CN104789352A - Comprehensive utilization method of cumin and prepared product - Google Patents
Comprehensive utilization method of cumin and prepared product Download PDFInfo
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- CN104789352A CN104789352A CN201510146145.7A CN201510146145A CN104789352A CN 104789352 A CN104789352 A CN 104789352A CN 201510146145 A CN201510146145 A CN 201510146145A CN 104789352 A CN104789352 A CN 104789352A
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- cumin
- extract
- comprehensive utilization
- obtains
- centrifugation
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- 229910000029 sodium carbonate Inorganic materials 0.000 description 1
- 239000011780 sodium chloride Substances 0.000 description 1
- 239000001509 sodium citrate Substances 0.000 description 1
- NLJMYIDDQXHKNR-UHFFFAOYSA-K sodium citrate Chemical compound O.O.[Na+].[Na+].[Na+].[O-]C(=O)CC(O)(CC([O-])=O)C([O-])=O NLJMYIDDQXHKNR-UHFFFAOYSA-K 0.000 description 1
- 239000007787 solid Substances 0.000 description 1
- 239000012086 standard solution Substances 0.000 description 1
- 239000008117 stearic acid Substances 0.000 description 1
- 239000013589 supplement Substances 0.000 description 1
- 235000019605 sweet taste sensations Nutrition 0.000 description 1
- 208000024891 symptom Diseases 0.000 description 1
- 229940126680 traditional chinese medicines Drugs 0.000 description 1
- 229910021642 ultra pure water Inorganic materials 0.000 description 1
- 239000012498 ultrapure water Substances 0.000 description 1
- 238000010792 warming Methods 0.000 description 1
- 235000020138 yakult Nutrition 0.000 description 1
Landscapes
- Preparation Of Compounds By Using Micro-Organisms (AREA)
- Peptides Or Proteins (AREA)
Abstract
The invention belongs to the field of deep processing of agricultural products, and provides a comprehensive utilization method of cumin. The comprehensive utilization method is characterized by comprising the following steps: using ultrasonic microwaves for assisting a steam distillation method and a enzymolysis-acetone combined extraction method to extract cumin essential oils and oleoresins, and then using a plurality of processes for extracting cumin polyphenols, cumin isolated proteins, cumin peptides and cumin dietary fibers from defatted cumin; using a high-pressure homogenization-assisted enzymolysis method for extracting dietary fibers from cumin straws; realizing the full utilization of seeds, branches and leaves as well as straws of cumin plants. According to the comprehensive utilization method of cumin, provided by the invention, the cumin essential oils, cumin oleoresins, cumin polyphenols, cumin isolated proteins, cumin peptides and cumin dietary fibers can be extracted from cumin seeds; the cumin straw dietary fibers can be extracted from the cumin straws, so as to effectively avoid the resource waste and environmental pollution and realize the full utilization of cumin.
Description
Technical field
The invention belongs to deep-processing technical field of agricultural products, be specifically related to a kind of method of comprehensive utilization of cumin.
Background technology
Cumin, also known as Cuminum cyminum L, cumin of resting in peace, fennel etc. of resting in peace, be the annual or 2 years raw herbaceous plant of umbelliferae cumin apium, fruit is Long Circle, has strong aromatic odour.Originate in Mediterranean Zone, extensively plant on India, Iran, Turkey and other places, Xinjiang of China and Gansu are the major production bases of cumin, and cultivated area and output account for 70% of national share.Cumin fruit is often using seed form or be ground into powder as foodstuff flavouring; Its fruit also can be used as medicine, and has restoring consciouness and promotes blood circulation, falls the effects such as fiery flat liver, also may be used for treatment maldigestion, stomach cold stomachache, the just symptom such as frequency of suffering from a deficiency of the kidney.
The food fibre of the essential oil containing 3%-4% in cumin seed, the fat of about 15%, the protein of about 20%, the total flavones of 4%-5% and 30%-40%, also contain the multiple amino acids of the mineral elements such as potassium, calcium, magnesium, iron and multivitamin and needed by human body, nutritive value is very high.At present, generally only have seed or powder to be used by as seasonings after cumin harvesting, product form is single, and price is lower, is unfavorable for increasing peasant income and enterprise's creation of value.
Meanwhile, cumin can produce a large amount of stalk when producing and gather in the crops, and except a little as except animal-feed, major part is used as waste and is directly lost or burn, cause the serious wasting of resources and environmental pollution, also become a great problem of puzzlement enterprise.If the compositions such as the food fibre in cumin stalk can be made full use of, greatly will improve cumin processing added value, thus advantageously promote the sound development of cumin processing industry.Therefore the method for comprehensive utilization of a kind of economical and efficient, eco-friendly cumin seed, branches and leaves and stalk is badly in need of.
Summary of the invention
For the weak point that this area exists, the object of the invention is the method for comprehensive utilization proposing the overall plant of a kind of cumin.
Another object of the present invention is the product proposing to obtain in method of comprehensive utilization.
Realizing above-mentioned purpose technical scheme of the present invention is:
A kind of method of comprehensive utilization of cumin, it is characterized in that, adopt ultrasonic microwave to assist steam distillation and enzymolysis-acetone combined extraction method to extract cumin essential oil and oleo-resinous, then adopt kinds of processes from degreasing cumin, extract cumin polyphenol, cumin protein isolate, cumin polypeptide and cumin food fibre; And adopt high-pressure homogeneous auxiliary enzymes solution to extract food fibre from cumin stalk; Realize the complete utilization of the seed of cumin plant, branches and leaves and stalk.
Described method, comprises the steps:
1) after cumin seed mixes with 1:3-1:5 mass volume ratio with water, at ultrasonic wave 30-50kHz, pre-treatment 10-30min under microwave 30-60W; Then the cumin seed through ultrasonic microwave process is carried out distillation method extraction, Extracting temperature is 96-100 DEG C, the oil content obtained is collected, is packed, obtains cumin essential oil.
