CN101838285B - Method for separating phospholipids in purified oil material - Google Patents
Method for separating phospholipids in purified oil material Download PDFInfo
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- CN101838285B CN101838285B CN 201010166413 CN201010166413A CN101838285B CN 101838285 B CN101838285 B CN 101838285B CN 201010166413 CN201010166413 CN 201010166413 CN 201010166413 A CN201010166413 A CN 201010166413A CN 101838285 B CN101838285 B CN 101838285B
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Abstract
The invention relates to a method for separating phospholipids in purified oil materials, which is characterized in that (1) soybeans, eggs, fish, oil materials with higher content of phospholipids and byproducts thereof are extracted twice by 95% of ethanol and are filtered, and the extracts are combined and steamed in a rotation mode to obtain crude fat containing the phospholipids; (2) silica gel is weighed and arranged in a chromatographic column, and the height of the silica gel is 2-5cm; (3) the crude fat sample is weighed and loaded, neutral fat and free fatty acid are eluted by ethylacetate firstly, then cephalin is eluted by absolute ethyl alcohol, and finally lecithin is eluted by 80-95% of ethanol after PE is eluted; and (4) the eluent is decompressed and concentrated to be dry. The invention separates PE, PC, neutral fat and free fatty acid in the lipid at one step via silica gel column chromatography by safe solvents, the equipment is simple, the dosage of silica is small, the reagent consumption is low, the cost is low, the application range is wide, and the oil materials containing the phospholipids can be flexibly applied according to specific circumstances.
Description
Technical field
The invention belongs to farming, secondary product of forestry deep processing field.
Background technology
Extract phosphatide and application from various oil plants, China has a lot of research and practices.Solvent extraction process, the most frequently used solvent is acetone, acetone can dissolve grease and free fatty acids but dissolved phosphorus fat not, thereby can extract phosphatide with dehydrated alcohol or aqueous ethanol again with acetone with the removals such as grease in the phosphatide.In the solvent lixiviate, lixiviate often, the time is long, and acetone is harmful, when the product of acetone treatment is used for food, has the residual of acetone.Supercutical fluid CO
2Extraction process, its ultimate principle is with CO
2Fluid charges in the particular pressure temperature device, make it from oil sample, optionally neutral fat to be extracted under above-critical state, remaining polarity fat part mainly is phosphatide, can add the entrainment agent extraction using alcohol comes out, but the requirement height to plant and instrument, production cost significantly improves, and the phosphatide that extracts still will further separate, as uses column chromatography.Column chromatography is stationary phase with the sorbent material, and the solute in the mobile phase is by stationary phase the time, thereby because their absorption and the difference of desorption ability reach the purpose of separating.Now Chang Yong sorbent material is silica gel, aluminum oxide, diatomite etc.; Elutriant often selects for use chloroform, acetone, methyl alcohol etc. that the organic solvent of certain toxicity is arranged, and the dissolvent residual in the product is the shortcoming of this method.Also have preparative thin layer chromatography, also there is the reagent safety problem in methods such as inorganic salt composite precipitation technology.
Summary of the invention
The purpose of this invention is to provide a kind of safe, effective, fast, silica gel column chromatography method comes the phosphatide in the separation and purification oil plant cheaply.
Concrete processing step of the present invention:
(1) with soybean, egg, fish, contain raw materials such as the higher oil plant of phosphatide and by product thereof with 95% extraction using alcohol 2 times, filter, united extraction liquid revolves and steams the crude fat that obtains containing phosphatide, wherein contains triglyceride level, neutral fats such as free fatty acids;
(2) take by weighing silica gel and be loaded in the chromatography column, the silica gel height is 2～5cm;
(3) take by weighing sample on the crude fat sample, use the eluent ethyl acetate triglyceride level earlier, neutral fat and free fatty acidies such as cholesterol are used a certain amount of dehydrated alcohol wash-out kephalin (PE) again, treat that the PE wash-out finishes, and use 95%～80% ethanol elution Yelkin TTS (PC) then;
(4) each elutriant of concentrating under reduced pressure is to doing, and the ethyl acetate of recovery, ethanol can recycling.
