CN107823137A - A kind of preparation method of injection refined fish oil - Google Patents
A kind of preparation method of injection refined fish oil Download PDFInfo
- Publication number
- CN107823137A CN107823137A CN201711336867.4A CN201711336867A CN107823137A CN 107823137 A CN107823137 A CN 107823137A CN 201711336867 A CN201711336867 A CN 201711336867A CN 107823137 A CN107823137 A CN 107823137A
- Authority
- CN
- China
- Prior art keywords
- fish oil
- lipase
- preparation
- injection
- refined fish
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Granted
Links
Classifications
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K9/00—Medicinal preparations characterised by special physical form
- A61K9/10—Dispersions; Emulsions
- A61K9/107—Emulsions ; Emulsion preconcentrates; Micelles
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K35/00—Medicinal preparations containing materials or reaction products thereof with undetermined constitution
- A61K35/56—Materials from animals other than mammals
- A61K35/60—Fish, e.g. seahorses; Fish eggs
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K9/00—Medicinal preparations characterised by special physical form
- A61K9/0012—Galenical forms characterised by the site of application
- A61K9/0019—Injectable compositions; Intramuscular, intravenous, arterial, subcutaneous administration; Compositions to be administered through the skin in an invasive manner
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
- C12P7/00—Preparation of oxygen-containing organic compounds
- C12P7/64—Fats; Fatty oils; Ester-type waxes; Higher fatty acids, i.e. having at least seven carbon atoms in an unbroken chain bound to a carboxyl group; Oxidised oils or fats
- C12P7/6409—Fatty acids
- C12P7/6427—Polyunsaturated fatty acids [PUFA], i.e. having two or more double bonds in their backbone
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
- C12P7/00—Preparation of oxygen-containing organic compounds
- C12P7/64—Fats; Fatty oils; Ester-type waxes; Higher fatty acids, i.e. having at least seven carbon atoms in an unbroken chain bound to a carboxyl group; Oxidised oils or fats
- C12P7/6436—Fatty acid esters
- C12P7/6445—Glycerides
Abstract
The invention discloses a kind of preparation method of injection refined fish oil.Using food grade fish oil as raw material, reaction is hydrolyzed using lipase as catalyst, is hydrolyzed using partial glyceride lipase and produces diglyceride and monoglyceride, the ester type refined fish oil of glycerol for injection three is prepared in alkali-refining deacidification, column chromatography removal of impurities.The product purification fish oil principal component content of triglyceride that the present invention obtains is high（More than 95%）, EPA content of fatty acid is that 18% ~ 30%, DHA content of fatty acid is 15% ~ 30%, and impurity content is low（Diglyceride and monoglyceride are less than 0.5%）, oxidation index is low（Acid number is less than 0.2, and peroxide value is less than 1.0, and anisidine value is less than 5.0）, product quality reaches injection refined fish oil standard.
Description
Technical field：
The invention belongs to technology of pharmaceutical engineering field, specifically, is related to a kind of preparation method of injection refined fish oil.
Background technology：
Fish oil is the natural oil refined in a kind of fat from fish, is formed rich in omega-3 unsaturated fatty acid sweet
Oily three esters.Docosahexaenoic acid in the serial polyunsaturated fatty acids of ω -3（DHA）And eicosapentaenoic acid（EPA）It is fish oil
Main composition, they be not only form meninx lipid main component, but also with prevention cardiovascular and cerebrovascular disease, adjust
Save blood ester, reduce blood viscosity, brain tonic and intelligence development, improvement and increase memory, enhancing human body antitumor immune function, suppress inflammation and
Improve eyesight and other effects.It is that ω -3 fish oil fats emulsion injection prepared by raw material can quickly increase carefully by injection refined fish oil
After birth omega-fatty acid content, anti-inflammatory and immunoregulation effect are clinically played, reduce complication, and for Critically ill
People, case fatality rate can be significantly reduced, reduce antibiotic dosage, shorten the hospital stays.
Its main component of the fish oil of natural origin is triglycerides, is practically free of diglyceride, monoglyceride, in the absence of fat
The artificial synthesized composition such as fat acetoacetic ester.The fish oil of natural origin reduces the techniques such as impurity and prepared by improving EPA, DHA content
As injection refined fish oil.Injection refined fish oil is as new function type fat dairy milk starting material, its natural attribute and quality mark
Standard has strict requirements, includes its source, the fatty acid species of principal component, content of fatty acid, diglyceride contains in product
Amount, the index such as monoglyceride content, peroxide value, anisidine value, acid number, iodine number.The how unsaturated of high content in fish oil
Aliphatic acid causes its oxidation index to be difficult to control, and is not easy to produce the qualified injection refined fish oil of oxidation index, simultaneously because
Production technology lack etc. reason, at present China can not produce injection refined fish oil, cause the country be difficult to carry out fish oil fat breast
Research, hinder China's Fat Emulsion industry development.
