CN109984963B - Preparation method of pine pollen-turmeric antioxidant skin care product - Google Patents
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- 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
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- A61K8/9767—Pinaceae [Pine family], e.g. pine or cedar
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- A61K8/9794—Liliopsida [monocotyledons]
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- 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
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Abstract
The invention discloses a preparation method of a pine pollen-turmeric antioxidant skin care product, which comprises the steps of preparing a pine pollen extract and a turmeric extract, and mixing the pine pollen extract and the turmeric extract to prepare the antioxidant skin care product. The invention has the beneficial effects that: the antioxidant skin care product prepared from natural extracts of pine pollen and turmeric does not contain harmful chemical additives, can effectively remove free radicals, reduce the ROS level, improve the activity of antioxidant enzyme, improve the skin elasticity and delay senility.
Description
Technical Field
The invention relates to the field of daily-use chemistry, in particular to a preparation method of a pine pollen-turmeric antioxidant skin care product.
Background
With the development of modern science, active oxygen free radical theory has been widely accepted in a plurality of theories of skin aging. In the normal physiological metabolism process of human body, a large amount of Reactive Oxygen Species (ROS) are generated under the influence of light, dust, radiation, ultraviolet rays, air pollution, fried food, pressure and the like. The human body is in a balanced state, and when the accumulation amount of ROS cannot be counteracted with the ability of the human body to remove free radicals, the balance is broken. The skin covers the whole body of people to isolate the inside of the human body from the external environment, is equivalent to a 'barrier' between the human body and the external environment, is directly exposed to the environment, is an important organ for protecting the human body and maintaining the stability and balance of the internal environment of the human body, more easily and more intuitively reflects the oxidative damage of the organism, and is expressed as skin aging or diseases such as photoaging, dermatitis, skin tumor and the like.
Because the antioxidant mechanism of the human body is not an independent system, but the result of the mutual coordination and combined action of various mechanisms of the human body. Therefore, the development and use of skin care products with antioxidant effect directly act on the body surface, can achieve the effect of delaying skin aging, and gradually become the main requirements of consumer groups. At present, an evaluation system for eliminating free radicals, reducing the level of ROS (reactive oxygen species), improving the antioxidant activity and other antioxidant effects is widely accepted and popularized, natural products from plants are sought, the use of chemical components is reduced, and antioxidant active components are obtained on the basis of health and safety, so that the evaluation system becomes a development hotspot for designing and developing anti-aging skin care products.
Disclosure of Invention
The invention provides a preparation method of a pine pollen-turmeric antioxidant skin care product, aiming at the problem that the existing antioxidant skin care product has non-natural chemical components.
The technical scheme for solving the technical problems is as follows: a preparation method of a pine pollen-turmeric antioxidant skin care product is characterized by comprising the following steps:
1) preparing a pine pollen extract: taking dried pollen of Pinus massoniana of Pinaceae, breaking cell wall, soaking in 95% ethanol solution for 12-15h for degreasing and decolorizing, centrifuging, taking precipitate, drying by blowing, adding distilled water with the mass being 12 times of that of the precipitate, stirring and leaching at 95 ℃ for 3h, centrifuging, taking precipitate, repeatedly leaching for 2 times, combining filtrates obtained by 3 times of leaching, decolorizing the filtrate by AB-8 macroporous resin, and concentrating under reduced pressure to 20% of the original volume to obtain pollen Pini extract solution;
2) preparing a turmeric extract: pulverizing dried Curcuma rhizome, sieving with 40 mesh sieve, collecting powder 500g, extracting with 8 times weight of 80% ethanol solution under heating and refluxing for three times (2 hr each time), filtering, mixing ethanol extractive solutions, concentrating under reduced pressure, and oven drying to obtain total curcumin compound Curcuma rhizome extract;
3) preparing a pine pollen-turmeric antioxidant skin care product: mixing the pine pollen extract solution obtained in the step 1) and the turmeric extract obtained in the step 2), adding the mixture into a solution of 1, 3-butanediol and purified water in a volume ratio of 3:2, and uniformly stirring to obtain the pine pollen-turmeric antioxidant skin care product.
