CN104397707A - Processing method for maca algae essence - Google Patents
Processing method for maca algae essence Download PDFInfo
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- CN104397707A CN104397707A CN201410768262.2A CN201410768262A CN104397707A CN 104397707 A CN104397707 A CN 104397707A CN 201410768262 A CN201410768262 A CN 201410768262A CN 104397707 A CN104397707 A CN 104397707A
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- agate coffee
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- marine alga
- algae
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Classifications
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2002/00—Food compositions, function of food ingredients or processes for food or foodstuffs
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
Landscapes
- Medicines Containing Plant Substances (AREA)
- Non-Alcoholic Beverages (AREA)
Abstract
The invention discloses a processing method for maca algae essence, and relates to maca. The processing method comprises the following steps: 1) performing enzymolysis on maca powder or/and decocting maca powder, centrifuging, taking supernatant, and concentrating to prepare maca juice; 2) performing enzymolysis on algae or/and decocting algae, centrifuging, taking supernatant, and concentrating to prepare alga juice; 3) uniformly mixing the maca juice prepared by the step 1) and the algae juice prepared by the step 2), and adding an auxiliary material; 4) concentrating the materials which are uniformly mixed in the step 3) to prepare the maca algae essence, canning, sterilizing and preserving. Through the processes of enzymolysis, decoction, concentration and the like; the nutritional components in the maca and the algae can be fully extracted; the nutritional essence of high-mountain maca and the nutritional essence of marine algae are fused; the auxiliary materials, such as longan juice or/and collagen peptide, are added to prepare the maca algae essence. An in-vitro anti-oxidation experiment proves that the obtained product has an anti-oxidation effect; a cell model experiment proves that the algae juice prepared by the processing method has an immunity regulating effect.
Description
Technical field
The present invention relates to a kind of agate coffee, particularly relate to a kind of processing method of agate coffee algae essence.
Background technology
Agate coffee (Lepidium meyenii), Cruciferae, Lepidium, is grown on South America Andes height above sea level more than 4000 meters regions, is a kind of rare plateau plant.The agate coffee root that Peru aborigines just utilized nature to dry before thousands of year supplements the nutrients and treatment various diseases (Ai Z, Cheng A F, Yu Y T, et al.Antidepressant-like behavioral, anatomical, and biochemical effects of petroleum ether extract from maca (Lepidium meyenii) in mice exposedto chronic unpredictable mild stress [J] .J Med Food, 2014,17 (5): 535-542).Agate coffee contains abundant nutrient, containing protein 10% ~ 16%, carbohydrate about 59%, dietary fiber about 8.5% and multiple free fatty, free amino acid and trace element in the agate coffee of natural air drying, and be rich in agate coffee alkene, macamide, glucosinolate and alkaloid (Gonzales G F.Ethnobiology and ethnopharmacology of Lepidium meyenii (maca), a plant fromthe Peruvian highlands [J] .Evid-Based Compl Alt, 2012,2012:1-10).People also pass through modernization means, continuous discovery also confirms that agate coffee has multiple physiological action, as antioxidation (the Shenghua Zha of maca polysaccharide, QingshengZhao, Jinjin Chen, et al.Extraction, purification and antioxidant activities of the polysaccharidesfrom maca (Lepidium meyenii) [J] .Carbohyd polym, 2014, 111:584-587), Improving memory power effect (the Rubio J of agate coffee water extract, Qiong W, Liu X, et al.Aqueous extract of black maca (Lepidium meyenii) onmemory impairment induced by ovariectomy in mice [J] .Evid-Based Compl Alt, 2011, 2011:1-7), prevention of prostatic hyperplasia effect (the Noratto G at pueraria root powder end, Condezo-Hoyos L, Gasco M, et al.Lepidiummeyenii (maca) consumption prevent benign prostatic hyperplasia [J] .Faseb J, 2013, 27) etc.
The abundant nutrition composition that the marine algas such as sea-tangle have because of it, as contained protein about 8.2% in dry sea-tangle, carbohydrate 56.2%, dietary fiber about 9.8%, and containing abundant iodine, calcium, phosphorus, iron, carrotene, thioflavin etc., and its output large (national algae total output reached 17.7 ten thousand tons in 2012).Research show algal polysaccharides have hypoglycemic, (imperial court is glad in reducing blood lipid, imperial court is auspicious, Pang Jiahong, etc. laminarin is hypoglycemic, the research of blood fat [J]. Journal of Nutrition, 2007,29 (1): 99-100), anti-oxidant (Chen Yiyong, Zhang Tianjun, Yang Jun, etc. the viscometric properties of Nantong Porphyra haitanensis polysaccharide and antioxidation activity [J] thereof. food research and development, 2013,34 (14): 77-80) physiological activity such as.
