A traditional Chinese medicine combination and its preparation method, and its application in the preparation of products with the effect of removing postpartum pigmentation spots.

Abstract

This invention relates to the field of biomedical technology, specifically disclosing a traditional Chinese medicine composition, its preparation method, and its application in the preparation of products with the effect of removing postpartum pigmentation spots. The traditional Chinese medicine composition comprises the following raw material components: Rehmannia glutinosa (processed), Cistanche deserticola, Miracle Fruit kernel, Morinda officinalis, Cornus officinalis, and Arctium lappa. Studies have shown that the traditional Chinese medicine composition has tyrosinase inhibitory activity; therefore, the traditional Chinese medicine composition of this invention can be used as an active ingredient in the preparation of cosmetics or drugs with tyrosinase inhibitory activity; in particular, further using the traditional Chinese medicine composition of this invention as an active ingredient in the preparation of cosmetics or drugs with the effect of removing postpartum pigmentation spots has significant application value.

Description

Technical Field

[0001] This invention relates to the field of biomedicine, specifically to a traditional Chinese medicine combination and its preparation method, and its application in the preparation of products that remove postpartum pigmentation spots. Background Technology

[0002] Pigmentation spots are caused by uneven distribution of melanin granules in the skin, resulting in spots and patches that are darker than the normal skin tone. External factors contributing to pigmentation include epidermal irritation from chemicals and ultraviolet radiation; internal factors include endocrine disorders.

[0003] Postpartum women are often prone to endocrine disorders, which can lead to the formation of pigmentation spots. Pigmentation spots can be removed through surgical and non-surgical methods. Due to the weakness of postpartum women and the need for breastfeeding, non-surgical methods are the best choice for removing postpartum pigmentation spots. Non-surgical methods typically involve the use of medications or cosmetics. Since pigmentation spots are caused by uneven distribution of melanin granules in the skin, screening for active ingredients that inhibit melanin production is crucial. These active ingredients can effectively reduce the melanin content in the skin, thus achieving the effect of removing pigmentation spots. Tyrosinase is a key enzyme regulating melanin synthesis, catalyzing the production of melanin and other pigments. Therefore, developing an active ingredient with tyrosinase inhibitory activity can effectively inhibit melanin production, thereby removing pigmentation spots.

[0004] Traditional Chinese medicine (TCM) is a natural medicine found in nature. Compared to chemically synthesized active ingredients or hormones, it is safer and more suitable for postpartum women. Therefore, developing compositions using TCM as raw materials to remove postpartum pigmentation spots has significant application value. Summary of the Invention

[0005] In order to overcome at least one of the technical problems existing in the prior art, the present invention provides a traditional Chinese medicine combination and its preparation method and its application in the preparation of a product with the effect of removing postpartum pigmentation spots.

[0006] The above-mentioned technical problem to be solved by the present invention is achieved through the following technical solution:

[0007] The present invention first provides a traditional Chinese medicine composition comprising the following raw material components: Rehmannia glutinosa (processed), Cistanche deserticola, Miracle Fruit kernel, Morinda officinalis, Cornus officinalis, and Arctium lappa.

[0008] This invention provides a novel traditional Chinese medicine composition made from Rehmannia glutinosa, Cistanche deserticola, Miracle Fruit kernel, Morinda officinalis, Cornus officinalis, and Arctium lappa. Studies have shown that the traditional Chinese medicine composition has tyrosinase inhibitory activity.

[0009] Preferably, the traditional Chinese medicine composition comprises the following raw material components in parts by weight:

[0010] Prepared Rehmannia root 5-10 parts; Cistanche deserticola 5-10 parts; Miracle fruit kernel 5-10 parts; Morinda officinalis 3-8 parts; Cornus officinalis 3-6 parts; Arctium lappa 3-6 parts.

[0011] Most preferably, the traditional Chinese medicine composition comprises the following raw material components in parts by weight:

[0012] Prepared Rehmannia root 8 parts; Cistanche deserticola 8 parts; Miracle fruit kernel 8 parts; Morinda officinalis 5 parts; Cornus officinalis 4 parts; Arctium lappa 4 parts.

[0013] This invention also provides a method for preparing the above-mentioned traditional Chinese medicine combination, which includes the following steps:

[0014] Prepared Rehmannia glutinosa, Cistanche deserticola, Miracle Fruit kernel, Morinda officinalis, Cornus officinalis, and Arctium lappa are mixed and pulverized to obtain Chinese medicine powder. The Chinese medicine powder is then extracted with an ethanol aqueous solution. After extraction, the extract is concentrated and dried to obtain the Chinese medicine alcohol extract. The Chinese medicine combination is obtained by taking the Chinese medicine alcohol extract.

