Application of snow rabbit extract in preparation of product with nicotinic acid inhibition function

Abstract

The invention belongs to the field of natural product chemistry, and particularly relates to application of a snow rabbit extract to preparation of a product with a nicotinic acid inhibiting function. The snow rabbit extract is found to be capable of remarkably removing nicotinic acid and relieving the irritation effect of the nicotinic acid on the skin, and can be used for preparing products with the function of inhibiting the nicotinic acid. A new path is provided for effective development and utilization of the snow rabbit extract, function maximization of limited resources is achieved, and the application prospect is wide.

Description

Technical Field

[0001] The present invention belongs to the field of natural product chemistry, and particularly relates to the application of extracts from Saussurea laniceps Hand.-Mazz. in preparing products with the function of inhibiting nicotinic acid, especially the application of extracts from callus of Saussurea laniceps Hand.-Mazz. in preparing products with the function of inhibiting nicotinic acid. Background Art

[0002] Nicotinamide is a derivative of vitamin B3 and is also a recognized skin anti-aging ingredient in the field of beauty dermatology. In recent years, it has been increasingly valued by people. Its most important effect in skin anti-aging is to reduce and prevent the dull, yellowish, and sallow skin color produced during the early aging process of the skin. It can also repair the damaged lipid barrier of the stratum corneum and improve skin resistance. Relevant literature shows that nicotinamide has the best stability in a solution with a pH of 6. When the pH value is lower or higher than 6, it will be partially hydrolyzed into nicotinic acid, and light and high-temperature conditions will accelerate the conversion of nicotinamide in cosmetics into nicotinic acid.

[0003] At the same time, the current traditional reaction technology for preparing nicotinamide is the chemical hydrolysis method, which uses alkaline substances as catalysts, and various hydrolysis processes are generated due to different catalysts and different recovery and separation methods. In this chemical hydrolysis process, when the pH value increases, the conversion rate increases, but the yield of the by-product nicotinic acid also increases. Although nicotinic acid and nicotinamide are considered to play the same role as B vitamins, relevant literature reports that nicotinic acid may cause skin discomfort.

[0004] For example, nicotinic acid can activate G protein-coupled receptors (such as GPR109A), leading to vasodilation and increased blood flow, thereby causing flushing, making the skin on the face, neck, and upper chest red, hot, and itchy, and at the same time, some people may experience rashes or skin allergic reactions.

[0005] Therefore, how to effectively remove nicotinic acid, slow down its irritation to the skin, and inhibit the production of nicotinic acid in products containing nicotinamide is a problem that needs to be solved by current nicotinamide cosmetics. Existing methods for removing nicotinic acid are as follows: 1. Acid-base neutralization method, adding an appropriate amount of alkaline solution, and part of the nicotinic acid is converted into salts; 2. Solvent extraction method, using a solvent with strong hydrophilicity or water to transfer nicotinic acid into the aqueous phase; 3. Distillation technology, since the boiling point of nicotinic acid is relatively low, it can be distilled and separated. Although the existing methods can remove nicotinic acid, they increase the process operation and may introduce new impurities, increasing the difficulty of quality control.

[0006] Saussurea laniceps, a perennial herb of the genus Saussurea DC. in the Compositae family, has a flowering and fruiting period from August to October, and its leaves and capitula are covered with thick wool. It mainly grows in alpine scree slopes at an altitude of 3200 - 5280 meters and is a very typical alpine plant. Research shows that Saussurea laniceps contains rich flavonoids, polyphenols, polysaccharides and secondary metabolites, and has functions such as scavenging free radicals, anti-aging, and analgesia. Summary of the Invention

[0007] The present invention discovers that the extract of Saussurea laniceps can effectively convert and remove nicotinic acid, slow down or inhibit the side effects of nicotinic acid, and at the same time does not affect the efficacy of the product added with the extract of Saussurea laniceps. Using the extract of Saussurea laniceps as a nicotinic acid inhibitor not only does not require adding an additional nicotinic acid removal process, nor does it introduce new impurities and increase the difficulty of quality control, but also further develops and utilizes the functions of the extract of Saussurea laniceps.

[0008] The present invention discovers that the callus extract of Saussurea laniceps can significantly remove nicotinic acid and slow down the irritating effect of nicotinic acid on the skin, and can be used to prepare products with the function of inhibiting nicotinic acid.

[0009] The present invention provides the application of the extract of Saussurea laniceps in preparing products with the function of inhibiting nicotinic acid. The products with the function of inhibiting nicotinic acid include at least one of the following:

[0010] (1) Additives;

[0011] (2) Cosmetics;

[0012] (3) Health products;

[0013] (4) Foods;

[0014] (5) Medicines.

[0015] The Saussurea laniceps mentioned above is the whole plant of Saussurea laniceps, and / or normal tissues, and / or callus. The normal tissues and / or callus of Saussurea laniceps are derived from at least one of roots, stems, leaves, flowers, fruits, and seeds, preferably at least one of roots, stems, and leaves; preferably the callus of Saussurea laniceps.