2) by step 1) extract cumin seed residue remaining after essential oil and dry or dry and pulverize after its moisture content is 8%-10%, the sodium acetate buffer getting powder and pH4.0-6.0 after pulverizing mixes with 1:10 mass volume ratio and shakes up, zytase and the cellulase mixed enzyme solution of 0.5%-5% is added by enzyme-to-substrate concentration ratio, 45-55 DEG C of enzymolysis 30-180min, centrifugation 4000-8000g (centrifugal force unit, represent with g) centrifugal 10-30min, gained precipitation is for extracting ziran oil resin, and supernatant liquor saves backup.
The precipitation obtained after enzymolysis is dried, (precipitation can be lumpd after drying to pulverize the rear 30-70 of mistake eye mesh screen, so need to pulverize), acetone extraction is added by solid-to-liquid ratio 1:2-1:5, then centrifugation, gained supernatant liquor adopts the method for evaporation that acetone is volatilized and removes, and gained deep green viscous liquid is ziran oil resin.
3) by step 2) add acetone after after the residue that obtains of centrifugation mixes with 1:10-1:20 ratio with 65%-70% ethanol, supersound extraction 10-60min, then centrifugation, collect supernatant liquor, residue repeats to extract 2-3 time as stated above, united extraction liquid, evaporative removal ethanol, obtains cumin polyphenolic extract.
4) by step 3) obtain cumin polyphenol after the polyphenolic extract freeze-drying that obtains and slightly carry powder, cumin polyphenol slightly being carried powder with the HCl solution of pH 3-4.5, to be formulated as polyphenol content be that (CAE/mL is polyphenol content unit to 0.2-2.0mg CAE/mL, be expressed as mg chlorogenic acid equivalent/mL sample solution) solution, then chromatography column is loaded to, AB-8 macroporous resin selected by filler, stripping liquid flow velocity is 0.25-2.50BV/h, adsorption temp is 25-35 DEG C, wash-out polyphenol solution is out collected, freeze-drying, obtains cumin polyphenol.
5) by step 3) extract residue 45-55 DEG C of oven dry remaining after polyphenol, pulverize after 30-70 eye mesh screen, then after mixing with 1:10-1:40 ratio with phosphate buffered saline buffer (pH 8), stir under 30-40 DEG C of condition and extract 1-3h, centrifugation, collect supernatant liquor, ultrafiltration and concentration, freeze-drying, obtains cumin protein isolate.
6) by step 5) the cumin protein isolate that obtains, by enzyme-to-substrate concentration ratio (1-6): 100 add proteolytic enzyme, be placed in extra-high tension unit, after 100-400MPa, 40-60 DEG C of process 5-15min, 10-15min is with the activity of passivation proteolytic enzyme in boiling water bath heating, centrifugation, collect supernatant liquor, 60-65 DEG C of rotary evaporation removes moisture, and the concentrated solution freeze-drying obtained, is cumin polypeptide.
7) by step 2) supernatant liquor that obtains and step 6) centrifugal after after the precipitation that obtains merges, add massfraction be 85-99%, be preheated to the ethanol of 60 DEG C leave standstill after centrifugal, collecting precipitation after 45-55 DEG C of oven dry, is pulverized, is obtained food fibre in baking oven.
8) 30-70 mesh sieve is crossed after cumin branches and leaves remaining when being gathered in the crops by cumin and crushed stalk, after mixing with 1:10-1:30 ratio with the phosphate buffered saline buffer of pH7.0-8.0, adopt high pressure homogenizer 100-350bar circulation homogeneous 1-5 time, then by enzyme-to-substrate concentration ratio (1-2.5): 100 add proteolytic enzyme, under 50-65 DEG C of condition after enzymolysis 120-240min, boiling water bath keeps 10-15min with the activity of passivation proteolytic enzyme, centrifugation, after gained precipitation 45-55 DEG C is dried, pulverize, be cumin stalk food fibre.
Wherein, step 1) in extraction time can be 30min.
Described mass volume ratio unit can be g/mL, kg/L, ton/m
3.
Wherein, step 2) in, zytase: cellulase=1:1-2:1.Step 2) in by step 1) extract cumin seed residue remaining after essential oil and dry or dry be crushed to grain size < 400 μm after its moisture content is 8%-10%.
Preferably, described step 2) in, cross 30-70 eye mesh screen by after the precipitation oven dry obtained after enzymolysis, pulverizing, add acetone by solid-to-liquid ratio 1:2-1:5, stir with rotating speed 100-150rpm under room temperature and extract the centrifugal 10-30min separation of 30-60min, 4000-8000g.
Wherein, described step 2) in, the temperature of oven dry is 45-55 DEG C.
Wherein, described step 3) in, cross 30-70 eye mesh screen after pulverizing after residue 45-55 DEG C of oven dry is 8%-10% to moisture content, then mix with ethanol, under ultrasonic wave 45-55kHz, microwave 100-150W, extract 10-60min.
Wherein, step 3), step 5), step 6) and step 8) condition of centrifugation is: centrifugation 10-30min under 4000-8000g.
Wherein, described step 4) in, the stripping liquid flow velocity of chromatography column is 0.25-2.50BV/h, and adsorption temp is 25-35 DEG C, and adsorption time is 1-3.5h; After absorption reaches balance, adopt the ethanolic soln of 65-75% to carry out desorb, stripping liquid flow velocity is 0.3-2.5BV/h, and stripping liquid consumption is 3-4BV.
Wherein, described step 6) in, cumin protein isolate add enzyme before adjust ph, the method for adjustment is mix with 1:10 mass volume ratio with the phosphate buffered saline buffer of pH 7.0-8.0.
Wherein, described step 6) in, after supernatant liquor and precipitation merge, add and be preheated to 60 DEG C, 95% ethanol, room temperature is centrifugal after leaving standstill 1h.
Preferably, described step 7) in supernatant liquor and precipitation are merged after, add massfraction be 85-99%, be preheated to the ethanolic soln of 60 DEG C leave standstill after centrifugal, volume ratio is ethanolic soln: (supernatant liquor+precipitation)=1:2-1:4.
