CN103361320B - Method of extracting SOD from milk - Google Patents
Method of extracting SOD from milk Download PDFInfo
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- CN103361320B CN103361320B CN201310319538.4A CN201310319538A CN103361320B CN 103361320 B CN103361320 B CN 103361320B CN 201310319538 A CN201310319538 A CN 201310319538A CN 103361320 B CN103361320 B CN 103361320B
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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 extracting SOD from milk.
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 extracting from fresh milk, its enzyme activity is high, and it has anti-oxidant, many effects such as delaying human body caducity.Propolis in formula has the blood fat of reduction and cholesterol, and strengthens vasodilation power, reduces vascular fragility, and active cells, improves the effects such as immunizing power.The two its functional effect after science compatibility improves greatly, and safety non-toxic is nuisanceless.
" purification and the qualification of superoxide-dismutase in cow's milk (SOD) " reported purification and the qualification of superoxide-dismutase (SOD) in cow's milk.Fresh milk is through ultracentrifugation, chloroform-Ethanol Treatment, and the steps such as acetone precipitation, dialysis, DEAE cellulose chromatography make wherein Cu-ZnSOD obtain purifying, and after purifying, the specific activity of enzyme is 149.5V/mg.(Sun Qinglin, etc. the purification of superoxide-dismutase in cow's milk (SOD) and qualification [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 has been sketched the discovery of superoxide-dismutase (SOD), has been distributed and current exploitation situation at organic sphere, type to contained SOD in cow's milk and content, measuring method, extraction and purifying and characteristic thereof have carried out briefly introducing simultaneously, and inquire into regard to the cow's milk of production enhancement SOD and the meaning of milk-product thereof and feasibility.The preparation method of this civilian cow's milk SOD is, uses (NH4) 2SO4 to saltout, and the throw out obtaining is dissolved in a small amount of SMUF again, crosses post, uses K
2hPO
4(pH7.4) wash-out, collects elutriant freeze-drying and obtains finished product.(Feng Qihao. cow's milk and superoxide-dismutase wherein [J]. China Dairy Industry, 1993 (2): 68-73).
Summary of the invention
The object 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 DEG C in advance, for subsequent use;
(2) mix according to the ratio of 2: 1 with milk being pre-chilled to 10 DEG C of following distilled water;
(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 above-mentioned solution is adjusted to 7-7.5, leaves standstill 5-8 hour;
(6) filter, collect filtrate, add the ethanol of 3 times of milk and distilled water total amounts, stir, leave 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, extraction process safe green is pollution-free, equipment is simple.
Embodiment
Below in conjunction with embodiment, to according to embodiment provided by the invention, details are as follows:
Embodiment 1
1. raw milk detects qualified rear filtration, purification and is pre-chilled to 4 DEG C, and distilled water and 95% ethanol are carried and is pre-chilled to 4 DEG C simultaneously,
For subsequent use;
2. get and be pre-chilled to the distilled water 200g of 10 DEG C and mix with 100g milk;
3. in above-mentioned mixed solution, add the copper gluconate of 40g and the Zinc Gluconate of 30g, the filtration that stirs, collects the stillness of night, for subsequent use;
4. the cold ethanol that adds 600g in the above-mentioned stillness of night, stirs;
5. with the sodium hydroxide solution of 30mol/L, the pH value of above-mentioned solution is adjusted to 7.4, leaves standstill 5 hours;
6. filter, collect filtrate, add the cold ethanol of 900g, stir, leave standstill 6 hours;
7. filter, collecting precipitation, cryodrying, is ground into fine powder;
8. surveying SOD activity with 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 DEG C, distilled water and 95% ethanol is carried and is pre-chilled to 8 DEG C simultaneously, for subsequent use;
2. get and be pre-chilled to the distilled water 500g of 8 DEG C and mix with 250g milk;
3. in above-mentioned mixed solution, add the copper gluconate of 100g and the Zinc Gluconate of 75g, the filtration that stirs, collects the stillness of night, for subsequent use;
4. the cold ethanol that adds 1500g in the above-mentioned stillness of night, stirs;
5. with the sodium hydroxide solution of 30mol/L, the pH value of above-mentioned solution is adjusted to 7.3, leaves standstill 6 hours;
6. filter, collect filtrate, add the cold ethanol of 2250g, stir, leave standstill 7 hours;
7. filter, collecting precipitation, cryodrying, is ground into fine powder;
8. surveying SOD activity with 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 DEG C, distilled water and 95% ethanol is carried and is pre-chilled to 10 DEG C simultaneously, for subsequent use;
2. get and be pre-chilled to the distilled water 2000g of 10 DEG C and mix with 1000g milk;
3. in above-mentioned mixed solution, add the copper gluconate of 400g and the Zinc Gluconate of 300g, the filtration that stirs, collects the stillness of night, for subsequent use;
4. the cold ethanol that adds 6000g in the above-mentioned stillness of night, stirs;
5. with the sodium hydroxide solution of 30mol/L, the pH value of above-mentioned solution is adjusted to 7.4, leaves standstill 7 hours;
6. filter, collect filtrate, add the cold ethanol of 9000g, stir, leave standstill 7.5 hours;
7. filter, collecting precipitation, cryodrying, is ground into fine powder;
8. surveying SOD activity with 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 is only preferred embodiment of the present invention, can not be considered to for limiting practical range of the present invention.All equalization variation and improvement etc. of doing according to the present patent application scope, within all should still belonging to patent covering scope of the present invention.
Claims (1)
1. a method of extracting SOD from milk, its step is as follows:
(1) raw milk, distilled water, 95% ethanol are pre-chilled to below 10 DEG C in advance, for subsequent use;
(2) mix according to the ratio of 2: 1 with milk being pre-chilled to 10 DEG C of following distilled water;
(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 above-mentioned solution is adjusted to 7-7.5, leaves standstill 5-8 hour;
(6) filter, collect filtrate, add the ethanol of 3 times of milk and distilled water total amounts, stir, leave standstill 5-8 hour;
(7) filter collecting precipitation, cryodrying, pulverizing;
(8) ethanol distillation reclaims.
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| CN201310319538.4A CN103361320B (en) | 2013-07-25 | 2013-07-25 | Method of extracting SOD from milk |
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| CN201310319538.4A CN103361320B (en) | 2013-07-25 | 2013-07-25 | Method of extracting SOD from milk |
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| CN103361320B true 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 (6)
| Title |
|---|
| 冯启浩.牛乳及其中的超氧化物歧化酶.《中国乳品工业》.1993,第21卷(第2期),68-73. |
| 奶中超氧化物歧化酶SOD活性的研究;王惠生等;《西北农业大学学报》;19950831;第23卷(第4期);99-101 * |
| 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》.1998,第19卷(第4期),34-36. |
| 牛乳中超氧化物歧化酶SOD的提纯与鉴定;孙庆林等;《内蒙古农牧学院学报》;19981231;第19卷(第4期);34-36 * |
| 牛乳及其中的超氧化物歧化酶;冯启浩;《中国乳品工业》;19930430;第21卷(第2期);68-73 * |
| 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》.1995,第23卷(第4期),99-101. |
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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 |