Summary of the invention
The object of this invention is to provide the processing method of advantages of simple, workable a kind of agate coffee algae essence.
The present invention includes following steps:
1) by pueraria root powder end enzymolysis or/and decoct, centrifuging and taking supernatant, concentrated, be prepared into agate coffee juice;
2) by marine alga enzymolysis or/and decoct, centrifuging and taking supernatant, concentrated, be prepared into marine alga juice;
3) by step 1) obtained agate coffee juice and step 2) marine alga juice that obtains mixes, and adds auxiliary material;
4) by step 3) in mixing material concentrate, be prepared into agate coffee algae essence, tinning, preserves after sterilization.
In step 1) in, the granularity at described pueraria root powder end can be 50 ~ 1000 orders; Described enzymolysis, pueraria root powder end can be 1 with the solid-liquid ratio of water: (5 ~ 20), wherein, pueraria root powder end is calculated in mass, and water is calculated by volume, and enzyme can be cellulase, and enzyme dosage is 7 ~ 35EGUg
-1marine alga, the temperature of enzymolysis can be 40 ~ 60 DEG C, and the time of enzymolysis can be 0.5 ~ 3.5h; Described decoction, pueraria root powder end can be 1 with the solid-liquid ratio of water: (5 ~ 20), and wherein, pueraria root powder end is calculated in mass, and water is calculated by volume, and decocting temperature is 75 ~ 100 DEG C, and decocting time can be 4 ~ 10h; Described centrifugal condition can be 3000 ~ 8000rmin
-1, 10 ~ 30min; Described concentrating can by supernatant concentration 5 ~ 15 times, and described concentrating can adopt Vacuum Concentration, and concentrated temperature can be 35 ~ 65 DEG C.
In step 2) in, described marine alga can be sea-tangle or laver etc.; Described enzymolysis, the solid-liquid ratio of marine alga and water can be 1: (2 ~ 15), and wherein, marine alga is calculated in mass, and water is calculated by volume, and enzyme can be cellulase, and enzyme dosage is 7 ~ 35EGUg
-1marine alga, the temperature of enzymolysis is 40 ~ 60 DEG C, and the time of enzymolysis is 0.5 ~ 3.5h; Described decoction, the solid-liquid ratio of marine alga and water can be 1: (2 ~ 15), and wherein, marine alga is calculated in mass, and water is calculated by volume, and the temperature of decoction can be 75 ~ 100 DEG C, and the time of decoction can be 1 ~ 5h; Described centrifugal condition can be 3000 ~ 8000rmin
-1, 10 ~ 30min; Described concentrating can by supernatant concentration 1 ~ 5 times, and described concentrating can adopt Vacuum Concentration, and concentrated temperature is 35 ~ 65 DEG C.
In step 3) in, the agate coffee in described agate coffee juice and marine alga juice and the mass percent of marine alga can be 50% ~ 80%: 20% ~ 50%, and wherein the amount of agate coffee and marine alga is scaled the quality of pueraria root powder end and marine alga before process; Described auxiliary material can be collagen peptide or/and longan juice, and the molecular weight of described collagen peptide can be below 5000Da, and described longan juice can by longan boiling gained, its solid-liquid ratio can be 1: (5 ~ 20), wherein, longan is calculated in mass, and water is calculated by volume; The addition of described auxiliary material can be 0 ~ 5% of agate coffee juice and marine alga juice total amount by mass percentage.
In step 4) in, described concentrated method can be Vacuum Concentration, and concentrated temperature is 35 ~ 65 DEG C, and concentrated multiple is 2 ~ 5 times; The mode of described sterilization can be pasteurize, and the condition of pasteurize is 60 ~ 80 DEG C, 20 ~ 40min; Described preservation can be preserved under low temperature (0 ~ 4 DEG C) or normal temperature.
The present invention mainly adopt pueraria root powder end and marine alga add water enzymolysis or/and decoct, add longan juice or/and collagen peptide and obtain a kind of agate coffee algae essence.The present invention passes through enzymolysis or/and decocting process, fully can extract the nutritional labeling in agate coffee and marine alga, originate natural, and by hexichol for bitter taste diazanyl radical scavenging activity, hydroxyl radical free radical Scavenging activity, ferrous ion sequestering power and reducing power determination and analysis, immunity regulatory cell model, proves that the agate coffee algae essence that the present invention obtains has anti-oxidant and immunoregulation effect.
