CN116747162A - Radix Potentillae Anserinae extract with tyrosinase inhibiting effect - Google Patents
Radix Potentillae Anserinae extract with tyrosinase inhibiting effect Download PDFInfo
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- 102000003425 Tyrosinase Human genes 0.000 title claims abstract description 28
- 108060008724 Tyrosinase Proteins 0.000 title claims abstract description 28
- 230000002401 inhibitory effect Effects 0.000 title claims abstract description 11
- 239000000284 extract Substances 0.000 title claims description 25
- 241001441970 Anserinae Species 0.000 title claims description 5
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims abstract description 50
- 244000308495 Potentilla anserina Species 0.000 claims abstract description 42
- 235000016594 Potentilla anserina Nutrition 0.000 claims abstract description 41
- 238000000034 method Methods 0.000 claims abstract description 24
- 238000005325 percolation Methods 0.000 claims abstract description 18
- 238000001035 drying Methods 0.000 claims abstract description 12
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 10
- 239000000843 powder Substances 0.000 claims description 8
- 238000001816 cooling Methods 0.000 claims description 4
- 239000002245 particle Substances 0.000 claims description 4
- 238000005086 pumping Methods 0.000 claims description 4
- 238000002791 soaking Methods 0.000 claims description 3
- 238000004140 cleaning Methods 0.000 claims description 2
- 238000011049 filling Methods 0.000 claims description 2
- 239000008187 granular material Substances 0.000 claims description 2
- 238000002156 mixing Methods 0.000 claims description 2
- 238000010298 pulverizing process Methods 0.000 claims description 2
- 238000002360 preparation method Methods 0.000 claims 1
- 238000004108 freeze drying Methods 0.000 abstract description 10
- 230000000694 effects Effects 0.000 abstract description 9
- 239000002994 raw material Substances 0.000 abstract description 3
- 239000002537 cosmetic Substances 0.000 abstract description 2
- 230000002087 whitening effect Effects 0.000 abstract description 2
- 239000007788 liquid Substances 0.000 abstract 1
- 229930003935 flavonoid Natural products 0.000 description 17
- 150000002215 flavonoids Chemical class 0.000 description 17
- 235000017173 flavonoids Nutrition 0.000 description 17
- 150000008442 polyphenolic compounds Chemical class 0.000 description 17
- 235000013824 polyphenols Nutrition 0.000 description 17
- 238000000605 extraction Methods 0.000 description 12
- 238000010992 reflux Methods 0.000 description 11
- 239000000243 solution Substances 0.000 description 11
- 238000006243 chemical reaction Methods 0.000 description 8
- 238000007602 hot air drying Methods 0.000 description 7
- 230000005764 inhibitory process Effects 0.000 description 6
- 238000002835 absorbance Methods 0.000 description 5
- WTDRDQBEARUVNC-LURJTMIESA-N L-DOPA Chemical compound OC(=O)[C@@H](N)CC1=CC=C(O)C(O)=C1 WTDRDQBEARUVNC-LURJTMIESA-N 0.000 description 4
- WTDRDQBEARUVNC-UHFFFAOYSA-N L-Dopa Natural products OC(=O)C(N)CC1=CC=C(O)C(O)=C1 WTDRDQBEARUVNC-UHFFFAOYSA-N 0.000 description 4
- XUMBMVFBXHLACL-UHFFFAOYSA-N Melanin Chemical compound O=C1C(=O)C(C2=CNC3=C(C(C(=O)C4=C32)=O)C)=C2C4=CNC2=C1C XUMBMVFBXHLACL-UHFFFAOYSA-N 0.000 description 4
- GAMYVSCDDLXAQW-AOIWZFSPSA-N Thermopsosid Natural products O(C)c1c(O)ccc(C=2Oc3c(c(O)cc(O[C@H]4[C@H](O)[C@@H](O)[C@H](O)[C@H](CO)O4)c3)C(=O)C=2)c1 GAMYVSCDDLXAQW-AOIWZFSPSA-N 0.000 description 4
