CN108514123A - The active peptides extracted from blood clam prepare compound method - Google Patents

The active peptides extracted from blood clam prepare compound method Download PDF

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Publication number
CN108514123A
CN108514123A CN201810169499.7A CN201810169499A CN108514123A CN 108514123 A CN108514123 A CN 108514123A CN 201810169499 A CN201810169499 A CN 201810169499A CN 108514123 A CN108514123 A CN 108514123A
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parts
active peptides
blood clam
added
compound
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陈雨诗
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Panan Send Biotechnology Co Ltd
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    • AHUMAN NECESSITIES
    • A23FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23LFOODS, 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
    • A23L33/00Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
    • A23L33/10Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
    • A23L33/17Amino acids, peptides or proteins
    • A23L33/18Peptides; Protein hydrolysates
    • AHUMAN NECESSITIES
    • A23FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23LFOODS, 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
    • A23L2/00Non-alcoholic beverages; Dry compositions or concentrates therefor; Their preparation
    • A23L2/52Adding ingredients
    • AHUMAN NECESSITIES
    • A23FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23VINDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
    • A23V2002/00Food compositions, function of food ingredients or processes for food or foodstuffs

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  • Life Sciences & Earth Sciences (AREA)
  • Health & Medical Sciences (AREA)
  • Nutrition Science (AREA)
  • Chemical & Material Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Food Science & Technology (AREA)
  • Polymers & Plastics (AREA)
  • Molecular Biology (AREA)
  • Proteomics, Peptides & Aminoacids (AREA)
  • Mycology (AREA)
  • Medicines Containing Material From Animals Or Micro-Organisms (AREA)

Abstract

The present invention discloses a kind of active peptides extracted from blood clam and prepares compound method, includes the extraction of blood clam active peptides, the preparation process of the preparation of compound, cooling preservation, compound is:The salt wormwood artemisia powder and cellulase after crushing are taken by weight, distilled water is added and adjusts pH value to 5.5 ~ 6.0, 30 35min are digested at 45 ~ 50 DEG C, filter to take enzymolysis liquid, absolute ethyl alcohol is added and antioxidant stirs 20 25min, heat drying obtains medicinal extract, it is spare, by milk, vitamin A, white sugar, grape juice, honey, blood clam active peptides and lemon juice dissolve to obtain base fluid, the water that medicinal extract is added to 2 2.3 times of volumes dissolves by heating and the base fluid of medicinal extract quality 45 ~ 48% is added, obtain compound, the compound of preparation has unique flavor, it is beneficial to oral health, nutriment is not oxidizable after the process of preservation and corkage, have the function of pre- preventing tumor and adjusts human immunity.

