JPH0310609B2 - - Google Patents

Description

[Detailed description of the invention]

The present invention purifies aloe sap selected from aloe vera, aloe aloe aloe, and aloe albopictus to produce a substance that is soluble in water, has no bitter taste, is colorless to pale yellow, and has physiologically active effects such as wound healing effects. This invention relates to a method for purifying aloe sap for obtaining. Conventionally, aloe vera or aloe extract has been widely used especially in the private sector to treat cracks, chaps, burns, cuts, etc. However, aloe vera or its extract has the odor and bitter taste characteristic of aloe, and its color is dark. In addition to being brown in color, it contains colored pigment components that easily discolor, and water-insoluble resin components that tend to ooze out in formulations, so aloe and aloe extract can be used as they are, or added to cosmetics and external preparations. However, it is not stable and has major problems such as odor, color, and the formation of precipitates, which limits the scope of its use and also limits the amount of it when incorporated into cosmetics. It was hot. For this reason, various methods for obtaining fractions from aloe have been proposed. For example, US Pat. No. 3,103,466 discloses a method of concentrating Aloe vera sap, adding alcohol to it, washing the resulting precipitate with ether, and drying and powdering it to obtain purified uronic acid. In addition, USP 3360511 discloses that after extracting the juice from aloe vera leaves and removing the precipitate with 2% H ₃ PO ₄ .12MoO ₃ , alcohol is added, and the resulting precipitate is bleached with perchloric acid. A method is disclosed in which a purified polysaccharide is obtained by washing the precipitate with alcohol and drying it into powder. Note that in both USP Nos. 3,103,466 and 3,360,511, only components insoluble in the water-alcohol mixed solvent are used as active ingredients. Further, USP 3,470,109 describes a method in which a jelly part is collected from an aloe leaf, the jelly part is homogenized and filtered, and then freeze-dried and powdered. Note that water-insoluble components and colored substances are not removed by this method. Furthermore,
USP 3892853 and USP 4178372 disclose that colored substances in aloe vera gel are decolorized with hydrogen peroxide solution,
A method has been proposed to obtain a stable aloe vera gel by adding a stabilizer, but this method does not remove water-insoluble components. Furthermore, in JP-A-54-151113, the sap of Kidachialoe is extracted with ethanol, the extract is concentrated to obtain a yellowish brown solid, and this is subjected to silica gel column chromatography to extract ethyl acetate. - It has been disclosed that effluent is carried out in a methanol mixed solvent system and a specific effluent is collected and used as an active ingredient for wound healing, but this method does not allow for colored pigments that easily change color. has not been completely eliminated. The present inventors have conducted various studies on fractions from aloe, and have found that the sap of aloe vera, aloe vera, aloe aloe, and aloe aloe aloe is treated with activated carbon by the batch method. In addition, before or during the activated carbon treatment, 50~
By performing heat treatment at 90℃ for 0.5 to 3 hours,
Contains no water-insoluble substances, is soluble in water and has no bitter taste.
A stable colorless to pale yellow fraction with a pleasant taste can be obtained through simple operations, and this fraction also has excellent bioactive effects such as wound healing and is highly safe for use. This knowledge led to the present invention. That is, the present invention provides 50% of the sap of aloe selected from aloe vera, chabon aloe, and kidachi aloe.
While or after heat treatment at ~90°C for 0.5 to 3 hours, activated carbon treatment is performed by a batch method using activated carbon in an amount of 0.1 to 20% by weight based on the weight of the sap, and then do,
Provided is a method for purifying aloe resin, which comprises collecting a liquid. The present invention will be explained in more detail below. In the method for purifying aloe sap according to the present invention,
Aloe vera, Syabon aloe, and Kidachi aloe are used as the raw materials. Also,
