CN104628812A - Method for purifying deer blood oligopeptides by macroporous adsorption resin - Google Patents
Method for purifying deer blood oligopeptides by macroporous adsorption resin Download PDFInfo
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- CN104628812A CN104628812A CN201310544988.3A CN201310544988A CN104628812A CN 104628812 A CN104628812 A CN 104628812A CN 201310544988 A CN201310544988 A CN 201310544988A CN 104628812 A CN104628812 A CN 104628812A
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Abstract
The invention discloses a method for purifying deer blood oligopeptides by macroporous adsorption resin. The method comprises the steps: with deer blood as a raw material, adsorbing a supernatant after enzymatic hydrolysis in a macroporous adsorption resin column, collecting the effluent, rinsing with an acid or an alkali, eluting the resin column with a solvent obtained by combining two or more of an organic solvent, water, an acid or an alkali, respectively concentrating the effluent, the acid or alkali wash liquid and the eluate under reduced pressure, and drying to obtain deer blood oligopeptide products with different purities. The content of the oligopeptides in the obtained blood deer oligopeptide products is 65-95%. The method has the advantages that the technology is simple, the preparation process is safe, the whole extraction process is suitable for industrialized production and the production cost is reduced.
Description
Technical field
The present invention relates to a kind of method adopting macroporous adsorbent resin technology purifying deer blood oligopeptide.
Background technology
Deer blood is traditional rare Chinese medicine, is exactly the treasure of imperial palace people of imperial lineage, high-level officials' disease curing and body-building since ancient times.The various functions of many scholars to deer blood are studied, and show that deer blood has and strengthen immunity, enrich blood, delay senility, antifatigue, raising sexual function, benefit essence, calm the nerves, promote the function such as metabolism and wound healing.Containing multiple effective active composition in deer blood, there is a lot of physiological function, as superoxide-dismutase, immunoglobulin (Ig), protoheme etc.
Modern scientific research shows, the utilization of the mankind to protein is become by protein digestion small molecules oligopeptide to absorb, instead of former thought to need breaks down proteins be amino acid.The maximum feature of oligopeptide mechanism of absorption does not need to digest direct absorption, do not need to consume body energy, and absorption rate is fast, and preferential absorption, in human body, comparatively amino acid is fast for the speed of synthetic protein.Therefore need be only the needs that oligopeptide can meet the mankind by proteolysis.
The utilization of deer blood resource, prior art is the direct utilization to the former blood of deer blood mostly, such as, with wine mixed preparing deer-blood wine, but deer-blood wine often quality is bad, have flocks to produce, and can sex change be there is in proteins and peptides, cause biological activity and nutritive ingredient partly or entirely to be lost.After Chinese patent (102199648 A) utilizes papain and Sumizyme MP to carry out enzymolysis to deer blood successively, adopt membrane technique purifying, obtain bioactive peptide, open up its application in biology field.But this patent utilization membrane technique, makes the cost producing bioactive peptide higher.
Summary of the invention
The object of the present invention is to provide a kind of method adopting purification with macroreticular resin deer blood oligopeptide.
The technical scheme that the present invention realizes above-mentioned purpose is as follows:
In order to widen the range of application of deer blood resource, its nutritive ingredient is absorbed and used better, obtain high purity oligopeptide accordingly, we utilize macroporous adsorbent resin from deer blood, prepare the processing method of oligopeptide.
Adopt the method for Chinese patent (102199648 A) to carry out enzymolysis to deer blood, the supernatant liquor after enzymolysis adsorbs according to series connection suction type on macroporous adsorptive resins.Upon adsorption saturated after, with solvent elution, elutriant concentrates, and obtains the oligopeptide product of different purity after drying.
The present invention with deer blood for raw material, supernatant liquor after enzymolysis adsorbs on macroporous adsorptive resins, collect effluent liquid, after acid or alkaline flushing, use organic solvent, water, acid or alkali according to two yuan or multiplexed combination solvent elution resin column again, effluent liquid, acid or alkaline wash and elutriant concentrating under reduced pressure respectively, obtains the deer blood oligopeptide product of different purity after drying.
Adopt a method for purification with macroreticular resin deer blood oligopeptide, it is characterized in that the method comprises the following steps:
After A, deer hemase solution, to be the purity of oligopeptide product be supernatant liquor 50% is adsorbed according to series connection suction type by single resin column or many resin columns, collects effluent liquid after absorption;
B, adsorb saturated after resin column 1 ~ 20 BV volumetric concentration be 1 ~ 20% acid or mass concentration be the alkaline flushing of 1 ~ 20%, collect acid or alkali elutriant; Then resin column uses the stripping liquid wash-out of 1 ~ 20 BV again, and flow velocity is 1 ~ 25 BV/h; Effluent liquid, acid or alkali elutriant and stripping liquid, after concentrating under reduced pressure recycling design, obtain deer blood oligopeptide after drying.