[0015] Preferably, the ethanol aqueous solution refers to an ethanol aqueous solution with a volume fraction of 40-60%.

[0016] Most preferably, the ethanol aqueous solution refers to an ethanol aqueous solution with a volume fraction of 50%.

[0017] Preferably, the ratio of the amount of Chinese herbal medicine powder to the ethanol aqueous solution is 1kg:10-20L;

[0018] Most preferably, the ratio of Chinese medicine powder to ethanol aqueous solution is 1 kg: 15 L.

[0019] Preferably, the preparation method of the traditional Chinese medicine combination further includes the following steps:

[0020] The ethanol extract of traditional Chinese medicine was loaded onto a macroporous resin column and first eluted with an ethanol aqueous solution of 12-14% (v / v) to remove impurities. Then, it was eluted again with an ethanol aqueous solution of 26-28% (v / v). The eluent obtained from the ethanol aqueous solution of 26-28% (v / v) was collected, concentrated, and dried to obtain the macroporous resin eluent of traditional Chinese medicine. The macroporous resin eluent of traditional Chinese medicine is the traditional Chinese medicine combination mentioned above.

[0021] Most preferably, the ethanol extract of the traditional Chinese medicine is loaded onto a macroporous resin column, and first eluted with a 13% (v / v) ethanol aqueous solution to remove impurities; then eluted with a 27% (v / v) ethanol aqueous solution, and the eluent obtained from the 27% (v / v) ethanol aqueous solution is collected, concentrated and dried to obtain the macroporous resin eluted fraction of the traditional Chinese medicine; the macroporous resin eluted fraction of the traditional Chinese medicine is taken to obtain the traditional Chinese medicine combination.

[0022] Preferably, the macroporous resin column is a D101 type macroporous resin column.

[0023] Further research by the inventors revealed that the macroporous resin eluted fraction obtained by further eluting the ethanol extract of traditional Chinese medicine under the macroporous resin column elution conditions described in this invention exhibits a significantly enhanced tyrosinase inhibitory effect compared to the ethanol extract. Therefore, using the macroporous resin eluted fraction as a traditional Chinese medicine composition demonstrates superior tyrosinase inhibitory activity compared to using the ethanol extract.

[0024] Furthermore, the inventors emphasize that the elution conditions of the macroporous resin column in this invention are crucial. Macroporous resin elution fractions prepared using different elution conditions exhibit completely different types and contents of active ingredients, leading to vastly different tyrosinase inhibitory activities. The inventors have surprisingly discovered in extensive research that only macroporous resin elution fractions prepared using the aforementioned macroporous resin column elution conditions of this invention show a significantly enhanced tyrosinase inhibitory effect compared to the ethanol extract of traditional Chinese medicine. However, macroporous resin elution fractions prepared using other macroporous resin column elution conditions do not show a significantly enhanced tyrosinase inhibitory effect compared to the ethanol extract.

[0025] Preferably, the preparation method of the traditional Chinese medicine combination further includes the following steps: loading the macroporous resin elution fraction of the traditional Chinese medicine onto a silica gel column; firstly, eluting and removing impurities with an organic solvent composed of cyclohexane and acetone in a volume ratio of 82-84:18-16; then eluting again with an organic solvent composed of cyclohexane and acetone in a volume ratio of 71-73:29-27, collecting the eluent eluted with the organic solvent composed of cyclohexane and acetone in a volume ratio of 71-73:29-27, concentrating and drying to obtain the silica gel elution fraction of the traditional Chinese medicine; taking the silica gel elution fraction of the traditional Chinese medicine yields the traditional Chinese medicine combination.

[0026] Most preferably, the preparation method of the traditional Chinese medicine combination further includes the following steps: loading the macroporous resin elution fraction of the traditional Chinese medicine onto a silica gel column; firstly, eluting and removing impurities with an organic solvent composed of cyclohexane and acetone in a volume ratio of 83:17; then eluting again with an organic solvent composed of cyclohexane and acetone in a volume ratio of 72:28, collecting the eluent eluted by the organic solvent composed of cyclohexane and acetone in a volume ratio of 72:28, concentrating and drying it to obtain the silica gel elution fraction of the traditional Chinese medicine; taking the silica gel elution fraction of the traditional Chinese medicine is the traditional Chinese medicine combination.