The invention has the advantages that: by enzymolysis, decoction, the PROCESS FOR TREATMENT such as concentrated, fully can extract the nutritional labeling in agate coffee and marine alga, merge the nutrition elite of high mountain agate coffee and marine algae, and add longan juice or/and auxiliary materials such as collagen peptides, prepare agate coffee algae essence.Products obtained therefrom of the present invention has antioxidation through antioxidation in vitro experiment proof, and proves that gained marine alga juice of the present invention has immunoregulation effect through cell model experiment.
Accompanying drawing explanation
Fig. 1 is pueraria root powder end.
Fig. 2 is marine algae extract powder.
Fig. 3 is agate coffee algae essence.
Fig. 4 is the cytotoxicity experiment result that laver extract immunoregulation effect is evaluated.
Fig. 5 is that the laver extract of laver extract immunoregulation effect evaluation is on the impact of macrophage RAW264.7 phagocytic activity.
Fig. 6 is that the laver extract of laver extract immunoregulation effect evaluation is on the impact of macrophage RAW264.7 release IL-6.
Fig. 7 is that the laver extract of laver extract immunoregulation effect evaluation is on the impact of macrophage RAW264.7 release NO.
In Fig. 4 ~ 7, * represents and control group significant difference, its variance analysis p<0.05.
Fig. 8 is the DPPH clearance rate curve that agate coffee algae essence oxidation resistance is evaluated.
Fig. 9 is the reducing power curve that agate coffee algae essence oxidation resistance is evaluated.
Figure 10 is the ferrous ion chelation percent curve that agate coffee algae essence oxidation resistance is evaluated.
Figure 11 is the OH clearance rate curve that agate coffee algae essence oxidation resistance is evaluated.
Detailed description of the invention
Below in conjunction with the drawings and specific embodiments, the present invention is further illustrated.
Embodiment 1 one kinds adds the processing method of the agate coffee algae essence of sea-tangle, and it comprises the following steps:
1) preparation of agate coffee juice: select agate coffee, section, oven dry, pulverizing.Get pueraria root powder end 80g, by solid-liquid ratio 1: 15, (w: v) add water 1.2L, 95 DEG C decoct 8h, 5000rmin
-1centrifugal 20min, gets supernatant, and rotary evaporation concentrates (75 DEG C) 5 times, is prepared into agate coffee juice 240mL.
2) preparation of kelp juice: get Fresh Laminaria Japonica 10g, cleans, smashs to pieces.By solid-liquid ratio 1: 2 (w: v) add water 20mL, by 14EGUg
-1sea-tangle adds cellulase 0.2g, and (enzyme activity is 700EGUg
-1), enzymolysis 2h under 40 DEG C of conditions.5000rmin
-1centrifugal 20min, gets supernatant, and rotary evaporation concentrates (40 DEG C) 2 times, is prepared into kelp juice 10mL.
3) preparation of longan juice: select the dried longan pulp 5g after peeling, stoning, cleans.By solid-liquid ratio 1: 15, (w: v) add water 75mL, 95 DEG C decoct 30min.Filter, get filtrate, rotary evaporation concentrates (75 DEG C) 15 times, is prepared into longan juice 5mL.
4) prepare burden: get above step 1), 2), 3) in prepared agate coffee juice 240mL, kelp juice 10mL, longan juice 5mL mix, then adds collagen peptide 5g, mix.
5) concentrated: concentrated (40 DEG C) 3 times of material rotary evaporation after mixing, obtain agate coffee algae essence.
6) tinning and sterilization: tinning, pasteurize (65 DEG C, 30min).
7) preserve: agate coffee marine alga is skillful in normal temperature and preserves.
8) agate coffee algae essence moisture determination: utilize electronics moisture determination instrument (Sai Duolisi, MA35, Germany) to measure agate coffee algae essence water content, result display agate coffee algae essence water content is 51.17 ± 1.00%.