Aloe sap is obtained by squeezing, crushing, crushing, etc. the whole plant or leaf part of these aloe plants, or the jelly part obtained by removing the leaf skin from the leaf part. In the present invention, the sap obtained by compressing, crushing, crushing, etc. the whole aloe plant, leaf part, or jelly part is used, but in this case, the sap obtained by compressing, crushing, crushing, etc. In other words, the sap mixed with solid matter may be passed on to the next process as is. However, it is recommended to remove solids by an appropriate separation method such as centrifugal filtration, filter press filtration, natural filtration using paper or cloth, etc., and use only the sap in the following steps for ease of operation. Desirable from this point of view. Note that if the solid matter is not removed at this stage, it will be removed at a later appropriate stage. In the method of the present invention, the aloe sap thus obtained is subjected to activated carbon treatment and heat treatment using a batch method. This batch method of activated carbon treatment reliably removes colored components that easily discolor, and reliably yields a colorless to pale yellow, non-bitter, water-soluble substance that has physiologically active effects such as wound healing. On the other hand, if activated carbon treatment is not performed, colored components and water-insoluble components are not removed well, and the object of the present invention cannot be achieved. In addition, heat treatment further reliably removes water-insoluble components, as well as components that become water-insoluble due to heat denaturation and components that become water-insoluble due to denaturation over time (inflammatory substances). Therefore, by performing heat treatment in addition to activated carbon treatment, water-insoluble substances and substances that denature with heat or over time (inflammatory substances) are removed, resulting in a stable physiological substance that does not change over time. An active substance is obtained. Furthermore, the heat treatment eliminates the viscosity of the aloe sap, making over-processing extremely easy, which is advantageous in terms of operability. In this case, the heat treatment and the activated carbon treatment by the batch method are such that the activated carbon treatment is performed after the heat treatment, or the heat treatment and the activated carbon treatment are performed simultaneously. When the activated carbon treatment is performed after the heat treatment, the time for the activated carbon treatment can be shortened and the amount of activated carbon used can be reduced compared to when the activated carbon treatment is performed first. Furthermore, by performing the heat treatment and the activated carbon treatment simultaneously, the overall manufacturing time can be shortened, and the heating time and amount of activated carbon used can also be reduced. Furthermore, as mentioned above, the heat treatment eliminates the viscosity of the aloe sap, making it easier to operate. The heat treatment is performed under normal pressure or reduced pressure,
It is carried out at a temperature of 50 to 90°C, more preferably 60 to 80°C, and as mentioned above, this heat treatment more reliably removes water-insoluble components and colored components, and
Components that become water-insoluble due to heat denaturation and denaturation over time (inflammatory substances) are removed. Also, the heating time is
It is 0.5 to 3 hours, more preferably 1 to 2 hours.
If the heating temperature is lower than 50°C or the heating time is shorter than 30 minutes, the effect of the heat treatment will not be sufficiently exhibited, and the above impurity components will not be sufficiently removed in the process of separating insoluble materials. . Also, the heating temperature is 90
If the temperature is higher than 0.degree. C., or if the heating time is longer than 3 hours, problems such as deterioration and fermentation of the active ingredients and decomposition and solubilization of insoluble materials occur. In the present invention, the heat treatment may be performed by simply heating the aloe sap under the above treatment conditions, but the aloe sap can be concentrated at the same time when performing this heat treatment (in this case, the aloe sap is 10 times or less, more preferably 2 to 5
(It is preferable to make a concentrated solution of about twice the size), and it is also possible to evaporate the water almost completely. If activated carbon treatment is performed after heat treatment, use the product as it is or after removing water-insoluble matter. In addition, when performing activated carbon treatment after heat treatment, if the sap is concentrated during this heat treatment process, water should preferably be added to the sap in an amount of 2 to 10 times the initial amount of sap.
It is preferable to add activated carbon to the concentrated solution. In the method of the present invention, the activated carbon treatment is carried out by employing the batch treatment method in which activated carbon is added to aloe sap and stirred, as described above. In this case, the use of activated carbon column chromatography requires the use of a relatively large amount of activated carbon and is difficult to industrialize. A high proportion of sugars and other substances are adsorbed and lost, resulting in a low yield, and because the proportion of active ingredients (organic substances) adsorbed is high, inorganic substances such as sodium chloride are present in the resulting fraction. The content is high, and therefore, when applied to a wound site, there is a risk of pain, which poses a problem in terms of usability. Moreover, activated carbon column chromatography is not employed in the present invention because water-insoluble components are not completely removed and inflammatory substances are included. On the other hand, by performing activated carbon treatment using the batch method, a relatively small amount of activated carbon can be used.