- 229930003944 flavone Natural products 0.000 description 4
- 150000002212 flavone derivatives Chemical class 0.000 description 4
- 235000011949 flavones Nutrition 0.000 description 4
- 150000007965 phenolic acids Chemical class 0.000 description 4
- VHBFFQKBGNRLFZ-UHFFFAOYSA-N vitamin p Natural products O1C2=CC=CC=C2C(=O)C=C1C1=CC=CC=C1 VHBFFQKBGNRLFZ-UHFFFAOYSA-N 0.000 description 4
- 238000010438 heat treatment Methods 0.000 description 3
- 239000008055 phosphate buffer solution Substances 0.000 description 3
- 238000011160 research Methods 0.000 description 3
- AZQWKYJCGOJGHM-UHFFFAOYSA-N 1,4-benzoquinone Chemical compound O=C1C=CC(=O)C=C1 AZQWKYJCGOJGHM-UHFFFAOYSA-N 0.000 description 2
- AHMIDUVKSGCHAU-UHFFFAOYSA-N Dopaquinone Natural products OC(=O)C(N)CC1=CC(=O)C(=O)C=C1 AHMIDUVKSGCHAU-UHFFFAOYSA-N 0.000 description 2
- 102000004190 Enzymes Human genes 0.000 description 2
- 108090000790 Enzymes Proteins 0.000 description 2
- AHMIDUVKSGCHAU-LURJTMIESA-N L-dopaquinone Chemical compound [O-]C(=O)[C@@H]([NH3+])CC1=CC(=O)C(=O)C=C1 AHMIDUVKSGCHAU-LURJTMIESA-N 0.000 description 2
- OUYCCCASQSFEME-QMMMGPOBSA-N L-tyrosine Chemical compound OC(=O)[C@@H](N)CC1=CC=C(O)C=C1 OUYCCCASQSFEME-QMMMGPOBSA-N 0.000 description 2
- 208000003351 Melanosis Diseases 0.000 description 2
- 210000004027 cell Anatomy 0.000 description 2
- 238000001914 filtration Methods 0.000 description 2
- 235000013305 food Nutrition 0.000 description 2
- LNTHITQWFMADLM-UHFFFAOYSA-N gallic acid Chemical compound OC(=O)C1=CC(O)=C(O)C(O)=C1 LNTHITQWFMADLM-UHFFFAOYSA-N 0.000 description 2
- 230000008099 melanin synthesis Effects 0.000 description 2
- 230000002018 overexpression Effects 0.000 description 2
- 230000003647 oxidation Effects 0.000 description 2
- 238000007254 oxidation reaction Methods 0.000 description 2
- 235000009048 phenolic acids Nutrition 0.000 description 2
- 235000001674 Agaricus brunnescens Nutrition 0.000 description 1
- 241000143060 Americamysis bahia Species 0.000 description 1
- JMGZEFIQIZZSBH-UHFFFAOYSA-N Bioquercetin Natural products CC1OC(OCC(O)C2OC(OC3=C(Oc4cc(O)cc(O)c4C3=O)c5ccc(O)c(O)c5)C(O)C2O)C(O)C(O)C1O JMGZEFIQIZZSBH-UHFFFAOYSA-N 0.000 description 1
- 206010008570 Chloasma Diseases 0.000 description 1
- 241000196324 Embryophyta Species 0.000 description 1
- 235000003417 Plumeria rubra f acutifolia Nutrition 0.000 description 1
- 244000040691 Plumeria rubra f. acutifolia Species 0.000 description 1
- 101710147108 Tyrosinase inhibitor Proteins 0.000 description 1
- 238000010521 absorption reaction Methods 0.000 description 1
- 238000009825 accumulation Methods 0.000 description 1
- 239000004480 active ingredient Substances 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 238000006555 catalytic reaction Methods 0.000 description 1
- 230000002596 correlated effect Effects 0.000 description 1
- 235000013399 edible fruits Nutrition 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- IVTMALDHFAHOGL-UHFFFAOYSA-N eriodictyol 7-O-rutinoside Natural products OC1C(O)C(O)C(C)OC1OCC1C(O)C(O)C(O)C(OC=2C=C3C(C(C(O)=C(O3)C=3C=C(O)C(O)=CC=3)=O)=C(O)C=2)O1 IVTMALDHFAHOGL-UHFFFAOYSA-N 0.000 description 1
- 239000000469 ethanolic extract Substances 0.000 description 1
- 239000000706 filtrate Substances 0.000 description 1
- 235000012055 fruits and vegetables Nutrition 0.000 description 1