The present invention can separate kephalin and Yelkin TTS fully through above-mentioned column chromatography.Detect PE purity through thin-layer chromatography (TLC) and reach more than 80%, PC purity reaches more than 90%.
Principal feature of the present invention is to utilize the high silicagel column of filling 2～5cm successfully to separate fully PE, PC and neutral fat.It is the strong polarity of utilizing phosphatide, come separating phospholipids with safety solvent ethanol, (general silica gel height all is greater than more than the 10cm in the prior art, even the height of filling gel is arranged on 6～10cm, all be difficult to PE is eluted), have only the silica gel height is dropped to 2～5cm as far as possible, could well elute PE; Because the group of phosphoric acid termination is tert-butylamine in the PC structure, can with the silica gel tight joint, under the situation of 2～5cm silica gel height, dehydrated alcohol still is difficult to wash-out PC, so just can separate PE, PC fully, after treating the complete wash-out of PE, strengthen the polarity of eluent again with 95%～80% ethanol, thereby the PC wash-out.This method can adopt as long as contain the oil plant of phosphatide, and can apply in a flexible way the applied range of raw material, and suitability is strong.What another characteristics of the present invention were that the separation and purification from fat extraction to last phosphatide uses all is safety solvent ethyl acetate, second alcohol and water, and can recycling, and the product that obtains with this purification technique is used for food, medicine and makeup, and is safe.
Advantage of the present invention is: 1, adopt safety solvent to separate the PE in the lipid, PC, neutral fat and free fatty acids once by silica gel column chromatography; 2, equipment is simple, and the silica gel consumption is few, and reagent consumption is few, and cost is low; 3, applied widely, can apply in a flexible way as the case may be to the oil plant that contains phosphatide.
Embodiment
The present invention will be described further by following examples.
Embodiment 1.
Take by weighing the 1.0113g yolk fat with 95% extraction using alcohol, be dissolved in the ethyl acetate, standby.Get gross porosity II silica gel 2.6g adding moving phase ethyl acetate and make pulpous state, wet method is loaded in (20mmx120mm) glass column, and the silica gel height is about 2cm in the post.The yolk fat sample for preparing is slowly poured in the post, at first added compositions such as 60mL moving phase eluent ethyl acetate triglyceride level, cholesterol, free fatty acids after sample introduction finishes; Use the 160mL dehydrated alcohol again, every 20mL collects effluent liquid step by step, the TLC qualitative analysis, and 20～140mL is rich in the PE part; Use 270mL 90% dehydrated alcohol at last, every 30mL collects step by step, the TLC qualitative analysis, and 60～240mL is rich in the PC component as the chromatography product.Merge identical component, get PE, PC and be respectively 0.0472g and 0.3372.
Embodiment 2.
Take by weighing the 0.4073g Poyang Lake Yellow catfish fat with 95% extraction using alcohol, be dissolved in the ethyl acetate, standby.Get gross porosity II silica gel 2.8g adding moving phase ethyl acetate and make pulpous state, wet method is loaded in (20mmx120mm) glass column.The fish fats sample for preparing is slowly poured in the post, at first added 30mL moving phase eluent ethyl acetate triglyceride level after sample introduction finishes, cholesterol, neutral fats such as free fatty acids; Collect effluent liquid step by step with the every 20mL of 140mL dehydrated alcohol again, the TLC qualitative analysis, 20～120mL is rich in the PE part; Collect step by step with the every 30mL of 240mL 90% dehydrated alcohol at last, the TLC qualitative analysis, 60～210mL is rich in the PC component as the chromatography product.Merge identical component, get PE, PC and be respectively 0.0182g and 0.0352.
Claims (2)
1. the method for phosphatide in the separation and purification oil plant is characterized in that taking by weighing the 1.0113g yolk fat with 95% extraction using alcohol, is dissolved in the ethyl acetate, and is standby; Get gross porosity II silica gel 2.6g adding moving phase ethyl acetate and make pulpous state, wet method is loaded in the 20mm x120mm glass column, and the silica gel height is about 2cm in the post; The yolk fat sample for preparing is slowly poured in the post, at first added 60mL moving phase eluent ethyl acetate triglyceride level, cholesterol, free fatty acids after sample introduction finishes; Use the 160mL dehydrated alcohol again, every 20mL collects effluent liquid step by step, the TLC qualitative analysis, and 20～140mL is rich in the PE part; Use the 270mL90% dehydrated alcohol at last, every 30mL collects step by step, the TLC qualitative analysis, and 60～240mL is rich in the PC component as the chromatography product, merges identical component, gets PE, PC and is respectively 0.0472g and 0.3372.