Not only can the amount of activated carbon used be reduced and costs reduced, but the yield can be improved by preventing the loss of active ingredients, especially amino acids and polysaccharides, and the content of inorganic salts can be reduced. It can improve pain during external application, and it can also almost completely remove water-insoluble substances, reliably remove inflammatory substances, and increase water solubility.
It is possible to reliably remove colored pigment components that easily change color, and reliably obtain a fraction that is free from bitterness and has good taste. The activated carbon used in this batch method activated carbon treatment may be animal charcoal, such as animal charcoal or bone charcoal, or vegetable charcoal, such as coconut shell charcoal, but vegetable charcoal, which has high adsorption power and is inexpensive, is more preferable. Activated carbon may be powdered or granular, but powder, especially 100%
~400 mesh is preferred. The amount of activated carbon used is 0.1 to 20% (weight %, same hereinafter), more preferably 1 to 10%, based on the weight of the original aloe sap. In the case of activated carbon treatment by the batch method, after adding the activated carbon, it is preferable to stir the liquid by means such as mechanical stirring, and in this case, the stirring time is 30 minutes to 1 hour. Note that the activated carbon treatment can be carried out at low temperature or room temperature, and when carried out simultaneously with the heat treatment, it is carried out under heating. After the activated carbon treatment, if other treatments are to be carried out afterwards, the activated carbon can be removed and then subjected to the prescribed treatment, and if the activated carbon treatment is also carried out as a final step, it can be separated as appropriate. Activated carbon is removed by a means, and the resulting liquid is collected as a target fraction. The target fraction obtained by the activated carbon treatment is almost colorless and transparent in the form of a treatment liquid. This target fraction has colored pigment components that easily change color and water-insoluble components removed, and includes polysaccharides, organic acids, glutamic acid, aspartic acid, serine, alanine, proline, histidine, valine, threosine, and leucine. , amino acids and proteins such as isoleucine, glycine, and lysine, Mg, Ca,
Contains water-soluble inorganic salts such as K and Na. It has a slightly sour taste and an amino acid-like flavor, is not bitter, and has excellent physiologically active effects such as wound healing. Therefore, it can be suitably used for various purposes as described below. Depending on the intended use, the treatment liquid can be used as it is as a target fraction, but it can be used by distilling off water by employing an appropriate drying method such as heat drying, spray drying, or freeze drying. It is preferable to collect it as a free-flowing white to pale yellow powder from the viewpoint of storage stability and wide-ranging usability. In the present invention, in addition to activated carbon treatment and heat treatment, extraction treatment with a water-soluble organic solvent can be performed, and resins and other poorly alcohol-soluble substances can be removed more reliably by this extraction treatment with the organic solvent. Can be done. This extraction treatment may be performed at any stage before or after the activated carbon treatment or heat treatment, but especially when performing the above three treatments, it may be performed before or after the activated carbon treatment after the heat treatment, Alternatively, it is preferable to perform the extraction treatment before or after the step of simultaneously applying the heat treatment and the activated carbon treatment, thereby reducing the amount of activated carbon used and shortening the treatment time. In addition, when performing extraction treatment with a water-soluble organic solvent prior to activated carbon treatment or heat treatment,
