CN103461645A - Preparation method of cottonseed protein - Google Patents
Preparation method of cottonseed protein Download PDFInfo
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- CN103461645A CN103461645A CN2013103828194A CN201310382819A CN103461645A CN 103461645 A CN103461645 A CN 103461645A CN 2013103828194 A CN2013103828194 A CN 2013103828194A CN 201310382819 A CN201310382819 A CN 201310382819A CN 103461645 A CN103461645 A CN 103461645A
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Abstract
The invention discloses a preparation method of cottonseed protein, which comprises the steps that (1) cottonseed meal is dephenolized; and (2) the cottonseed protein is extracted from the dephenolized cottonseed meal, wherein dephenolization of the cottonseed meal is to leach the cottonseed meal with a mixed solvent of alcohol and n-hexane, and a volume ratio of alcohol to n-hexane is (1-5):5. The dephenolized cottonseed meal is leached by a mixed aqueous solution of sodium hydroxide and sodium sulfite, and the cottonseed protein is obtained by acid precipitation of a leaching solution. With the adoption of the method, a removed rate of gossypol can reach above 80%; free gossypol (FG) is less than or equal to 400mg/kg; a yield of the cottonseed protein reaches above 60%; and the purity is greater than 90%.
Description
Technical field
The invention belongs to the cottonseed processing field, be specifically related to the preparation method of cottonseed protein.
Background technology
China is the cotton state of the large product of the first in the world, and annual cottonseed output reaches 900 * 10
4more than t, its main producing region is in the North China Plain and Plain, northwest.Cottonseed is not only important vegetable oil material, but also the protein that contains 20% left and right, be important oil plant and the protein crop that is only second to soybean in the world today, therefore for countries in the world, the byproduct Cottonseed Meal that cottonseed is got after oil becomes important plant protein resource.Due to the existence that gossypol is arranged in Cottonseed Meal, existing Cottonseed Meal processing method generally comprises two steps: Cottonseed Meal dephenolize (also referred to as detoxification), then from the dephenolize cotton dregs, extract cottonseed protein.At present, to the main employing chemical method of removing of gossypol in Cottonseed Meal, biological fermentation process, solvent extraction and liquid-liquid-solid three phase extraction method; To the extraction of cottonseed protein mainly adopt the molten acid of alkali sink method, salt is carried or zymolysis technique.
(1) cotton dregs dephenolize technology
Chemical method removes gossypol in Cottonseed Meal, and the chemical property of its technology utilization gossypol makes under certain conditions its destruction or is bonding state.Commonly used is to add sulfate (ferrous sulfate, copper sulphate), and its metal ion and dregs of rice Free Gossypol formation complex compound lose activity aldehyde radical and hydroxyl in gossypol, reach the detoxification purpose; Also have the urea of interpolation and gossypol to form the schiff bases addition product, salt adding makes gossypol oxidation in neutral environment.Its shortcoming is that formed complex compound still is retained in cotton dregs, and the palatability of the dregs of rice is poor, is difficult for digested.
Biological fermentation process is to adopt yeast that gossypol is converted into to other materials, to reach the purpose of detoxification.Because transformation mechanism is not clear, the control difficulty of process conditions is large, and detoxification efficiency can't guarantee; Cotton dregs after hot moulding leaches, in pretreatment and dregs of rice drying course, protein is sex change, and its nutritive value also can't be guaranteed; In addition, investment is also a defect of this method greatly.
Solvent extraction is to utilize the dissolubility of gossypol in solvent, usually adopts methyl alcohol, acetone etc. to be extracted, and because the dissolvent residual problem has increased the harmfulness of product, has limited the scope of application of the standby cottonseed protein of this legal system simultaneously.
(2) extractive technique of cottonseed protein
Though the heavy method of the molten acid of alkali is a kind of effective ways that prepare cottonseed protein, produces a large amount of spent acid, alkali waste water in preparation process; Although the lipidated protein of salt formulation preparation is high, extraction rate of protein less than 35%; The Enzymatic Extraction good product quality, but enzymolysis process control difficulty is large, and production cost is high.
Summary of the invention
The objective of the invention is to solve the problems referred to above in existing cottonseed protein production, the preparation method of the cottonseed protein that a kind of gossypol removal efficiency is high is provided.