[0027] In further research, the inventors discovered that the tyrosinase inhibitory effect of the silica gel elution fraction of traditional Chinese medicine obtained by further eluting the macroporous resin elution fraction of traditional Chinese medicine under the silica gel column elution conditions described in this invention is significantly improved compared to that of the macroporous resin elution fraction of traditional Chinese medicine; thus, the prepared traditional Chinese medicine composition has excellent tyrosinase inhibitory effect.

[0028] Similarly, the inventors need to emphasize that the silica gel column elution conditions of this invention are crucial. Silica gel elution fractions prepared using different silica gel column elution conditions result in completely different types and contents of active ingredients, leading to entirely different tyrosinase inhibitory activities. The inventors surprisingly discovered in extensive research that only silica gel elution fractions prepared using the aforementioned silica gel column elution conditions of this invention exhibit significantly enhanced tyrosinase inhibitory activity compared to those eluted from macroporous resins of traditional Chinese medicine. However, silica gel elution fractions prepared using other silica gel column elution conditions do not show a significantly enhanced tyrosinase inhibitory activity compared to those eluted from macroporous resins of traditional Chinese medicine.

[0029] This invention provides an application of the above-mentioned traditional Chinese medicine combination in the preparation of products with tyrosinase inhibitory effects.

[0030] This invention provides an application of the above-mentioned traditional Chinese medicine combination in the preparation of a product with the effect of removing postpartum pigmentation spots.

[0031] Preferably, the product is a cosmetic or a pharmaceutical.

[0032] Beneficial Effects: This invention provides a novel traditional Chinese medicine composition made from Rehmannia glutinosa, Cistanche deserticola, Miracle Fruit kernel, Morinda officinalis, Cornus officinalis, and Arctium lappa. Studies have shown that the traditional Chinese medicine composition has tyrosinase inhibitory activity. Therefore, the traditional Chinese medicine composition of this invention can be used as an active ingredient to prepare cosmetics or drugs with tyrosinase inhibitory activity. In particular, further using the traditional Chinese medicine composition of this invention as an active ingredient to prepare cosmetics or drugs with the effect of removing postpartum pigmentation spots has important application value. Detailed Implementation

[0033] The present invention will be further explained below with reference to specific embodiments, but the embodiments do not limit the present invention in any way.

[0034] Example 1: Preparation of the Traditional Chinese Medicine Composition

[0035] Raw material composition by weight: 8 parts Rehmannia glutinosa (processed); 8 parts Cistanche deserticola; 8 parts Miracle Fruit kernel; 8 parts Morinda officinalis

[0036] sky

[0037] 5 parts; Cornus officinalis 4 parts, Arctium lappa 4 parts;

[0038] Preparation of the traditional Chinese medicine composition: Prepared Rehmannia glutinosa, Cistanche deserticola, Miracle Fruit kernel, Morinda officinalis, Cornus officinalis, and Arctium lappa are mixed and pulverized, then passed through a 100-mesh sieve to obtain traditional Chinese medicine powder. The powder is then heated to boiling with an ethanol aqueous solution and refluxed for 1 hour. After extraction, the extract is concentrated and dried to obtain the alcohol extract of traditional Chinese medicine. The traditional Chinese medicine composition is obtained from the alcohol extract.

[0039] The ethanol-water solution mentioned above refers to an ethanol-water solution with a volume fraction of 50%; the ratio of the amount of Chinese medicine powder to the ethanol-water solution is 1 kg: 15 L.

[0040] Example 2: Preparation of the Traditional Chinese Medicine Composition

[0041] Raw material composition by weight: 8 parts Rehmannia glutinosa (processed); 8 parts Cistanche deserticola; 8 parts Miracle Fruit kernel; 8 parts Morinda officinalis

[0042] sky

[0043] 5 parts; Cornus officinalis 4 parts, Arctium lappa 4 parts;

[0044] Preparation of Traditional Chinese Medicine Compositions:

[0045] (1) Take Rehmannia glutinosa, Cistanche deserticola, Miracle Fruit kernel, Morinda officinalis, Cornus officinalis and Arctium lappa, mix them together and pulverize them, pass them through a 100-mesh sieve to obtain Chinese medicine powder. Then, heat the Chinese medicine powder with an ethanol aqueous solution to boiling point and reflux for 1 hour. After extraction, concentrate and dry the extract to obtain Chinese medicine alcohol extract; wherein, the ethanol aqueous solution refers to an ethanol aqueous solution with a volume fraction of 50%; the ratio of Chinese medicine powder to ethanol aqueous solution is 1kg:15L;