[0016] The extract of Saussurea laniceps mentioned above is the extract of the whole plant of Saussurea laniceps, and / or normal tissues, and / or callus, further the water extract and / or alcohol extract of the whole plant of Saussurea laniceps, and / or normal tissues, and / or callus, further the water extract of the whole plant of Saussurea laniceps, and / or normal tissues, and / or callus, and more preferably the water extract of the callus of Saussurea laniceps.

[0017] The preparation method of the extract of Saussurea laniceps includes: extracting the whole plant, and / or normal tissues, and / or callus of Saussurea laniceps with ethanol and / or water. The ethanol is preferably 60%-80% ethanol, and more preferably 65%-75% ethanol. Further, the whole plant, and / or normal tissues and / or callus of Saussurea laniceps are extracted with water; more preferably, the callus of Saussurea laniceps is extracted with water. Before extraction, the ethanol and / or water mixed with the whole plant, and / or normal tissues, and / or callus of Saussurea laniceps are ground and crushed. The obtained extract is centrifuged and / or filtered, and then dried. The extraction method is maceration extraction.

[0018] The callus described in the present invention includes the newly formed tissues after wounding stimulation and / or the subculture of the newly formed tissues.

[0019] The present invention induces the callus of Saussurea laniceps by the following method: taking the root, stem or leaf of Saussurea laniceps, cutting it and inoculating it on an induction medium. The induction medium is an MS medium containing plant growth hormones. Preferably, the induction medium further contains 20-40 g / L sucrose. The plant growth hormones are at least one of 6-BA and / or NAA. Preferably, the content of 6-BA is 0.4-0.6 mg / L, and the content of NAA is 0.2-0.4 mg / L.

[0020] The culture method of the callus of Saussurea laniceps includes the following steps: taking the callus of Saussurea laniceps and culturing it in a subculture medium for 10-35 days. The subculture medium is based on the MS medium and also contains plant growth hormones. Preferably, the subculture medium further contains 20-40 g / L sucrose and / or 20-40 g / L potato powder. Preferably, the plant growth hormones are at least one of 6-BA and / or NAA. Preferably, the content of 6-BA is 0.4-0.6 mg / L, and the content of NAA is 0.2-0.4 mg / L.

[0021] The present invention also provides the application of the extract of Saussurea laniceps in the preparation of products containing niacinamide or products reducing niacin stimulation. The products include additives, cosmetics, health products, foods or drugs.

[0022] In an embodiment of the present invention, when the mass ratio of the extract of Saussurea laniceps to niacin is 1:10, the irritation of niacin can be effectively reduced, especially the irritation to the skin. The present invention provides at least one of the following products with the function of inhibiting niacin, containing the extract of Saussurea laniceps:

[0023] (1) Additives;

[0024] (2) Cosmetics;

[0025] (3) Health products;

[0026] (4) Food;

[0027] (5) Medicine.

[0028] The Saussurea laniceps is the whole plant of Saussurea laniceps, and / or normal tissue, and / or callus. The normal tissue and / or callus of Saussurea laniceps is derived from at least one of root, stem, leaf, flower, fruit, and seed, preferably at least one of root, stem, and leaf; preferably the callus of Saussurea laniceps.

[0029] The extract of Saussurea laniceps is the extract of the whole plant of Saussurea laniceps, and / or normal tissue, and / or callus, further the water extract and / or alcohol extract of the whole plant of Saussurea laniceps, and / or normal tissue, and / or callus, further the water extract of the whole plant of Saussurea laniceps, and / or normal tissue, and / or callus, and more preferably the water extract of the callus of Saussurea laniceps.

[0030] The preparation method of the extract of Saussurea laniceps includes: extracting the whole plant of Saussurea laniceps, and / or normal tissue, and / or callus with ethanol and / or water. The ethanol is preferably 60%-80% ethanol, more preferably 65%-75% ethanol. Further, the whole plant of Saussurea laniceps, and / or normal tissue, and / or callus is extracted with water; more preferably, the callus of Saussurea laniceps is extracted with water. Before extraction, the ethanol and / or water mixed with the whole plant of Saussurea laniceps, and / or normal tissue, and / or callus is ground and crushed. The obtained extract is centrifuged and / or filtered, and then dried. The extraction method is maceration extraction.

[0031] The callus described in the present invention includes the newly formed tissue after wounding stimulation and / or the subculture of the newly formed tissue.

[0032] The present invention induces the callus of Saussurea laniceps by the following method: taking the root, stem or leaf of Saussurea laniceps, cutting it and inoculating it on an induction medium. The induction medium is an MS medium containing plant growth hormones. Preferably, the induction medium further contains 20-40 g / L sucrose. The plant growth hormones are at least one of 6-BA and / or NAA. Preferably, the content of 6-BA is 0.4-0.6 mg / L, and the content of NAA is 0.2-0.4 mg / L.