The present invention realizes that the technical scheme that above-mentioned purpose adopts is as follows:
A kind of preparation method of cottonseed protein, cottonseed protein is extracted in the dephenolize and (2) that comprise (1) Cottonseed Meal from the Cottonseed Meal of dephenolize, it is characterized in that, the dephenolize of Cottonseed Meal is the mixed solvent lixiviate Cottonseed Meal with ethanol and normal hexane, wherein, the volume ratio of described ethanol and normal hexane is (1～5): 5.
Further, the volume ratio of described ethanol and normal hexane is (1～4): 5.
Further, during the lixiviate dephenolize, the solid-liquid ratio of Cottonseed Meal and mixed solvent is 1:(3～6).
Further, during the lixiviate dephenolize, extraction temperature is 40～50 ℃.
The optimal conditions of Cottonseed Meal dephenolize is: the volume ratio of ethanol and normal hexane is 4:5, and the solid-liquid ratio of Cottonseed Meal and mixed solvent is 1:4, and extraction temperature is 50 ℃.
Further, the method for extracting cottonseed protein from the Cottonseed Meal of dephenolize is: with the mixed aqueous solution of NaOH and sodium sulfite, the Cottonseed Meal of dephenolize is carried out to lixiviate, leaching liquor acid is sunk and is obtained cottonseed protein.
Further, the mass ratio of described NaOH and sodium sulfite is 5:(2～5).
Further, during the lixiviate cottonseed protein, the total concentration of NaOH and sodium sulfite is 25～35g/L.
Further, while using the mixed aqueous solution lixiviate of NaOH and sodium sulfite, extraction time is 0.5～1.5 hour.
Further, while using the mixed aqueous solution lixiviate of NaOH and sodium sulfite, extraction temperature is 50～70 ℃.
PH when further, leaching liquor acid is heavy is 4～6.
The optimal conditions that cottonseed protein extracts is: the mass ratio of NaOH and sodium sulfite is 1:1, and the total concentration of NaOH and sodium sulfite is 30g/L, solid-liquid ratio 1:15, and extraction time is 1 hour, extraction temperature is 60 ℃.
The accompanying drawing explanation
The influence curve of the volume ratio that Fig. 1 is ethanol and normal hexane in mixed solvent to the gossypol removal efficiency.
Fig. 2 is the influence curve of solid-liquid ratio to the gossypol removal efficiency.
Fig. 3 is the influence curve of extraction temperature to the gossypol removal efficiency.
The influence curve of the mass ratio that Fig. 4 is NaOH and sodium sulfite in immersion liquid to cottonseed protein yield and purity.
Fig. 5 is the influence curve of extraction time to cottonseed protein yield and purity.
Fig. 6 is for extracting the influence curve of temperature to cottonseed protein yield and purity.
The specific embodiment
Below in conjunction with embodiment and accompanying drawing, the present invention is described in further details.
The preparation method of cottonseed protein is as follows:
(1) the Cottonseed Meal raw material is pulverized;
(2) the mixed solvent lixiviate of ethanol and normal hexane for the Cottonseed Meal of pulverizing, remove the gossypol in Cottonseed Meal;
(3) Cottonseed Meal of dephenolize, with after mixed solvent separates, obtains the Cottonseed Meal of dephenolize;
(4) Cottonseed Meal of dephenolize adds water, then adds NaOH and sodium sulfite, extracts cottonseed protein;
(5) the dephenolization cottonseed dregs of rice, after lixiviate, are isolated leaching liquor, the pH to 4 of acid adding adjusting leaching liquor～6, and acid is heavy, and precipitation is cottonseed protein.
The inventive method has no particular limits Cottonseed Meal, can be the residue of cottonseed after milling process is got oil, can be also the residue adopted after the extraction method is got oil, in following instance, the Cottonseed Meal raw material be the residue after No. six extraction methods are carried oil, and gossypol content is 2230mg/kg.