[0046] (2) The ethanol extract of traditional Chinese medicine was loaded onto a macroporous resin column (the macroporous resin column was filled with D101 type macroporous resin packing material with a volume fraction of 50 times the weight of the ethanol extract of traditional Chinese medicine). First, it was eluted with 5 column volumes of 13% ethanol aqueous solution to remove impurities. Then, it was eluted with 5 column volumes of 27% ethanol aqueous solution. The eluent obtained from the 27% ethanol aqueous solution was collected, concentrated and dried to obtain the macroporous resin eluted fraction of traditional Chinese medicine. The macroporous resin eluted fraction of traditional Chinese medicine was taken to obtain the traditional Chinese medicine combination.

[0047] Example 3: Preparation of the Traditional Chinese Medicine Composition

[0048] Raw material composition by weight: 8 parts Rehmannia glutinosa (processed); 8 parts Cistanche deserticola; 8 parts Miracle Fruit kernel; 8 parts Morinda officinalis

[0049] sky

[0050] 5 parts; Cornus officinalis 4 parts, Arctium lappa 4 parts;

[0051] Preparation of Traditional Chinese Medicine Compositions:

[0052] (1) Take Rehmannia glutinosa, Cistanche deserticola, Miracle Fruit kernel, Morinda officinalis, Cornus officinalis and Arctium lappa, mix them together and pulverize them, pass them through a 100-mesh sieve to obtain Chinese medicine powder. Then, heat the Chinese medicine powder with an ethanol aqueous solution to boiling point and reflux for 1 hour. After extraction, concentrate and dry the extract to obtain Chinese medicine alcohol extract; wherein, the ethanol aqueous solution refers to an ethanol aqueous solution with a volume fraction of 50%; the ratio of Chinese medicine powder to ethanol aqueous solution is 1kg:15L;

[0053] (2) The Chinese herbal alcohol extract was loaded onto a macroporous resin column (the macroporous resin column was filled with D101 type macroporous resin packing material with a volume fraction of 50 times the weight of the Chinese herbal alcohol extract). First, it was eluted with 5 column volumes of 13% ethanol aqueous solution to remove impurities. Then, it was eluted with 5 column volumes of 27% ethanol aqueous solution. The eluent eluted with 27% ethanol aqueous solution was collected, concentrated and dried to obtain the Chinese herbal macroporous resin eluted fraction.

[0054] (3) The macroporous resin elution fraction of the Chinese medicine was loaded onto a silica gel column (the silica gel column was filled with 200-300 mesh silica gel, which was 30 times the weight of the macroporous resin elution fraction of the Chinese medicine); first, it was eluted with an organic solvent composed of cyclohexane and acetone in a volume ratio of 83:17, which was 3 times the column volume; then, it was eluted with an organic solvent composed of cyclohexane and acetone in a volume ratio of 72:28, which was 6 times the column volume. The eluent eluted with the organic solvent composed of cyclohexane and acetone in a volume ratio of 72:28 was collected, concentrated and dried to obtain the silica gel elution fraction of the Chinese medicine; the silica gel elution fraction of the Chinese medicine was taken to obtain the Chinese medicine combination.

[0055] Preparation of Traditional Chinese Medicine Composition in Comparative Example 1

[0056] Raw material composition by weight: 8 parts Rehmannia glutinosa (processed); 8 parts Cistanche deserticola; 8 parts Miracle Fruit kernel; 8 parts Morinda officinalis

[0057] sky

[0058] 5 parts; Cornus officinalis 4 parts, Arctium lappa 4 parts;

[0059] Preparation of Traditional Chinese Medicine Compositions:

[0060] (1) Take Rehmannia glutinosa, Cistanche deserticola, Miracle Fruit kernel, Morinda officinalis, Cornus officinalis and Arctium lappa, mix them together and pulverize them, pass them through a 100-mesh sieve to obtain Chinese medicine powder. Then, heat the Chinese medicine powder with an ethanol aqueous solution to boiling point and reflux for 1 hour. After extraction, concentrate and dry the extract to obtain Chinese medicine alcohol extract; wherein, the ethanol aqueous solution refers to an ethanol aqueous solution with a volume fraction of 50%; the ratio of Chinese medicine powder to ethanol aqueous solution is 1kg:15L;

[0061] (2) The ethanol extract of traditional Chinese medicine is loaded onto a macroporous resin column (the macroporous resin column is filled with D101 type macroporous resin packing material with a weight of 50 times that of the ethanol extract of traditional Chinese medicine). First, it is eluted with 5 column volumes of water to remove impurities. Then, it is eluted with 5 column volumes of 20% ethanol aqueous solution. The eluent obtained from the 20% ethanol aqueous solution is collected, concentrated and dried to obtain the macroporous resin eluted fraction of traditional Chinese medicine. The macroporous resin eluted fraction of traditional Chinese medicine is taken to obtain the traditional Chinese medicine combination.