- 235000004515 gallic acid Nutrition 0.000 description 1
- 229940074391 gallic acid Drugs 0.000 description 1
- BEJNERDRQOWKJM-UHFFFAOYSA-N kojic acid Chemical compound OCC1=CC(=O)C(O)=CO1 BEJNERDRQOWKJM-UHFFFAOYSA-N 0.000 description 1
- 229960004705 kojic acid Drugs 0.000 description 1
- WZNJWVWKTVETCG-UHFFFAOYSA-N kojic acid Natural products OC(=O)C(N)CN1C=CC(=O)C(O)=C1 WZNJWVWKTVETCG-UHFFFAOYSA-N 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- 238000005259 measurement Methods 0.000 description 1
- 210000002752 melanocyte Anatomy 0.000 description 1
- 230000003061 melanogenesis Effects 0.000 description 1
- 239000000203 mixture Substances 0.000 description 1
- 108010058393 monophenolase Proteins 0.000 description 1
- 150000002989 phenols Chemical class 0.000 description 1
- 238000003672 processing method Methods 0.000 description 1
- 239000000047 product Substances 0.000 description 1
- FDRQPMVGJOQVTL-UHFFFAOYSA-N quercetin rutinoside Natural products OC1C(O)C(O)C(CO)OC1OCC1C(O)C(O)C(O)C(OC=2C(C3=C(O)C=C(O)C=C3OC=2C=2C=C(O)C(O)=CC=2)=O)O1 FDRQPMVGJOQVTL-UHFFFAOYSA-N 0.000 description 1
- 230000001105 regulatory effect Effects 0.000 description 1
- IKGXIBQEEMLURG-BKUODXTLSA-N rutin Chemical compound O[C@H]1[C@H](O)[C@@H](O)[C@H](C)O[C@@H]1OC[C@H]1[C@H](O)[C@@H](O)[C@H](O)[C@@H](OC=2C(C3=C(O)C=C(O)C=C3OC=2C=2C=C(O)C(O)=CC=2)=O)O1 IKGXIBQEEMLURG-BKUODXTLSA-N 0.000 description 1
- ALABRVAAKCSLSC-UHFFFAOYSA-N rutin Natural products CC1OC(OCC2OC(O)C(O)C(O)C2O)C(O)C(O)C1OC3=C(Oc4cc(O)cc(O)c4C3=O)c5ccc(O)c(O)c5 ALABRVAAKCSLSC-UHFFFAOYSA-N 0.000 description 1
- 235000005493 rutin Nutrition 0.000 description 1
- 229960004555 rutoside Drugs 0.000 description 1
- 229930182490 saponin Natural products 0.000 description 1
- 235000017709 saponins Nutrition 0.000 description 1
- 150000007949 saponins Chemical class 0.000 description 1
- 238000007789 sealing Methods 0.000 description 1
- 210000003491 skin Anatomy 0.000 description 1
- 208000017520 skin disease Diseases 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
- 229960004441 tyrosine Drugs 0.000 description 1
- 235000013311 vegetables Nutrition 0.000 description 1
Classifications
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K8/00—Cosmetics or similar toiletry preparations
- A61K8/18—Cosmetics or similar toiletry preparations characterised by the composition
- A61K8/96—Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution
- A61K8/97—Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution from algae, fungi, lichens or plants; from derivatives thereof
- A61K8/9783—Angiosperms [Magnoliophyta]
- A61K8/9789—Magnoliopsida [dicotyledons]
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L3/00—Preservation of foods or foodstuffs, in general, e.g. pasteurising, sterilising, specially adapted for foods or foodstuffs
- A23L3/34—Preservation of foods or foodstuffs, in general, e.g. pasteurising, sterilising, specially adapted for foods or foodstuffs by treatment with chemicals
- A23L3/3454—Preservation of foods or foodstuffs, in general, e.g. pasteurising, sterilising, specially adapted for foods or foodstuffs by treatment with chemicals in the form of liquids or solids
- A23L3/3463—Organic compounds; Microorganisms; Enzymes
- A23L3/3472—Compounds of undetermined constitution obtained from animals or plants
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61Q—SPECIFIC USE OF COSMETICS OR SIMILAR TOILETRY PREPARATIONS
