CN101603061A - The enriching method of conjugated linolic acid in a kind of Fructus Maydis oil - Google Patents
The enriching method of conjugated linolic acid in a kind of Fructus Maydis oil Download PDFInfo
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- CN101603061A CN101603061A CNA2009100725429A CN200910072542A CN101603061A CN 101603061 A CN101603061 A CN 101603061A CN A2009100725429 A CNA2009100725429 A CN A2009100725429A CN 200910072542 A CN200910072542 A CN 200910072542A CN 101603061 A CN101603061 A CN 101603061A
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- linolic acid
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Abstract
The enriching method of conjugated linolic acid in a kind of Fructus Maydis oil, it relates to a kind of enriching method of conjugated linolic acid.The invention solves the low problem of content of conjugated linolic acid in the Fructus Maydis oil of existing method preparation.Method: one, with chitosan magnetic micro-sphere as carrier fixation of microbe lipase; Two, Fructus Maydis oil and methyl alcohol are added in the there-necked flask, and heating is also stirred, and temperature adds the immobilization microbial lipase when being raised to 52 ℃, the reaction that picks up counting, and reaction proceeds to 10h and 20h adds methyl alcohol respectively, stops when proceeding to 26h; Three, use the peristaltic pump discharging after reaction stops, feed liquid being put into got supernatant liquid after separating funnel staticly settles then; Four, supernatant liquid is transferred in the distilling flask after distillation, processed and promptly finishes.Enriching method of the present invention is simple, and the content height of conjugated linolic acid in the gained Fructus Maydis oil for containing 8.10~10.51mg in every gram Fructus Maydis oil, can satisfy needed by human body after the enrichment.
Description
Technical field
The present invention relates to a kind of enriching method of conjugated linolic acid.
Background technology
Contain rich nutrient substances in the Fructus Maydis oil, except that being rich in polyunsaturated fatty acid, also contain the carotene, VA, the VD etc. that are called as vitamin source.It is unsaturated fatty acids that the lipid acid of Fructus Maydis oil is formed most of (80%), is representative with oleic acid (30%) and linolic acid (LA).Recent study is found, conjugated linolic acid (the Conjugated linoleic acid that also exists in the Fructus Maydis oil, hereinafter to be referred as CLA), it is linoleic isomers, having and remove free radical, strengthen antioxidant capacity of human body and immunological competence, promotion are grown, regulated blood cholesterol levels and triglyceride level, prevent atherosclerosis, promote fats oxidn to decompose, promote that human body protein is synthetic, human body is carried out being used as of comprehensive optimum adjusting, is popular third generation functional food material the most in the world at present.But the content of conjugated linolic acid is low in the Fructus Maydis oil of existing method preparation, for only containing 0.15~0.42mg in every gram Fructus Maydis oil, can not satisfy needed by human body.
Summary of the invention
The present invention is for the low problem of content of conjugated linolic acid in the Fructus Maydis oil that solves existing method preparation, and the enriching method of conjugated linolic acid in a kind of Fructus Maydis oil is provided.
The enriching method of conjugated linolic acid is realized according to the following steps in the Fructus Maydis oil: one, with chitosan magnetic micro-sphere as carrier fixation of microbe lipase, the immobilization temperature is 30~40 ℃, the pH value is 6~9, microbial lipase concentration 800~1200U/ml, fix behind 10~20h immobilized microorganism lipase; Two, 300g Fructus Maydis oil and 20ml methyl alcohol are joined in the 500ml there-necked flask, the thermostat water bath that places then heats, and stir with the speed of 180~220r/min, when being elevated to 52 ℃, temperature adds 10g immobilized microorganism lipase again, reaction picks up counting, reaction proceeds to 10h and 20h adds 10ml methyl alcohol respectively, stops when proceeding to 26h; Three, use the peristaltic pump discharging after reaction stops, feed liquid being put into got supernatant liquid after the 1000ml separating funnel staticly settles 3~5h then; Four, supernatant liquid is transferred in the 500ml distilling flask, place on the rotatory evaporator then, in temperature is that 65 ℃, pressure are to distill 5min under the condition of 200mmHg, discard and heat up in a steamer head, the receiving trap that renews then, continue heating up, is that 72 ℃, pressure are distillation and collect cut under the condition of 50mmHg in temperature, promptly finishes the enrichment of conjugated linolic acid in the Fructus Maydis oil.