Step (2) is 40 ℃ of extraction temperatures, extraction time 1h, solid-liquid ratio 1:3(w/v, g/ml) time, the volume ratio of ethanol and normal hexane on the impact of gossypol removal efficiency as shown in Figure 1, as can be seen from Figure 1, the more little removal efficiency that more is conducive to gossypol of ethanol and normal hexane ratio value, while as ethanol and normal hexane volume ratio, being 5:5, the gossypol removal efficiency can reach more than 80%, removal efficiency during for 5:1 only has 70%, therefore, during lixiviate, the volume ratio of ethanol and normal hexane is controlled at (1～5): 5.
Step (2) is 40 ℃ of temperature, time 1h, mixed solvent (ethanol: in the time of n-hexane=5:5), on the impact of gossypol removal efficiency as shown in Figure 2, as can be seen from Figure 2, solid-liquid ratio is 1:3(g/ml to solid-liquid ratio) the gossypol removal efficiency can reach more than 80%, by comparison, when solid-liquid ratio is 1:1, the gossypol removal efficiency will hang down 10%, and therefore, the solid-liquid ratio of Cottonseed Meal and mixed solvent is controlled at 1:(3～6).
Step (2) is at time 1h, during mixed solvent (ethanol: n-hexane=5:5), solid-liquid ratio 1:3(g/ml), extraction temperature on the impact of gossypol removal efficiency as shown in Figure 3, as can be seen from Figure 3, extraction temperature can make the gossypol removal efficiency reach more than 80% at 40～50 ℃.
The Cottonseed Meal of step (3) dephenolize can adopt prior art to carry out with separating of mixed solvent, as squeezing, filtration, distillation, centrifugation etc.
When step (4) cottonseed protein extracts, NaOH and sodium sulfite total concentration can be controlled in 25～35g/L.
Step (4) is at solid-liquid ratio 1:10, NaOH and sodium sulfite total concentration are 30g/L, 20 ℃ of temperature, during time 1h, NaOH and sodium sulfite different quality than under on the impact of cottonseed protein yield and purity as shown in Figure 4, as can be seen from Figure 4, the impact of different quality comparison cottonseed protein yield and purity is just in time contrary, in general, the mass ratio of NaOH and sodium sulfite is controlled at 5:(2～5), can obtain high yield and purity.
Step (4) is 20 ℃ of temperature, NaOH and sodium sulfite total concentration are 30g/L, the mass ratio of NaOH and sodium sulfite is 5:5, and during solid-liquid ratio 1:10, extraction time on the impact of cottonseed protein yield and purity as shown in Figure 5, as can be seen from Figure 5, although extraction time is longer, the cottonseed protein yield is higher, and cottonseed protein purity is fallen after rising on the contrary, therefore, extraction time is easy to control at 0.5～1.5 hour.
Step (4) is 1h in extraction time, NaOH and sodium sulfite total concentration are 30g/L, the mass ratio of NaOH and sodium sulfite is 5:5, during solid-liquid ratio 1:10, extracts temperature on the impact of cottonseed protein yield and purity as shown in Figure 6, as can be seen from Figure 6, temperature is comparatively complicated on the impact of the purity of cottonseed protein, and the higher yield of humidity is corresponding also higher, therefore, in the situation that guarantee purity, extract temperature and be controlled at 50～70 ℃.
When step (5) leaching liquor is adjusted pH, available machine acid or the inorganic acid of common are, example hydrochloric acid, sulfuric acid, nitric acid, acetic acid etc.
the mensuration of gossypol content
By " in animal feed free with assay method total gossypol " ISO 6866-1985, undertaken.
Take 1-2g sample (being accurate to 0.001 g), be placed in 250ml tool plug conical flask, add 20 beades, with pipette, accurately add the 50mL solvent orange 2 A, the jam-pack bottle stopper, put into oscillator vibration 1h (120 left and right per minute).Filter with dry quantitative filter paper, add a cover a watch crystal during filtration to reduce solvent evaporates on funnel, discard initial several filtrates, collect filtrate in 100mL tool plug conical flask.
Draw in equivalent double filtrate 5-10mL (every part of gossypol that approximately contains 50-100 μ g) the brown volumetric flask a of respectively to two 25mL and b with pipette, if necessary, with solvent orange 2 A, be supplemented to 10 mL.
With isopropyl alcohol-n-hexane mixed solvent dilution bottle a, to scale, shake up, this solution is as the reference solution of Specimen Determination liquid.