Description

The active peptides extracted from blood clam prepare compound method
Technical field
The invention belongs to technical field of beverage preparation, and in particular to a kind of active peptides from blood clam extraction prepare compound Method.
Background technology
Blood clam (Arca subcrenata Lischke), Mollusca, lamellibranchiata, blood clam mesh, blood clam section, blood clam category are main It is coastal to be distributed in China, Korea and Japan.The nutrition of blood clam and medical value are higher, and shell and meat all have medical value.In The effect of doctor thinks that its is warm-natured sweet in flavor, returns spleen, Liver Channel, the stomach invigorating of tool middle benefit gas, nourishing yin and supplementing blood, clinically cures mainly anaemia, physically weak etc., hair Blood clam polypeptide extract can significantly inhibit the proliferation of more plants of tumour cells, but the extraction to blood clam active peptides in the prior art Rate is not high and its application field is not extensive.
Beverage is the important consumer goods of people, and currently on the market, drink quantity and beverage types are rich and varied, each to have Characteristic.However, in addition to packing drinking water, Other Drinks, such as soda, fruit-vegetable juice beverage, tea beverage, usual sugar content It is higher.Studies have shown that the high food containing sugared, especially high containing sucrose of long-term consumption, having become the diseases such as induced Diabetic, obesity The cause of disease of disease.
The prior art provides more health drink, such as Chinese invention granted patent document, Authorization Notice No. CN 103054035 B, the invention are related to a kind of health drink, disclose a kind of throat cooling beverage and preparation method, the beverage is with Pu public affairs The raw materials such as English extract, chrysanthemum extract, peppermint oil, sugar, sugar alcohol, non-sugar sweetener, acid pass through allotment, hot canning, height The processing technologys such as warm instantaneous sterilizing and obtain.The invention can reduce usage amount sugared in beverage, and by reasonable compatibility, respectively Kind dispensing effect is complementary, plays the functions such as the good relieving sore-throat that wets one's whistle, the preventing decayed tooth that cleans the teeth to cooperate with, but the beverage can after corkage Can be in a short time in beverage nutriment can aoxidize and reduce its health-care efficacy.
Invention content
The purpose of the present invention is to provide a kind of active peptides extracted from blood clam to prepare compound method, the compounding of preparation Object has unique flavor, is beneficial to oral health, and nutriment is not oxidizable after the process of preservation and corkage, has and prevents to swell Tumor and the effect for adjusting human immunity.
The technical solution that the present invention is taken to achieve the above object is:The active peptides extracted from blood clam prepare compound Method, includes the extraction of blood clam active peptides, and the preparation process of the preparation of compound, cooling preservation, compound is:By weight Meter takes the salt wormwood artemisia powder and cellulase after crushing, and distilled water is added and adjusts pH value to 5.5 ~ 6.0, solid-to-liquid ratio 1g:10- 17ml digests 30-35min at 45 ~ 50 DEG C, filters to take enzymolysis liquid, and absolute ethyl alcohol is added and antioxidant stirs 20-25min, Heat drying obtains medicinal extract, spare, and milk, vitamin A, white sugar, grape juice, honey, blood clam active peptides and lemon juice are dissolved Base fluid is obtained, the water that medicinal extract is added to 2-2.3 times of volume dissolves by heating and be added the base fluid of medicinal extract quality 45 ~ 48%, obtains compound. Selecting salt wormwood artemisia to be main material makes the main mouthfeel in the compound of preparation be salt wormwood artemisia flavor and add the ingredients such as grape juice, honey Further enriching the mouthfeel of compound reduces the usage amount of white sugar simultaneously and keeps compound suitable for people of all ages, using former based on salt wormwood artemisia powder Material, which prepares gained compound, has good heat-clearing, disperse accumulation, the work(for expanding blood vessel, preventing heart disease and enhancing human immunity Effect, i.e. compound have good health-care effect, and blood clam active peptides, vitamin A, milk are added in compound preparation process Equal ingredients make its collaboration play a role improve compound it is anti-oxidant, wet one's whistle, the effect of oral health, and further promoted multiple Mouthfeel with object enhances its fragrance, while covering compound bitterness sense that may be present, mistake of the people in long-term drinking compound Cheng Zhong, compound can promote the non-activated lymphopoiesis of human body, play the effect of pre- preventing tumor.