The deer blood oligopeptide that the present invention obtains is white or faint yellow solid powder, purity 65 ~ 95%.
In step A, described resin refers to the polystyrene macroporous adsorbent resin of Semi-polarity or polarity.
In step A, macroporous adsorbent resin and deer blood mass ratio are 0.5:1 ~ 10:1, and resin column blade diameter length ratio is 1:4 ~ 1:30, and column temperature is 25 ~ 50 DEG C.
In step B, described stripping liquid refers to that the 3rd class organic solvent, water, food-grade acid or alkali are according to two yuan or multiplexed combination solvent, adopts isocratic elution or gradient elution during wash-out.
Product of the present invention measures with reference to liquid phase chromatography in GB GB/T 22729-2008.
Chromatographic condition:
Chromatographic column: of the same type other that TSK gel G2000 SWXL 300 mm × 7.8 mm or performance are close is therewith applicable to the gel column measuring proteins and peptides.
Moving phase: acetonitrile: water: trifluoroacetic acid=45:55:0.1 (v:v:v); Flow velocity: 0.5 mL/min; Determined wavelength: 220 nm; Reference wavelength: 360 nm; Column temperature: 30 oC, sample size: 20 μ L.
The present invention is compared with prior art method, feature there are provided a kind of technique utilizing macroporous adsorbent resin technology to prepare oligopeptide from deer blood, its production cost is low, simple to operate, be easy to realize suitability for industrialized production, high, the good water solubility of oligopeptide purity of preparation, without fishy smell, the industries such as medicine, healthcare products, food, makeup can be widely used in.
Embodiment
Following examples are used for further illustrating the present invention, but do not mean that this any limitation of the invention.
Embodiment 1
Take HPD750 and LSD001 resin two parts, every part of 300 g, wet method is respectively charged in the glass column (A post and B post) of two internal diameter 3 cm, high 66 cm of post, and two post series connection, carry adsorption liquid with infusion pump simultaneously.Supernatant liquor (5 L) after deer hemase solution is flow through this series connection resin column successively, and adsorb, flow velocity 10 BV/h, resin column temperature is 50 DEG C, collects effluent liquid.Effluent liquid is through concentrating under reduced pressure, and it is the oligopeptide of 90% that normal pressure 60 DEG C of dryings obtain 1000 g purity.Add 5% acetum wash-out with 6 BV/h flow velocitys respectively in the A post adsorbed, B post, consumption is 12 BV, collects sour elutriant.Elutriant, through concentrating under reduced pressure, obtains in 50 DEG C of vacuum decompression dryings the oligopeptide product that 200 g purity are 80%.Then use ethanol-10% oxalic acid (3:7, v/v) to carry out wash-out to A post, B post, flow velocity 15 BV/h, elutriant consumption 10 BV, the elutriant obtained is after concentrating under reduced pressure recycling design, and ultra red ray drying obtains the oligopeptide product that 50 g purity are 66%.
Embodiment 2
Take LX-28 and DM 301 resin two parts, every part of 200 g, wet method is respectively charged in the glass column (A post and B post) of two internal diameter 5 cm, high 80 cm of post, and two post series connection, carry adsorption liquid with infusion pump simultaneously.Supernatant liquor (1 L) after deer hemase solution is flow through this series connection resin column successively, and adsorb, flow velocity 5 BV/h, resin column temperature is 30 DEG C, collects effluent liquid.Effluent liquid is through concentrating under reduced pressure, and it is the oligopeptide of 93% that vacuum lyophilization obtains 220 g purity.Add 10% sodium hydrogen carbonate solution wash-out with 8 BV/h flow velocitys respectively in the A post adsorbed, B post, consumption is 8 BV, collects elutriant.The elutriant obtained is through concentrating under reduced pressure, and 60 DEG C of constant pressure and dries obtain the oligopeptide product that 45 g purity are 85%.Then ethanol-20% sodium bicarbonate (5:5, v/v) is used to carry out wash-out to A post, B post, flow velocity 25 BV/h, elutriant consumption 8 BV.Elutriant is after concentrating under reduced pressure recycling design, and ultra red ray drying obtains the oligopeptide product that 8 g purity are 70%.