Extraction is performed by adding an organic solvent to the aloe sap, and the extract is collected and the organic solvent is distilled off, or the organic solvent is distilled off and water is added to it for the next process (activated carbon treatment process, heating). processing process). When extraction treatment with a water-soluble organic solvent is performed after activated carbon treatment, extraction is performed by adding an organic solvent to the treated liquid obtained by removing activated carbon, preferably the concentrated liquid or dried product, and heat treatment. If extraction is performed after heat treatment, an organic solvent is added to the sap after heat treatment, preferably the concentrate or heat-dried product (by performing the extraction treatment on the concentrate or dry product in this way, the amount of solvent used can be reduced). After the extraction, the extract, its concentrate, or the solvent distilled off is collected as described above. In addition, when extraction treatment is performed as the final step, it is preferable to distill off the solvent from the obtained extract and collect it as the desired fraction, but depending on the purpose of use, distilling off the solvent may be necessary. Otherwise (particularly when ethanol or isopropanol is used as the water-soluble solvent), the extract or its concentrate can be used as is. In this extraction process, water-soluble solvents such as ethanol, methanol, isopropanol, acetone, etc. are used as organic solvents for extraction, and the extraction process is performed using these water-soluble solvents at a concentration of 20 to 80%, more preferably 25 to 40%. It is preferable to use a mixed solvent with water containing . In this case, the water-soluble solvent may be used alone or in combination of two or more. Also, the amount of organic solvent (water-soluble solvent, especially mixed solvent of water-soluble solvent and water) used should be determined based on the amount of the substance to be extracted in the organic solvent.
The amount is preferably set to a concentration of 0.1 to 30%, more preferably 1 to 10%. The extraction conditions preferably include a temperature of 0 to 25° C. and an extraction time of 1 hour to several days, more preferably 3 to 48 hours. After the extraction process, the extract is collected. The target fraction obtained by performing this extraction treatment with a water-soluble organic solvent is one in which resins and poorly alcohol-soluble substances are reliably removed. Therefore, the target fraction of the present invention obtained by the above method exhibits a very high healing promoting effect on wounds such as cuts and burns, has an excellent tissue activating effect, and contains physiologically active substances. External skin medicine for abrasions, cuts, xerokeratosis, cracks, chapped skin, rough skin, chilblains, frostbite, burns, rashes, eczema, heat rash, athlete's foot, hemorrhoid medicine, peptic ulcer treatment,
Various wound healing products such as eye disease treatments, intractable ulcer treatments, aftershave medicated creams and lotions, scalp and hair cosmetics, skin cosmetics, lip creams and sticks, sun care products, cleaning agents, etc.
It can be added to products aimed at preventing rough skin and revitalizing tissues. In this case, the target fraction (physiologically active substance) obtained by the present invention does not contain colored pigment components that easily change color or water-insoluble components, and is colorless to pale yellow, and does not contain water-insoluble components due to heat denaturation or denaturation over time. (inflammatory substances) are reliably removed, stable and discolored,
There is no deterioration in quality, and those extracted using water-soluble organic solvents do not contain alcohol-insoluble substances, so they can be added to various products without damaging their appearance. There is no risk of problems with product quality. In addition, this physiologically active substance is water-soluble and can be dissolved in water at almost any ratio, and can also be dissolved in mixed solvents of water and ethanol, isopropanol, glycerin, propylene glycol, etc., and is especially soluble in water-soluble organic solvents. The target fraction obtained by the extraction process has high solubility in these water-containing organic solvents, so it is easy to use when blended into products, such as emulsion ointments, oil ointments, alcoholic lotions, etc. It can be easily prepared into molds. Moreover,
The physiologically active substance of the present invention is highly safe and non-irritating when applied to the skin, and is therefore suitable for use as external preparations, cosmetics, and the like. In addition, when the physiologically active substance of the present invention is incorporated into external preparations, cosmetics, etc., it can be prepared in an appropriate dosage form, and the amount incorporated is preferably 0.01 to 30% of the total product. In addition, the physiologically active substances of the present invention include polysaccharides, organic acids,
Contains amino acids and proteins such as glutamic acid, aspartic acid, serine, alanine, proline, histidine, valine, threonine, leucine, isoleucine, glycine, and lysine, and water-soluble inorganic salts such as Mg, Ca, K, and Na, and is not drinkable. It is non-toxic, has a slightly sour taste, has an amino acid-like umami flavor, and has no bitterness, so it can be used as a health drink, medicinal liquor, etc.