- A61Q19/00—Preparations for care of the skin
- A61Q19/02—Preparations for care of the skin for chemically bleaching or whitening the skin
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2800/00—Properties of cosmetic compositions or active ingredients thereof or formulation aids used therein and process related aspects
- A61K2800/74—Biological properties of particular ingredients
- A61K2800/78—Enzyme modulators, e.g. Enzyme agonists
- A61K2800/782—Enzyme inhibitors; Enzyme antagonists
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2800/00—Properties of cosmetic compositions or active ingredients thereof or formulation aids used therein and process related aspects
- A61K2800/80—Process related aspects concerning the preparation of the cosmetic composition or the storage or application thereof
- A61K2800/805—Corresponding aspects not provided for by any of codes A61K2800/81 - A61K2800/95
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2800/00—Properties of cosmetic compositions or active ingredients thereof or formulation aids used therein and process related aspects
- A61K2800/80—Process related aspects concerning the preparation of the cosmetic composition or the storage or application thereof
- A61K2800/84—Products or compounds obtained by lyophilisation, freeze-drying
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- Health & Medical Sciences (AREA)
- General Health & Medical Sciences (AREA)
- Engineering & Computer Science (AREA)
- Chemical & Material Sciences (AREA)
- Botany (AREA)
- Microbiology (AREA)
- Veterinary Medicine (AREA)
- Public Health (AREA)
- Animal Behavior & Ethology (AREA)
- Mycology (AREA)
- Epidemiology (AREA)
- Birds (AREA)
- Dermatology (AREA)
- Biotechnology (AREA)
- Chemical Kinetics & Catalysis (AREA)
- General Chemical & Material Sciences (AREA)
- Nutrition Science (AREA)
- Food Science & Technology (AREA)
- Polymers & Plastics (AREA)
- Cosmetics (AREA)
- Medicines Containing Plant Substances (AREA)
- Coloring Foods And Improving Nutritive Qualities (AREA)
Abstract
The invention relates to a method for preparing a cosmetic with whitening effect, which comprises the steps of taking fresh potentilla anserina as a raw material, drying the fresh potentilla anserina by a freeze-drying method, extracting the dried potentilla anserina by a percolation method with 75% ethanol, recovering the ethanol from the percolation liquid at a low temperature, and freeze-drying the obtained dry product, wherein the dried product has the effect of inhibiting tyrosinase.
Description
(II) Specification
Technical Field
The invention relates to an extract with tyrosinase inhibiting effect, which is prepared from fresh potentilla anserina by freeze drying and percolating with 75% ethanol. Belongs to the technical field of daily chemical industry.
The background technology is as follows:
in the human body, the amount of melanin determines the color of the skin. Melanin production is mainly regulated by Tyrosinase (TY) in melanocytes, and excessive TY or excessive activity thereof can lead to excessive accumulation of melanin, and hyperpigmented skin diseases such as freckle, chloasma, etc. appear.
In addition, the over-expression of tyrosinase can lead to the catalysis of phenolic compounds in fruits, vegetables and foods into quinone to cause browning reaction. Blackening of shrimps and the like are all related to TY overexpression.