The present invention adopts highdensity magnetic microsphere immobilized microbial lipase, carries out transesterify between catalysis Fructus Maydis oil and the methyl alcohol, catalyzed reaction finish after distill, processed, make the Fructus Maydis oil of high cla levels; Because of having adopted enzyme immobilization technology, thereby reduced the restraining effect of methyl alcohol to enzyme.And separating technology is simple to operate, the rate of recovery is high after the catalyzed reaction, and the immobilized microorganism lipase that reclaims can reuse, and reduces production costs, and simplifies production technique.
The enriching method of conjugated linolic acid is simple in the Fructus Maydis oil of the present invention, and cost is low; The content height of conjugated linolic acid in the gained Fructus Maydis oil after the enrichment among the present invention, for containing 8.10~10.51mg in every gram Fructus Maydis oil, needed by human body can be satisfied, promptly abundant conjugated linolic acid can be absorbed by the edible corn germ oil, edible good for health for a long time.
Embodiment
Embodiment one: the enriching method of conjugated linolic acid is realized according to the following steps in the present embodiment Fructus Maydis oil: one, with chitosan magnetic micro-sphere as carrier fixation of microbe lipase, the immobilization temperature is 30~40 ℃, the pH value is 6~9, microbial lipase concentration 800~1200U/ml, fix behind 10~20h immobilized microorganism lipase; Two, 300g Fructus Maydis oil and 20ml methyl alcohol are joined in the 500ml there-necked flask, the thermostat water bath that places then heats, and stir with the speed of 180~220r/min, when being elevated to 52 ℃, temperature adds 10g immobilized microorganism lipase again, reaction picks up counting, reaction proceeds to 10h and 20h adds 10ml methyl alcohol respectively, stops when proceeding to 26h; Three, use the peristaltic pump discharging after reaction stops, feed liquid being put into got supernatant liquid after the 1000ml separating funnel staticly settles 3~5h then; Four, supernatant liquid is transferred in the 500ml distilling flask, place on the rotatory evaporator then, in temperature is that 65 ℃, pressure are to distill 5min under the condition of 200mmHg, discard and heat up in a steamer head, the receiving trap that renews then, continue heating up, is that 72 ℃, pressure are distillation and collect cut under the condition of 50mmHg in temperature, promptly finishes the enrichment of conjugated linolic acid in the Fructus Maydis oil.
The preparation of chitosan magnetic micro-sphere in the present embodiment step 1: adopt chemical coprecipitation to prepare magnetic fluid-Fe
3O
4Hydrate adopts inverse suspension polymerization method synthesizing magnetic chitosan microball then, and the volumetric concentration 1.0%~5.0% of chitosan, the iron-holder of magnetic fluid are 40wt%~70wt%, and the volumetric concentration of glutaraldehyde is 10%~25%.
After the enrichment of conjugated linolic acid was finished in the present embodiment Fructus Maydis oil, immobilized microorganism lipase can be stayed in the there-necked flask, directly adds feed liquid, carried out the next batch reaction; Also can take out immobilized microorganism lipase, reclaim the washing back, and washings is that ethanol and ether mix gained solution by 1: 1 volume ratio.
Embodiment two: what present embodiment and embodiment one were different is that the immobilization temperature is 32~38 ℃ in the step 1, and the pH value is 6.5~8.5, and microbial lipase concentration 850~1150U/ml fixes 12~18h.Other step and parameter are identical with embodiment one.
Embodiment three: what present embodiment and embodiment one were different is that the immobilization temperature is 30 ℃ in the step 1, and the pH value is 6, microbial lipase concentration 800U/ml, fixedly 10h.Other step and parameter are identical with embodiment one.
Embodiment four: what present embodiment and embodiment one were different is that the immobilization temperature is 40 ℃ in the step 1, and the pH value is 9, microbial lipase concentration 1200U/ml, fixedly 20h.Other step and parameter are identical with embodiment one.
Embodiment five: what present embodiment and embodiment one were different is that the immobilization temperature is 35 ℃ in the step 1, and the pH value is 7, microbial lipase concentration 900U/ml, fixedly 12h.Other step and parameter are identical with embodiment one.
Embodiment six: what present embodiment and embodiment one were different is that the immobilization temperature is 38 ℃ in the step 1, and the pH value is 8, microbial lipase concentration 1100U/ml, fixedly 16h.Other step and parameter are identical with embodiment one.
Embodiment seven: what present embodiment and embodiment two, three, four, five or six were different is that the speed with 190~210r/min stirs in the step 2.Other step and parameter are identical with embodiment two, three, four, five or six.
Embodiment eight: what present embodiment and embodiment seven were different is that the speed with 180r/min stirs in the step 2.Other step and parameter are identical with embodiment seven.
Embodiment nine: what present embodiment and embodiment seven were different is that the speed with 220r/min stirs in the step 2.Other step and parameter are identical with embodiment seven.