[0033] The method for culturing Saussurea laniceps callus comprises the following steps: taking Saussurea laniceps callus and culturing it in a subculture medium for 10 - 35 days. The subculture medium is based on MS medium and also contains plant growth hormones. Preferably, the subculture medium further contains 20 - 40 g / L sucrose and / or 20 - 40 g / L potato powder. Preferably, the plant growth hormone is at least one of 6 - BA and / or NAA. Preferably, the content of 6 - BA is 0.4 - 0.6 mg / L and the content of NAA is 0.2 - 0.4 mg / L.

[0034] The present invention also provides a composition containing niacinamide. This composition contains Saussurea laniceps extract as a protective agent, thereby being able to inhibit the production of nicotinic acid. The mass ratio of Saussurea laniceps extract to niacinamide is at least 0.0002:1, preferably at least 0.0005:1, preferably at least 0.001:1, preferably at least 0.005:1, more preferably at least 0.01:1, further preferably at least 0.02:1, further preferably at least 0.1:1, further preferably at least 0.2:1, still further preferably at least 0.3:1; as an embodiment, it is preferably 0.0005 - 0.4:1, more preferably 0.001 - 0.35:1, more preferably 0.005 - 0.35:1, more preferably 0.01 - 0.35:1, more preferably 0.02 - 0.35:1, further preferably 0.1 - 0.35:1, still further preferably 0.2 - 0.35:1; the concentration of Saussurea laniceps extract is 0.01 - 0.5%, and in one embodiment of the present invention, it is 0.1%. The composition is at least one of the following products:

[0035] (1) Additive;

[0036] (2) Cosmetic;

[0037] (3) Health care product;

[0038] (4) Food;

[0039] (5) Medicine.

[0040] The Saussurea laniceps is the whole plant of Saussurea laniceps, and / or normal tissue, and / or callus. The normal tissue and / or callus of Saussurea laniceps is derived from at least one of root, stem, leaf, flower, fruit, and seed, preferably at least one of root, stem, and leaf; preferably it is Saussurea laniceps callus.

[0041] The Saussurea laniceps extract is an extract of the whole plant of Saussurea laniceps, and / or normal tissues, and / or callus, further an aqueous extract and / or alcohol extract of the whole plant of Saussurea laniceps, and / or normal tissues, and / or callus, further an aqueous extract of the whole plant of Saussurea laniceps, and / or normal tissues, and / or callus, and still further an aqueous extract of the callus of Saussurea laniceps.

[0042] The method for preparing the Saussurea laniceps extract includes: extracting the whole plant of Saussurea laniceps, and / or normal tissues, and / or callus with ethanol and / or water. The ethanol is preferably 60%-80% ethanol, and further preferably 65%-75% ethanol. Further, the whole plant of Saussurea laniceps, and / or normal tissues, and / or callus is extracted with water; still further, the callus of Saussurea laniceps is extracted with water. Before extraction, the ethanol and / or water mixed with the whole plant of Saussurea laniceps, and / or normal tissues, and / or callus is ground and pulverized. The obtained extract is centrifuged and / or filtered, and then dried. The extraction method is maceration extraction.

[0043] The callus described in the present invention includes newly formed tissues after trauma stimulation and / or subculture of newly formed tissues.

[0044] The present invention induces callus of Saussurea laniceps by the following method: taking the root, stem or leaf of Saussurea laniceps, cutting it and inoculating it on an induction medium. The induction medium is an MS medium containing plant growth hormones. Preferably, the induction medium further contains 20-40 g / L sucrose. The plant growth hormone is at least one of 6-BA and / or NAA. Preferably, the content of 6-BA is 0.4-0.6 mg / L, and the content of NAA is 0.2-0.4 mg / L.

[0045] The culture method of the callus of Saussurea laniceps includes the following steps: taking the callus of Saussurea laniceps and culturing it in a subculture medium for 10-35 days. The subculture medium is based on the MS medium and also contains plant growth hormones. Preferably, the subculture medium further contains 20-40 g / L sucrose and / or 20-40 g / L potato powder. Preferably, the plant growth hormone is at least one of 6-BA and / or NAA. Preferably, the content of 6-BA is 0.4-0.6 mg / L, and the content of NAA is 0.2-0.4 mg / L.

[0046] The present invention also provides an operation method for removing or inhibiting nicotinic acid, and the steps include: adding the Saussurea laniceps extract, or the above product with the function of inhibiting nicotinic acid, or the above composition for inhibiting nicotinic acid to a sample.

[0047] If the sample contains nicotinic acid, the mass ratio of the extract of Saussurea laniceps to nicotinic acid is at least 0.02:1, preferably at least 0.05:1, preferably at least 0.1:1, preferably at least 0.5:1, preferably 0.05 - 40:1, more preferably 0.1 - 35:1, more preferably 0.5 - 35:1; further preferably 1 - 35:1, more preferably 2 - 20:1; the concentration of the extract of Saussurea laniceps is 0.01 - 0.5 mg / mL, and in one embodiment of the present invention, it is 0.1 mg / mL.