9) agate coffee algae essence total sugar content measures: adopt anthrone-sulphuric acid method to survey extract total sugar content.Get the glucose powder 1.000g of drying to constant weight, after dissolving completely with appropriate distilled water, be all transferred in 1L volumetric flask, and be settled to graduation mark with distilled water, be mixed with 1.00mgmL
-1dextrose standard sample, then to dilute successively with distilled water be 0.60,0.40,0.30,0.20,0.15,0.10mgmL
-1standard items test fluid, for subsequent use.Take anthrone powder 0.400g, first with a small amount of concentrated sulfuric acid dissolution, then be settled to 200mL with the concentrated sulfuric acid, make the By Anthrone Sulphuric acid solution of 0.2%, for subsequent use.Get agate coffee algae essence 1.000g, be mixed with concentration for 1.0mgmL with distilled water
-1agate coffee algae essence solution, for subsequent use.During mensuration, standard items test fluid or the agate coffee algae essence solution of getting each concentration of 5mL are respectively placed in 25mL colorimetric cylinder, add 20mL anthrone sulfuric acid solution, separately replace standard items as negative control using distilled water.Mix rear 100 DEG C of water-bath 10min, complement to 25mL with distilled water after ice-water bath cooling, go out to measure absorbance in 630nm after mixing.Take absorbance as abscissa, dextrose standard sample concentration is ordinate drawing standard curve, and fit curve equation.Gained calibration curve equation is: y=0.1738x-0.0143, and wherein: y is concentration, unit is mgmL
-1; X is absorbance, and unit is 1; Matched curve R
2=0.9962, show that this curve is at 0.1 ~ 0.6mgmL
-1relation between absorbance and concentration can be represented in concentration range; Calculating agate coffee algae essence total sugar content is thus 36.77%.
Embodiment 2 one kinds adds the processing method of the agate coffee algae essence of laver, and it comprises the following steps:
1) preparation of agate coffee juice: select agate coffee, section, oven dry, pulverizing.Get pueraria root powder end 160g, by solid-liquid ratio 1: 15, (w: v) add water 2.4L, 95 DEG C decoct 6h, 5000rmin
-1centrifugal 15min, gets supernatant, and rotary evaporation concentrates (75 DEG C) 5 times, is prepared into agate coffee juice 480mL.
2) preparation of laver juice: get laver powder 20g, by solid-liquid ratio 1: 15, (w: v) add water 300mL, 95 DEG C decoct 4h.Centrifugal (8000rmin
-1, 15min), get supernatant, rotary evaporation concentrates (75 DEG C) 5 times, is prepared into laver juice 20mL.
3) preparation of longan juice: the dried longan pulp 10g selecting peeling, stoning, cleans.By solid-liquid ratio 1: 15, (w: v) add water 150mL, 95 DEG C decoct 50min.Filter, get filtrate, rotary evaporation concentrates (75 DEG C) 15 times, is prepared into longan juice 10mL.
4) prepare burden: get above step 1), 2), 3) in prepared agate coffee juice 480mL, laver juice 20mL, longan juice 10mL, then add collagen peptide 10g, mixing.
5) concentrated: concentrated (35 DEG C) 3 times of material rotary evaporation after mixing, are prepared into agate coffee algae essence.
6) tinning and sterilization: tinning, pasteurize (65 DEG C, 30min).
7) preserve: agate coffee marine alga is skillful in normal temperature and preserves.
Embodiment 3 laver extract immunoregulation effect is evaluated, and it comprises the following steps:
1) laver extract preparation: by embodiment 2 step 2) in method prepare laver juice, freeze drying obtains laver extract powder.
2) laver extract total sugar content measures: adopt embodiment 1 step 9) in anthrone-sulphuric acid method measure total sugar content in laver extract.In result display laver extract, total sugar content is 38.60%.
3) cell chulture: cell line is mouse macrophage RAW264.7, used medium is the high glucose medium (DMEM) containing 10% hyclone (FBS), and condition of culture is 37 DEG C, 5%CO
2, every 1 ~ 2 day replaced medium, go down to posterity when cell aggregation degree reaches 80 ~ 90%.
4) Cytotoxic evaluation: the cytotoxicity adopting mtt assay detection of drugs.Specific as follows: to get 1.000g laver extract powder, be mixed with 1.00mgmL with the DMEM culture medium containing 10%FBS
-1laver extract mother liquor, and successively dilution be 0.60,0.40,0.30,0.20,0.15,0.10mgmL
-1laver extract test fluid.Macrophage RAW264.7 is inoculated in 96 orifice plates, 3 parts.Inoculum density is 5 × 10
4individual mL
-1, inoculum concentration is 100 μ L, cultivates (37 DEG C, 5%CO
2) 1 ~ 2h, after cell attachment, absorb old nutrient solution, then add variable concentrations test fluid 100 μ L, and do negative control with the nutrient solution not containing laver extract.Cultivate (37 DEG C, 5%CO respectively
2) 12,24, after 36h, every hole adds 5mgmL
-1mTT solution 20 μ L, continue to cultivate after 4h, every hole adds 100 μ L tri-liquid and (gets dodecyl sodium sulfate 10g, isopropyl alcohol 5mL, 10molL
-1hydrochloric acid 0.1mL, dissolves with distilled water and to be settled to 100mL formulated) overnight incubation, to dissolve first a ceremonial jade-ladle, used in libation.Finally survey light absorption value in 550nm wavelength place.Result as shown in Figure 4, at 100 ~ 800 μ gmL
-1in concentration range, laver extract is to cytotoxic, and laver extract stimulates 24h significantly can promote cell proliferation, improves cell viability.