[0062] The difference between Comparative Example 1 and Example 2 is that the elution conditions of the silica gel column are different.

[0063] Preparation of Traditional Chinese Medicine Composition in Comparative Example 2

[0064] Raw material composition by weight: 8 parts Rehmannia glutinosa (processed); 8 parts Cistanche deserticola; 8 parts Miracle Fruit kernel; 8 parts Morinda officinalis

[0065] 5 parts; Cornus officinalis 4 parts, Arctium lappa 4 parts;

[0066] Preparation of Traditional Chinese Medicine Compositions:

[0067] (1) Take Rehmannia glutinosa, Cistanche deserticola, Miracle Fruit kernel, Morinda officinalis, Cornus officinalis and Arctium lappa, mix them together and pulverize them, pass them through a 100-mesh sieve to obtain Chinese medicine powder. Then, heat the Chinese medicine powder with an ethanol aqueous solution to boiling point and reflux for 1 hour. After extraction, concentrate and dry the extract to obtain Chinese medicine alcohol extract; wherein, the ethanol aqueous solution refers to an ethanol aqueous solution with a volume fraction of 50%; the ratio of Chinese medicine powder to ethanol aqueous solution is 1kg:15L;

[0068] (2) The ethanol extract of traditional Chinese medicine was loaded onto a macroporous resin column (the macroporous resin column was filled with D101 type macroporous resin packing material with a volume fraction of 50 times the weight of the ethanol extract of traditional Chinese medicine). First, it was eluted with 5 column volumes of 20% ethanol aqueous solution to remove impurities. Then, it was eluted with 5 column volumes of 40% ethanol aqueous solution. The eluent obtained from the 40% ethanol aqueous solution was collected, concentrated and dried to obtain the macroporous resin eluent of traditional Chinese medicine. The macroporous resin eluent of traditional Chinese medicine was taken to obtain the traditional Chinese medicine combination.

[0069] The difference between Comparative Example 2 and Example 2 is that the elution conditions of the silica gel column are different.

[0070] Preparation of the traditional Chinese medicine composition in Comparative Example 3

[0071] Raw material composition by weight: 8 parts Rehmannia glutinosa (processed); 8 parts Cistanche deserticola; 8 parts Miracle Fruit kernel; 8 parts Morinda officinalis

[0072] sky

[0073] 5 parts; Cornus officinalis 4 parts, Arctium lappa 4 parts;

[0074] Preparation of Traditional Chinese Medicine Compositions:

[0075] (1) Take Rehmannia glutinosa, Cistanche deserticola, Miracle Fruit kernel, Morinda officinalis, Cornus officinalis and Arctium lappa, mix them together and pulverize them, pass them through a 100-mesh sieve to obtain Chinese medicine powder. Then, heat the Chinese medicine powder with an ethanol aqueous solution to boiling point and reflux for 1 hour. After extraction, concentrate and dry the extract to obtain Chinese medicine alcohol extract; wherein, the ethanol aqueous solution refers to an ethanol aqueous solution with a volume fraction of 50%; the ratio of Chinese medicine powder to ethanol aqueous solution is 1kg:15L;

[0076] (2) The ethanol extract of traditional Chinese medicine was loaded onto a macroporous resin column (the macroporous resin column was filled with D101 type macroporous resin packing material with a volume fraction of 50 times the weight of the ethanol extract of traditional Chinese medicine). First, it was eluted with 5 column volumes of 40% ethanol aqueous solution to remove impurities. Then, it was eluted with 5 column volumes of 60% ethanol aqueous solution. The eluent obtained from the 60% ethanol aqueous solution was collected, concentrated and dried to obtain the macroporous resin eluted fraction of traditional Chinese medicine. The macroporous resin eluted fraction of traditional Chinese medicine was taken to obtain the traditional Chinese medicine combination.

[0077] The difference between Comparative Example 3 and Example 2 is that the silica gel column elution conditions are different.