The domestic and international not report on injection refined fish oil preparation technology at present, is refined on food-grade
The technique report of fish oil.Currently in order to raising EPA and DHA content, the method for preparing the ester refined fish oil of food grade glycerin three are main
Including enzyme process and chemical method.Chinese patent CN101255380B discloses one kind ethyl ester type fish oil, glycerine, 435 types fat
Enzyme passes through the triglyceride type fish oil that is prepared of ester exchange reaction, the fatty-acid ethyl ester of this method products obtained therefrom, diglyceride,
The indexs such as monoglyceride, acid number, peroxide value, anisidine value control without effective, and prepared fish oil and non-natural
Fish oil.Chinese patent CN103242969B disclose one kind under the conditions of the base catalysts such as sodium hydroxide, ethyl ester type fish oil and
Glycerine, by the method for transesterification chemical conversion triglyceride type fish oil, is that a kind of prepare synthesizes fish oil under 155 DEG C ~ 165 DEG C high temperature
Method.Patented method high temperature processing can cause the generation of aldoketones oxidation material, cause oxidation index such as aminoanisole
The rise of value, and products obtained therefrom contains the impurity such as substantial amounts of fatty-acid ethyl ester, diglyceride, monoglyceride residual, has had no
Effect control.
The Master's thesis that Song Shijun delivers at it《The research of enzymatic clarification EPA, DHA triglycerides》In make use of RMIM enzymes
Alcoholysis has been carried out to fish oil, after molecular distillation, ester exchange is carried out using 435 enzymes and ethyl ester type EPA and DHA, is recycled inclined
Glycerin fatty enzyme hydrolysis diglyceride therein.The technical process produces abundant fatty acid second during first step alcoholysis
Ester, triglycerides are only left 8.3%, prepared subsequently after ester exchange reaction and inclined glycerin fatty enzyme hydrolysis diglyceride
The triglyceride type fish oil arrived is synthesis fish oil, and has fatty-acid ethyl ester residual.The technique also 100 DEG C of high temperature -160 DEG C it
Between used multiple-grade molecular distillation so that its product oxidation number is higher（Methyl oxyaniline value reaches 13.7）, do not efficiently control
Oxidation index.
Chinese patent CN105821088A discloses one kind and utilizes lipase hydrolysis fish oil, by molecular distillation, ester exchange
Method be prepared for triglyceride type fish oil, the fish oil that same this method is produced can remain EPA and DHA-EE, and it is
Synthesize fish oil.High temperature molecular distillation during this can also cause the oxidation index of product to raise, and not controlled subsequently
System.
The fish oil that existing patent family is refining to obtain is the fish oil of synthesis, and without control aliphatic acid second well
The oxidation index such as the impurity such as ester, diglyceride, monoglyceride and acid number, peroxide value, anisidine value.
The content of the invention：
Present invention aim to address above mentioned problem, exploitation one kind can keep refined fish oil product natural attribute, improve simultaneously
The method of the impurity such as EPA, DHA content, control diglyceride, monoglyceride and the injection refined fish oil of peroxidating index.
To achieve these goals, the present invention adopts the following technical scheme that：
A kind of preparation method of injection refined fish oil, using food grade fish oil as raw material, enter water-filling using lipase as catalyst
Solution reaction, hydrolyzed using partial glyceride lipase and produce diglyceride and monoglyceride, alkali-refining deacidification, column chromatography removal of impurities is prepared into
To the ester type refined fish oil of glycerol for injection three.
As preferable, in the preparation method of above-mentioned injection refined fish oil, the lipase is Sn-1,3 spies
Different in nature lipase.
As preferable, in the preparation method of above-mentioned injection refined fish oil, the Sn-1,3 specific fats
Enzyme is TLIM or RMIM lipase.
As preferable, in the preparation method of above-mentioned injection refined fish oil, weight in Lipase catalyzed hydrolysis reaction
Amount proportioning is fish oil 100, lipase 1 ~ 15, water 1 ~ 10.