Preferably, compound is made of following component and parts by weight:60-180 parts of salt wormwood artemisia powder, steams 1-3 parts of cellulase 500-600 parts of distilled water, 20-24 parts of absolute ethyl alcohol, 0.06-0.12 parts of antioxidant, 2-7 parts of milk, vitamin A 0.01-0.03 Part, 15-30 parts of white sugar, 27-38 parts of grape juice, 45-50 parts of honey, 3-7 parts of blood clam active peptides, 27-30 parts of lemon juice.It selects Salt wormwood artemisia, which is main material, makes the main mouthfeel in the compound of preparation be salt wormwood artemisia flavor and add the ingredients such as grape juice, honey into one The usage amount that the mouthfeel of the abundant compound of step reduces white sugar simultaneously keeps compound suitable for people of all ages, is main material system using salt wormwood artemisia powder Standby gained compound has effects that good heat-clearing, disperse accumulation, expands blood vessel, prevents heart disease and enhance human immunity, Antioxidant is added in compound can effectively extend the effective time of nutritional ingredient in compound, when extending the preservation of compound Between, when compound is opened, since the addition of antioxidant can effectively reduce the oxidation rate of compound, keep compound Effect.
Preferably, antioxidant is made of the vitamin C of 3-5 parts by weight and the mixture of 0.1-0.6 parts by weight, mixture For L- leucylglycines and D- leucylglycines, the two mass ratio is 47 ~ 53:0.5-0.8.Antioxidant can be with O2·、 HOO and OH fast reactions generate half hydroascorbic acid, remove singlet oxygen, restore Thiyl Radical, can also make oxidation VEReduced form is reverted to, plays the role of indirect antioxidant, and synergy is played to vitamin C by the mixture of addition and is made With, mixture can be passivated catalysed oxidn, while the hydrogen ion generated can be such that antioxidant regenerates with complexing of metal ion, Realize permanent oxidation resistant effect.
Preferably, the extraction step of blood clam active peptides is:Fresh blood clam decladding is taken, distilled water is added, is homogenized, removal of impurities, Filtering, obtains filtrate A, and acetic acid solution stirring extraction is then added in filtrate A, and centrifugation takes supernatant, NaCl is added in supernatant It is 2.5mol/L to solution ultimate density, while flocculent deposit is precipitated, centrifuges, obtain blood clam active peptides, pass through above method institute The purity of the blood clam active peptides obtained is high, and purity is 92% or more, and recovery rate is between 78% ~ 82.5%, the polypeptide molecular weight of extraction Between 3000 ~ 6000Da, anticancer experiment in vitro shows inhibitory rate to 57 ~ 59%, strong to body immunity adjustment effect, Especially in terms of lymphproliferation response effect significantly, non-activated lymphopoiesis can be promoted, Con can also be promoted A activates T or B lymphopoiesis.
Preferably, in distilled water D- mandelic acids and L- mandelic acids containing 0.08-0.1wt% mixture, D- hardship apricots Benevolence acid and L- mandelic acid weight ratios are 52 ~ 57:0.3 ~ 0.9, the mixing of D- mandelic acids and L- mandelic acids is added in distilled water The hydrophily of blood clam cell tissue can be improved in body, so that it is dissolved in distilled water and forms aperture under cell membrane, promotes intracellular Content leaks, realize improve blood clam active peptides yield improve.
Preferably, except miscellaneous operation is:Homogenate is used to the NaOH's and 0.1-0.15mol/L of 0.15-0.25mol/L respectively EDTA-2Na solution impregnates, and filtering obtains filtrate A, realizes removing foreign protein, pigment and mineral matter and other components, avoids mentioned component Influence to the extraction of blood clam active peptides.
Preferably, compound cooling preservation temperature is 2-5 DEG C, and the work of blood clam active peptides is effectively ensured by Cord blood Property, it is conducive to the assimilation effect of active peptides and compound remaining ingredient in human body stomach.