It can be suitably used in foods and beverages such as soft drinks, and in oral products such as toothpaste, ointments for gingivitis and periodontal diseases, mouthwashes, mouth fresheners, oral drops, troches, and mouthwashes. can also be suitably used. EXAMPLES Hereinafter, the present invention will be explained in more detail by showing Examples and Comparative Examples. [Example 1] 10 kg of Kidachi aloe leaves were crushed in a mixer,
After filtration with cotton cloth and natural filtration with paper, 7.5 kg of slightly opaque yellow-green sap was obtained. Next, 7.5 kg of this sap was heated at 70°C for 30 minutes under a reduced pressure of 300 mmHg to obtain 3.7 kg of concentrated sap. 300g for this
powdered activated carbon was added, and after stirring at room temperature for about 30 minutes, it was suctioned off using paper, and then the activated carbon was washed with water and the washing solution was added to the solution. This liquid was concentrated under reduced pressure using an aspirator at a bath temperature of 40°C, and further vacuum-dried to form a white powder (physiologically active substance).
172.0g was obtained. [Example 2] 10 kg of aloe vera leaves were processed in the order of food slicer, pulper, and finisher to produce green sap.
Obtained 7.6Kg. 760 g of powdered activated carbon was added to this sap, stirred for about 1 hour while heating to 70°C, allowed to cool to room temperature, and suctioned through paper. The activated carbon was then washed with water and the washing solution was added to the solution. This liquid was spray-dried to obtain 72.2 g of white powder (physiologically active substance). [Example 3] 10 kg of Syabon aloe leaves were crushed in a mixer, filtered through cotton cloth, and then naturally filtered through paper to obtain 6.6 kg of yellow-green, slightly opaque sap.
This sap was freeze-dried to obtain 237.6 g of yellowish brown powder. Add 33% ethanol aqueous solution 2 to this and
After stirring for a minute, the mixture was allowed to stand at room temperature for 1 day. The precipitated pale yellow precipitate was filtered through paper to obtain a reddish brown liquid. This liquid was concentrated to dryness while heating at 70°C to obtain 133.0 g of a yellowish brown extract. to this
Add 2200 ml of water and 180 g of powdered activated carbon, stir at room temperature for about 30 minutes, and then filter through paper with suction.
The activated carbon was then washed with water and the washing solution was added to the solution. This liquid was freeze-dried to obtain 101.0 g of pale yellow powder (physiologically active substance). [Example 4] Using 10 kg of Syabon aloe leaves, first cut the leaves into approximately 5 kg.
The leaves were cut into cm pieces, the leaf skin was removed, and the jelly part was collected. This was crushed using a mixer to obtain 5.1 kg of slightly yellow viscous sap. Add acetone 5 to this sap,
After stirring for 30 minutes, the mixture was allowed to stand at room temperature for 1 day. The precipitated pale yellow precipitate was suction-filtered using paper to obtain a pale yellow liquid. This liquid was concentrated to dryness under reduced pressure using an aspirator at a bath temperature of 40°C, and then mixed with water 2 and powdered activated carbon.
After adding 50 g and stirring for about 30 minutes while heating to 70°C, the activated carbon was filtered using paper, the activated carbon was then washed with water, and the washing liquid was added to the liquid. This liquid is freeze-dried and made into a pale yellow powder (physiologically active substance).
30.3g was obtained. [Example 5] 10 kg of Kidachi aloe leaves were crushed in a mixer,
After filtration with cotton cloth and natural filtration with paper, 7.5 kg of slightly opaque yellow-green sap was obtained. Next, this sap was heated to 70°C and concentrated to about 1/3 of its volume to obtain 2.5 kg of concentrate. 260 for this
After adding g of powdered activated carbon and stirring for about 30 minutes,
The activated carbon was then washed with water and the washing solution was added to the solution. 1.3 to this liquid
of methanol was added, stirred for about 30 minutes, and then allowed to stand at room temperature for about 1 day. The precipitated white precipitate was suctioned and filtered using paper to obtain a colorless and transparent liquid.
This liquid was concentrated under reduced pressure using an aspirator to distill off methanol, and then spray-dried to obtain 90.7 g of a white powder (physiologically active substance). [Example 6] 10 kg of aloe leaves were processed using a food slicer, a pulper, and a finisher in this order to obtain 7.7 g of green sap. 385 g of powdered activated carbon was added to this sap, stirred for about 30 minutes while heating to 70°C, then allowed to cool to room temperature, filtered with suction using paper, then the activated carbon was washed with water, and the washing liquid was added to the liquid. This liquid was freeze-dried to obtain 246 g of pale yellow powder.