As preferable, in the preparation method of above-mentioned injection refined fish oil, the bar of Lipase catalyzed hydrolysis reaction
Part is：Stirring, vacuum -0.05 ~ -0.1MPa, 30 ~ 70 DEG C of temperature, time 2h ~ 6h.
As preferable, in the preparation method of above-mentioned injection refined fish oil, the partial glyceride lipase is
Lipase G50 or Lipase SMG1 lipase.
As preferable, in the preparation method of above-mentioned injection refined fish oil, partial glyceride lipase hydrolysing step
Middle weight proportion is fish oil 100, partial glyceride lipase 0.1 ~ 10, water 0.5 ~ 10.
As preferable, in the preparation method of above-mentioned injection refined fish oil, partial glyceride lipase hydrolysing step
Condition be：Stirring, vacuum -0.05 ~ -0.1MPa, 30 ~ 70 DEG C of temperature, time 2h ~ 6h.
As preferable, in the preparation method of above-mentioned injection refined fish oil, the alkali used in the alkali-refining deacidification is
Solid base, it is calcium hydroxide or calcium oxide；The step of alkali-refining deacidification is：It is fish oil 100 to add solid base proportioning, solid base 6 ~
12, in vacuum -0.05 ~ -0.1MPa, 40 ~ 70 DEG C of temperature, stir depickling under conditions of filtering after 2h ~ 6h.
As preferable, in the preparation method of above-mentioned injection refined fish oil, the column chromatography removal of impurities is used to fill out
Expect for silica gel or glycol-based silica gel, the consumption proportion of column chromatography removal step is fish oil 100, silica gel 5 ~ 40.
Compared with prior art, beneficial effects of the present invention：
1. high content EPA and DHA is prepared by steps such as lipase hydrolysis, partial glyceride lipase hydrolysis in the present invention
（33%-60%）Refined fish oil, need not move through the ethyl esterified artificial lactate synthesis of grade, keep the native configurations of fish oil triglycerides with
Characteristic.
2. refined fish oil principal component content of triglyceride prepared by the present invention is high（More than 95%）, EPA content of fatty acid is
18% ~ 30%, DHA content of fatty acid are 15% ~ 30%, and impurity content is low（Diglyceride and monoglyceride are less than 0.5%）, oxidation refers to
Mark low（Acid number is less than 0.2, and peroxide value is less than 1.0, and anisidine value is less than 5.0）, product quality reaches injection and refines
Fish oil standard.
3. production operation of the present invention is simple, and used enzyme recoverable, technical process produces without using organic solvent
Waste water is few, energy-conserving and environment-protective.The commercially viable production of the technique obtains the qualified products for meeting injection refined fish oil, fills up domestic empty
In vain, break up monopoly, be advantageous to the development of domestic fatty dairy milk starting material and preparation industry.
Embodiment
Acid number, peroxide value, anisidine value, EPA content, DHA content are according to Europe in following examples products obtained therefrom
" FISH OIL, RICH IN OMEGA-3 ACIDS " standard methods describeds are detected in the version of continent pharmacopeia 8.0.
Fatty-acid ethyl ester, monoglyceride and diglyceride, content of triglyceride utilize high effective liquid chromatography for measuring：Will
100mg fish oil samples are placed in 10mL volumetric flasks, are added n-hexane dissolution and constant volume, are filtrated to get need testing solution.Testing conditions：
Utilize Agilent high performance liquid chromatograph, chromatographic column C18, 250 mm × 4.6 mm, 5 μm, mobile phase acetonitrile/isopropanol is from 0
40min, which becomes, turns to 40%/60% 0%/100%, and flow velocity 1mL/min, detector is evaporation photodetector.
Embodiment 1：
Fish oil raw material 1Kg is taken, 10g TLIM lipase and 10g water is added, inflated with nitrogen evacuation of air, is 30 DEG C in reaction temperature,
Vacuum -0.05 ~ -0.1MPa, 2h is reacted under conditions of stirring frequency 150r/min, is filtered after reaction.1g is added into oil strain
Partial glyceride lipase Lipase G50 and 5g water, inflated with nitrogen evacuation of air, it is 30 DEG C in reaction temperature, vacuum -0.05 ~ -
2h is reacted under conditions of 0.1MPa, stirring frequency 150r/min, is centrifuged after reaction, oil phase is filtered.60g hydrogen is added into oil strain
Calcium oxide, inflated with nitrogen evacuation of air, it is 40 DEG C in temperature, vacuum -0.05 ~ -0.1MPa, stirring frequency 150r/min condition
Lower absorption 2h, is filtered after absorption.The silicagel column for loading 50g silica gel carries out column chromatography, is prepared into refined fish oil finished product 786g.Finished product
Indices are shown in Table 1.