[0078] Preparation of the traditional Chinese medicine composition in Comparative Example 4

[0079] Raw material composition by weight: 8 parts Rehmannia glutinosa (processed); 8 parts Cistanche deserticola; 8 parts Miracle Fruit kernel; 8 parts Morinda officinalis

[0080] sky

[0081] 5 parts; Cornus officinalis 4 parts, Arctium lappa 4 parts;

[0082] Preparation of Traditional Chinese Medicine Compositions:

[0083] (1) Take Rehmannia glutinosa, Cistanche deserticola, Miracle Fruit kernel, Morinda officinalis, Cornus officinalis and Arctium lappa, mix them together and pulverize them, pass them through a 100-mesh sieve to obtain Chinese medicine powder. Then, heat the Chinese medicine powder with an ethanol aqueous solution to boiling point and reflux for 1 hour. After extraction, concentrate and dry the extract to obtain Chinese medicine alcohol extract; wherein, the ethanol aqueous solution refers to an ethanol aqueous solution with a volume fraction of 50%; the ratio of Chinese medicine powder to ethanol aqueous solution is 1kg:15L;

[0084] (2) The Chinese herbal alcohol extract was loaded onto a macroporous resin column (the macroporous resin column was filled with D101 type macroporous resin packing material with a volume fraction of 50 times the weight of the Chinese herbal alcohol extract). First, it was eluted with 5 column volumes of 13% ethanol aqueous solution to remove impurities. Then, it was eluted with 5 column volumes of 27% ethanol aqueous solution. The eluent eluted with 27% ethanol aqueous solution was collected, concentrated and dried to obtain the Chinese herbal macroporous resin eluted fraction.

[0085] (3) Load the macroporous resin elution fraction of the Chinese medicine onto a silica gel column (the silica gel column is filled with 200-300 mesh silica gel, which is 30 times the weight of the macroporous resin elution fraction of the Chinese medicine); first, elute and remove impurities with an organic solvent composed of cyclohexane and acetone in a volume ratio of 99:1, which is 3 times the column volume; then, elute again with an organic solvent composed of cyclohexane and acetone in a volume ratio of 90:10, which is 6 times the column volume. Collect the eluent eluted by the organic solvent composed of cyclohexane and acetone in a volume ratio of 90:10, concentrate and dry it to obtain the silica gel elution fraction of the Chinese medicine; take the silica gel elution fraction of the Chinese medicine to obtain the Chinese medicine combination.

[0086] The difference between Comparative Example 4 and Example 3 is that the elution conditions of the silica gel column are different.

[0087] Preparation of the traditional Chinese medicine composition in Comparative Example 5

[0088] Raw material composition by weight: 8 parts Rehmannia glutinosa (processed); 8 parts Cistanche deserticola; 8 parts Miracle Fruit kernel; 8 parts Morinda officinalis

[0089] sky

[0090] 5 parts; Cornus officinalis 4 parts, Arctium lappa 4 parts;

[0091] Preparation of Traditional Chinese Medicine Compositions:

[0092] (1) Take Rehmannia glutinosa, Cistanche deserticola, Miracle Fruit kernel, Morinda officinalis, Cornus officinalis and Arctium lappa, mix them together and pulverize them, pass them through a 100-mesh sieve to obtain Chinese medicine powder. Then, heat the Chinese medicine powder with an ethanol aqueous solution to boiling point and reflux for 1 hour. After extraction, concentrate and dry the extract to obtain Chinese medicine alcohol extract; wherein, the ethanol aqueous solution refers to an ethanol aqueous solution with a volume fraction of 50%; the ratio of Chinese medicine powder to ethanol aqueous solution is 1kg:15L;

[0093] (2) The Chinese herbal alcohol extract was loaded onto a macroporous resin column (the macroporous resin column was filled with D101 type macroporous resin packing material with a volume fraction of 50 times the weight of the Chinese herbal alcohol extract). First, it was eluted with 5 column volumes of 13% ethanol aqueous solution to remove impurities. Then, it was eluted with 5 column volumes of 27% ethanol aqueous solution. The eluent eluted with 27% ethanol aqueous solution was collected, concentrated and dried to obtain the Chinese herbal macroporous resin eluted fraction.

[0094] (3) The macroporous resin elution fraction of the Chinese medicine was loaded onto a silica gel column (the silica gel column was filled with 200-300 mesh silica gel, which was 30 times the weight of the macroporous resin elution fraction of the Chinese medicine); first, it was eluted with an organic solvent composed of cyclohexane and acetone in a volume ratio of 70:30, which was 3 times the column volume; then, it was eluted with an organic solvent composed of cyclohexane and acetone in a volume ratio of 60:40, which was 6 times the column volume. The eluent eluted with the organic solvent composed of cyclohexane and acetone in a volume ratio of 60:40 was collected, concentrated and dried to obtain the silica gel elution fraction of the Chinese medicine; the silica gel elution fraction of the Chinese medicine was taken to obtain the Chinese medicine combination.