[0048] The Saussurea laniceps is the whole plant of Saussurea laniceps, and / or normal tissue, and / or callus. The normal tissue and / or callus of Saussurea laniceps is derived from at least one of roots, stems, leaves, flowers, fruits, and seeds, preferably at least one of roots, stems, and leaves; preferably the callus of Saussurea laniceps.

[0049] The extract of Saussurea laniceps is the extract of the whole plant of Saussurea laniceps, and / or normal tissue, and / or callus, further the water extract and / or alcohol extract of the whole plant of Saussurea laniceps, and / or normal tissue, and / or callus, further the water extract of the whole plant of Saussurea laniceps, and / or normal tissue, and / or callus, and still further the water extract of the callus of Saussurea laniceps.

[0050] The preparation method of the extract of Saussurea laniceps includes: extracting the whole plant of Saussurea laniceps, and / or normal tissue, and / or callus with ethanol and / or water. The ethanol is preferably 60% - 80% ethanol, more preferably 65% - 75% ethanol. Further, extracting the whole plant of Saussurea laniceps, and / or normal tissue, and / or callus with water; still further, extracting the callus of Saussurea laniceps with water. Before extraction, the ethanol and / or water mixed with the whole plant of Saussurea laniceps, and / or normal tissue, and / or callus is ground and crushed. The obtained extract is centrifuged and / or filtered, and then dried. The extraction method is maceration extraction.

[0051] The callus in the present invention includes the newly formed tissue after wounding stimulation and / or the subculture of the newly formed tissue.

[0052] The present invention induces the callus of Saussurea laniceps by the following method: taking the roots, stems or leaves of Saussurea laniceps, cutting them and inoculating them on an induction medium. The induction medium is an MS medium containing plant growth hormones. Preferably, the induction medium further contains 20 - 40 g / L sucrose. The plant growth hormones are at least one of 6 - BA and / or NAA. Preferably, the content of 6 - BA is 0.4 - 0.6 mg / L, and the content of NAA is 0.2 - 0.4 mg / L.

[0053] The culture method of Saussurea laniceps callus comprises the following steps: taking the Saussurea laniceps callus and culturing it in a subculture medium for 10 - 35 days. The subculture medium is based on the MS medium and also contains plant growth hormones. Preferably, the subculture medium further contains 20 - 40 g / L sucrose and / or 20 - 40 g / L potato powder. Preferably, the plant growth hormone is at least one of 6 - BA and / or NAA. Preferably, the content of 6 - BA is 0.4 - 0.6 mg / L, and the content of NAA is 0.2 - 0.4 mg / L.

[0054] The present invention also provides a method for detecting nicotinic acid, and the steps include: performing liquid chromatography - mass spectrometry detection on a sample. The chromatographic conditions are as follows: using a C18 chromatographic column, and the elution program: mobile phase A: water - formic acid (100:0.1), mobile phase B: acetonitrile, A:B = 90:10, isocratic elution for 10 min; the mass spectrometry conditions are as follows: using a triple quadrupole mass detector, MRM mode, negative ion mode (-), electrospray ionization source (ESI), drying gas temperature 220 °C, drying gas flow rate 10 L / min, sheath gas temperature 350 °C, sheath gas flow rate 11 L / min, nebulizer pressure 35 psi, capillary voltage 3500 V (-).

[0055] The products of the present invention with the function of inhibiting nicotinic acid include, but are not limited to, additives, cosmetics, health products, foods, drugs, etc., and the presentation states include, but are not limited to, liquid, solid, oily, powdery, milky, pasty, creamy, gel - like, aerosol - like, powder - mist - like, etc.

[0056] The cosmetics described in the present invention refer to daily chemical industrial products that are applied to the human body surface such as the skin, hair, nails, and lips by means of rubbing, spraying, or other similar methods for the purposes of cleaning, protecting, beautifying, and modifying. When used as cosmetics, they can be classified into cleansing cosmetics (used to wash the skin), skin care cosmetics (such as cleansing creams, facial cleansers, bath agents, hair shampoos and conditioners, shaving creams, etc.), basic cosmetics (basic treatments for the face and hair before makeup, such as various creams, lotions, makeup waters, facial masks, hair creams, hair gels, etc. styling products), beauty cosmetics (products for beautifying the face and hair, such as rouge, lipsticks, eyeshadows, hair dyeing, perming, hairstyling, fixing products, etc.), and therapeutic cosmetics (daily chemical products between drugs and cosmetics, such as cooling, deodorizing, freckle-removing, sunscreen, body shaping, hair growth, hair removal, hair dyeing, insect repellent, olive essence, etc.). According to their external forms, they can be divided into liquids (such as facial cleansers, bath liquids, hair shampoos, makeup waters, perfumes, facial cleansing lotions, makeup removers, essence liquids, stock solutions, etc.), lotions (such as lotions, milks, hair conditioners, essence lotions), creams and ointments (such as moisturizing creams, foundation creams, hair creams, concealers, hair straightening creams, essence creams, makeup primers), powders (such as face powders, talcum powders, loose powders, facial cleansing powders, rice powders), blocks (such as powder compacts, blush powders, lipsticks, hair waxes), and oils (such as makeup remover oils, skin moisturizing oils, hair conditioning oils, essence oils).