Embodiment ten: what present embodiment and embodiment seven were different is that the speed with 190r/min stirs in the step 2.Other step and parameter are identical with embodiment seven.
Embodiment 11: what present embodiment and embodiment seven were different is that the speed with 200r/min stirs in the step 2.Other step and parameter are identical with embodiment seven.
Embodiment 12: what present embodiment and embodiment eight, nine, ten or 11 were different is that methyl alcohol is chemical pure in the step 2.Other step and parameter are identical with embodiment eight, nine, ten or 11.
Embodiment 13: the enriching method of conjugated linolic acid is realized according to the following steps in the present embodiment Fructus Maydis oil: one, with chitosan magnetic micro-sphere as carrier fixation of microbe lipase, the immobilization temperature is 35 ℃, the pH value is 7, microbial lipase concentration 1000U/ml fixedly gets immobilized microorganism lipase behind the 15h; Two, 300g Fructus Maydis oil and 20ml methyl alcohol are joined in the 500ml there-necked flask, the thermostat water bath that places then heats, and stir with the speed of 180r/min, when being elevated to 52 ℃, temperature adds 10g immobilized microorganism lipase again, reaction picks up counting, reaction proceeds to 10h and 20h adds 10ml methyl alcohol respectively, stops when proceeding to 26h; Three, use the peristaltic pump discharging after reaction stops, feed liquid being put into got supernatant liquid after the 1000ml separating funnel staticly settles 3h then; Four, supernatant liquid is transferred in the 500ml distilling flask, place on the rotatory evaporator then, in temperature is that 65 ℃, pressure are to distill 5min under the condition of 200mmHg, discard and heat up in a steamer head, the receiving trap that renews then, continue heating up, is that 72 ℃, pressure are distillation and collect cut under the condition of 50mmHg in temperature, promptly finishes the enrichment of conjugated linolic acid in the Fructus Maydis oil.
The content height of conjugated linolic acid in the gained Fructus Maydis oil after the enrichment for containing 8.10mg in every gram Fructus Maydis oil, can satisfy needed by human body in the present embodiment.
Embodiment 14: the enriching method of conjugated linolic acid is realized according to the following steps in the present embodiment Fructus Maydis oil: one, with chitosan magnetic micro-sphere as carrier fixation of microbe lipase, the immobilization temperature is 30 ℃, the pH value is 6, microbial lipase concentration 800U/ml fixedly gets immobilized microorganism lipase behind the 10h; Two, 300g Fructus Maydis oil and 20ml methyl alcohol are joined in the 500ml there-necked flask, the thermostat water bath that places then heats, and stir with the speed of 190r/min, when being elevated to 52 ℃, temperature adds 10g immobilized microorganism lipase again, reaction picks up counting, reaction proceeds to 10h and 20h adds 10ml methyl alcohol respectively, stops when proceeding to 26h; Three, use the peristaltic pump discharging after reaction stops, feed liquid being put into got supernatant liquid after the 1000ml separating funnel staticly settles 4h then; Four, supernatant liquid is transferred in the 500ml distilling flask, place on the rotatory evaporator then, in temperature is that 65 ℃, pressure are to distill 5min under the condition of 200mmHg, discard and heat up in a steamer head, the receiving trap that renews then, continue heating up, is that 72 ℃, pressure are distillation and collect cut under the condition of 50mmHg in temperature, promptly finishes the enrichment of conjugated linolic acid in the Fructus Maydis oil.
The content height of conjugated linolic acid in the gained Fructus Maydis oil after the enrichment for containing 8.56mg in every gram Fructus Maydis oil, can satisfy needed by human body in the present embodiment.
Embodiment 15: the enriching method of conjugated linolic acid is realized according to the following steps in the present embodiment Fructus Maydis oil: one, with chitosan magnetic micro-sphere as carrier fixation of microbe lipase, the immobilization temperature is 40 ℃, the pH value is 8, microbial lipase concentration 1200U/ml fixedly gets immobilized microorganism lipase behind the 10h; Two, 300g Fructus Maydis oil and 20ml methyl alcohol are joined in the 500ml there-necked flask, the thermostat water bath that places then heats, and stir with the speed of 200r/min, when being elevated to 52 ℃, temperature adds 10g immobilized microorganism lipase again, reaction picks up counting, reaction proceeds to 10h and 20h adds 10ml methyl alcohol respectively, stops when proceeding to 26h; Three, use the peristaltic pump discharging after reaction stops, feed liquid being put into got supernatant liquid after the 1000ml separating funnel staticly settles 4h then; Four, supernatant liquid is transferred in the 500ml distilling flask, place on the rotatory evaporator then, in temperature is that 65 ℃, pressure are to distill 5min under the condition of 200mmHg, discard and heat up in a steamer head, the receiving trap that renews then, continue heating up, is that 72 ℃, pressure are distillation and collect cut under the condition of 50mmHg in temperature, promptly finishes the enrichment of conjugated linolic acid in the Fructus Maydis oil.