[0095] The difference between Comparative Example 5 and Example 3 is that the silica gel column elution conditions are different.

[0096] Experiment Example 1: Tyrosinase Inhibitory Activity Test

[0097] Add 50 μL of L-DOPA solution, 50 μL of the sample solution to be tested, and / or 50 μL of tyrosinase solution to a 96-well plate; mix well and react in a water bath (30℃) for 25 min; then measure the absorbance A at 490 nm. Each experiment was repeated 3 times and the average value was taken; calculate the tyrosinase inhibition rate; the test results are shown in Table 1.

[0098] The L-DOPA solution contains 1 g / L of L-DOPA and is prepared using a 0.2 mol / L phosphate buffer solution with a pH of 6.8. The test sample solution contains 1 g / L of the test sample and is prepared using a 0.2 mol / L phosphate buffer solution with a pH of 6.8. The tyrosinase solution contains 100 U / mL and is prepared using a 0.2 mol / L phosphate buffer solution with a pH of 6.8. The test sample is the traditional Chinese medicine composition prepared in Examples 1-3 and Comparative Examples 1-5.

[0099] Tyrosinase inhibition rate = [1-(A X1 -A X2 ) / (A Y1 -A Y2 )]x100%; where, A X1 The absorbance was measured under conditions of adding tyrosinase solution and adding the sample solution; A X2 The absorbance measurements are taken without tyrosinase solution (replaced with an equal volume of phosphate buffer solution) and with the sample solution. A Y1 The absorbance measured under conditions where tyrosinase solution is added but the sample solution is not added (replaced with an equal volume of phosphate buffer solution); A Y2 The absorbance values ​​were measured without tyrosinase solution and without the sample solution (replaced with an equal volume of phosphate buffer solution).

[0100] Table 1. Results of tyrosinase inhibitory activity test of the traditional Chinese medicine composition of the present invention

[0101] Tyrosinase inhibition rate The traditional Chinese medicine composition prepared in Example 1 66.9％ The traditional Chinese medicine composition prepared in Example 2 83.7％ The traditional Chinese medicine composition prepared in Example 3 95.1％ The traditional Chinese medicine composition prepared in Comparative Example 1 52.6％ The traditional Chinese medicine composition prepared in Comparative Example 2 70.3％ The traditional Chinese medicine composition prepared in Comparative Example 3 42.4％ The traditional Chinese medicine composition prepared in Comparative Example 4 60.1％ The traditional Chinese medicine composition prepared in Comparative Example 5 80.6％

[0102] As can be seen from the experimental results in Table 1, the traditional Chinese medicine compositions prepared by this invention all have tyrosinase inhibitory effects.

[0103] As can be seen from the experimental results in Table 1, the tyrosinase inhibition rate of the traditional Chinese medicine composition prepared in Example 2 is significantly higher than that of the traditional Chinese medicine composition prepared in Example 1. This indicates that the macroporous resin elution fraction obtained by further eluting the traditional Chinese medicine alcohol extract using the macroporous resin column elution conditions described in this invention exhibits a significantly improved tyrosinase inhibition effect compared to the alcohol extract. Therefore, using the macroporous resin elution fraction as the traditional Chinese medicine composition results in a superior tyrosinase inhibition effect compared to using the alcohol extract.

[0104] As can be seen from the experimental results in Table 1, the tyrosinase inhibition rate of the traditional Chinese medicine compositions prepared in Comparative Examples 1 and 3 is lower than, or even significantly lower than, that of the traditional Chinese medicine composition prepared in Example 1. While the tyrosinase inhibition rate of the traditional Chinese medicine composition prepared in Comparative Example 2 is higher than that of the traditional Chinese medicine composition prepared in Example 1, the increase is not significant and is much smaller than that of the traditional Chinese medicine composition prepared in Example 2. This indicates that the elution conditions of the macroporous resin column in this invention are crucial. The types and contents of active ingredients in the macroporous resin eluted fractions prepared using different macroporous resin column elution conditions are completely different, leading to completely different tyrosinase inhibitory activities. The above experimental results show that only when the ethanol extract of traditional Chinese medicine is further eluted using the macroporous resin column elution conditions described in this invention can the tyrosinase inhibition effect of the macroporous resin eluted fraction be significantly improved compared to the ethanol extract of traditional Chinese medicine. However, the tyrosinase inhibition effect of the macroporous resin eluted fractions prepared by further eluting the ethanol extract of traditional Chinese medicine using other macroporous resin column elution conditions cannot be significantly improved compared to the ethanol extract of traditional Chinese medicine.

