CN103361320A - Method of extracting SOD from milk - Google Patents
Method of extracting SOD from milk Download PDFInfo
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- CN103361320A CN103361320A CN2013103195384A CN201310319538A CN103361320A CN 103361320 A CN103361320 A CN 103361320A CN 2013103195384 A CN2013103195384 A CN 2013103195384A CN 201310319538 A CN201310319538 A CN 201310319538A CN 103361320 A CN103361320 A CN 103361320A
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Abstract
The invention provides a method of extracting SOD from milk. The method comprises following steps: cooling fresh milk, distilled water, and 95% ethanol below the temperature of 10 DEG C in advance, mixing the cooled distilled water and milk according to a ratio of 2:1, adding cooper gluconate with a weight of 40% of milk weight and zinc gluconate with a weight of 30% of milk weight, evenly stirring, filtering, collecting the clear liquid, adding ethanol with a weight two times of that of milk and distilled water in the clear liquid, stirring; adjusting the pH value of the solution to 7 to 7.5 with a sodium hydroxide solution of 30 mol/L, collecting the filtrate, adding ethanol with a weight three times of that of milk and distilled water in the filtrate, evenly stirring, standing, filtering, collecting the deposition, drying in low temperature, grinding, and recycling ethanol through distillation. The method of extracting SOD from milk has the advantages of high extraction rate, high activity, low cost, safe, green and non-polluted extraction technology, and simple equipment.
Description
Technical field
The present invention relates to field of food, especially a kind of method of from milk, extracting SOD.
Background technology
" a kind of SOD prozyme and preparation method thereof ", this invention relates to a kind of healthcare products, particularly relates to a kind of SOD prozyme and preparation method thereof, the albumosease type SOD that from fresh milk, extracts, its enzyme activity is high, and it has anti-oxidant, many effects such as delaying human body caducity.Propolis in the prescription has blood fat reducing and cholesterol, and strengthens vasodilation power, reduces vascular fragility, and active cells improves the effects such as immunizing power.The two its functional effect behind the science compatibility improves greatly, and safety non-toxic is nuisanceless.
" purification and the evaluation of superoxide-dismutase in the cow's milk (SOD) " reported purification and the evaluation of superoxide-dismutase (SOD) in the cow's milk.Fresh milk is through ultracentrifugation, and chloroform-Ethanol Treatment, the steps such as acetone precipitation, dialysis, DEAE cellulose chromatography make wherein that Cu-ZnSOD has obtained purifying, and the specific activity of enzyme is 149.5V/mg after purifying.(Sun Qinglin, etc. the purification of superoxide-dismutase in the cow's milk (SOD) and evaluation [J]. Nei Mongol Agriculture and Animal Husbandry College journal, 1998 (4): 34-36).
" cow's milk and superoxide-dismutase wherein " reported cow's milk and superoxide-dismutase wherein.This literary composition sketched superoxide-dismutase (SOD) discovery, distribute and present development and use situation at organic sphere, simultaneously the type of contained SOD in the cow's milk and content, measuring method, extraction and purifying and characteristic thereof have been carried out introducing briefly, and inquired into regard to the cow's milk of production enhancement SOD and meaning and the feasibility of milk-product thereof.The preparation method of this civilian cow's milk SOD is, uses (NH4) 2SO4 to saltout, and the throw out that obtains is dissolved among a small amount of SMUF again, crosses post, uses K
2HPO
4(pH7.4) wash-out is collected the elutriant freeze-drying and is got finished product.(Feng Qihao. cow's milk and superoxide-dismutase [J] wherein. China Dairy Industry, 1993 (2): 68-73).
Summary of the invention
The objective of the invention is the deficiency for existing SOD extractive technique, a kind of method of extracting SOD from milk is provided, form active higher SOD product.