To this was added 2.4 g of 33% ethanol, stirred for 30 minutes, and allowed to stand at room temperature for about 1 day. The precipitated pale yellow precipitate was suctioned and filtered using paper to obtain a colorless and transparent liquid. This was concentrated under reduced pressure using an aspirator at a bath temperature of 40°C, and further vacuum-dried to obtain 115.5 g of a white powder (physiologically active substance). [Example 7] Add 10 kg of aloe vera leaves to 10 kg of water, crush with a mixer, heat at 70°C for 1 hour, filter naturally using paper, and then distill off the water under reduced pressure with an aspirator. 150.0 g of powder was obtained. To this was added 2.25% of a 33% ethanol aqueous solution, and after stirring for about 1 hour, it was allowed to stand at room temperature for about 1 day. Next, the precipitated pale yellow precipitate was removed using paper to obtain a reddish brown liquid. This liquid was concentrated to dryness under reduced pressure using an aspirator to obtain 112.0 g of a yellowish brown extract. Add water and 300g of activated carbon from step 3 to this, stir for about 30 minutes, then
It was filtered through a paper centrifuge to obtain a colorless and transparent solution. Next, this solution was concentrated under reduced pressure using an aspirator at a bath temperature of 40°C, and further vacuum-dried to obtain 81.0 g of a white to pale yellow powder (physiologically active substance). [Comparative Example 1] 10 kg of Kidachi Aloe leaves were crushed in a mixer,
After filtration with cotton cloth and natural filtration with paper, 7.5 kg of slightly opaque yellow-green sap was obtained. This sap is freeze-dried and turned into a yellowish brown powder (comparative product).
Obtained 210g. [Comparative Example 2] 10 kg of Kidachi Aloe leaves were crushed in a mixer,
After natural filtration with cotton cloth and paper, 7.5 kg of slightly opaque yellow-green sap was obtained. 70% of this sap
℃ for 30 minutes, filtered through paper, and freeze-dried the liquid to form a dark brown powder (comparative product).
Obtained 198g. [Comparative Example 3] 10 kg of aloe leaves were processed in the order of food slicer, pulper, and finisher to obtain 7.7 kg of green sap. This sap was heated to 70°C and concentrated to about 1/5 of its volume to obtain 1.6 kg of concentrate. This was naturally filtered using paper, 0.8% of ethanol was added to the liquid, stirred for 30 minutes, and left to stand at room temperature for about 1 day. Obtained. This liquid was concentrated to dryness under reduced pressure using an aspirator at a bath temperature of 40°C to obtain 154 g of a yellowish brown extract (comparative product). [Comparative Example 4] 10 kg of freshly collected Aloe leaves were ground in a mixer to obtain 7.9 kg of sap. Ethanol 32 was added to this sap, and the mixture was thoroughly stirred at room temperature and left to stand overnight, after which the precipitate that formed was removed. Next, the supernatant ethanol extract was concentrated under reduced pressure at 40 to 50°C to obtain 120 g of a yellowish brown solid. This was dissolved in 10 g of water and subjected to chromatography using about 2 kg of activated carbon (mesh size 60-80). Further, 10 ml of water was further eluted, and the resulting aqueous solution total 20 was spray-dried to obtain about 58 g of a white comparative substance (comparative product). Next, the physiologically active substances obtained in the above examples and comparative examples, and the physiologically active effects of comparative products will be explained using the following experimental examples. [Experiment 1] Male Wistar rats (125 to 155 g) were divided into groups of 5 to 7 animals. Under Nembutal anesthesia, the hair on the back was extensively shaved, sterilized with alcohol, and the hair was sterilized along the midline with a scalpel. A 4 cm sharp incision was made, and sutured at 3 points at 1 cm intervals using a Miyutsuher needle. After wound creation, the physiologically active substances obtained in Examples 1 to 7 at ~40 mg/kg were each dissolved in physiological saline and administered subcutaneously once a day for 7 consecutive days. On the 7th day after wound creation, the rats were killed with chloroform, the wound skin was peeled off, the subcutaneous tissue was removed, and three 1 cm wide strips of skin perpendicular to the wound line were removed from each rat.
Individual patches were prepared, and the tensile strength required to separate the wound skin pieces was measured using an Instron tension measuring machine, and the average value was used as an index of the degree of wound healing. As a control, 2 ml/Kg of physiological saline was subcutaneously administered to the rat, and the tension was measured in the same manner. The results are shown in Tables 1 and 2.