[0057] The health products described in the present invention can be prepared into powders, tablets, capsules, liquid preparations, gels, etc., and can also be prepared into tea bags, beverages, candies, etc.

[0058] The drugs described in the present invention can be classified according to the dispersion system into solution type (a homogeneous dispersion system in which the drug is dispersed in the dispersion medium in the form of molecules or ions (the diameter of the particles is less than 1 nm), also known as low molecular solution, such as aromatic waters, solutions, syrups, glycerins, spirits, injections, etc.), colloidal solution type (a homogeneous dispersion system mainly formed by macromolecules (the diameter of the particles is 1 - 100 nm) dispersed in the dispersion medium, also known as macromolecular solution, such as mucilage, collodion, film-forming agents, etc.), emulsion type (a non-homogeneous dispersion system in which oily drugs or drug oil solutions are dispersed in the dispersion medium in the form of droplets, such as oral emulsions, intravenous emulsions, some liniments, etc. Suspension type: a non-homogeneous dispersion system in which solid drugs are dispersed in the dispersion medium in the form of fine particles, such as mixtures, lotions, suspensions, etc.), gas dispersion type (a dispersion system in which liquid or solid drugs are dispersed in a gas dispersion medium in the form of fine particles, such as aerosols), particulate dispersion type (the drug is dispersed in a liquid or solid state in the form of different-sized particles, such as microsphere preparations, microcapsule preparations, nanocapsule preparations, etc.), and solid dispersion type (a dispersion system in which solid drugs exist in the form of aggregates, such as tablets, powders, granules, capsules, pills, etc.).

[0059] The advantages of the present invention are as follows: The present invention discovers that the callus extract of Saussurea laniceps can effectively transform and remove nicotinic acid, inhibit or alleviate the irritation of nicotinic acid, slow down or inhibit the side effects of by-product nicotinic acid while realizing the efficacy of products (especially products containing niacinamide), and can inhibit the generation of nicotinic acid in products containing niacinamide, improve the product quality and stability, provide a new path for the effective development and utilization of products for skin care and beauty (especially products containing niacinamide), realize the maximization of the functions of limited resources, and have broad application prospects. Description of the Drawings

[0060] Figure 1 When performing liquid chromatography-mass spectrometry detection on the nicotinic acid group (nicotinic acid - 0.1 mg), the callus extract group of Saussurea laniceps (Saussurea laniceps - 1 mg), and the combination group of nicotinic acid and the callus extract of Saussurea laniceps (nicotinic acid - 0.1 mg + Saussurea laniceps - 1 mg), the chromatogram of nicotinic acid ion (m / z = 122, M 烟酸 -H + ) was extracted in the negative ion mode, and the chromatographic peak of nicotinic acid ion appeared at about 1.738 min.

[0061] Figure 2 The mass spectrometry peak diagram in the range of 1.589 - 1.924 min for the nicotinic acid group (nicotinic acid - 0.1 mg) is shown. It can be seen that the mass spectrometry peak of nicotinic acid (m / z = 122.1, M 烟酸 -H + ) appears.

[0062] Figure 3 When performing liquid chromatography-mass spectrometry detection on the callus extract group of Saussurea laniceps (Sample 1, 0.5% callus extract of Saussurea laniceps), the niacinamide group (Sample 2, 1% niacinamide), and the combination group of niacinamide and the callus extract of Saussurea laniceps (Sample 3, 0.5% callus extract of Saussurea laniceps + 1% niacinamide), the chromatogram of niacinamide ion (m / z = 123, M 烟酰胺 +H + ) was extracted in the positive ion mode, and the chromatographic peak of niacinamide ion appeared at about 1.746 min.

[0063] Figure 4 The mass spectrometry peak diagram in the range of 1.667 - 1.879 min for Sample 2 (1% niacinamide) is shown. It can be seen that the mass spectrometry peak of niacinamide (m / z = 123, M 烟酰胺 +H + ) appears.

[0064] Figure 5When performing liquid chromatography - mass spectrometry (LC - MS) detection on the Saussurea laniceps callus extract group (Sample 1, 0.5% Saussurea laniceps callus extract), the nicotinamide group (Sample 2, 1% nicotinamide), and the combination group of nicotinamide and Saussurea laniceps callus extract (Sample 3, 0.5% Saussurea laniceps callus extract + 1% nicotinamide), the chromatogram peak of nicotinic acid ion (m / z = 122, M 烟酸 -H + ) was extracted under the negative ion mode.

[0065] Figure 6 Figure 2 is the mass spectrometry peak diagram of Sample 2 (1% nicotinamide) in the range of 1.718 - 1.914 min. It can be seen that the mass spectrometry peak diagram of nicotinic acid (m / z = 122.1, M 烟酸 -H + ) is shown. Detailed Implementation Modes

[0066] The present invention will be described in detail below with reference to the accompanying drawings and specific embodiments. This embodiment is implemented on the premise of the technical solution of the present invention, and detailed implementation methods and specific operation processes are given. However, the protection scope of the present invention is not limited to the following embodiments.