Therefore, the tyrosinase inhibitor with better action is found to have important significance.
The existing researches show that flavonoid components and polyphenol components in plants have better tyrosinase inhibition effect. The research shows that the potentilla anserina extract has better tyrosinase inhibiting effect, and the inhibiting effect has obvious relation with the processing method and the extraction process of the potentilla anserina.
(III) summary of the invention:
the invention is obtained by the following technical scheme.
1. Removing fresh root tuber of Potentilla anserina Potentilla anserina L, cleaning, and pulverizing into coarse granule (particle diameter of 2-4 mm);
2. placing the crude particles of the fresh potentilla anserina into a drying box of a freeze dryer, cooling to-30 ℃ to-40 ℃ and maintaining for 2-4 hours; the condenser is cooled to-60 to-70 ℃. Vacuum pumping is started, the drying box is slowly heated, the temperature is raised to 0 ℃ and controlled to be 12-16 hours, the temperature is continuously raised to 35-40 ℃ and maintained for 6-8 hours, and the product is obtained. The water content of the coarse powder of the dry potentilla anserina is controlled below 5 percent.
3. The prepared freeze-dried potentilla anserina coarse powder is extracted by a 75% ethanol percolation method, and the steps are as follows: adding 75% ethanol into freeze-dried potentilla anserina coarse powder, uniformly mixing, moistening for 30min, filling into a percolator, adding 3 times of 75% ethanol, soaking for 12 hours, and starting percolating at a percolating speed of 3-5 ml/kg.min. And (3) receiving 8 times of percolate, and recovering ethanol from the percolate at low temperature to obtain concentrated solution.
3. Placing the concentrated solution in a drying box of a freeze dryer, cooling to-30 ℃ to-40 ℃ and maintaining for 2-4 hours; the condenser is cooled to-60 to-70 ℃. Vacuum pumping is started, the drying oven is slowly heated, the temperature is raised to 0 ℃ and controlled to be 12-16 hours, the temperature is continuously raised to 35-40 ℃ and maintained for 6-8 hours, and then the extract with the tyrosinase inhibiting effect is obtained. The water content of the extract is controlled below 5%.
The invention has the following advantages
The freeze-dried potentilla anserina extract can be used as a raw material of cosmetics with whitening effect and a raw material for fruit and vegetable fresh-keeping.
(IV) detailed description of the invention
The present invention will be further illustrated with reference to the following examples, but the present invention is not limited to the following examples.
Example 1 the lyophilized water extract of potentilla anserina contained more active ingredient than the sun-dried water extract of potentilla anserina.
The potentilla anserina contains saponins, flavonoids and multi-classified components. More and more researches indicate that flavonoid components and polyphenol components have the effect of inhibiting tyrosinase. For this purpose, the content of total flavonoids and the content of total polyphenols in the water extract of freeze-dried potentilla anserina and sun-dried potentilla anserina were compared, and the specific method is as follows:
taking 100g of potentilla anserina with two drying methods, respectively adding 75% ethanol for reflux extraction, adding 10 times of 75% ethanol for the first time, refluxing for 2 hours, adding 8 times of 75% ethanol for the second time, refluxing for 1 hour, combining the two 75% ethanol reflux extracts, filtering, recovering ethanol from filtrate until no alcohol smell exists, and adding water to constant volume to 100ml to obtain the potentilla anserina. The content of total flavonoids in the extracting solution is measured according to the method under the pagodatree flower item of the Chinese pharmacopoeia of 2020 edition, and the result is shown in table 1; the total phenolic acid content of the extract was determined according to the method for determining total phenolic acid in foods, and the results are shown in Table 1.