The product that the method for comprehensive utilization that the present invention proposes prepares.
Beneficial effect of the present invention is:
1) the present invention can extract cumin essential oil, ziran oil resin, cumin polyphenol, cumin protein isolate, cumin polypeptide, cumin food fibre from cumin seed, extract from cumin stalk and obtain cumin stalk food fibre, effectively prevent the wasting of resources and environmental pollution, achieve the full use of cumin;
2) 7 kinds of products that the present invention obtains not only can enrich market product, and the added value of cumin can be improved, increase Business Economic Benefit, improve farmers' income, certain promoter action is also served to the structural adjustment of cumin plant husbandry and industrialization development simultaneously;
3) method therefor of the present invention and operation of equipment simply, are easy to realize suitability for industrialized production;
4) product that the present invention obtains serves many purposes: cumin essential oil can be applied to food service industry as antioxidant, such as, added in meat industry by cumin essential oil and not only can play the effect of rendering palatable, can also suppress the oxidation of meat product; Cumin essential oil also can be used as functional ingredient and adds in medicine effects such as playing prevention stomach cold stomachache, control diarrhoea to; In addition cumin essential oil also can play the effect such as protection against rodents, bactericidal;
Ziran oil resin can add in food as seasonings, such as, roast goods, refrigerated products, puffed food and food flavouring etc.;
Cumin polyphenol is a kind of natural antioxidant, has the effects such as antibacterial, anti-oxidant;
Cumin protein isolate is rich in multiple amino acids, and containing 8 kinds of indispensable amino acids, nutritious, not containing cholesterol, there are good emulsifying character, hydration, foaminess etc., in addition, also there is good anti-oxidant activity and alpha-amylase activity rejection ability;
Cumin polypeptide can be applied to food service industry, makes protein peptide drink, both can enrich market, has again anti-oxidant, supplementary amino acid, Bearberry Extract;
Cumin and stalk dietary fiber particles less, volume is fluffy, has good water-holding power, water retention capacity and fat absorbing ability, can be added in food, as water-holding agent, oil protecting agent and leavening agent etc.; In addition, can add in protective foods as functional ingredient, there is the effects such as hypoglycemic, hypotensive, reducing blood-fat, minimizing inflammation.
Accompanying drawing explanation
Fig. 1 is cumin and stalk comprehensive utilization schema.
Fig. 2 is cumin essential oil Typical gas chromatograph-mass spectrum (GC-MS).
Fig. 3 is the fatty acid profile (GC) of ziran oil resin.
Fig. 4 is cumin and stalk comprehensive utilization product (7 kinds) photo, in Fig. 4, (a): cumin essential oil, (b): ziran oil resin, (c): cumin polyphenol, (d): cumin protein isolate, (e): cumin polypeptide, (f): cumin food fibre, (g): cumin stalk food fibre.
Fig. 5 is the biscuit made of cumin food fibre and steamed bun photo, in Fig. 5 (a): saline taste biscuit, and (b): sweet taste biscuit, (c): steamed bun.
Embodiment
Below in conjunction with drawings and Examples, the specific embodiment of the present invention is described in further detail.Those skilled in the art should know, and scope of the present invention is not limited only to specific embodiment, can carry out various modification and change without departing from the spirit of the present invention.
In embodiment, Sumizyme MP used is Sumizyme MP (Alcalase 2.4L), and lot number PLN05355, enzyme work >=3000U/ml, be purchased from Novozymes company; Zytase used is inscribe-Isosorbide-5-Nitrae-beta-xylanase, and lot number X2629, enzyme work >=1000U/g, be purchased from Sigma company; Described cellulase is R-10, lot number 130918-01, and enzyme work >=12000U/g is purchased from Yakult company.Other reagent are analytical pure, are purchased from Beijing traditional Chinese medicines group.
If not otherwise specified, the means adopted in embodiment are the technique means of this area routine.
The extraction of embodiment 1, cumin essential oil
The cumin of results is removed impurity through selection by winnowing, obtains the cumin seed of cleanliness >=98%, after then being mixed with 1:5g/mL with water by cumin seed, put into ultrasonic microwave extractor, pre-treatment 25min under ultrasonic frequency vibratory 35kHz, microwave power 45W; Then the cumin seed through ultrasonic microwave process is transferred to extraction of essential oil unit (producer: Shanghai Ju Yuan mechanical means company limited, model: JYT5L) distillation basket in, add water in retort, the height of water is no more than distillation basket, extracts by wet distillation.Temperature in setting extractor is 100 DEG C, and extraction time is 30min.The oil content obtained is collected, packed, obtains cumin essential oil.Outward appearance is shown in Fig. 4 it (a).
The extraction of embodiment 2, ziran oil resin
Cumin seed slag baking oven remaining after embodiment 1 being extracted essential oil is dried at 50 DEG C, its moisture content is made to reach about 8%, grain size < 400 μm is crushed to Universalpulverizer, the sodium acetate buffer getting powder and pH 4.5 after pulverizing mixes with 1:10g/mL ratio and shakes up, zytase and the cellulase mixed enzyme solution (zytase: cellulase=1:1) of 3.5% is added by enzyme-to-substrate concentration ratio, 50 DEG C of enzymolysis 120min, the centrifugal 20min of 7500g, gained precipitation is for extracting ziran oil resin, supernatant liquor saves backup.
The precipitation obtained after enzymolysis is centrifugal 50 DEG C oven dry, 40 eye mesh screens are crossed after pulverizing, acetone is added by solid-to-liquid ratio 1:3g/mL, under room temperature, 120rpm stirs and extracts 45min, the centrifugal 20min of 7500g (centrifugal rear remaining solid precipitation is used for embodiment 3), gained supernatant liquor adopts vacuum rotary evaporator at 45 DEG C, make acetone volatilize and removes, and gained deep green viscous liquid is ziran oil resin.Outward appearance is shown in Fig. 4 it (b).