[0067] I. Preparation of Saussurea laniceps Callus Extract

[0068] 1. Induction and Sub - culture of Saussurea laniceps Plant Callus

[0069] In a sterile laminar flow hood, use a scalpel to cut the roots, stems or leaves of Saussurea laniceps into segments with a length of 0.5 cm - 1.0 cm, and inoculate them into the culture medium formula of MS + 6 - BA 0.5 mg / L + NAA 0.3 mg / L + sucrose 30 g / L + gellan gum 3.5 g / L to induce callus.

[0070] Transfer a part of the Saussurea laniceps callus to the culture medium formula of MS + 6 - BA 0.5 mg / L + NAA 0.5 mg / L + potato powder 30 g / L + sucrose 30 g / L + gellan gum 3.5 g / L for sub - culture. After 25 - 30 days, harvest the Saussurea laniceps callus, denoted as "Saussurea laniceps callus", and store it frozen for later use.

[0071] 2. Water Extract of Saussurea laniceps Plant Callus

[0072] Take the frozen - stored Saussurea laniceps callus for thawing, add deionized water, then grind and homogenize it for 10 - 20 min, and extract for 3 hours. Then centrifuge the extracted liquid at 8000 rpm - 10000 rpm for 40 min - 60 min. After centrifugation, take the supernatant, filter it through a 0.45 uM filter membrane, and take the filtrate. Freeze - dry the filtrate at low temperature for 10 - 14 days, and the obtained powder is the Saussurea laniceps callus extract.

[0073] II. Suppression of nicotinic acid in nicotinamide by the callus extract of Saussurea laniceps

[0074] 1. Instruments and reagents

[0075] Triple quadrupole liquid chromatography - mass spectrometry (Agilent G6470B), UPLC column (Agilent ZORBAX RRHDEclipse Plus C18: column length 100 mm, diameter 3.0 mm, particle size 1.8 μm), electronic balance (Mettler XS105DU), acetonitrile (Aladdin LCMS grade), formic acid (Aladdin HPLC grade), ultrapure water.

[0076] 2. Information of reference substances and samples

[0077] Nicotinic acid: Anpel, batch number 20210118, content 99.8%.

[0078] Cultures of Saussurea involucrata: Purchased from Dalian Prukang Biotechnology Co., Ltd., extract of cultures of Saussurea involucrata.

[0079] Callus extract of Saussurea laniceps: The preparation method is shown in "I. Preparation of the callus extract of Saussurea laniceps".

[0080] 3. Analytical method

[0081] Using a triple quadrupole liquid chromatography - mass spectrometry (LC - MS / MS),

[0082] Chromatographic conditions:

[0083] Chromatographic column: Agilent ZORBAX RRHD Eclipse Plus C18: column length 100 mm, diameter 3.0 mm, particle size 1.8 μm;

[0084] Mobile phase: Mobile phase A: water - formic acid (100:0.1), mobile phase B: acetonitrile;

[0085] Flow rate: 0.4 mL per minute;

[0086] Column temperature: 30 °C;

[0087] Injection volume: 2 μL;

[0088] Elution program: Mobile phase A:B = 90:10, isocratic elution for 10 min;

[0089] Detector: Triple quadrupole mass detector;

[0090] Ion source: Electrospray ionization source (ESI);

[0091] Mass spectrometry parameters: drying gas temperature: 220 °C, drying gas flow rate: 10 L / min, sheath gas temperature: 350 °C,

[0092] sheath gas flow rate: 11 L / min, nebulizer pressure: 35 psi, capillary voltage: 3500 V (-);

[0093] Triple quadrupole liquid chromatography-mass spectrometry (LC-MS / MS), MRM mode, negative ion mode (-), To MS

[0094]

[0095] Using water as a diluent, prepare the test solution, nicotinic acid standard stock solution, test sample and nicotinic acid composition solution. The operation process is as follows. The amounts taken during the preparation process can be adjusted according to the actual experimental needs and are not limited to the specific values in the following operations:

[0096] Test solution: Take about 10 mg of the test sample, accurately weigh it, place it in a 10 mL volumetric flask, and dilute it to the mark with the diluent; then take 1 mL and place it in a 10 mL volumetric flask, and dilute it to the mark with the diluent. Its concentration is 0.1 mg / mL. Transfer out the solution as the test solution.

[0097] Nicotinic acid standard stock solution: Accurately weigh 10 mg of nicotinic acid standard in a 10 mL volumetric flask, dilute it to the mark with the diluent, and its concentration is 1 mg / mL; transfer out the solution as the standard stock solution.