Compared with prior art, beneficial effects of the present invention are:Selecting salt wormwood artemisia to be main material makes in the compound of preparation Main mouthfeel is salt wormwood artemisia flavor and adds the ingredients such as grape juice, honey and further enrich the mouthfeel of compound while reducing white sugar Usage amount keep compound suitable for people of all ages, using salt wormwood artemisia powder be main material prepare gained compound have good heat-clearing, disappear The effect of accumulating, expanding blood vessel, prevent heart disease and enhance human immunity, i.e. compound have good health-care effect, multiple Its collaboration is set to play a role with the ingredients such as the very high blood clam active peptides of recovery rate, vitamin A, milk are added in object preparation process Improve compound it is anti-oxidant, wet one's whistle, the effect of oral health, and further promoted compound mouthfeel, enhance its fragrance, Compound bitterness sense that may be present is covered simultaneously, for people during long-term drinking compound, compound can promote human body Non-activated lymphopoiesis plays the effect of pre- preventing tumor.
Specific implementation mode
The present invention is described in further detail with reference to embodiments:
Embodiment 1:
The active peptides extracted from blood clam prepare compound method, include the extraction of blood clam active peptides, the preparation of compound is cold But it preserves, steps are as follows for specific preferred operations:
The extraction step of blood clam active peptides is:Fresh blood clam decladding is taken, distilled water, homogenate is added, homogenate solid-to-liquid ratio is 1: 1.3, the mixture of D- mandelic acids and L- mandelic acids containing 0.08wt%, D- mandelic acids and L- semen armeniacae amaraes in distilled water Sour weight ratio is 55:0.6, use the EDTA-2Na solution of the NaOH and 0.1mol/L of 0.17mol/L to impregnate respectively, filtering must filter Then liquid A presses solid-to-liquid ratio 1 in filtrate A:8 are added the acetic acid solution stirring extraction of 0.12mol/L, mixing time 6h, stirring Speed is 50r/min, and 10min is centrifuged under the conditions of 5000r/min, takes supernatant, and it is final to solution that NaCl is added in supernatant A concentration of 2.5mol/L, while flocculent deposit is precipitated, 9000r/min centrifuges 12min under the conditions of 3 DEG C, obtains blood clam active peptides;
The preparation process of compound is:1 part of 120 parts of salt wormwood artemisia powder and cellulase after crushing are taken by weight, are added and are steamed 540 parts of distilled water simultaneously adjusts pH value to 5.8, solid-to-liquid ratio 1g:14ml digests 32min at 48 DEG C, filters to take enzymolysis liquid, and nothing is added 0.08 part of stirring 24min of 22 parts of water-ethanol and antioxidant, heat drying obtains medicinal extract, spare, by 5 parts of milk, vitamin A 0.02 Part, 16 parts of white sugar, 28 parts of grape juice, 47 parts of honey, 4 parts of blood clam active peptides and 28 parts of lemon juice dissolve to obtain base fluid, by medicinal extract The water that 2.2 times of volumes are added dissolves by heating and is added the base fluid of medicinal extract quality 46%, obtains compound, antioxidant is by 4 parts by weight Vitamin C and the mixture of 0.4 parts by weight composition, mixture are L- leucylglycines and D- leucylglycines, the two matter Amount is than being 48:0.7;
High-temperature short-time sterilization:Bottling after compound is continued into 10s high-temperature short-time sterilizations under the conditions of 103 DEG C;
Cooling and inspection:Check that bottle has an abnormal phenomenon such as crack-free, it is labelled after by compound in 4 DEG C of Cord bloods.
Embodiment 2:
The active peptides extracted from blood clam prepare compound method, include the extraction of blood clam active peptides, the preparation of compound is cold But it preserves, concrete operation step is as follows:
The extraction step of blood clam active peptides is:Fresh blood clam decladding is taken, distilled water, homogenate is added, homogenate solid-to-liquid ratio is 1: 1.5, the mixture of D- mandelic acids and L- mandelic acids containing 0.09wt%, D- mandelic acids and L- semen armeniacae amaraes in distilled water Sour weight ratio is 52:0.7, use the EDTA-2Na solution of the NaOH and 0.13mol/L of 0.2mol/L to impregnate respectively, filtering must filter Then liquid A presses solid-to-liquid ratio 1 in filtrate A:10 are added the acetic acid solution stirring extraction of 0.13mol/L, and mixing time 7h is stirred It is 45r/min to mix speed, and 11min is centrifuged under the conditions of 5000r/min, takes supernatant, NaCl is added in supernatant to solution most Final concentration of 2.5mol/L, while flocculent deposit is precipitated, 9000r/min centrifuges 12min under the conditions of 2 DEG C, and it is more to obtain blood clam activity Peptide;