2. the method for phosphatide in the separation and purification oil plant is characterized in that taking by weighing the 0.4073g Poyang Lake Yellow catfish fat with 95% extraction using alcohol, is dissolved in the ethyl acetate, and is standby; Get gross porosity II silica gel 2.8g adding moving phase ethyl acetate and make pulpous state, wet method is loaded in the 20mmx120mm glass column; The fish fats sample for preparing is slowly poured in the post, at first added 30mL moving phase eluent ethyl acetate triglyceride level after sample introduction finishes, cholesterol, free fatty acids; Collect effluent liquid step by step with the every 20mL of 140mL dehydrated alcohol again, the TLC qualitative analysis, 20～120mL is rich in the PE part; Collect step by step with the every 30mL of 240mL90% dehydrated alcohol at last, the TLC qualitative analysis, 60～210mL is rich in the PC component as the chromatography product, merges identical component, gets PE, PC and is respectively 0.0182g and 0.0352.
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| Application Number | Priority Date | Filing Date | Title |
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| CN 201010166413 CN101838285B (en) | 2010-05-07 | 2010-05-07 | Method for separating phospholipids in purified oil material |
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| CN 201010166413 CN101838285B (en) | 2010-05-07 | 2010-05-07 | Method for separating phospholipids in purified oil material |
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| CN101838285A CN101838285A (en) | 2010-09-22 |
| CN101838285B true CN101838285B (en) | 2013-09-04 |
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Families Citing this family (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102977136A (en) * | 2012-12-07 | 2013-03-20 | 江南大学 | Technique for simultaneously extracting soybean lecithin and pigment from soybean oil |
| CN104370954B (en) * | 2014-09-26 | 2017-01-25 | 福建农林大学 | Method for separating and purifying lecithin in fish roe through silica gel column chromatography |
| CN109011698B (en) * | 2018-08-25 | 2020-12-01 | 江苏曼氏生物科技股份有限公司 | On-line cleaning and storing method for silica gel filler separation column |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US4425276A (en) * | 1980-12-13 | 1984-01-10 | A. Nattermann & Cie Gmbh | Process for the separation of oil and/or phosphatidylethanolamine from alcohol soluble phosphatidylcholine products containing the same |
| CN1740180A (en) * | 2005-08-26 | 2006-03-01 | 浙江大学 | Simultaneous prepn process of high-purity egg yolk lecithin and cephalin |
| CN1243760C (en) * | 2003-12-09 | 2006-03-01 | 武汉大学 | Method for separation and purification of lecithin and cephalin |
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2010
- 2010-05-07 CN CN 201010166413 patent/CN101838285B/en active Active
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US4425276A (en) * | 1980-12-13 | 1984-01-10 | A. Nattermann & Cie Gmbh | Process for the separation of oil and/or phosphatidylethanolamine from alcohol soluble phosphatidylcholine products containing the same |
| CN1243760C (en) * | 2003-12-09 | 2006-03-01 | 武汉大学 | Method for separation and purification of lecithin and cephalin |
| CN1740180A (en) * | 2005-08-26 | 2006-03-01 | 浙江大学 | Simultaneous prepn process of high-purity egg yolk lecithin and cephalin |
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Inventor after: Zou Xinhua Inventor before: Li Jing Inventor before: Li Guangyan Inventor before: Deng Zeyuan Inventor before: Liu Rong Inventor before: Fan Yawei Inventor before: Hu Jiangning Inventor before: Lei Lin |
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Effective date of registration: 20170406 Address after: 528143 Foshan Sanshui Industrial Zone, Guangdong, Datang Park, No. 48-6 Patentee after: FOSHAN BOEN BIOTECHNOLOGY CO., LTD. Address before: 999 No. 330031 Jiangxi province Nanchang Honggutan University Avenue Patentee before: Nanchang University |
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