Embodiment 3
Take LSA-40 resin 3000 g, wet method loads in the glass column of internal diameter 20 cm, high 100 cm of post, carries adsorption liquid with infusion pump simultaneously.Supernatant liquor (20 L) after deer hemase solution is flow through this resin column successively, and adsorb, flow velocity 15 BV/h, resin column temperature is 20 DEG C, collects effluent liquid.Effluent liquid is through concentrating under reduced pressure, and it is the oligopeptide of 90% that vacuum lyophilization obtains 8000 g purity.Add 10% acetum wash-out in the resin column completed to absorption with 1 BV/h flow velocity, consumption is 12 BV, collects sour elutriant.Elutriant is through concentrating under reduced pressure, and 50 DEG C of constant pressure and dries obtain the oligopeptide product that 3000 g purity are 75%.Then use ethanol-15% citric acid (2:8, v/v) to carry out wash-out to A post, B post, flow velocity 8 BV/h, elutriant consumption 6 BV, the elutriant obtained is after concentrating under reduced pressure recycling design, and 40 DEG C of vacuum decompression dryings obtain the oligopeptide product that 500 g purity are 68%.
Claims (4)
1. adopt a method for purification with macroreticular resin deer blood oligopeptide, it is characterized in that the method comprises the following steps:
After A, deer hemase solution, to be the purity of oligopeptide product be supernatant liquor 50% is adsorbed according to series connection suction type by single resin column or many resin columns, collects effluent liquid after absorption;
B, adsorb saturated after resin column 1 ~ 20 BV volumetric concentration be 1 ~ 20% acid or mass concentration be the alkaline flushing of 1 ~ 20%, collect acid or alkali elutriant; Then resin column uses the stripping liquid wash-out of 1 ~ 20 BV again, and flow velocity is 1 ~ 25 BV/h; Effluent liquid, acid or alkali elutriant and stripping liquid, after concentrating under reduced pressure recycling design, obtain deer blood oligopeptide after drying.
2. the method for claim 1, is characterized in that: in step A, and described resin refers to the polystyrene macroporous adsorbent resin of Semi-polarity or polarity.
3. the method for claim 1, is characterized in that: in step A, and macroporous adsorbent resin and deer blood mass ratio are 0.5:1 ~ 10:1, and resin column blade diameter length ratio is 1:4 ~ 1:30, and column temperature is 25 ~ 50 DEG C.
4. the method for claim 1, is characterized in that: in step B, and described stripping liquid refers to that the 3rd class organic solvent, water, food-grade acid or alkali are according to two yuan or multiplexed combination solvent, adopts isocratic elution or gradient elution during wash-out.
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| Application Number | Priority Date | Filing Date | Title |
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| CN201310544988.3A CN104628812A (en) | 2013-11-07 | 2013-11-07 | Method for purifying deer blood oligopeptides by macroporous adsorption resin |
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| CN201310544988.3A CN104628812A (en) | 2013-11-07 | 2013-11-07 | Method for purifying deer blood oligopeptides by macroporous adsorption resin |
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105146668A (en) * | 2015-10-14 | 2015-12-16 | 中国科学院兰州化学物理研究所 | Deer blood polypeptide solid beverage |
| CN105795462A (en) * | 2016-03-29 | 2016-07-27 | 吉林省国鹿生物科技有限公司 | Preparation method of deer blood tablet polypeptide dry powder |
| CN107699599A (en) * | 2017-10-27 | 2018-02-16 | 中国科学院兰州化学物理研究所 | A kind of hunchbacked blood polypeptide with hypotensive and effect for reducing blood fat |
| CN114983852A (en) * | 2022-04-20 | 2022-09-02 | 赵宏强 | Anti-aging cosmetic with whitening and moisturizing effects and preparation method thereof |
-
2013
- 2013-11-07 CN CN201310544988.3A patent/CN104628812A/en active Pending
Non-Patent Citations (2)
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| 刘毅等: "马鹿血蛋白的酶解及多肽的分离纯化", 《食品科学》 * |
| 娄嵩: "大孔吸附树脂的吸附机理", 《化学进展》 * |
Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105146668A (en) * | 2015-10-14 | 2015-12-16 | 中国科学院兰州化学物理研究所 | Deer blood polypeptide solid beverage |
| CN105795462A (en) * | 2016-03-29 | 2016-07-27 | 吉林省国鹿生物科技有限公司 | Preparation method of deer blood tablet polypeptide dry powder |
| CN107699599A (en) * | 2017-10-27 | 2018-02-16 | 中国科学院兰州化学物理研究所 | A kind of hunchbacked blood polypeptide with hypotensive and effect for reducing blood fat |
| CN107699599B (en) * | 2017-10-27 | 2021-08-03 | 中国科学院兰州化学物理研究所 | Camel blood polypeptide with blood pressure and blood fat reducing effects |
| CN114983852A (en) * | 2022-04-20 | 2022-09-02 | 赵宏强 | Anti-aging cosmetic with whitening and moisturizing effects and preparation method thereof |
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