Wherein the content of total flavonoids in the pine pollen extract obtained in the step 1) is not less than 0.18 mg/mL; the content of the total curcumin in the turmeric extract obtained in the step 2) is not lower than 0.20 mg/mL; the mass ratio of the pine pollen extract solution to the total curcumin compound extract in the step 3) is (3-8) to 1.
The pine pollen is fresh yellow or light yellow fine powder, and has sweet and mild taste and no toxicity. The history of the pine pollen used as both medicine and food is over a thousand years, the pine pollen is one of two kinds of pollen collected in ancient Chinese medical Lao-Shi, is the only pollen variety used as both food and medicine in the medical treasury of China, the record of the pine pollen in the first pharmacopoeia of China, Shennong Bencao Jing, is listed as the superior; the pharmacopoeia of the Tang Dynasty "Xin Xiu Ben Cao" also includes it; ming dynasty Li Shizhen is better called in Ben Cao gang mu: flos Pini, sweet and warm, has no toxicity, and can moisten heart and lung, invigorate qi, remove wind, stop bleeding, and also be used for brewing wine. As an ancient medicine source and a nutrient source, pine pollen is confirmed to be a new resource food by the ministry of health of China in recent years and listed as common food management, and is received in the 2005 edition of Chinese pharmacopoeia. The pine pollen contains all nutrient components and life active substances of pine plants, wherein the pine pollen contains a plurality of proteins, more than 20 amino acids (including 8 kinds of essential amino acids for human bodies), 15 vitamins, more than 30 mineral elements, 11% fat, nearly hundred kinds of enzymes and coenzymes, as well as nucleic acid, unsaturated fatty acid, lecithin, flavonoid, monosaccharide, polysaccharide, cellulose and the like, the total amount reaches more than 200 kinds, and the pine pollen is reasonably matched, can completely supplement and balance the nutrition required for the human bodies, has the effects of improving immunity, resisting aging, resisting fatigue, regulating organism metabolism, reducing blood fat, beautifying and the like, and is widely applied to the fields of health care products, medicines, cosmetics, feed additives and the like at home and abroad.
Turmeric (Curcuma longa L.) which is a curcumin plant of Zingiberaceae is a traditional botanical drug widely used in countries of Asia, and is collected in the Law edition of pharmacopoeia of the people's republic of China. Curcumin is derived from dried rhizome of Curcuma longa (Curcuma longaL) belonging to Curcuma of Zingiberaceae, and is one of effective components of Curcuma longa (Curcuma Longae). Has various good biological activities and pharmacological effects of resisting inflammation, resisting oxidation, etc. In traditional Chinese medicine, the traditional Chinese medicine has the advantages of obvious effect and low toxicity, and is more and more valued at home and abroad, and is used for dispelling wind, activating blood, stimulating the menstrual flow, relieving pain and the like. In modern research, aiming at the structural characteristics of plant polyphenol, a novel derivative or analogue is generated by means of synthesis, dehydrogenation, condensation and the like, and the novel derivative or analogue is widely applied to the field of daily chemicals by utilizing the oxidation resistance, free radical removal, anti-inflammation and antibacterial effects of the novel derivative or analogue, such as tyrosinase inhibitors, antioxidants and the like.
The total curcumin compound prepared in the step 2) mainly comprises three phenolic substances, which are respectively as follows: curcumin (curcumin, C)21H20O6Formula I), Demethoxycurcumin (Demethoxycurcumin, C)20H18O5Formula II) and Bisdemethoxycurcumin (Bisdeomethoxycurcumin, C)19H16O4Formula III).
The invention has the beneficial effects that: the antioxidant skin care product prepared from natural extracts of pine pollen and turmeric does not contain harmful chemical additives, can effectively remove free radicals, reduce the ROS level, improve the activity of antioxidant enzyme, improve the skin elasticity and delay senility.
Detailed Description
The present invention is described below with reference to examples, which are provided for illustration only and are not intended to limit the scope of the present invention.