The preparation of embodiment 3, cumin polyphenolic extract
Example 2 is pulverized with Universalpulverizer after the centrifugal residue obtained 50 DEG C oven dry after adding acetone, (precipitation can be lumpd after drying to cross 40 eye mesh screens, so will pulverize and sieve), cumin residue powder after sieving mixes with 1:10g/mL with 70% ethanol, puts into ultrasonic microwave assisted extraction device, and setting hyperacoustic vibrational frequency is 51kHz, microwave power is 130W, after extracting 10min, the centrifugal 20min of 7500g, collects supernatant liquor; Then adopt aforesaid method to repeat extraction 2 times precipitation, merge the extracting solution of 3 times, 45 DEG C of rotary evaporations remove ethanol, obtain cumin polyphenolic extract.(extract remaining precipitation and be used for embodiment 5).
The purifying of embodiment 4, cumin polyphenolic extract
Obtain cumin polyphenol after polyphenolic extract freeze-drying embodiment 3 obtained and slightly carry powder, then adopt AB-8 macroporous resin to carry out purifying, concrete steps are as follows:
1) AB-8 macroporous resin treatment and dress post: the AB-8 macroporous resin getting certain mass, with 95% alcohol solution dipping 24 hours of 4 times of volumes, distilled water repeatedly cleans to elutant and clarifies; The NaOH solution adding 4 times of volume 2mol/L soaks 4-6h, is washed till neutrality, adds the HCl solution of the 2mol/L of 4 times of volumes with distilled water, soaks 4-6h, is washed till neutrality with distilled water, and suction filtration loads in the chromatography column that column volume is 10mL after removing moisture;
2) damping fluid cumin polyphenol slightly being carried powder pH3.5 is made into the solution that concentration is 1.0mgCAE/mL, draw the above-mentioned solution of 2mL and add chromatography column top to, carry out dynamic adsorption with distilled water, the elution speed of distilled water is 1.0BV/h, adsorption temp is 25 DEG C, and adsorption time is 2.5h;
3) step 2) absorption reach balance after, with 70% ethanol for stripping liquid carries out dynamic desorption, stripping liquid flow velocity is 1BV/h, and stripping liquid consumption is 3BV.
4) by step 3) in wash-out polyphenol solution out collect, freeze-drying, obtains cumin polyphenol.Outward appearance is shown in Fig. 4 it (c).
The extraction of embodiment 5, cumin protein isolate
1) pulverizing extracting in embodiment 3 after residue 50 DEG C remaining after polyphenol is dried with Universalpulverizer, crossing 40 eye mesh screens;
2) by step 1) pulverize after after powder mix with 1:25 ratio with phosphate buffered saline buffer (pH value is 8), stirring extraction 2.5h at 35 DEG C;
3) by step 2) centrifugal 20min under the mixed solution 7500g condition that obtains, collect supernatant liquor, ultrafiltration and concentration, freeze-drying, obtains cumin protein isolate.Outward appearance is shown in Fig. 4 it (d).
The extraction of embodiment 6, cumin polypeptide
1) after being mixed with 1:10 with the phosphate buffered saline buffer of pH 7.7 by the cumin protein isolate obtained in embodiment 5, add Sumizyme MP Alcalase2.4L by enzyme-to-substrate concentration ratio 4:100, put into sealed bag after above-mentioned sample mix is even for subsequent use;
2) by step 1) in sample be placed in extra-high tension unit, 300MPa, process 8min at 55 DEG C;
3) by step 2) enzymolysis solution that obtains heats 15min, the activity of deactivating enzyme in boiling water bath;
4) by step 3) deactivating enzyme live after enzymolysis solution centrifugal 20min under 7500g condition of obtaining, collect supernatant liquor, 65 DEG C of rotary evaporations remove moisture, and the concentrated solution freeze-drying obtained is cumin polypeptide.Outward appearance is shown in Fig. 4 it (e).
The extraction of embodiment 7, cumin food fibre
After the precipitation obtained after centrifugal in the supernatant liquor obtained in embodiment 2 and embodiment 6 is merged, add and be preheated to 60 DEG C, 95% ethanol, ethanol: (supernatant liquor+precipitation)=1:3 (volume ratio).Room temperature is centrifugal after leaving standstill 1h, and collecting precipitation after 50 DEG C of oven dry, is pulverized, obtained cumin diet fiber product in baking oven.Outward appearance is shown in Fig. 4 it (f).The biscuit that the cumin food fibre obtained by the present embodiment method is made and steamed bun photo Fig. 5.
The extraction of embodiment 8, cumin stalk food fibre
1) by cumin stalk branches and leaves with after pulverizing with Universalpulverizer, 40 eye mesh screens are crossed;
2) by step 1) after the cumin straw powder that obtains mixes with 1:20 ratio with the phosphoric acid buffer of pH7.7, adopt high pressure homogenizer to circulate under 250bar condition homogeneous 3 times, obtain cumin stalk suspension;
3) by step 2) the cumin stalk suspension that obtains is placed in Erlenmeyer flask, and be placed in 55 DEG C of isothermal vibration water-baths, add Sumizyme MP Alcalase 2.4L by enzyme-to-substrate concentration ratio 1.5:100, enzymolysis 150min;
4) by step 3) enzymolysis solution that obtains heats 15min, the activity of deactivating enzyme in boiling water bath;
5) by step 4) deactivating enzyme live after enzymolysis solution centrifugal 15min under 7500g condition of obtaining, collecting precipitation, is placed in 55 DEG C of baking ovens and dries, and pulverizes, and packaging, obtains cumin stalk food fibre.Outward appearance is shown in Fig. 4 it (g).
Embodiment 1-8 overall flow is as Fig. 1.