5) cytophagy merit rating: adopt neutral red test method to detect laver extract to the impact of macrophage RAW264.7 phagocytic activity.Specific as follows: macrophage RAW264.7 is inoculated in 96 orifice plates, inoculum density is 5 × 10
4individual mL
-1, inoculum concentration is 100 μ L, cultivates (37 DEG C, 5%CO
2) 1 ~ 2h, after cell attachment, absorb old nutrient solution, then add variable concentrations laver extract test fluid 100 μ L, and do negative control with the nutrient solution not containing laver extract.Cultivate (37 DEG C, 5%CO
2) 12h, discard cells and supernatant, clean 2 times with PBS (pH=7.4), add dimethyl diaminophenazine chloride (dissolving with PBS, now with the current) the 100 μ L of 0.5%, cultivate (37 DEG C, 5%CO
2) 4h, absorb supernatant, clean 2 times with PBS, add cell pyrolysis liquid (V
absolute ethyl alcohol: V
acetic acid: V
ultra-pure water=1: 1: 2) 100 μ L, shake 10min under room temperature, survey light absorption value in 550nm wavelength place.Result as shown in Figure 5, at 100 ~ 800 μ gmL
-1in concentration range, laver extract significantly can strengthen the phagocytic activity of macrophage RAW264.7.Phagocytic activity is an important indicator (Wang Changlu of macrophage immunity ability, Cui Haiyan et al.Bidirectional immunomodulatory activities of polysaccharides purified frompleurotus nebrodensis [J] .Inflammation, 2014,34 (1): 83-93), experimental result shows that laver extract is by strengthening the phagocytic activity of macrophage, thus strengthens its immunocompetence.
6) detection of interleukin-6 (IL-6) and nitric oxide (NO): macrophage RAW264.7 is inoculated in 48 orifice plates, and inoculum density is 5 × 10
4individual mL
-1, inoculum concentration is 200 μ L, cultivates (37 DEG C, 5%CO
2) 1 ~ 2h, after cell attachment, absorb old nutrient solution, then add variable concentrations laver extract test fluid 200 μ L, and do negative control with the nutrient solution not containing laver extract.Cultivate again (37 DEG C, 5%CO
2) 24h, draw cells and supernatant, detect the secretory volume of IL-6 and NO respectively by IL-6 detection kit and Griess method.IL-6 result as shown in Figure 6, at 100 ~ 800 μ gmL
-1in concentration range, laver extract significantly can promote that macrophage RAW264.7 discharges IL-6.NO result as shown in Figure 7, at 100 ~ 800 μ gmL
-1in concentration range, laver extract significantly can promote that macrophage RAW264.7 discharges, and its facilitation effect increases with laver extract concentrations and significantly strengthens.IL-6 be considered to amboceptor,immune main in body (O ' shea, J.J, P.J.Murray.Cytokine signaling modules in inflammatory responses [J] .Immunity, 2008,28:477-487), the IL-6 produced in subrange effectively can strengthen the immunocompetence of body.And the immunocompetence of the immunocytes such as the adjustable phagocyte of NO, thus strengthen the immunocompetence (Apetoh of body, L., F.Ghiringhelli, A.Tesniere, et al.2007.Toll-likereceptor 4-dependent contribution ofthe immune system to anticancer chemotherapy andradiotherapy [J] .Nat Med, 2007,13 (9): 1050 – 1059).Result shows, and laver extract can promote the propagation of RAW264.7 and can strengthen its phagocytic activity, stimulates its secretory immune cell factor, strengthens the immunocompetence of macrophage.
Embodiment 4 agate coffee algae essence determination oxidative, it comprises the following steps:
1) agate coffee algae essence preparation: prepare agate coffee algae essence according to step in embodiment 1, freeze drying is prepared into agate coffee algae essence powder.Get agate coffee algae essence powder, be mixed with 50.00mgmL with distilled water
-1solution, and be diluted to 10.00 successively, 8.00,6.00,4.00,3.00,2.00,1.50,1.00,0.75,0.50mgmL
-1test fluid, for subsequent use.