Preparation work:
1) preparing a pine pollen extract: taking dry pollen of pinus massoniana of Pinaceae, breaking cell wall, soaking in 95% ethanol solution for 12-15h for degreasing and decoloring, centrifuging, taking precipitate, drying by blowing, adding distilled water with the mass being 12 times of that of the precipitate, stirring and leaching at 95 ℃ for 3h, centrifuging, taking precipitate, repeatedly leaching for 2 times, combining filtrates obtained by 3 times of leaching, decoloring the filtrates by AB-8 macroporous resin, concentrating under reduced pressure to 20% of the original volume to obtain a pollen pini extract solution, preparing a rutin standard solution, drawing a linear regression curve, determining the content of total flavonoids to be 0.25mg/mL by an ultraviolet-spectrophotometry, detecting the appearance, pH, the total number of bacterial colonies, mould, saccharomycetes, heat-resistant coliform group bacteria and other microbial indexes, and testing for later use after the total flavonoids are qualified;
2) preparing a turmeric extract: pulverizing dried Curcuma rhizome, sieving with 40 mesh sieve, collecting powder 500g, extracting with 8 times weight of 80% ethanol solution under heating and refluxing for three times, each for 2 hr, filtering, mixing ethanol extractive solutions, concentrating under reduced pressure, oven drying to obtain Curcuma rhizome extract as total curcumin compound, detecting appearance, pH, total bacterial count, mold and yeast, heat-resisting coliform group bacteria and other microorganism indexes by thin layer chromatography with total curcumin content of 0.28mg/mL, and checking.
Examples 1-3 are methods for preparing a pollen pini-turmeric antioxidant skin care product, respectively, the same operations being: mixing the pollen pini extract solution and the curcuma extract, adding the mixture into a solution of 1, 3-butanediol and purified water in a volume ratio of 3:2, and uniformly stirring, wherein the difference is that the proportions of the pollen pini extract solution and the curcuma extract in examples 1-3 are respectively 5:1, 8:1 and 3: 1.
DPPH clearance test
1. Test materials: absolute ethyl alcohol, analytically pure; 1, 1-diphenyl-2-trinitrophenylhydrazine (DPPH); dimethyl sulfoxide (DMSO);
2. the main test instrument: varioskan Flash enzyme-linked immunosorbent assay (ELISA); Milli-Q ultrapure water; an ultraviolet-visible spectrophotometer (TU-1810); a Brand pipette; a 96-hole enzyme label plate;
3. positive control drug: vc solution;
4. the test steps are as follows: accurately weighing 20mg of DPPH, and diluting to 250mL with absolute ethyl alcohol to obtain a solution with a concentration of 2X 10- 4A solution of DPPH in mol/L. The characteristic purple red chromogen 517nm absorption peak of DPPH solution is utilized, the decrease of A517 absorption after the test substance is added is measured by an enzyme-linked immunosorbent assay to show the scavenging capacity of the enzyme-linked immunosorbent assay (ELISA) instrument on organic free radicals, the test sample is diluted to different concentrations, 190 mu L DPPH and 10 mu L of sample are added into a 96-hole ELISA plate, and the total reaction volume is 200 mu L. The blank control group is 190 μ L DPPH +10 μ L DMSO, the tinfoil paper is wrapped well after being mixed uniformly and then is shaken in the dark for reaction for 30min, and the change of the absorbance value under the wavelength of 517nm is measured;
5. data processing: the inhibition rate of the antioxidant substance was calculated according to the following formula,
inhibition [ ([ 1- ([ DPPH. ] t/[ DPPH. ] t ═ 0) ] × 100%,
in the formula: [ DPPH. t ═ 0-initial concentration of 1, 1-diphenyl-2-trinitrophenylhydrazine free radical in the blank system,
[ DPPH. t-concentration of 1, 1-diphenyl-2-trinitrophenylhydrazine radical after addition of the drug to be tested.
The measured results are shown in table 1, the pine pollen extract and the curcumin extract have obvious antioxidant effect compared with a positive control Vc, when the mass ratio of the pine pollen extract to the curcumin extract is 3:1, the in-vitro antioxidant effect of the composition has obvious antioxidant effect compared with the single use of the pine pollen extract and the curcumin extract, and the in-vitro antioxidant effect of the composition has a synergistic effect.