Experimental example 1, cumin essential oil basal component are analyzed
Analyze the basal component of cumin essential oil in embodiment 1, concrete steps are as follows:
Cumin essential oil normal hexane dilutes 10 times, adopts gas-chromatography to analyze.Analytical conditions for gas chromatography: initial temperature 60 DEG C, retains 1min, with 5 DEG C/min ramp to 160 DEG C, then with 20 DEG C/min ramp to 220 DEG C, retains 8min; Injector temperature 250 DEG C; Sample size 0.5 μ l; Carrier gas (helium) flow 1ml/min; Splitting ratio 100:1.Mass spectroscopy condition: ionization mode EI; Ionization voltage 70eV; Ion source temperature 260 DEG C; Quality of scanning scope 30-600amu.Component resolving: reference standard mass-spectrometric data storehouse is resolved, adopts ionic current areas of peak normalization method to calculate each composition percentage contents.The results are shown in Figure 2 and table 1.
This composition analysis of table 1 cumin (essence) oil base
As can be seen from Table 1, the cumin essential oil adopting ultrasonic microwave to assist extraction by steam distillation mainly containing 20 kinds of compositions, wherein content account for more than 10% be respectively cumylene (33.28%), 2-carene (22.03%), γ-terpinene (16.56%) and left-handed-beta-pinene (10.41%).
Fatty acid analysis in experimental example 2, ziran oil resin
Analyze the lipid acid of the ziran oil resin that embodiment 2 obtains, concrete steps are as follows:
1) methyl esterification of fatty acid: add 6ml hydrochloric acid-methanol (1:0.85, v/v) solution in 0.5g ziran oil resin, and at 85 DEG C water-bath 10min.Secondly, add 2ml normal hexane, make fatty acid methyl ester transfer in organic phase, rear adding distil water, to bottleneck, takes out supernatant, then uses 1ml normal hexane extraction once, merges supernatant liquor, to be measured after filtering.
2) fatty acid determination: adopt Varian 450 – GC type gas chromatograph to carry out, detector is hydrogen flame ionization detector (FID), and capillary column is CP-Wax 52CB type (30m × 320 μm × 0.5 μm, Agilent, California, the U.S.).Operational condition is as follows: initial temperature be 150 DEG C keep 1min, after be warming up to 230 DEG C with 10 DEG C/min speed, keep 15min; Sampler temperature is 220 DEG C; Detector temperature is 300 DEG C; Carrier gas is nitrogen; Flow rate of carrier gas is 1mL/min.The results are shown in Figure 3 and table 2.
Table 2 ziran oil resin fatty acid analysis
Table 2 result shows, adopt the ziran oil resin that enzymolysis-acetone combined extraction technology extracts to comprise 9 kinds of lipid acid, be respectively tetradecanoic acid (C14:0), palmitinic acid (C16:0), Zoomeric acid (C16:1), stearic acid (C18:0), oleic acid (C18:1), linolic acid (C18:2), alpha-linolenic acid (C18:3), eicosanoic acid (C20:0) and erucic acid (C22:1).Wherein, linolic acid (C18:2) and oleic acid (C18:1) are main lipid acid moietys, are respectively 39.40% and 32.40%.The unsaturated fatty acid content of ziran oil resin is higher, is 77.82%.Have report to point out, unsaturated fatty acids is at reduction blood cholesterol levels and blood fat, and preventing cardiovascular disease, protects the meaning in brain and neural system very large.
Total phenol content analysis in experimental example 3, cumin polyphenol
Cumin polyphenol 0.5g adding distil water in Example 4 after purifying is mixed with 0.05% polyphenol solution, then the forint phenol reagent of 1.0mL10% (v/v) is added, 30min is reacted in 30 DEG C of water-baths, then the sodium carbonate solution of 2.0mL10% (w/v) is added, in 30 DEG C of water-baths, react 30min after abundant mixing, under 736nm, measure absorption photometric value immediately.Adopt the chlorogenic acid standard solution Criterion curve of 0.02,0.04,0.06,0.08 and 0.10mg/mL, obtain equation of linear regression y=8.7671x+0.0068, R2=0.9994.The total phenol content of sample solution is expressed as mg chlorogenic acid equivalent (chlorogenic acid equivalent, CAE)/mL sample solution.
Polyphenol content and purity in table 3 cumin
As can be seen from Table 3,100g degreasing (essential oil and oleo-resinous) cumin powder, after extraction purification, can obtain 6.89g cumin polyphenol, be red-brown powder; The purity of cumin polyphenol is 88.42 ± 1.03%.
The amino acid composition analysis of experimental example 4, cumin protein isolate and cumin polypeptide
Cumin protein isolate in Example 5 and embodiment 6 and cumin polypeptides freeze-dry powder 75mg, with 10mL 6M HCl enzymolysis 24h at 110 DEG C.After hydrolysis, mixed solution is poured in 50mL volumetric flask, use ultrapure water constant volume, be then evaporated to dry under 60 DEG C of vacuum conditions.Dry sample being dissolved in pH value is in the sodium citrate buffer solution of 2.2, and adjustment amino acid concentration is 50-250nmol/mL, and then loading carries out determined amino acid to Hitachi L-8800 amino acidanalyser.
Table 4 cumin protein isolate and polypeptide amino acid compositional analysis (g amino-acid residue/100g butt sample)
As can be seen from Table 4, in cumin protein isolate and cumin polypeptide all containing 9 kinds of non-essential amino acid and 8 kinds of indispensable amino acids.In cumin proteins and peptides, the content of Glu and Asp is higher, and this two seed amino acid has very strong radical scavenging activity; The fragment comprising Glu and Pro has hypoglycemic activity.In addition, in cumin proteins and peptides, the content of Methionin is higher, is respectively 4.59% and 4.51%, can make up the shortage of Methionin in the foods such as cereal.Above result shows that cumin protein isolate and cumin polypeptide are good natural products.