TABLE 1 content of total flavonoids and total polyphenols in 75% ethanol extracts of lyophilized and sun-dried Potentilla anserina
| Group of | Total flavone mg/g | Total polyphenol mg/g |
| Freeze-dried potentilla anserina | 7.89 | 4.65 |
| Sun-dried potentilla anserina | 4.62 | 2.02 |
Note that: the total flavone content refers to the amount of rutin equivalent per 1g of dry potentilla anserina (without calculating moisture); total polyphenols refer to the amount of comparable gallic acid per 1g of dry potentilla anserina (without accounting for moisture)
As can be seen from table 1, the content of total flavonoids and total phenolic acids in lyophilized potentilla anserina is significantly higher than in sun-dried potentilla anserina, which may be related to both aspects; on the one hand, the freeze drying can freeze water in the cells of the fresh potentilla anserina, increase the volume, lead the cells to swell and split, and is beneficial to the extraction of the components; on the other hand, it may be associated with instability of the total flavonoids and total phenolic acids, and freeze-drying cannot destroy the components.
Example 2 lyophilized potentilla anserina was extracted with 75% ethanol by percolation with greater amounts of total flavonoids and total polyphenols than by heated reflux. The specific method comprises the following steps:
taking 100g of freeze-dried potentilla anserina coarse powder and 2 parts. Extracting with 75% ethanol under reflux and percolating respectively, and extracting with reflux method as described in example 1. The percolation extraction method comprises the following steps: freeze-drying radix Potentillae Anserinae coarse powder 100g, adding 75% ethanol 100ml, sealing, moistening for 30min, percolating, and compacting layer by layer; adding 300ml of 75% ethanol according to the operation of a percolation method, soaking for 12 hours, starting percolation at a percolation speed of 5ml/min.kg, collecting 800ml of percolate, recovering ethanol until no alcohol smell exists, adding water to a constant volume of 100ml, and measuring the total flavonoids and total polyphenols under the condition of the item 1. The results are shown in Table 2.
Table 2 comparison of total flavonoids and total polyphenols extracted from lyophilized potentilla anserina by percolation and by thermal reflux
| Extraction method | Total flavone mg/g | Total polyphenol mg/g |
| Percolation method | 10.96 | 8.89 |
| Heating reflux method | 7.82 | 4.18 |
As can be seen from Table 2, the percolation method of extraction retains more flavonoid and polyphenol components due to the short heating time (heating only when recovering ethanol under reduced pressure).
Example 3 the 75% ethanol percolation extract of lyophilized potentilla anserina is lyophilized to better retain total flavonoids and total polyphenols
Preparing various extracting solutions according to the method, concentrating under reduced pressure, and respectively adopting hot air drying (the temperature is 60-70 ℃); the lyophilization conditions are the conditions of claim 2. The amounts of total flavonoids and total polyphenols in the dried material were measured and converted to the amounts in the unit potentilla anserina, the results are shown in Table 3.
TABLE 3 amounts of Total Flavonoids and Total Polyphenols in Potentilla anserina extracts by different extraction and drying methods
| Extraction method | Decoction and hot air drying method | Reflux and hot air drying method | Percolation and hot air drying method | Percolating, freeze drying |
| Total flavone | 4.10mg/g | 5.13mg/g | 6.04mg/g | 10.89mg/g |
| Total polyphenols | 2.06mg/g | 2.86mg/g | 3.07mg/g | 8.61mg/g |
Table 3 shows that the total flavonoids and total polyphenols in the freeze-dried potentilla anserina can be better preserved by percolation extraction and freeze-drying.
Example 4 lyophilized potentilla anserina 75% ethanol percolation extract inhibited tyrosinase and compared to other extracts of alternative extraction methods. The method comprises the following steps:
preparing a solution:
mushroom tyrosinase (Shanghai Aladin Co.), L-dopa and kojic acid (Nanjing Ordofovir Biotech Co., ltd.) were prepared with 0.05mol/LpH 6.8.6.8 phosphate buffer solution respectively to 1.43X10 -5 mol/L、4.8×10 -3 mol/L and 4.1X10 -3 mol/L。
Preparing potentilla anserina extract solution: respectively dissolving the three dry extracts in phosphate buffer solution to obtain enzyme 1ml solution equivalent to 1g potentilla anserina, and filtering.