[0098] Test sample and nicotinic acid composition solution (10:1): Take 1 mL of the test sample stock solution (1 mg / mL) and 100 μL of the nicotinic acid standard stock solution, place them in the same 10 mL volumetric flask, and dilute it to the mark with the diluent. Obtain a composition solution with a mass ratio of test sample to nicotinic acid of 10:1. In other proportion compositions, the final concentration of the test sample remains unchanged at 0.1 mg / mL, and the concentration of nicotinic acid is diluted proportionally in turn, and so on.

[0099] 4. Linearity and range

[0100] 4.1 Solution preparation

[0101] Accurately weigh the nicotinic acid standard and prepare a series of standard curve working solutions respectively. Using the mass concentration of nicotinic acid as the abscissa (X) and the peak area response value result as the ordinate (Y), perform linear regression.

[0102] 4.2 Experimental results

[0103] Precisely measure 2 μL of each linear test solution for injection analysis. Starting from the limit of quantitation, calculate the linear relationship between the peak area and the concentration. The linear regression equation is y = 0.240578x + 28557.441013, R 2= 0.9986, see Table 1.

[0104] Table 1. Results of the linearity and range test of nicotinic acid

[0105] Theoretical concentration (ng / mL) 250 500 1000 2000 10000 10000000 Peak area 3149 6288 11743 22405 102576 2434273

[0106] 4.3 Inhibitory rate of Saussurea laniceps callus extraction on nicotinic acid at different mass ratios

[0107] As Figure 1-2 shown, in the nicotinic acid group (03-YS-0.1mg, 0.1mg), the Saussurea laniceps callus extract group (04-Saussurea laniceps-1mg, 1mg), and the group of nicotinic acid and Saussurea laniceps callus extract (05-YS-0.1mg + Saussurea laniceps-1mg, nicotinic acid 0.1mg + Saussurea laniceps callus extract 1mg), as Figure 1 and Figure 2 shown, only the nicotinic acid group has chromatographic peaks and mass spectrometry peaks (m / z = 122, M 烟酸 -H + ), indicating that the Saussurea laniceps callus has a transformation, removal, and inhibitory effect on nicotinic acid.

[0108] In the combined solution formed by mixing the Saussurea laniceps callus extract and nicotinic acid, the final mass concentration of the Saussurea laniceps callus extract is 0.1 mg / mL, and the mass ratio of the Saussurea laniceps callus extract to nicotinic acid is shown in Table 2.

[0109] Table 2. Nicotinic acid inhibitory rate at different mass ratios of Saussurea laniceps callus extract to nicotinic acid

[0110]

[0111] 4.4 Saussurea laniceps callus can relieve the irritation caused by nicotinic acid

[0112] Nicotinic acid can cause a certain degree of irritation. Ten subjects were selected to evaluate the use of a single nicotinic acid aqueous solution (Comparative Example 2), a nicotinic acid aqueous solution added with Saussurea laniceps callus extract (Example 1), and the Saussurea laniceps callus extract alone (Comparative Example 1). The formulation of each group is shown in Table 3, and the irritation degree after evaluation is shown in Table 4.

[0113] Table 3. Experimental grouping list and mass ratio

[0114] Example 1 Comparative Example 1 Comparative Example 2 Extract of Saussurea laniceps Hand.-Mazz. callus 0.01 0.01 0 Nicotinic acid 0.1 0 0.1 Deionized water 99.89 99.99 99.9

[0115] Table 4. Skin feel test results

[0116] Subjects Example 1 Comparative Example 1 Comparative Example 2 1 The stinging sensation is weakened and there is a slight cool feeling No discomfort Stinging starts after about 10 seconds 2 No discomfort No discomfort It feels very cool and a bit stimulating after application 3 No discomfort No discomfort There is a slight stinging and heat sensation after application 4 No discomfort No discomfort It feels very cool and slightly stimulating after application 5 No discomfort No discomfort It feels very cool and slightly stimulating after application 6 There is a soothing feeling No discomfort There is stinging 7 No discomfort No discomfort There is stinging, itching and skin redness 8 The stinging sensation is weakened and there is a cool feeling No discomfort There is a slight stinging 9 No discomfort No discomfort There is a heat sensation 10 There is a continuous cool feeling No discomfort There is stinging

[0117] As can be seen from the test results in Table 4, the subjects using Comparative Example 2 had strong stinging and discomfort, manifested as stinging, heat sensation and itching, while there was no obvious feeling in Comparative Example 1. At the same time, Example 1 with the addition of the callus extract of Saussurea gossypiphora and niacin was significantly milder compared to Comparative Example 2. Two subjects indicated that the stinging sensation was weakened with a bit of cool feeling. Seven subjects had no obvious discomfort or a slight sense of relief. The remaining one subject showed that after adding the callus of Saussurea gossypiphora, there was some cool feeling compared to the stinging sensation of the pure niacin group. In summary, the callus of Saussurea gossypiphora can significantly reduce the irritation caused by niacin.

[0118] 4.5 Inhibitory effect of the callus extract of Saussurea gossypiphora on the production of nicotinic acid from nicotinamide

[0119] Sample 1: 0.5% callus extract of Saussurea gossypiphora (5 mg / mL).