The basal component analysis of experimental example 5, cumin food fibre, cumin stalk food fibre
Determination and analysis is carried out to the moisture of 2 kinds of food fibres that embodiment 7 and embodiment 8 obtain, protein, fat, starch, ash content, total dietary fiber content:
1, moisture determination: moisture determination adopts GB5009.3-2010.Get clean aluminum weighing bottle, be placed in 101 DEG C of-105 DEG C of loft drier, bottle cap tiltedly props up in bottle limit, heating 1.0h, and taking-up is built, and puts cooling 0.5h in moisture eliminator, weighs, and repeats to be dried to that front and back twice are of poor quality is no more than 2mg, is constant weight.Take four kinds of food fibre 3g-5g (being accurate to 0.0001g) respectively, put in weighing bottle, sample thickness is no more than 5mm, add a cover, after precision weighing, put in 101 DEG C ~ 105 DEG C loft drier, bottle cap tiltedly props up in bottle limit, after dry 2h-4h, build taking-up, weigh after putting into moisture eliminator cooling 0.5h.And then put into 100 DEG C of dry about 1h of-105 DEG C of loft drier, take out, weigh again after putting into moisture eliminator cooling 0.5h.And repeat above to be operated to that front and back twice are of poor quality is no more than 2mg, be constant weight.
Result calculates
Moisture content (%)=100 × (m
1-m
2)/(m
1– m
3)
In formula:
M
1---the quality of weighing bottle and sample, g;
M
2---the quality after weighing bottle and samples dried, g;
M
3---the quality of weighing bottle, g.
During moisture content >=1g/100g, calculation result retains three position effective digitals; During moisture content < 1g/100g, result retains two position effective digitals.
Note: during twice constant weight value in the end calculates, gets last weighing value.
2, protein content determination: take 0.50g tetra-kinds of food fibres respectively and put into alimentary canal, add the vitriol oil (concentration 98%) 10mL, digestion temperature 420 DEG C, time 1.5h, measures the protein content (Foss company of Sweden KIELTEC ANALYSISER kjeldahl apparatus) in four kinds of food fibres with kjeldahl apparatus.
3, fat test: take 1.0g tetra-kinds of food fibres respectively and be placed in clean paper sleeve, add a small amount of absorbent cotton, in lixiviate beaker, add 80mL sherwood oil, extract fat in sample with the automatic fatty detector of Foster Kato company Soxtec Avanti 2050.After lixiviate terminates, take out extraction cup, and extraction cup is placed in 100 DEG C of loft drier 30min, cool in moisture eliminator and weigh again, calculate lipid content.
Lipid content (%)=W
2× 100%/W
1
W
1example weight before-lixiviate, g;
W
2fat weight after-lixiviate drying, g.
4, starch test: measure according to the method for AOAC996.11.Get four kinds of food fibres (10mg) respectively and join (16*120mm) in glass test tube, rap test tube, to guarantee that all samples are all fallen bottom test tube; Add in 0.2mL80% ethanol to sample and increase its solvability, mix with turbine mixer; Add the Thermostable α-Amylase (100U/mL) of 3mL immediately, in boiling water bath, hatch 6min (the 2nd, 4,6min shakes test tube energetically); Add 0.1mL starch glucolase (3300U/mL), with turbine mixer mixing, water-bath 30min at 50 DEG C; Transferred to by the test tube of total Test in 100mL volumetric flask, clean with wash bottle cleaning down, use distilled water constant volume, mixing, decile solution is centrifugal 10min under 3000r; Diluting soln after transfer decile (0.1mL) is in glass test tube; Add glucose oxidase (Glucose oxidase plus peroxidase, the GOPOD) reagent of 3mL to (comprise D-Glucose control group and blank group) in each test tube, water-bath 20min at 50 DEG C; D-Glucose control group comprises 0.1mLD-glucose solution and 3.0mLGOPOD reagent, and blank group comprises 0.1mL hydration 3.0mLGOPOD reagent; The absorbancy of working sample, D-Glucose control group and blank group under 510nm.Calculate by following formula:
Starch content (%)=(A
1-A
2) * (F/W) * FV*0.9
A
1the absorbancy of-sample;
A
2the absorbancy of-blank group;
The absorbancy of F-100/ control group;
W-example weight, g;
The volume of FV-final constant volume, mL.
5, determination of ash: determination of ash is with reference to the method for GB 5009.4-2010.Concrete steps are: the porcelain crucible getting suitable size is put in retort furnace, and calcination 0.5h at 550 DEG C ± 25 DEG C, is cooled to about 200 DEG C, take out, put into moisture eliminator and cool 30min, precise.Repeat calcination to front and back twice weighing difference to be no more than 0.5mg be constant weight.Then, take 3g-10g (being accurate to 0.0001g) four kinds of food fibres respectively and be placed in porcelain crucible, first on hot plate, with little fire heating, sample is fully carbonized to smokelessly, be then placed in retort furnace, at 550 DEG C ± 25 DEG C calcination 4h.Be cooled to about 200 DEG C, take out, put into moisture eliminator and cool 30min, time before weighing as found that ignition residue has a carbon granule, little water should be instilled in sample moistening, making blocking loosening, evaporating water again calcination to namely representing that without carbon granule ashing completely, can weigh.Repeat calcination to front and back twice weighing difference to be no more than 0.5mg be constant weight.Be calculated as follows.
X
1＝100×(m
1-m
2)/(m
3-m
2)
X
1ash oontent in-sample, g/100g;
M
1the quality of-crucible and ash content, g;
M
2the quality of-crucible, g;
M
3the quality of-crucible and sample, g.
Note: the absolute difference of twice that obtains under repeated condition independent measurement result must not exceed 5% of arithmetical av.
6, dietary fiber content measures: carry out with reference to AOAC 991.43 method.
