treated with base cream (p < 0.05) at 120 min and 180 min after the application.

Figure 4 shows the corneometry mean value (indication of skin moisture) of the skin areas of different treatment, showing superior moisture of skin treated with test product containing 5 % peat extract. In the Figure, time in minutes is presented on the abscisse and the mean capacitancy is presented on the ordinate. The squares positioned on their apex illustrate the results for the test product, the squares the results of the placebo and the triangles the results of the control areas.

Figures 5-9 show the difference between the baseline value and the values after treatments. Compared with the control and the base cream, product containing 5 % peat extract gave superior moisture to skin over all time points measured and for area size under the curves.

Figure 5 shows the boxplots of the differences in moisturisation of skin between the baseline corneometry values and the corneometry values obtained at 60 min after treatment with the test product A, the base cream B, and untreated control C.

Figure 6 shows the boxplots of the differences in moisturisation of skin between the baseline corneometry values and the corneometry values obtained at 120 min after treatment with the test product D, the base cream E, and untreated control F.

Figure 7 shows the boxplots of the differences in moisturisation of skin between the baseline corneometry values and the corneometry values obtained at 180 min after treatment with the test product G, the base cream H, and untreated control I.

Figure 8 shows the boxplots of the differences in moisturisation of skin between the baseline corneometry values and the corneometry values obtained at 360 min. after treatment with the test product J, the base cream K, and untreated control L.

Figure 9 shows the boxplots of the values of the areas under the curves of the differences between the baseline and the corneometry values obtained along the time for different treatments, with the test product M, the base cream N, and untreated control O.

Example 6 The effect of peat extract on skin roughness and wrinkles was studied.

Methods Six female subjects, age of 30 to 60 year with wrinkles on face, were selected. For each subject, half of the face was treated with base cream, whereas the other half was treated with test product containing base cream and 1 % peat extract, twice a

day for 8 weeks. The base cream and the peat extract were identical to those used in Example 4.

The sides of face and the application of product on each side were randomized.

Before and after treatment, several cutaneous parameters related to skin wrinkling were instrumentally analysed of facial skin. Cutaneous elasticity was determined using Cutometer SEM 575. Cutaneous hydration level was determined with Cornemeter CM 825 Combi 3 Courage & Khazaka. Cutanous roughness was measured by cutaneous replica and image analysis using Quantilines, Monaderm.

The statistical data analysis was carried out with software SAS system for Windows version 8.0.

Results Effect of peat extract on hydration level of skin The area treated with the product showed a highly statistically significant increase in hydration values (an increase of 21. 4 %, p < 0.01).

Product containing peat extract showed superior hydration values compared with the base cream (an increase of 15.2 %), as can be seen in the histogram in Figure 10 wherein the cornemetry value is presented on the ordinate. The left-most couple of bars represents the results before treatment, the middle couple of bars represents the results after treatment and the right-most couple of bars represents the changes after treatment. In the bar couples, the left bar represents the result for the test product, and the right bar the results for the placebo (base cream only).

Effects of peat extract on skin elasticity Ro value obtained from cutometer defines the maximal deformation of skin. The lower the value is, the higher the skin elasticity.

After two months of treatment, the area treated with the test product containing 1 % peat extract showed a highly statistically significant decrease (29 %) in Ro values (p=0.01). This is superior to the result for the area treated with base cream only, which showed a decrease of 14 % in Ro values. The results are given in the histogram of Figure 11 wherein the Ro values (maximal deformation) is presented on the ordinate. The order of the bars is identical to that of Figure 10.

In the test of overall elasticity, the product containing base cream and 1 % peat extract increased the overall elasticity, whereas the base cream decreased the overall elasticity. The histogram in Figure 12 shows the R2 values (overall elasticity) in ordinate for the test. The order of the bars is identical to that of Figure 10.

Effects of peat extract on skin surface roughness Ra value (means roughness) The treatment with a cream containing 1 % peat extract decreased the mean roughness of the skin by 16.5 %, whereas the base cream decreased mean roughness only by 3.5 %, as shown in the histogram of Figure 13, wherein the Ra (mean roughness of skin) is presented on the ordinate. The order of the bars is identical to that of Figure 10.

Rz value (Maximum roughness value) The treatment with cream containing 1 % peat extract decreased the maximum roughness by 7.4 %, whereas the base cream only led to a decrease of 1.3 %, as

illustrated in the histogram of Figure 14, wherein the Rz values (maximum roughness of skin) is presented on the ordinate. The order of the bars is identical to that of Figure 10.

A cream containing a peat extract according to the present invention thus shows significant improvement in skin surface smoothness and reduction of wrinkles.

Without wishing to be bound by a theory, the applicants believe that this is due to the lipid extract from peat.

It will be appreciated that the present invention can be incorporated in the form of a variety of embodiments, only a few of which are disclosed herein. It will be apparent for the specialist in the field that other embodiments exist and do not depart from the spirit of the invention. Thus, the described embodiments are illustrative and should not be construed as restrictive.

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Mora, M. P. ; Tourne-Peteilh, C.; Charveron, M.; Fabre, B.; Milon, A.; Muller, I.

(1999) Optimization of plant sterols incorporation in human keratinocyte plasma membrane and modulation of membrane fluidity. Chemistry and Physics of Lipids, 101 (2), 255-265.

Fagernas, L. (1986) Extraction of peat lipids with supercritical carbon dioxide. Acta Chemica Scandinavica, A 40,538-545.