2) agate coffee algae essence powder moisture measures: utilize electronics moisture determination instrument to measure agate coffee algae essence powder water content, it is 28.83% that result shows its water content.
3) hexichol is for bitter taste diazanyl free radical (DPPH) Scavenging activity: assay method is with reference to Wu (Wu Shao-Chi, PanChorng-Liang.Preparation of algal-oligosaccharide mixtures by bacterial agarases and theirantioxidative properties [J] .Fisheries Sci, 2004,70 (6): 1164-1173) method of people such as, and slightly make an amendment, specific as follows: in 2mL centrifuge tube, to add test fluid 250 μ L, 50mmolL successively
-1trishydroxymethylaminomethane-hydrochloric acid (Tris-HCl) cushioning liquid (pH=7.4) 250 μ L, 0.1mol/L DPPH
2ethanolic solution 1mL.After mixing, lucifuge reaction 30min, utilizes ultraviolet-uisible spectrophotometer (platinum Ai Ermo, lambda-35, the U.S.) to measure absorbance in 517nm place.Separately test fluid is replaced to do negative control with distilled water.According to following formulae discovery DPPH clearance rate: DPPH clearance rate (%)=[(A
contrast-A
test fluid)/A
contrast] × 100%, A in formula
contrastfor negative control group reactant liquor absorbance, A
test fluidfor test fluid group reaction liquid absorbance, with DPPH clearance rate for abscissa, concentration is that ordinate draws clearance rate curve, fit regression curve model, calculates EC
50value (EC
50value represents the test fluid concentration needed for DPPH of removing 50%, and unit is mgmL
-1.EC
50its DPPH Scavenging activity of less expression is stronger).Found that agate coffee algae essence powder DPPH Scavenging activity EC
50for 8.95mgmL
-1, EC after deduction moisture
50value is 6.37mgmL
-1(as Fig. 8).
4) reducing power measures: assay method is with reference to Wu (Wu Shao-Chi, Pan Chorng-Liang.Preparation ofalgal-oligosaccharide mixtures by bacterial agarases and their antioxidative properties [J] .FisheriesSci, 2004, 70 (6): 1164-1173) method of people such as, and slightly make an amendment, specific as follows: in centrifuge tube, to add sample 500 μ L and 200mmol/L PBS (pH=6.6) the solution 250 μ L containing 1% potassium ferricyanide, in 50 DEG C of water-bath 20min after mixing, then add 10% trichloroacetic acid 250 μ L, mixing is with cessation reaction.The centrifugal 10min of 12000r/min, gets supernatant 250 μ L, and adds distilled water 250 μ L and 0.1% ferric chloride solution 250 μ L, measures absorbance after mixing in 700nm place.Test fluid is substituted as negative control using distilled water.According to following formulae discovery reducing power: reducing power=A
sample-A
cloudy, A in formula
cloudy: negative control reaction system absorbance, A
sample: example reaction system absorbance, set up matched curve model between reducing power and agate coffee algae essence powder concn, calculate its AC
0.5value (AC
0.5value represents A
sample-A
cloudytest fluid concentration when=0.5, unit is mgmL
-1).Result shows its AC
0.5value is 4.64mgmL
-1, its AC after deduction moisture
0.5value is 3.30mgmL
-1(as Fig. 9).
5) ferrous ion (Fe
2+) sequestering power mensuration: assay method is with reference to Wu (Wu Shao-Chi, Pan Chorng-Liang.Preparation of algal-oligosaccharide mixtures by bacterial agarases and their antioxidativeproperties [J] .Fisheries Sci, 2004,70 (6): 1164-1173) method of people such as, and slightly make an amendment, specific as follows: in centrifuge tube, to add methyl alcohol 700 μ L and 400 μm olL
-1feSO
4solution 50 μ L, adds sample solution 50 μ L again after mixing, mixing, finally adds 2mmolL
-1luxuriant and rich with fragrance Lip river piperazine 25 μ L, reacts 10min under room temperature, measures absorbance in 562nm place.Replace test fluid as positive control using the ethylenediamine tetra-acetic acid of same concentrations (EDTA), replace test fluid as negative control using distilled water.According to following formulae discovery Fe
2+chelation percent: chelation percent (%)=[(A
cloudy-A
sample)/A
cloudy] × 100%, A in formula
contrastfor negative control group reactant liquor absorbance, A
test fluidfor test fluid group reaction liquid absorbance, by setting up matched curve model between chelation percent and concentration, calculate its EC
50value (EC
50value represents the Fe of chelating 50%
2+required test fluid concentration, unit is mgmL
-1).Result shows its EC
50value is 2.55mgmL
-1, its EC after deduction moisture
50value is 1.82mgmL
-1(as Figure 10).