Concrete grammar is: take four kinds of food fibre 1.000 ± 0.005g (being accurate to 0.1mg) respectively in 100mL beaker, add 40mL MES-TRIS (2-(N-morpholino) sulfonic group ethane-three hydroxyl (methylol) aminomethane) damping fluid, pH8.2, is stirred to and is uniformly dispersed; Add 50 μ L heat-resistant alpha-amylase liquid, magnetic stirring apparatus stirring at low speed, and hatch 30min in boiling water bath after, be cooled to 60 DEG C, residue on 10mL distilled water flushing beaker inwall; Add the HCl of 5mL 0.561M, and constantly stir, NaOH or HCl of rear 1M at 60 DEG C adjust ph to 4.0-4.7; Add 100 μ L amyloglucosidase solution, fully mix, oscillation incubation 30min at 60 DEG C; Add 100uL protein enzyme solution, fully mix, oscillation incubation 30min at 60 DEG C; In beaker, add 95% ethanol (95% ethanol and mixeding liquid volume to be measured are than 4:1) that 225mL is preheated to 60 DEG C, under room temperature, precipitate 1h; Enzymolysis solution after alcohol settling is transferred in crucible, with residue in 78% ethanol purge beaker, proceed to suction filtration in crucible in the lump, use 78% ethanol, 95% ethanol and acetone cleaning crucible 2 times more respectively, then crucible being placed in 105 DEG C of baking ovens placements spends the night to constant weight, record crucible and residue weight (W
2).Measure the content of protein, ash content in residue, its weight is designated as P, A respectively.
Result calculates:
Dietary fiber content (%)=100 × (W
2-W
1)/W-P-A
W-example weight, g;
W
1-crucible and diatomaceous weight, g;
W
2the weight of-crucible, diatomite and residue, g;
The content of protein in P-residue, g/100g;
The content of ash content in A-residue, g/100g.
Note: the absolute difference of twice that obtains under repeated condition independent measurement result must not exceed 5% of arithmetical av.
7, the mensuration of mineral element: carry out with reference to GB19644-2010 method.
Concrete grammar: claim 2-3g cumin food fibre and cumin stalk food fibre in crucible, be placed on electric furnace and carry out concentrated and carbonization by suitable temperature, until cumin food fibre and cumin stalk food fibre become black completely, no longer produce thick smoke, namely now carbonization is complete.Be put in muffle furnace by crucible and carry out ashing, 650 DEG C of calcination 3-4h, until the powder of the solid-state grizzle of black, then ashing is complete.Take out crucible and be cooled to room temperature, add 5mL (1:3) hydrochloric acid, boil on electric furnace after abundant dissolving, filter with quantitative paper, move in 25mL volumetric flask, constant volume, liquid now in volumetric flask is transparent liquid, for measuring the content of iron, then use 25 times of diluent determining calcium, with 500 times of diluent determining potassium and magnesium.The standard series of selected potassium, calcium, iron, magnesium, by changing the instrument conditions such as acetylene flow, lamp current, combustion head height, surveying its absorbancy, determining the best instrument condition of each element determination.The standard series of selected potassium, calcium, iron, magnesium, under above-mentioned best instrument and test conditions, the linearity range of mensuration potassium, calcium, iron, magnesium and detectability.
Table 5 cumin food fibre and cumin stalk food fibre basal component analyze (butt)
Table 5 result shows, in cumin food fibre and cumin stalk food fibre, total dietary fiber content is respectively 84.18% and 81.74%; In addition, two kinds of food fibres, all containing mineral elements such as higher potassium, calcium, magnesium, iron, therefore, can supplement the new source of mineral substance as human body.
Implementation result
Cumin as conventional seasonings, containing this several functions nutritive ingredient such as a kind of characteristic chemical constituent and food fibre, polyphenol and albumen of essential oil.The cumin essential oil made according to the method in embodiment 1 has the distinctive fragrance of cumin, and containing 20 kinds of volatile components.This essential oil can be applied to food service industry as antioxidant, such as, added in meat industry by cumin essential oil and not only can play the effect of rendering palatable, can also suppress the oxidation of meat product; Essential oil also can be used as functional ingredient and adds in medicine effects such as playing prevention stomach cold stomachache, control diarrhoea to; In addition cumin essential oil also can play the effect such as protection against rodents, bactericidal.
The ziran oil resin made according to the method in embodiment 2 can add in food as seasonings, such as, roast goods, refrigerated products, puffed food and food flavouring etc.
The cumin polyphenol made according to the method in embodiment 3 and 4 is red-brown powder, has stronger oxidation-resistance, can be added in protective foods.
The cumin protein isolate made according to the method in embodiment 5 is yellow powder, has good whipability and emulsifying property etc., can be applicable to food service industry; In addition, the anti-oxidant activity of cumin protein isolate and α-amylase rejection ability are also comparatively strong, therefore can be applied to field of health care products.
The cumin polypeptide made according to the method in embodiment 6 is buff powder, and anti-oxidant activity is comparatively strong, can develop cumin protein peptide drink, plays the effects such as anticancer, anti-cancer, immunity moderation.
The cumin food fibre made according to the method for embodiment 7 and embodiment 8 and stalk dietary fiber particles less, volume is fluffy, there is good water-holding power, water retention capacity and fat absorbing ability, can be added in food, as water-holding agent, oil protecting agent and leavening agent etc.; In addition, can add in protective foods as functional ingredient, there is the effects such as hypoglycemic, hypotensive, reducing blood-fat, minimizing inflammation.
Claims (10)
1. the method for comprehensive utilization of a cumin, it is characterized in that, adopt ultrasonic microwave to assist steam distillation and enzymolysis-acetone combined extraction method to extract cumin essential oil and oleo-resinous, then adopt kinds of processes from degreasing cumin, extract cumin polyphenol, cumin protein isolate, cumin polypeptide and cumin food fibre; And adopt high-pressure homogeneous auxiliary enzymes solution to extract food fibre from cumin stalk; Realize the complete utilization of the seed of cumin plant, branches and leaves and stalk.