The preparation process of compound is:1 part of 75 parts of salt wormwood artemisia powder and cellulase after crushing are taken by weight, and distillation is added 540 parts of water simultaneously adjusts pH value to 5.8, solid-to-liquid ratio 1g:14ml digests 32min at 48 DEG C, filters to take enzymolysis liquid, is added anhydrous 0.09 part of stirring 22min of 22 parts of ethyl alcohol and antioxidant, heat drying obtains medicinal extract, spare, by 7 parts of milk, vitamin A 0.01 Part, 20 parts of white sugar, 30 parts of grape juice, 48 parts of honey, 4 parts of blood clam active peptides and 27 parts of lemon juice dissolve to obtain base fluid, by medicinal extract The water that 2 times of volumes are added dissolves by heating and is added the base fluid of medicinal extract quality 46%, obtains compound, antioxidant by 4 parts by weight dimension The mixture composition of raw element C and 0.4 parts by weight, mixture are L- leucylglycines and D- leucylglycines, the two quality Than being 53:0.5.
High-temperature short-time sterilization:Bottling after compound is continued into 15s high-temperature short-time sterilizations under the conditions of 103 DEG C;
Cooling and inspection:Check that bottle has an abnormal phenomenon such as crack-free, it is labelled after by compound in 4 DEG C of Cord bloods.
Embodiment 3:
The blood clam active peptides for selecting embodiment 1 to extract buy mouse lymph from Zhongshan University's Experimental Animal Center daughter cell library Tumor cell strain YAC-1 buys SPF grade Kunming mouses by Zhejiang Province's Experimental Animal Center, male, weight 18-20 g, general It is raised under logical clean environment, 17-20 DEG C of temperature, animal freely can ingest and drink water.
It is 5 × 107/mL to take mouse lymphoma cell, adjustment cell density, 20 porocyte culture plates is added, often 1 mL of hole is separately added into 1 mL of the active peptides of blood clam containing various concentration liquid per hole, makes the blood clam active peptides liquid final concentration be respectively 20,100,500 μ g/mL, and set positive and negative (5 μ g/mL Con A of final concentration and isometric cell culture fluid) and compare Hole is 2 mL per hole solution final volume, and every group is all provided with 3 multiple holes, sets 37 DEG C, 5% CO218 h, 2000 r/ are cultivated in incubator Min centrifuges 10 min, discards culture solution, and 65% pre-cooled ethanol is resuspended, mixing, is placed in 2 DEG C of fixations overnight.It is thin after fixation 2000r/min centrifuges 10 min, discards liquid, each pipe plus PI dye liquors (PI 2. 5mg, RNase 5 mg, Triton X- 100 15 μ L are dissolved in 40 mL of PBS, and filtering, 2 DEG C are kept in dark place) 250 μ L, avoid light place 18min after mixing, warp 340 mesh nylon net filters, the flow cytomery cell cycle, calculate mouse lymphoma cell enters the cell percentages of S phases Rate is 42.8%.
Embodiment 4:
The subjective appreciation of compound prepared by the active peptides extracted from blood clam:
Please all certain sensory's experiences of 10 product quality inspections lab technician to embodiment 1, embodiment 2 and commercially available health care Drink control group carries out subjective appreciation, and standards of grading are shown in Table 1, and each index is all made of ten point system, and full marks are 50 points, calculate each finger After marking score, it is added to get subjective appreciation score value after being multiplied by weighted value.Results of sensory evaluation is:Implement 1 as 48.1, embodiment 2 For 48.8, control group 44.1, it follows that compounding prepared by the active peptides of the slave blood clam of embodiment 1 and embodiment 2 extraction The subjective appreciation score of object is higher than control group, it is easier to be esthetically acceptable to the consumers.
The sensory evaluation scores standard of compound prepared by the active peptides that table 1 is extracted from blood clam
Routine operation in above-described embodiment is well known to those skilled in the art, herein without repeating.
Technical scheme of the present invention is described in detail in embodiment described above, it should be understood that the above is only For specific embodiments of the present invention, it is not intended to restrict the invention, all any modifications made in the spirit of the present invention, Supplement or similar fashion replacement etc., should all be included in the protection scope of the present invention.