[0105] As can be seen from the experimental results in Table 1, the tyrosinase inhibition rate of the traditional Chinese medicine composition prepared in Example 3 is significantly higher than that of the traditional Chinese medicine composition prepared in Example 2. This indicates that the tyrosinase inhibition effect of the traditional Chinese medicine silica gel elution fraction prepared by further eluting the macroporous resin elution fraction of the traditional Chinese medicine under the silica gel column elution conditions described in this invention is further significantly improved compared to that of the macroporous resin elution fraction of the traditional Chinese medicine. This results in the prepared traditional Chinese medicine composition having excellent tyrosinase inhibition effect.

[0106] As can be seen from the experimental results in Table 1, the tyrosinase inhibition rate of the traditional Chinese medicine compositions prepared in Comparative Examples 4 and 5 is lower than, and even significantly lower than, that of the traditional Chinese medicine composition prepared in Example 2. This indicates that the silica gel column elution conditions of this invention are crucial. Silica gel elution fractions prepared using different silica gel column elution conditions have completely different types and contents of active ingredients, leading to completely different tyrosinase inhibitory activities. The above experimental results show that only silica gel elution fractions prepared using the silica gel column elution conditions described in this invention can achieve a significantly higher tyrosinase inhibition rate compared to those eluted with macroporous resin. However, silica gel elution fractions prepared using other silica gel column elution conditions do not show a significantly higher tyrosinase inhibition rate compared to those eluted with macroporous resin.

Claims

1. A traditional Chinese medicine composition, characterized in that, It is made from the following raw materials in parts by weight: 8 parts Rehmannia glutinosa; 8 parts Cistanche deserticola; 8 parts Miracle Fruit kernel; 5 parts Morinda officinalis; 4 parts Cornus officinalis; 4 parts Arctium lappa. The preparation method of the aforementioned traditional Chinese medicine combination includes the following steps: (1) Take Rehmannia glutinosa, Cistanche deserticola, Miracle Fruit kernel, Morinda officinalis, Cornus officinalis and Arctium lappa, mix them and crush them, pass them through a 100-mesh sieve to obtain Chinese medicine powder. Then, heat the Chinese medicine powder with an ethanol aqueous solution to boiling point and reflux for 1 hour. After extraction, concentrate and dry the extract to obtain Chinese medicine alcohol extract. The ethanol aqueous solution refers to an ethanol aqueous solution with a volume fraction of 50%. The ratio of Chinese medicine powder to ethanol aqueous solution is 1 kg: 15 L. (2) The ethanol extract of Chinese medicine was loaded onto a macroporous resin column. The macroporous resin column was filled with D101 type macroporous resin packing material with a weight of 50 times that of the ethanol extract of Chinese medicine. First, it was eluted with 5 column volumes of 13% ethanol aqueous solution to remove impurities. Then, it was eluted with 5 column volumes of 27% ethanol aqueous solution. The eluent eluted with 27% ethanol aqueous solution was collected, concentrated and dried to obtain the macroporous resin eluted part of Chinese medicine. (3) The macroporous resin elution fraction of the Chinese medicine was loaded onto a silica gel column, and the silica gel column was filled with 200-300 mesh silica gel, which was 30 times the weight of the macroporous resin elution fraction of the Chinese medicine. First, the elution was carried out with an organic solvent composed of cyclohexane and acetone in a volume ratio of 83:17, which was 3 times the column volume. Then, the elution was carried out with an organic solvent composed of cyclohexane and acetone in a volume ratio of 72:28, which was 6 times the column volume. The eluent eluted by the organic solvent composed of cyclohexane and acetone in a volume ratio of 72:28 was collected, concentrated and dried to obtain the silica gel elution fraction of the Chinese medicine. The Chinese medicine combination was obtained by taking the silica gel elution fraction of the Chinese medicine.

2. The application of the traditional Chinese medicine combination according to claim 1 in the preparation of products with tyrosinase inhibitory effects.

3. The application of the traditional Chinese medicine combination according to claim 1 in the preparation of a product with the effect of removing postpartum pigmentation spots.

4. The application according to claim 3, characterized in that, The product in question is a cosmetic or a pharmaceutical.