The brown volumetric flask a of solvent orange 2 A respectively to two 25mL with 2 parts of 10mL of pipette, extract
0and b
0in.
Supplement bottle a with isopropyl alcohol-n-hexane mixed solvent
0to scale, shake up, this solution is as the reference solution of blank determination liquid.
Add 2.0mL aniline in volumetric flask b and b
0in, heating 30min colour developing on boiling water bath.
Be cooled to room temperature, with isopropyl alcohol-n-hexane mixed solvent constant volume, shake up and standing 1h.
Use the 10mm colorimetric pool, at wavelength 440nm place, use spectrophotometer with a
0for reference solution is measured blank determination liquid b
0absorbance, a of take is that reference solution measures the absorbance of Specimen Determination liquid b, deducts the absorbance of blank determination liquid from the absorbance of Specimen Determination liquid, obtains proofreading and correct absorbance A.
Computing formula:
In formula: X--free gossypol content, mg/kg;
A--proofreaies and correct absorbance;
The m--sample mass, g;
V--measures the volume with filtrate, mL;
The a--mass-absorption coefficient, free gossypol is 62.5cm
-1g
-1l.
Recording total free gossypol content in the Cottonseed Meal raw material is: 2230mg/kg.
The gossypol removal efficiency: i.e. dephenolize gossypol efficiency, the larger removal effect of its value is better.
Computing formula:
In formula: Y---gossypol removal efficiency in Cottonseed Meal
M
always---total gossypol content in cottonseed
M
sample---gossypol content after the sample dephenolize.
Solvent orange 2 A compound method: measure about 500mL isopropyl alcohol, n-hexane mixed solvent (n-hexane: isopropyl alcohol=6:4(v:v)), 2 mL 3-amino-1-propyl alcohol, 8 mL glacial acetic acids and 50 mL water, in the volumetric flask of 1000 mL, then are settled to scale with n-hexane-isopropyl alcohol mixed solvent.
(1) Cottonseed Meal is pulverized;
(2) the mixed solvent lixiviate of ethanol and normal hexane for the Cottonseed Meal of pulverizing, remove the gossypol in Cottonseed Meal, wherein, the volume ratio of ethanol and normal hexane is 4:5, and the solid-liquid ratio of Cottonseed Meal and mixed solvent is 1:4(w/v, g/ml), extraction temperature is 50 ℃, 1 hour time, the gossypol removal efficiency is 89.60%, dephenolize cotton dregs Free Gossypol (FG)≤400mg/kg;
(3) Cottonseed Meal of dephenolize, with after mixed solvent separates, obtains the Cottonseed Meal of dephenolize;
(4) Cottonseed Meal of dephenolize adds water, add again NaOH and sodium sulfite, extract cottonseed protein, wherein, the solid-liquid ratio of dephenolize cotton dregs and immersion liquid is 1:15, and temperature is 60 ℃, and extraction time is 1h, in immersion liquid, the mass ratio of NaOH and sodium sulfite is 1:1, and the total concentration of NaOH and sodium sulfite is 30g/L;
(5) the dephenolization cottonseed dregs of rice, after lixiviate, are isolated leaching liquor (supernatant), and acid adding is regulated the pH to 4.8 of leaching liquor, the heavy 10min of acid, and centrifugal, precipitation is cottonseed protein, and the cotton seed protein yield is 61.44%, and purity is 91.5%.
Claims (10)
1. the preparation method of a cottonseed protein, cottonseed protein is extracted in the dephenolize and (2) that comprise (1) Cottonseed Meal from the Cottonseed Meal of dephenolize, it is characterized in that, the dephenolize of Cottonseed Meal is the mixed solvent lixiviate Cottonseed Meal with ethanol and normal hexane, wherein, the volume ratio of described ethanol and normal hexane is (1～5): 5.
2. the preparation method of cottonseed protein according to claim 1, it is characterized in that: the volume ratio of described ethanol and normal hexane is (1～4): 5.
3. the preparation method of cottonseed protein according to claim 1, it is characterized in that: during dephenolize, the solid-liquid ratio of Cottonseed Meal and mixed solvent is 1:(3～6).
4. the preparation method of cottonseed protein according to claim 1, it is characterized in that: during dephenolize, extraction temperature is 40～50 ℃.