Claims (7)

1. the active peptides extracted from blood clam prepare compound method, include the extraction of blood clam active peptides, the preparation of compound, Cooling preservation, it is characterised in that:The preparation process of the compound is:The salt wormwood artemisia powder and fiber after crushing are taken by weight Plain enzyme is added distilled water and adjusts pH value to 5.5 ~ 6.0, solid-to-liquid ratio 1g:10-17ml digests 30-35min at 45 ~ 50 DEG C, Enzymolysis liquid is filtered to take, absolute ethyl alcohol is added and antioxidant stirs 20-25min, heat drying obtains medicinal extract, spare, by milk, dimension Raw element A, white sugar, grape juice, honey, blood clam active peptides and lemon juice dissolve to obtain base fluid, by medicinal extract 2-2.3 times of volume of addition Water dissolves by heating and is added the base fluid of medicinal extract quality 45 ~ 48%, obtains compound.
2. the active peptides according to claim 1 extracted from blood clam prepare compound method, it is characterised in that:It is described multiple It is made of following component and parts by weight with object:60-180 parts of salt wormwood artemisia powder, 1-3 parts of cellulase, 500-600 parts of distilled water, nothing 20-24 parts of water-ethanol, 0.06-0.12 parts of antioxidant, 2-7 parts of milk, 0.01-0.03 parts of vitamin A, 15-30 parts of white sugar, Portugal 27-38 parts of grape juice, 45-50 parts of honey, 3-7 parts of blood clam active peptides, 27-30 parts of lemon juice.
3. the active peptides according to claim 1 extracted from blood clam prepare compound method, it is characterised in that:It is described anti- Oxidant is made of the vitamin C of 3-5 parts by weight and the mixture of 0.1-0.6 parts by weight, and the mixture is that L- leucyls are sweet Propylhomoserin and D- leucylglycines, the two mass ratio are 47 ~ 53:0.5-0.8.
4. the active peptides according to claim 1 extracted from blood clam prepare compound method, it is characterised in that:The hair The extraction step of blood clam active peptides is:Fresh blood clam decladding is taken, distilled water is added, is homogenized, is cleaned, filtering obtains filtrate A, then Acetic acid solution stirring extraction is added in filtrate A, centrifugation takes supernatant, and NaCl is added in supernatant to solution ultimate density and is 2.5mol/L, while flocculent deposit is precipitated, it centrifuges, obtains blood clam active peptides.
5. the active peptides according to claim 4 extracted from blood clam prepare compound method, it is characterised in that:Distilled water In D- mandelic acids and L- mandelic acids containing 0.08-0.1wt% mixture, D- mandelic acids and L- mandelic acid weight Than being 52 ~ 57:0.3~0.9.
6. the active peptides according to claim 4 extracted from blood clam prepare compound method, it is characterised in that:Removal of impurities behaviour As:Homogenate is used respectively the EDTA-2Na solution of the NaOH and 0.1-0.15mol/L of 0.15-0.25mol/L impregnate, mistake Filter, obtains filtrate A.
7. the active peptides according to claim 1 extracted from blood clam prepare compound method, it is characterised in that:It is described multiple It it is 2-5 DEG C with object cooling preservation temperature.
CN201810169499.7A 2018-02-28 2018-02-28 The active peptides extracted from blood clam prepare compound method Withdrawn CN108514123A (en)

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Publication number Priority date Publication date Assignee Title
CN110432262A (en) * 2019-07-10 2019-11-12 浙江海洋大学 A kind of preparation of waxed plant leaves sample and store method

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Publication number Priority date Publication date Assignee Title
JPH07285873A (en) * 1994-04-14 1995-10-31 Kunio Tsuji Metabolism activator and external preparation for skin and bathing agent containing the same
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Publication number Priority date Publication date Assignee Title
CN110432262A (en) * 2019-07-10 2019-11-12 浙江海洋大学 A kind of preparation of waxed plant leaves sample and store method

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