For realizing above-mentioned goal of the invention, the embodiment that the present invention takes is as follows:
(1) raw milk, distilled water, 95% ethanol are pre-chilled to below 10 ℃ in advance, for subsequent use;
The distilled water that (2) will be pre-chilled to below 10 ℃ mixes according to 2: 1 ratio with milk;
(3) in above-mentioned mixed solution, add the copper gluconate of milk amount 40% and the Zinc Gluconate of milk amount 30%, stir, filter, collect clear liquid, for subsequent use;
(4) in above-mentioned clear liquid, add the ethanol of 2 times of milk and distilled water total amounts, stir;
(5) with the sodium hydroxide solution of 30mol/L the pH value of mentioned solution is transferred to 7-7.5, left standstill 5-8 hour;
(6) filter, collect filtrate, add the ethanol of 3 times of milk and distilled water total amounts, stir, left standstill 5-8 hour;
(7) filter collecting precipitation, cryodrying, pulverizing;
(8) ethanol distillation reclaims.
The invention has the beneficial effects as follows: SOD product of the present invention, extraction yield is high, activity is high, cost is low, the extraction process safe green is pollution-free, equipment is simple.
Embodiment
Below in conjunction with embodiment, to details are as follows according to embodiment provided by the invention:
Embodiment 1
1. raw milk detects qualified rear filtration, purification and is pre-chilled to 4 ℃, and distilled water and 95% ethanol are carried be pre-chilled to 4 ℃ simultaneously,
For subsequent use;
2. getting the distilled water 200g that is pre-chilled to 10 ℃ mixes with 100g milk;
3. add the copper gluconate of 40g and the Zinc Gluconate of 30g in above-mentioned mixed solution, the stillness of night is collected in the filtration that stirs, and is for subsequent use;
4. in the above-mentioned stillness of night, add the cold ethanol of 600g, stir;
5. with the sodium hydroxide solution of 30mol/L the pH value of mentioned solution is transferred to 7.4, left standstill 5 hours;
6. filter, collect filtrate, add the cold ethanol of 900g, stir, left standstill 6 hours;
7. filter, collecting precipitation, is ground into fine powder at cryodrying;
8. surveying the SOD activity with the pyrogallol Autoxidation Method is 122859U/g;
9. ethanol distillation reclaims.
Embodiment 2
1. raw milk detects qualified rear filtration, purification and is pre-chilled to 8 ℃, distilled water and 95% ethanol is carried be pre-chilled to 8 ℃ simultaneously, and is for subsequent use;
2. getting the distilled water 500g that is pre-chilled to 8 ℃ mixes with 250g milk;
3. add the copper gluconate of 100g and the Zinc Gluconate of 75g in above-mentioned mixed solution, the stillness of night is collected in the filtration that stirs, and is for subsequent use;
4. in the above-mentioned stillness of night, add the cold ethanol of 1500g, stir;
5. with the sodium hydroxide solution of 30mol/L the pH value of mentioned solution is transferred to 7.3, left standstill 6 hours;
6. filter, collect filtrate, add the cold ethanol of 2250g, stir, left standstill 7 hours;
7. filter, collecting precipitation, is ground into fine powder at cryodrying;
8. surveying the SOD activity with the pyrogallol Autoxidation Method is 125015U/g;
9. ethanol distillation reclaims.
Embodiment 3
1. raw milk detects qualified rear filtration, purification and is pre-chilled to 10 ℃, distilled water and 95% ethanol is carried be pre-chilled to 10 ℃ simultaneously, and is for subsequent use;
2. getting the distilled water 2000g that is pre-chilled to 10 ℃ mixes with 1000g milk;
3. add the copper gluconate of 400g and the Zinc Gluconate of 300g in above-mentioned mixed solution, the stillness of night is collected in the filtration that stirs, and is for subsequent use;
4. in the above-mentioned stillness of night, add the cold ethanol of 6000g, stir;
5. with the sodium hydroxide solution of 30mol/L the pH value of mentioned solution is transferred to 7.4, left standstill 7 hours;
6. filter, collect filtrate, add the cold ethanol of 9000g, stir, left standstill 7.5 hours;
7. filter, collecting precipitation, is ground into fine powder at cryodrying;
8. surveying the SOD activity with the pyrogallol Autoxidation Method is 126236U/g;
9. ethanol distillation reclaims.