2. method of comprehensive utilization according to claim 1, is characterized in that, comprises the steps:
1) after cumin seed mixes with 1:3-1:5 mass volume ratio with water, at ultrasonic wave 30-50kHz, pre-treatment 10-30min under microwave 30-60W; Then the cumin seed through ultrasonic microwave process is carried out distillation method extraction, Extracting temperature is 96-100 DEG C, the oil content obtained is collected, is packed, obtains cumin essential oil;
2) by step 1) extract cumin seed residue remaining after essential oil and dry or dry be crushed to grain size < 400 μm after its moisture content is 8%-10%, the sodium acetate buffer getting powder and pH4.0-6.0 after pulverizing mixes with 1:10 mass volume ratio and shakes up, zytase and the cellulase mixed enzyme solution of 0.5%-5% is added by enzyme-to-substrate concentration ratio, 45-55 DEG C of enzymolysis 30-180min, centrifugation, gained precipitation is for extracting ziran oil resin, and supernatant liquor saves backup;
The precipitation obtained after enzymolysis is added acetone extraction by solid-to-liquid ratio 1:2-1:5, then centrifugation, gained supernatant liquor adopts the method for evaporation that acetone is volatilized and removes, and gained deep green viscous liquid is ziran oil resin;
3) by step 2) add acetone after after the residue that obtains of centrifugation mixes with 1:10-1:20 ratio with 65%-70% ethanol, supersound extraction 10-60min, then centrifugation, collect supernatant liquor, residue repeats to extract 2-3 time as stated above, united extraction liquid, evaporative removal ethanol, obtains cumin polyphenolic extract;
4) by step 3) obtain cumin polyphenol after the polyphenolic extract freeze-drying that obtains and slightly carry powder, with the HCl solution of pH 3-4.5, cumin polyphenol is slightly carried powder and be formulated as the solution that polyphenol content is 0.2-2.0mg CAE/mL, then chromatography column is loaded to, AB-8 macroporous resin selected by filler, stripping liquid flow velocity is 0.25-2.50BV/h, and adsorption temp is 25-35 DEG C, is collected by wash-out polyphenol solution out, freeze-drying, obtains cumin polyphenol;
5) by step 3) extract residue remaining after polyphenol in 45-55 DEG C of oven dry, pulverize after 30-70 eye mesh screen, then after mixing with 1:10-1:40 ratio with phosphate buffered saline buffer, stir under 30-40 DEG C of condition and extract 1-3h, centrifugation, collect supernatant liquor, ultrafiltration and concentration, freeze-drying, obtains cumin protein isolate;
6) by step 5) the cumin protein isolate that obtains, by enzyme-to-substrate concentration ratio (1-6): 100 add proteolytic enzyme, be placed in extra-high tension unit, after 100-400MPa, 40-60 DEG C of process 5-15min, 10-15min is with the activity of passivation proteolytic enzyme in boiling water bath heating, centrifugation, collect supernatant liquor, 60-65 DEG C of rotary evaporation removes moisture, and the concentrated solution freeze-drying obtained, is cumin polypeptide;
7) by step 2) supernatant liquor that obtains and step 6) centrifugal after after the precipitation that obtains merges, add massfraction be 85-99%, be preheated to the ethanol of 60 DEG C leave standstill after centrifugal, collecting precipitation after 45-55 DEG C of oven dry, is pulverized, is obtained food fibre in baking oven;
8) 30-70 mesh sieve is crossed after cumin branches and leaves remaining when being gathered in the crops by cumin and crushed stalk, after mixing with 1:10-1:30 ratio with the phosphate buffered saline buffer of pH7.0-8.0, adopt high pressure homogenizer 100-350bar circulation homogeneous 1-5 time, then by enzyme-to-substrate concentration ratio (1-2.5): 100 add proteolytic enzyme, under 50-65 DEG C of condition after enzymolysis 120-240min, boiling water bath keeps 10-15min with the activity of passivation proteolytic enzyme, centrifugation, after gained precipitation 45-55 DEG C is dried, pulverize, be cumin stalk food fibre.
3. method of comprehensive utilization according to claim 2, it is characterized in that, described step 2) in, 30-70 eye mesh screen is crossed by after the precipitation oven dry obtained after enzymolysis, pulverizing, acetone is added by solid-to-liquid ratio 1:2-1:5, stir with rotating speed 100-150rpm under room temperature and extract the centrifugal 10-30min separation of 30-60min, 4000-8000g.
4. the method for comprehensive utilization according to Claims 2 or 3, is characterized in that, described step 2) in, the temperature of oven dry is 45-55 DEG C.
5. method of comprehensive utilization according to claim 2, it is characterized in that, described step 3) in, residue 45-55 DEG C of oven dry is pulverized after moisture content is 8%-10%, after mixing with ethanol, under ultrasonic wave 45-55kHz, microwave 100-150W, extract 10-60min.
6. method of comprehensive utilization according to claim 2, is characterized in that, described step 3), step 5), step 6) and step 8) in the condition of centrifugation be: centrifugation 10-30min under 4000-8000g.
7. method of comprehensive utilization according to claim 2, is characterized in that, described step 4) in, the stripping liquid flow velocity of chromatography column is 0.25-2.50BV/h, and adsorption temp is 25-35 DEG C, and adsorption time is 1-3.5h; After absorption reaches balance, adopt the ethanolic soln of 65-75% to carry out desorb, stripping liquid flow velocity is 0.3-2.5BV/h, and stripping liquid consumption is 3-4BV.
8. method of comprehensive utilization according to claim 2, is characterized in that, described step 6) in, cumin protein isolate add enzyme before adjust ph, the method for adjustment is mix with 1:10 mass volume ratio with the phosphate buffered saline buffer of pH7.0-8.0.
9. method of comprehensive utilization according to claim 2, is characterized in that, described step 6) in, after supernatant liquor and precipitation merge, add and be preheated to 60 DEG C, 95% ethanol, room temperature is centrifugal after leaving standstill 1h.
10. the product for preparing of the arbitrary described method of comprehensive utilization of claim 1-9.
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| CN108004022A (en) * | 2017-12-12 | 2018-05-08 | 湖北中烟工业有限责任公司 | Preparation method for the Cuminum cyminum extract of quick-fried pearl base oil |
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