6) hydroxyl radical free radical (OH) Scavenging activity: assay method is with reference to fourth profit monarch (Ding Lijun, Zhou Guodong. the extraction of lotus seeds water-soluble sugar and the research [J] to radical scavenging activity thereof. Food Science, 2002,23 (8): 252-254) method of people such as, and slightly make an amendment, specific as follows: in centrifuge tube, to add sample solution 200 μ L, 9mmolL
-1feSO
4solution 200 μ L, 9mmolL
-1salicylic acid-ethanolic solution 200 μ L and 8.8mmolL
-1h
2o
2solution 200 μ L, 37 DEG C of reaction 10min, measure absorbance in 510nm place.Replace test fluid as positive control using the vitamin C of same concentrations (Vc), replace test fluid as negative control using distilled water.According to following formulae discovery OH clearance rate: clearance rate (%)=[(A
contrast-A
test fluid)/A
contrast] × 100%, A in formula
contrastfor negative control group reactant liquor absorbance, A
test fluidfor test fluid group reaction liquid absorbance, set up Regression curve between sample concentration and clearance rate, calculate EC
50value (EC
50value represents the test fluid concentration needed for OH of removing 50%, and unit is mgmL
-1).Result shows its OH Scavenging activity EC
50value is 9.47mgmL
-1, EC after deduction moisture
50value is 6.74mgmL
-1(as Figure 11).
During reactive oxygen species (ROS) excessive concentration in body, the immune response of possible induction of lymphocyte Secondary cases, thus the rapid cleavage of active cell or programmed death, body immunity is caused to reduce (Ke Chunlin, Qiao Deliang, Zeng Xiaoxiong. streptococcus zooepidemicus capsular polysaccharide is to the anti-oxidant of immunosupress mouse and immunoregulatory activity research [J]. Chinese microecology impurity, 2011,23 (8): 684-688).Analyze through above four indices, show that the agate coffee algae essence that the present invention obtains has antioxidation.Meanwhile, the marine alga (polysaccharide) of using in the present invention has immunoregulation effect, can strengthen body immunity.
The present invention relates to a kind of processing method of agate coffee algae essence, it comprises the following steps: the 1) preparation of agate coffee juice; 2) preparation of marine alga juice; 3) prepare burden; 4) concentrated; 5) tinning and sterilization; 6) finished product is preserved.Key of the present invention is by enzymolysis or/and decoct, the process such as to concentrate, nutritional labeling in abundant extraction agate coffee and marine alga, and by adding longan juice or/and the natural auxiliary material such as collagen peptide, prepare the agate coffee algae essence combining high mountain and sea-plant elite.The inventive method has the features such as technique is simple, easy to operate, and products obtained therefrom has the nutritional labeling of agate coffee, marine alga, longan, collagen peptide simultaneously.Analyzed by macrophage RAW264.7, prove that the present invention marine alga (polysaccharide) used has immunoregulation effect; Through the determination and analysis of hexichol for bitter taste diazanyl radical scavenging activity, hydroxyl radical free radical Scavenging activity, ferrous ion sequestering power and reducing power, prove that gained agate coffee algae essence of the present invention has antioxidation.
Claims (10)
1. a processing method for agate coffee algae essence, is characterized in that comprising the following steps:
1) by pueraria root powder end enzymolysis or/and decoct, centrifuging and taking supernatant, concentrated, be prepared into agate coffee juice;
2) by marine alga enzymolysis or/and decoct, centrifuging and taking supernatant, concentrated, be prepared into marine alga juice;
3) by step 1) obtained agate coffee juice and step 2) marine alga juice that obtains mixes, and adds auxiliary material;
4) by step 3) in mixing material concentrate, be prepared into agate coffee algae essence, tinning, preserves after sterilization.
2. the processing method of a kind of agate coffee algae essence as claimed in claim 1, is characterized in that in step 1) in, the granularity at described pueraria root powder end is 50 ~ 1000 orders.
3. the processing method of a kind of agate coffee algae essence as claimed in claim 1, it is characterized in that in step 1) in, described enzymolysis, pueraria root powder end is 1 with the solid-liquid ratio of water: (5 ~ 20), wherein, pueraria root powder end is calculated in mass, and water is calculated by volume, enzyme can be cellulase, and enzyme dosage is 7 ~ 35EGUg
-1marine alga, the temperature of enzymolysis is 40 ~ 60 DEG C, and the time of enzymolysis is 0.5 ~ 3.5h.