TABLE 1 DPPH clearance results of examples 1-3
Clearance rate test of di-hydroxyl free radical
1. Test materials: phosphate buffer at pH 7.4, 0.2 mol/L; 5.0mmol/L of phenanthroline; dimethyl sulfoxide (DMSO); FeCl2；H2O2；
2. The main test instrument: varioskan Flash enzyme-linked immunosorbent assay (ELISA); Milli-Q ultrapure water; an ultraviolet-visible spectrophotometer (TU-1810); a Brand pipette; a 96-hole enzyme label plate; a constant temperature water bath device;
3. positive control drug: vc solution.
4. The test steps are as follows: taking 40 mu L of phosphate buffer solution with pH 7.4 and 0.2mol/L and 160 mu L of ultrapure water as blanks; taking 40 mu L of phosphate buffer solution, 30 mu L of phenanthroline and 30 mu L of FeCl2(7.5mmol/L) and 110. mu.L of ultrapure water as A were intact; taking 40 mu L of phosphate buffer solution, 30 mu L of phenanthroline and 20 mu L of FeCl290 μ L of ultrapure water and 20 μ L of Vc solution (0.1%) as a lesion; taking 40 mu L of phosphate buffer solution, 30 mu L of phenanthroline and 20 mu L of FeCl280 μ L of ultrapure water, 10 μ L of drugs with different concentrations and 20 μ L of Vc solution are used as A samples, the total volume of the reaction is 200 μ L, the above groups of samples are shaken up and then placed in a constant temperature water tank, the temperature is kept at 37 ℃ for 60min, the samples are placed at a wavelength of 510nm, and the measured absorbance (A) value is shown in Table 2.
TABLE 2 Absorbance (A) assay record
5. Data processing: the clearance of hydroxyl radicals was calculated according to the following formula:
hydroxyl radical clearance ═ x 100% (sample a-a damaged)/(a undamaged-a damaged) ];
the results are shown in table 3, and the pollen pini-curcuma longa antioxidant skin care product obtained in examples 1-3 has a significant effect of removing hydroxyl radicals.
TABLE 3 results of hydroxyl radical scavenging rates for examples 1-3
The above description is only for the purpose of illustrating the preferred embodiments of the present invention and is not to be construed as limiting the invention, and any modifications, equivalents, improvements and the like that fall within the spirit and principle of the present invention are intended to be included therein.
Claims (3)
1. A preparation method of a pine pollen-turmeric antioxidant skin care product is characterized by comprising the following steps:
1) preparing a pine pollen extract: taking dried pollen of Pinus massoniana of Pinaceae, breaking cell wall, soaking in 95% ethanol solution for 12-15h for degreasing and decolorizing, centrifuging, taking precipitate, drying by blowing, adding distilled water with the mass being 12 times of that of the precipitate, stirring and leaching at 95 ℃ for 3h, centrifuging, taking precipitate, repeatedly leaching for 2 times, combining filtrates obtained by 3 times of leaching, decolorizing the filtrate by AB-8 macroporous resin, and concentrating under reduced pressure to 20% of the original volume to obtain pollen Pini extract solution;
2) preparing a turmeric extract: pulverizing dried Curcuma rhizome, sieving with 40 mesh sieve, collecting powder 500g, extracting with 8 times weight of 80% ethanol solution under heating and refluxing for three times (2 hr each time), filtering, mixing ethanol extractive solutions, concentrating under reduced pressure, and oven drying to obtain total curcumin compound Curcuma rhizome extract;
3) preparing a pine pollen-turmeric antioxidant skin care product: mixing the pine pollen extract solution obtained in the step 1) and the turmeric extract obtained in the step 2), adding the mixture into a solution of 1, 3-butanediol and purified water in a volume ratio of 3:2, and uniformly stirring to obtain a pine pollen-turmeric antioxidant skin care product; the ratio of the pollen pini extract solution to the curcumin extract is 3:1, 5:1 or 8: 1.
2. The method for preparing the pine pollen-turmeric antioxidant skin care product according to claim 1, wherein the content of total flavonoids in the pine pollen extract obtained in step 1) is not less than 0.18 mg/mL.
3. The method for preparing the pine pollen-turmeric antioxidant skin care product according to claim 1, wherein the total curcumin content in the turmeric extract obtained in step 2) is not less than 0.20 mg/mL.
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