The measurement principle is as follows:
tyrosinase activity was determined by tyrosinase dopa-rate oxidation. During melanogenesis, tyrosinase catalyzes the oxidation of L-tyrosine to L-dopa first with monophenolase activity and then the conversion of L-dopa to dopaquinone with diphenolase activity. Dopaquinone is a colored intermediate for melanin synthesis, and has a strong absorption at 475nm, and therefore the absorbance of the system is measured at 475nm to determine its tyrosinase activity. The reaction system is shown in Table 4.
TABLE 4 reaction system composition
| Name of the name | A1 | A2 | A3 | A4 |
| Phosphate buffer solution/. Mu.l | 80 | 160 | 0 | 0 |
| Enzyme/. Mu.l | 80 | 0 | 80 | 0 |
| Sample/. Mu.l | 0 | 0 | 80 | 80 |
| L-Dopa/μl | 80 | 80 | 80 | 80 |
A1: absorbance measured at 475nm of the reaction solution to which tyrosinase was added without adding the sample;
a2: absorbance measured at 475nm of the reaction solution without sample and without tyrosinase;
a3: absorbance measured at 475nm of the reaction solution with the sample and tyrosinase added;
a4: the absorbance of the reaction solution with the sample and without tyrosinase was measured at 475 nm.
As a result, the inhibition ratios measured are shown in Table 5
TABLE 5 inhibition of tyrosinase by different potentilla anserina extracts
| Extraction method | Decoction and hot air drying method | Reflux and hot air drying method | Percolation and hot air drying method | Percolating, freeze drying |
| Inhibition rate | 48.9％ | 60.2％ | 68.1％ | 92.3％ |
Table 5 can be seen. The extract prepared by the percolation method extraction cold drying method has very obvious effect of inhibiting tyrosinase, and the inhibition rate is obviously higher than that of the extracts prepared by other methods. The inhibition rate of the extract on tyrosinase is positively correlated with the amount of total flavonoids and total polyphenols.
Claims (3)
1. A preparation method of radix Potentillae Anserinae extract with tyrosinase inhibiting effect is provided. The method is characterized by comprising the following steps of:
(1) Digging fresh radix Potentillae Anserinae tuberous root, removing fibrous root, cleaning, and pulverizing into coarse granule (particle diameter of 2-4 mm);
(2) Placing the crude particles of the fresh potentilla anserina into a drying box of a freeze dryer, cooling to-30 ℃ to-40 ℃ and maintaining for 2-4 hours; the condenser is cooled to-60 to-70 ℃. Vacuum pumping is started, the drying box is slowly heated, the temperature is raised to 0 ℃ and controlled to be 12-16 hours, the temperature is continuously raised to 35-40 ℃ and maintained for 6-8 hours, and the product is obtained. The water content of the coarse powder of the dry potentilla anserina is controlled below 5 percent.
2. According to the requirement of claim 1, the prepared freeze-dried potentilla anserina coarse powder is extracted by a 75% ethanol percolation method, and the steps are as follows: adding 75% ethanol into freeze-dried potentilla anserina coarse powder, uniformly mixing, moistening for 30min, filling into a percolator, adding 3 times of 75% ethanol, soaking for 12 hours, and starting percolating at a percolating speed of 3-5 ml/kg.min. And (3) receiving 7-10 times of percolate, and recovering ethanol from the percolate at low temperature to obtain concentrated solution.
3. According to the requirement of claim 2, placing the concentrated solution in a drying box of a freeze dryer, cooling to-30 ℃ to-40 ℃ and maintaining for 2 to 4 hours; the condenser is cooled to-60 to-70 ℃. Vacuum pumping is started, the drying oven is slowly heated, the temperature is raised to 0 ℃ and controlled to be 12-16 hours, the temperature is continuously raised to 35-40 ℃ and maintained for 6-8 hours, and then the extract with the tyrosinase inhibiting effect is obtained. The water content of the extract is controlled below 5%.
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