[0120] Sample 2: 1% nicotinamide (10 mg / mL), stored at 4 °C. The content of nicotinic acid detected in the nicotinamide sample: 0.96%.

[0121] Sample 3: 0.5% callus extract of Saussurea gossypiphora (5 mg / mL) + 1% nicotinamide (10 mg / mL).

[0122] Method: The same as "3. Analytical method".

[0123] The LC-MS / MS spectra of Sample 1, Sample 2 and Sample 3 are as Figure 3 and Figure 5 shown. Extract m / z = 123 (M 烟酰胺 +H + ). Nicotinamide is contained in Sample 2 and Sample 3, and the peak areas are quite similar. The content of nicotinamide does not change after mixing with the callus extract of Saussurea gossypiphora (see Figure 3 ). Extract m / z = 122 (M 烟酸 -H + ). Only nicotinic acid is contained in Sample 2 (see Figure 5 ), and there is no nicotinic acid in Sample 1 and Sample 3. It shows that after mixing the callus extract of Saussurea gossypiphora and nicotinamide, the nicotinic acid in nicotinamide is converted and removed (see Figure 5 ).

[0124] No nicotinic acid was detected in Sample 3 after it was placed in a transparent container at room temperature for 14 days.

[0125] 4.6 Inhibitory effect of the extract of Saussurea involucrata culture on nicotinic acid

[0126] In the combined solution obtained by mixing the extract of the Saussurea involucrata culture of Purecare with nicotinic acid, the mass ratio of the Saussurea involucrata culture extract to nicotinic acid is as shown in Table 5, and the final concentration of the Saussurea involucrata culture in the combined solution is 0.1 mg / mL. The nicotinic acid inhibition rate of the commercial Saussurea involucrata culture was evaluated, and the results are shown in Table 5.

[0127] Table 5. Nicotinic acid inhibition rate of other commercial Saussurea involucrata

[0128]

[0129] As can be seen from the results in Table 5, both the extract of the callus of Saussurea laniceps and the Saussurea involucrata culture of Purecare have a certain effect of inhibiting nicotinic acid, but the inhibitory effect of the callus of Saussurea laniceps on nicotinic acid is significantly better.

Claims

1. Use of extracts of Saussurea laniceps in the preparation of at least one of the following products having a function of inhibiting nicotinic acid: (1) Additives; (2) Cosmetics; (3) Health products; (4) Foods; (5) Pharmaceuticals.

2. Use of extracts of Saussurea laniceps in the preparation of products containing nicotinamide or products reducing nicotinic acid irritation.

3. At least one of the following products having a function of inhibiting niacin, characterized in that, Containing extracts of Saussurea laniceps: (1) Additives; (2) Cosmetics; (3) Health products; (4) Foods; (5) Pharmaceuticals.

4. The product according to claim 3, characterized in that, The extracts of Saussurea laniceps are extracts of the whole plant of Saussurea laniceps, and / or normal tissues, and / or callus, and the normal tissues and / or callus are derived from at least one of roots, stems, leaves, flowers, fruits, and seeds.

5. The product according to claim 4, characterized in that, The preparation method of the extracts of Saussurea laniceps includes extracting the whole plant of Saussurea laniceps, and / or normal tissues, and / or callus with ethanol and / or water.

6. The product according to claim 5, wherein, The induction method of the callus includes: taking the roots, stems or leaves of Saussurea laniceps, cutting them and inoculating them on an induction medium; The culture method of the callus includes: culturing the callus of Saussurea laniceps in a subculture medium for 10 - 35 days.

7. A composition containing niacinamide, characterized in that, Containing extracts of Saussurea laniceps, wherein the mass ratio of the extracts of Saussurea laniceps to nicotinamide is at least 0.0002:

1.

8. Use of extracts of Saussurea laniceps in removing or inhibiting nicotinic acid.

9. A method for suppressing or removing nicotinic acid, characterized in that, The steps include: Adding extracts of Saussurea laniceps, or the product according to any one of claims 3 - 6, or the composition according to claim 7 to the sample.

10. The method according to claim 9, characterized in that If the sample contains nicotinic acid, the mass ratio of the extracts of Saussurea laniceps to nicotinic acid is at least 0.02:

1.

11. The method according to claim 7 or 8, characterized in that The concentration of the extracts of Saussurea laniceps in the sample is 0.01 - 0.5 mg / mL.

12. A method for detecting nicotinic acid, the steps comprising: Performing liquid chromatography - mass spectrometry detection on the sample. The chromatographic conditions are: using a C18 chromatographic column, elution program: mobile phase A: water - formic acid (100:0.1), mobile phase B: acetonitrile, A:B = 90:10, isocratic elution for 10 min; the mass spectrometry conditions are: using a triple quadrupole mass detector, MRM mode, negative ion mode (-), electrospray ionization source (ESI), drying gas temperature 220°C, drying gas flow rate 10 L / min, sheath gas temperature 350°C, sheath gas flow rate 11 L / min, nebulizer pressure 35 psi, capillary voltage 3500 V (-).

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