Above one embodiment of the present of invention are had been described in detail, but described content only is preferred embodiment of the present invention, can not be considered to be used to limiting practical range of the present invention.All equalizations of doing according to the present patent application scope change and improve etc., all should still belong within the patent covering scope of the present invention.
Claims (1)
1. method of from milk, extracting SOD, its step is as follows:
(1) raw milk, distilled water, 95% ethanol are pre-chilled to below 10 ℃ in advance, for subsequent use;
The distilled water that (2) will be pre-chilled to below 10 ℃ mixes according to 2: 1 ratio with milk;
(3) in above-mentioned mixed solution, add the copper gluconate of milk amount 40% and the Zinc Gluconate of milk amount 30%, stir, filter, collect clear liquid, for subsequent use;
(4) in above-mentioned clear liquid, add the ethanol of 2 times of milk and distilled water total amounts, stir;
(5) with the sodium hydroxide solution of 30mol/L the pH value of mentioned solution is transferred to 7-7.5, left standstill 5-8 hour;
(6) filter, collect filtrate, add the ethanol of 3 times of milk and distilled water total amounts, stir, left standstill 5-8 hour;
(7) filter collecting precipitation, cryodrying, pulverizing;
(8) ethanol distillation reclaims.
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201310319538.4A CN103361320B (en) | 2013-07-25 | 2013-07-25 | Method of extracting SOD from milk |
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| Application Number | Priority Date | Filing Date | Title |
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| CN201310319538.4A CN103361320B (en) | 2013-07-25 | 2013-07-25 | Method of extracting SOD from milk |
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| CN103361320A true CN103361320A (en) | 2013-10-23 |
| CN103361320B CN103361320B (en) | 2014-06-18 |
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Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101240267A (en) * | 2008-03-12 | 2008-08-13 | 许棚举 | SOD complex enzyme and preparation method thereof |
| CN101338305A (en) * | 2008-08-28 | 2009-01-07 | 新疆维吾尔自治区药物研究所 | Process for extracting superoxide dismutase and reduction glutathion from animal blood |
| CN102181411A (en) * | 2011-04-01 | 2011-09-14 | 黑龙江宝迪肉类食品有限公司 | Method for extracting superoxide dismutase (SOD) from porcine blood red cells |
-
2013
- 2013-07-25 CN CN201310319538.4A patent/CN103361320B/en active Active
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101240267A (en) * | 2008-03-12 | 2008-08-13 | 许棚举 | SOD complex enzyme and preparation method thereof |
| CN101338305A (en) * | 2008-08-28 | 2009-01-07 | 新疆维吾尔自治区药物研究所 | Process for extracting superoxide dismutase and reduction glutathion from animal blood |
| CN102181411A (en) * | 2011-04-01 | 2011-09-14 | 黑龙江宝迪肉类食品有限公司 | Method for extracting superoxide dismutase (SOD) from porcine blood red cells |
Non-Patent Citations (3)
| Title |
|---|
| 冯启浩: "牛乳及其中的超氧化物歧化酶", 《中国乳品工业》 * |
| 孙庆林等: "牛乳中超氧化物歧化酶SOD的提纯与鉴定", 《内蒙古农牧学院学报》 * |
| 王惠生等: "奶中超氧化物歧化酶SOD活性的研究", 《西北农业大学学报》 * |
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| CN103361320B (en) | 2014-06-18 |
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Inventor after: Zhao Yongquan Inventor after: Zhang Lijuan Inventor after: Cai Yuehua Inventor after: Hu Zhaobo Inventor before: Zhao Yongquan Inventor before: Zhang Lijuan Inventor before: Cai Yuehua Inventor before: Hu Zhaobo |