4. the processing method of a kind of agate coffee algae essence as claimed in claim 1, it is characterized in that in step 1) in, described decoction, pueraria root powder end is 1 with the solid-liquid ratio of water: (5 ~ 20), wherein, pueraria root powder end is calculated in mass, and water is calculated by volume, decocting temperature is 75 ~ 100 DEG C, and decocting time can be 4 ~ 10h.
5. the processing method of a kind of agate coffee algae essence as claimed in claim 1, is characterized in that in step 1) in, described centrifugal condition is 3000 ~ 8000rmin
-1, 10 ~ 30min; Described concentrating can by supernatant concentration 5 ~ 15 times, and described concentrating can adopt Vacuum Concentration, and concentrated temperature can be 35 ~ 65 DEG C.
6. the processing method of a kind of agate coffee algae essence as claimed in claim 1, is characterized in that in step 2) in, described marine alga is sea-tangle or laver; Described enzymolysis, the solid-liquid ratio of marine alga and water can be 1: (2 ~ 15), and wherein, marine alga is calculated in mass, and water is calculated by volume, and enzyme can be cellulase, and enzyme dosage is 7 ~ 35EGUg
-1marine alga, the temperature of enzymolysis is 40 ~ 60 DEG C, and the time of enzymolysis is 0.5 ~ 3.5h; Described decoction, the solid-liquid ratio of marine alga and water can be 1: (2 ~ 15), and wherein, marine alga is calculated in mass, and water is calculated by volume, and the temperature of decoction can be 75 ~ 100 DEG C, and the time of decoction can be 1 ~ 5h.
7. the processing method of a kind of agate coffee algae essence as claimed in claim 1, is characterized in that in step 2) in, described centrifugal condition is 3000 ~ 8000rmin
-1, 10 ~ 30min; Described concentrating can by supernatant concentration 1 ~ 5 times, and described concentrating can adopt Vacuum Concentration, and concentrated temperature is 35 ~ 65 DEG C.
8. the processing method of a kind of agate coffee algae essence as claimed in claim 1, it is characterized in that in step 3) in, agate coffee in described agate coffee juice and marine alga juice and the mass percent of marine alga are 50% ~ 80%: 20% ~ 50%, and wherein the amount of agate coffee and marine alga is scaled the quality of pueraria root powder end and marine alga before process.
9. the processing method of a kind of agate coffee algae essence as claimed in claim 1, it is characterized in that in step 3) in, described auxiliary material is that collagen peptide is or/and longan juice, the molecular weight of described collagen peptide can be below 5000Da, described longan juice can by longan boiling gained, and its solid-liquid ratio can be 1: (5 ~ 20), wherein, longan is calculated in mass, and water is calculated by volume; The addition of described auxiliary material can be 0 ~ 5% of agate coffee juice and marine alga juice total amount by mass percentage.
10. the processing method of a kind of agate coffee algae essence as claimed in claim 1, is characterized in that in step 4) in, described concentrated method is Vacuum Concentration, and concentrated temperature is 35 ~ 65 DEG C, and concentrated multiple is 2 ~ 5 times; The mode of described sterilization can be pasteurize, and the condition of pasteurize is 60 ~ 80 DEG C, 20 ~ 40min; Described preservation can be preserved under 0 ~ 4 DEG C or normal temperature.
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| CN105367677A (en) * | 2015-12-01 | 2016-03-02 | 中国药科大学 | Maca polysaccharide of immunoregulatory activity and preparation method therefor |
| CN106722929A (en) * | 2016-11-23 | 2017-05-31 | 丽江雪康源生物科技有限公司 | A kind of processing method of fine maca tablets |
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| CN102499415A (en) * | 2011-11-17 | 2012-06-20 | 深圳市凯比特生物资源与环境研究所 | Chlorella essence nutrient fluid and preparation method thereof |
| CN104095897A (en) * | 2014-06-24 | 2014-10-15 | 伏思思 | Maca dunaliella salina compound preparation |
| CN104187679A (en) * | 2014-09-13 | 2014-12-10 | 昆明元正生物科技有限公司 | Haematococcus pluvialis maca health product and preparation method thereof |
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| CN105367677A (en) * | 2015-12-01 | 2016-03-02 | 中国药科大学 | Maca polysaccharide of immunoregulatory activity and preparation method therefor |
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