The content height of conjugated linolic acid in the gained Fructus Maydis oil after the enrichment for containing 9.28mg in every gram Fructus Maydis oil, can satisfy needed by human body in the present embodiment.
Embodiment 16: the enriching method of conjugated linolic acid is realized according to the following steps in the present embodiment Fructus Maydis oil: one, with chitosan magnetic micro-sphere as carrier fixation of microbe lipase, the immobilization temperature is 35 ℃, the pH value is 9, microbial lipase concentration 800U/ml fixedly gets immobilized microorganism lipase behind the 20h; Two, 300g Fructus Maydis oil and 20ml methyl alcohol are joined in the 500ml there-necked flask, the thermostat water bath that places then heats, and stir with the speed of 210r/min, when being elevated to 52 ℃, temperature adds 10g immobilized microorganism lipase again, reaction picks up counting, reaction proceeds to 10h and 20h adds 10ml methyl alcohol respectively, stops when proceeding to 26h; Three, use the peristaltic pump discharging after reaction stops, feed liquid being put into got supernatant liquid after the 1000ml separating funnel staticly settles 4h then; Four, supernatant liquid is transferred in the 500ml distilling flask, place on the rotatory evaporator then, in temperature is that 65 ℃, pressure are to distill 5min under the condition of 200mmHg, discard and heat up in a steamer head, the receiving trap that renews then, continue heating up, is that 72 ℃, pressure are distillation and collect cut under the condition of 50mmHg in temperature, promptly finishes the enrichment of conjugated linolic acid in the Fructus Maydis oil.
The content height of conjugated linolic acid in the gained Fructus Maydis oil after the enrichment for containing 9.83mg in every gram Fructus Maydis oil, can satisfy needed by human body in the present embodiment.
Embodiment 17: the enriching method of conjugated linolic acid is realized according to the following steps in the present embodiment Fructus Maydis oil: one, with chitosan magnetic micro-sphere as carrier fixation of microbe lipase, the immobilization temperature is 38 ℃, the pH value is 7, microbial lipase concentration 1100U/ml fixedly gets immobilized microorganism lipase behind the 18h; Two, 300g Fructus Maydis oil and 20ml methyl alcohol are joined in the 500ml there-necked flask, the thermostat water bath that places then heats, and stir with the speed of 200r/min, when being elevated to 52 ℃, temperature adds 10g immobilized microorganism lipase again, reaction picks up counting, reaction proceeds to 10h and 20h adds 10ml methyl alcohol respectively, stops when proceeding to 26h; Three, use the peristaltic pump discharging after reaction stops, feed liquid being put into got supernatant liquid after the 1000ml separating funnel staticly settles 5h then; Four, supernatant liquid is transferred in the 500ml distilling flask, place on the rotatory evaporator then, in temperature is that 65 ℃, pressure are to distill 5min under the condition of 200mmHg, discard and heat up in a steamer head, the receiving trap that renews then, continue heating up, is that 72 ℃, pressure are distillation and collect cut under the condition of 50mmHg in temperature, promptly finishes the enrichment of conjugated linolic acid in the Fructus Maydis oil.
The content height of conjugated linolic acid in the gained Fructus Maydis oil after the enrichment for containing 10.51mg in every gram Fructus Maydis oil, can satisfy needed by human body in the present embodiment.
Claims (8)
1, the enriching method of conjugated linolic acid in a kind of Fructus Maydis oil, it is characterized in that the enriching method of conjugated linolic acid is realized according to the following steps in the Fructus Maydis oil: one, with chitosan magnetic micro-sphere as carrier fixation of microbe lipase, the immobilization temperature is 30~40 ℃, the pH value is 6~9, microbial lipase concentration 800~1200U/ml, fix behind 10~20h immobilized microorganism lipase; Two, 300g Fructus Maydis oil and 20ml methyl alcohol are joined in the 500ml there-necked flask, the thermostat water bath that places then heats, and stir with the speed of 180~220r/min, when being elevated to 52 ℃, temperature adds 10g immobilized microorganism lipase again, reaction picks up counting, reaction proceeds to 10h and 20h adds 10ml methyl alcohol respectively, stops when proceeding to 26h; Three, use the peristaltic pump discharging after reaction stops, feed liquid being put into got supernatant liquid after the 1000ml separating funnel staticly settles 3~5h then; Four, supernatant liquid is transferred in the 500ml distilling flask, place on the rotatory evaporator then, in temperature is that 65 ℃, pressure are to distill 5min under the condition of 200mmHg, discard and heat up in a steamer head, the receiving trap that renews then, continue heating up, is that 72 ℃, pressure are distillation and collect cut under the condition of 50mmHg in temperature, promptly finishes the enrichment of conjugated linolic acid in the Fructus Maydis oil.