【table】

[Experiment 2]

A group of 6 male Wistar rats (120g to 130g) were anesthetized with Nembutal, their backs were shaved, the hair on the skin surface was removed with hair removal cream, and then disinfected with alcohol. One circular burn wound was created in each rat under conditions of 100° C., 50 g load, 1 second, and a stainless steel head with a diameter of 1 cm. Physiologically active substances obtained in Examples 2, 6, and 7
were mixed into a hydrophilic base at a concentration of 1% and 5%, respectively, and applied twice a day for 6 consecutive days starting from the time of creating a thermal wound.
It was applied for days. The site was collected on the 6th day after the thermal wound was created, fixed in formalin, and histological sections were prepared using a conventional method.
The stained specimens were subjected to histological evaluation in five stages: -, ±, +, , to evaluate the degree of healing of the burn wound. As a control, a sample coated with only the base was evaluated in the same manner. The results are shown in Table 3.

[Experiment 3]

Slc: 10 male ddy mice (6 weeks old) were used per group.
Physiologically active substances in the footpads and comparative product 4
0.01 ml of % physiological saline solution was injected subcutaneously. As a control, saline 0.01 was applied to the contralateral footpad.
ml was administered in the same manner. Five hours after administration, both legs were amputated at the ankles, the weights of both left and right legs were measured, and the inflammatory potential was evaluated based on the rate of increase in leg weight due to drug administration compared to the control. The results are shown in Table 4.

[Experiment 4]

Using a group of 8 male Wistar rats (6 weeks old), the backs were extensively dehaired, and a 5% LAS (linear sodium alkylbenzene sulfonate) aqueous solution was applied to one side of 4 cm ² of skin on each side of the midline. On the other side, the same 5% LAS aqueous solution containing 5% of the physiologically active substance was applied, and the degree of redness was observed 24 hours later. The same animal was operated for 3 consecutive days, and judgments were made up to 72 hours later. The results are shown in Table 5. The evaluation criteria are as follows. Evaluation criteria Redness 0 points: No change observed 〃 1 point: Mild redness with unclear boundaries 〃 2 points: Strong redness with clear boundaries 〃 3 points: Formation of blisters and edema Results are cumulative of 8 specimens Indicated by dots.

【table】

[Experiment 5]

After creating rough skin with soap on the skin of the left and right forearms of a person for 4 days, Examples 2, 3,
A 5% blend of each of the physiologically active substances obtained in step 4 in a hydrophilic base was applied once a day for 3 days starting from the 4th day, and the impedance was used as an index to measure the degree of improvement in rough skin within 24 hours of application. Evaluation was made by measuring skin moisture content. A total of 40 subjects, 10 each, applied the base containing the physiologically active substance to one forearm, and the base alone to the other forearm, serving as a control. The results are shown in Figure 1. Note that the results are shown as the average value of electrical conductivity (μ). In Figure 1, , are treated with a base containing a physiologically active substance, , respectively;
C is a control. From the results shown in FIG. 1, it was confirmed that the physiologically active substance obtained by the method of the present invention enhances the moisturizing properties of the skin. [Experiment 6] The physiologically active substances obtained in Examples 2 and 5 and the comparative products obtained in Comparative Examples 1 and 4 were mixed into a hydrophilic base at a concentration of 5%, and the usability test was conducted. I did this. The panel selected 14 patients with cracked and chapped hands, and tested the five samples mentioned above three times. The test interval was one week. The drug was applied to the affected areas on the left and right hands, and symptoms were observed for 30 minutes after application. Judgment is based on the panel's sense of irritation, onset of pain, and degree of redness.
The reaction was carried out in four stages, including no reaction. The results are shown in Table 6.

[Experiment 7]