5. the preparation method of cottonseed protein according to claim 1 is characterized in that: with the mixed aqueous solution of NaOH and sodium sulfite, the Cottonseed Meal of dephenolize is carried out to lixiviate, the heavy cottonseed protein that obtains of leaching liquor acid.
6. the preparation method of cottonseed protein according to claim 5, it is characterized in that: the mass ratio of described NaOH and sodium sulfite is 5:(2～5).
7. the preparation method of cottonseed protein according to claim 6, it is characterized in that: during lixiviate, the total concentration of NaOH and sodium sulfite is 25～35g/L.
8. the preparation method of cottonseed protein according to claim 5 is characterized in that: during with the mixed aqueous solution lixiviate of NaOH and sodium sulfite, extraction time is 0.5～1.5 hour.
9. the preparation method of cottonseed protein according to claim 5 is characterized in that: during with the mixed aqueous solution lixiviate of NaOH and sodium sulfite, extraction temperature is 50～70 ℃.
10. the preparation method of cottonseed protein according to claim 5, it is characterized in that: pH when leaching liquor acid is heavy is 4～6.
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Cited By (8)
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| CN103875999A (en) * | 2014-03-07 | 2014-06-25 | 袁峻峰 | Cottonseed meal puffing and detoxifying method |
| CN104402967A (en) * | 2014-12-03 | 2015-03-11 | 环境保护部南京环境科学研究所 | Method for extracting protein from single-cottonseed cotton batting quickly |
| CN106306329A (en) * | 2015-06-25 | 2017-01-11 | 南通海辰蛋白科技有限公司 | Extraction technology of cottonseed proteins |
| CN107011411A (en) * | 2017-04-28 | 2017-08-04 | 济南中棉生物科技有限公司 | A kind of cottonseed protein production method that can extract a variety of byproducts |
| CN110606866A (en) * | 2019-10-25 | 2019-12-24 | 仲恺农业工程学院 | Method for refining cottonseed protein |
| CN113768043A (en) * | 2021-08-09 | 2021-12-10 | 武汉轻工大学 | Method for reducing content of free gossypol in cottonseed meal and feed |
| CN115651760A (en) * | 2022-09-21 | 2023-01-31 | 新疆冠农果茸股份有限公司 | Cottonseed mixed oil refining method |
| CN116235895A (en) * | 2023-03-14 | 2023-06-09 | 西南科技大学 | Method for preparing food emulsifier based on cottonseed protein isolate |
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Cited By (10)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103875999A (en) * | 2014-03-07 | 2014-06-25 | 袁峻峰 | Cottonseed meal puffing and detoxifying method |
| CN104402967A (en) * | 2014-12-03 | 2015-03-11 | 环境保护部南京环境科学研究所 | Method for extracting protein from single-cottonseed cotton batting quickly |
| CN104402967B (en) * | 2014-12-03 | 2018-02-06 | 环境保护部南京环境科学研究所 | One kind rapid extraction method of protein from simple grain cottonseed cotton-wool |
| CN106306329A (en) * | 2015-06-25 | 2017-01-11 | 南通海辰蛋白科技有限公司 | Extraction technology of cottonseed proteins |
| CN107011411A (en) * | 2017-04-28 | 2017-08-04 | 济南中棉生物科技有限公司 | A kind of cottonseed protein production method that can extract a variety of byproducts |
| CN107011411B (en) * | 2017-04-28 | 2020-12-18 | 济南中棉生物科技有限公司 | Cottonseed protein production method capable of extracting multiple byproducts |
| CN110606866A (en) * | 2019-10-25 | 2019-12-24 | 仲恺农业工程学院 | Method for refining cottonseed protein |
| CN113768043A (en) * | 2021-08-09 | 2021-12-10 | 武汉轻工大学 | Method for reducing content of free gossypol in cottonseed meal and feed |
| CN115651760A (en) * | 2022-09-21 | 2023-01-31 | 新疆冠农果茸股份有限公司 | Cottonseed mixed oil refining method |
| CN116235895A (en) * | 2023-03-14 | 2023-06-09 | 西南科技大学 | Method for preparing food emulsifier based on cottonseed protein isolate |
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