2, the enriching method of conjugated linolic acid in the Fructus Maydis oil according to claim 1 is characterized in that the immobilization temperature is 32~38 ℃ in the step 1, and the pH value is 6.5~8.5, and microbial lipase concentration 850~1150U/ml fixes 12~18h.
3, the enriching method of conjugated linolic acid in the Fructus Maydis oil according to claim 1 is characterized in that the immobilization temperature is 35 ℃ in the step 1, and the pH value is 7, microbial lipase concentration 900U/ml, fixedly 12h.
4, the enriching method of conjugated linolic acid in the Fructus Maydis oil according to claim 1 is characterized in that the immobilization temperature is 38 ℃ in the step 1, and the pH value is 8, microbial lipase concentration 1100U/ml, fixedly 16h.
5,, it is characterized in that the speed with 190~210r/min stirs in the step 2 according to the enriching method of conjugated linolic acid in claim 2, the 3 or 4 described Fructus Maydis oils.
6, the enriching method of conjugated linolic acid in the Fructus Maydis oil according to claim 5 is characterized in that the speed with 190r/min stirs in the step 2.
7, the enriching method of conjugated linolic acid in the Fructus Maydis oil according to claim 5 is characterized in that the speed with 200r/min stirs in the step 2.
8, according to the enriching method of conjugated linolic acid in claim 6 or the 7 described Fructus Maydis oils, it is characterized in that methyl alcohol is chemical pure in the step 2.
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Cited By (5)
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CN102839045A (en) * | 2012-09-10 | 2012-12-26 | 东北农业大学 | Method of treating soybean germ flakes by compound enzyme membrane |
CN102994491A (en) * | 2012-07-12 | 2013-03-27 | 浙江大学 | Immobilization method of Thermus lipase |
CN103849660A (en) * | 2014-03-28 | 2014-06-11 | 大连医诺生物有限公司 | Method for preparing conjugated linoleic acid through coupling method using immobilized lipase as catalyst |
CN109134237A (en) * | 2018-04-18 | 2019-01-04 | 邯郸晨光珍品油脂有限公司 | A kind of production method of conjugated linoleic acid |
CN110338222A (en) * | 2019-08-23 | 2019-10-18 | 李跃东 | A kind of high-purity conjugated linoleic acid yak yoghourt and preparation method thereof |
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2009
- 2009-07-20 CN CNA2009100725429A patent/CN101603061A/en active Pending
Cited By (9)
Publication number | Priority date | Publication date | Assignee | Title |
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CN102994491A (en) * | 2012-07-12 | 2013-03-27 | 浙江大学 | Immobilization method of Thermus lipase |
CN102839045A (en) * | 2012-09-10 | 2012-12-26 | 东北农业大学 | Method of treating soybean germ flakes by compound enzyme membrane |
CN102839045B (en) * | 2012-09-10 | 2014-07-16 | 东北农业大学 | Method of treating soybean germ flakes by compound enzyme membrane |
CN103849660A (en) * | 2014-03-28 | 2014-06-11 | 大连医诺生物有限公司 | Method for preparing conjugated linoleic acid through coupling method using immobilized lipase as catalyst |
CN103849660B (en) * | 2014-03-28 | 2016-01-06 | 大连医诺生物有限公司 | A kind of with immobilized lipase be catalyzer be coupled the method for legal system for conjugated linolic acid |
CN109134237A (en) * | 2018-04-18 | 2019-01-04 | 邯郸晨光珍品油脂有限公司 | A kind of production method of conjugated linoleic acid |
CN109134237B (en) * | 2018-04-18 | 2021-07-06 | 邯郸晨光珍品油脂有限公司 | Production method of conjugated linoleic acid |
CN110338222A (en) * | 2019-08-23 | 2019-10-18 | 李跃东 | A kind of high-purity conjugated linoleic acid yak yoghourt and preparation method thereof |
CN110338222B (en) * | 2019-08-23 | 2023-03-14 | 李跃东 | High-purity conjugated linoleic acid yak yoghourt and preparation method thereof |
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