The physiologically active substance obtained in Example 2 and the comparative products obtained in Comparative Examples 1, 2, and 3 were blended into a hydrophilic base at a concentration of 1.0%, and heated at 20°C.
The sample was stored for 3 months, and the degree of discoloration during that time was measured using a photoelectric whiteness meter. The results (changes in hunter whiteness over time) are shown in Figure 2. In addition, in Fig. 2, the physiologically active substances obtained by the present invention are shown, and ′, ′, and ′ are comparative products, respectively.
, B represents a base. Although the ointment containing the physiologically active substance of the present invention showed no change over time even after being stored at 20°C for 3 months, the ointment containing the comparative product that had not been treated with activated carbon showed significant discoloration. [Experiment 8] An acute toxicity test was conducted using ICR male mice. A 25% suspension of the physiologically active substances obtained in Examples 1 to 7 and a comparative product obtained in Comparative Examples 1 to 4 and a 2.5% suspension of gum arabic were orally administered. The results showed that LD ₅₀ >10000 mg/Kg mouse body weight. Regarding the comparison product, ., loose stools were observed at high doses, and severe diarrhea symptoms were observed at high doses. [Experiment 9] Hair was removed from the back of a male white rabbit (2.3-3.5 kg),
1 hour, 3 hours, 6 hours, 24 hours, 48 hours after sample application
Skin symptoms at 72 hours, 96 hours, 168 hours, and 336 hours were scored based on the Draize method. Ointments and aqueous solutions containing physiologically active substances were used as samples, and the concentrations of the ointments were 5% and 10%, and the aqueous solutions were 5%, 10%, and 20%. In addition, samples coated with an ointment base and distilled water were used as controls. As a result, no skin irritation was observed in any of the substances of the present invention. [Experiment 10] The mutagenicity of physiologically active substances was investigated using the Ames method (pre-incubation method). Mutagenicity is Salmonella typhimurium TA 98 and
Three species, TA100 and E.Coli WP 2 uvrA, were used as test bacteria. As a result, no mutagenicity was observed in the substance of the present invention. Next, examples of blending the substances of the present invention will be shown. [Formulation example 1] Hydrophilic ointment white petrolatum 250g Stearyl alcohol 220g Propylene glycol 120g Sodium lauryl sulfate 15g Ethyl paraoxybenzoate 0.25g Propyl paraoxybenzoate 0.15g Physiologically active substance of Example 1 50g Purified water Appropriate amount Total amount 1000g [Formulation Example 2] Macrogol ointment Macrogol 4000 500g Macrogol 400 470g Physiologically active substance of Example 2 30g Total amount 1000g [Formulation example 3] Drink vitamin B ₆ 5mg Nicotinic acid amide 20mg Anhydrous caffein 50mg Honey 2500mg Citric acid 50mg Vitamin C 100mg Add purified water to 1000mg or more of the physiologically active substance of Example 1 to make a total volume of 100ml. [Formulation Example 4] Soft drink sucrose 10g Citric acid 50mg 0.1g of the physiologically active substance of Example 3 Add carbonated water to make a total volume of 100ml. In addition, the drinks of Formulation Examples 3 and 4 are both free from bitterness and odor peculiar to aloe, and have a unique taste.
It was easy to drink, and no purgative effect was observed. [Formulation example 5] Toothpaste carboxymethylcellulose 0.3g Carrageenan 0.8g Sorbit 20.0g Propylene glycol 2.0g Dibasic calcium phosphate 45.0g Silicic anhydride 2.0g Sodium lauryl sulfate 1.5g Physiologically active substance from Example 4 0.8g Flavor Appropriate amount Add purified water to make a total volume of 100g. [Formulation example 6] Aftershave lotion Glycerin 4.0g Boric acid 1.0g Ethanol 20.0g Menthol 0.1g Physiologically active substance of Example 5 0.5g Fragrance Appropriate amount Purified water 74.4g [Formulation example 7] Suntan lotion carboxypolymer Methylene 1.5g Polyethylene glycol 600 5.0g Dihydroxyacetone 1.0g Diisopropanolamine appropriate amount Diisopropanolamine appropriate amount Physiologically active substance of Example 7 1.0g Fragrance appropriate amount Purified water Total amount 100g [Formulation Example 8] Shampoo lauryl sulfate triethanolamine Salt 15g Coconut oil fatty acid triethanolamide 2g Salt 1.5g Physiologically active substance from Example 6 2g Fragrance, pigment Appropriate amount Purified water Total amount 100g [Formulation example 9] Hair tonic alcohol 59.0g Glycerin 5.0g Salicylic acid 0.3g Cation activator 1.0g Physiologically active substance of Example 5 1.0g Flavor Appropriate amount Purified water Total amount 100g

[Brief explanation of the drawing]

Figure 1 is a graph showing the degree of improvement in rough skin when a hydrophilic base containing a physiologically active substance obtained by the method of the present invention is applied to rough skin. It is a graph showing a change over time in Hunter whiteness of a base when blended with a hydrophilic base.

Claims (1)

[Claims] 1. Add aloe sap selected from Aloe Vera, Syabon Aloe, and Kidachi Aloe at 50 to 90℃.
While or after heat treatment for 0.5 to 3 hours, 0.1 to the weight of the sap above
A method for purifying aloe sap, which comprises performing activated carbon treatment by a batch method using ~20% by weight of activated carbon, and then filtering to collect the liquid.