Embodiment 2：
Fish oil raw material 1Kg is taken, 60g RMIM lipase and 50g water is added, inflated with nitrogen evacuation of air, is 45 DEG C in reaction temperature,
Vacuum -0.05 ~ -0.1MPa, 3h is reacted under conditions of stirring frequency 150r/min.Filtered after reaction, 50g is added into oil strain
Partial glyceride lipase Lipase SMG1 and 35g water, inflated with nitrogen evacuation of air, it is 45 DEG C in reaction temperature, vacuum -0.05
3h is reacted under conditions of ~ -0.1MPa, stirring frequency 150r/min, is centrifuged after reaction, oil phase is filtered.90g is added into oil strain
Calcium oxide, inflated with nitrogen evacuation of air, it is 50 DEG C in reaction temperature, vacuum -0.05 ~ -0.1MPa, stirring frequency 150r/min's
Under the conditions of adsorb 3h, filtered after absorption.Load 100g glycol-based silica gel silicagel column carry out column chromatography, be prepared into refined fish oil into
Product 593g.Finished product indices are shown in Table 1.
Embodiment 3：
Fish oil raw material 1Kg is taken, 150g TLIM lipase and 100g water is added, inflated with nitrogen evacuation of air, is 70 in reaction temperature
DEG C, vacuum -0.05 ~ -0.1MPa, 6h is reacted under conditions of stirring frequency 150r/min.Filter after reaction, added into oil strain
100g partial glyceride lipase Lipase G50 and 50g water, inflated with nitrogen evacuation of air, it is 70 DEG C in reaction temperature, vacuum-
6h is reacted under conditions of 0.05 ~ -0.1MPa, stirring frequency 150r/min, is centrifuged after reaction, oil phase is filtered.Add into oil strain
Enter 120g calcium hydroxides, inflated with nitrogen evacuation of air, be 70 DEG C in reaction temperature, vacuum -0.05 ~ -0.1MPa, stirring frequency
4h is adsorbed under conditions of 150r/min, is filtered after absorption.The silicagel column for loading 150g silica gel carries out column chromatography, is prepared into refined fish
Oily finished product 504g.Finished product indices are shown in Table 1.
Embodiment 4：
Fish oil raw material 5Kg is taken, 350g TLIM lipase and 300g water is added, inflated with nitrogen evacuation of air, is 50 in reaction temperature
DEG C, vacuum -0.05 ~ -0.1MPa, 3h is reacted under conditions of stirring frequency 180r/min.Filter after reaction, added into oil strain
300g partial glyceride lipase Lipase G50 and 200g water, inflated with nitrogen evacuation of air, it is 50 DEG C in reaction temperature, vacuum-
3h is reacted under conditions of 0.05 ~ -0.1MPa, stirring frequency 150r/min, is centrifuged after reaction, oil phase is filtered.Add into oil strain
Enter 400g calcium hydroxides, inflated with nitrogen evacuation of air, be 50 DEG C in reaction temperature, vacuum -0.05 ~ -0.1MPa, stirring frequency
3h is adsorbed under conditions of 150r/min, is filtered after absorption.The silicagel column for loading 450g silica gel carries out column chromatography, is prepared into refined fish
Oily finished product 3.54Kg.Finished product indices are shown in Table 1.
Embodiment 5：
Fish oil raw material 10Kg is taken, 700g TLIM lipase and 650g water is added, inflated with nitrogen evacuation of air, is 50 in reaction temperature
DEG C, vacuum -0.05 ~ -0.1MPa, 3h is reacted under conditions of stirring frequency 180r/min.Filter after reaction, added into oil strain
600g partial glyceride lipase Lipase G50 and 450g water, inflated with nitrogen evacuation of air, it is 50 DEG C in reaction temperature, vacuum-
Centrifuged after 3h reactions are reacted under conditions of 0.05 ~ -0.1MPa, stirring frequency 180r/min, oil phase is filtered.Added into oil strain
800g calcium hydroxides, inflated with nitrogen evacuation of air, it is 50 DEG C in reaction temperature, vacuum -0.05 ~ -0.1MPa, stirring frequency
3h is adsorbed under conditions of 180r/min, is filtered after absorption.The silicagel column for loading 950g silica gel carries out column chromatography, is prepared into refined fish
Oily finished product 7.14Kg.Finished product indices are shown in Table 1.
Embodiment 6：
Fish oil raw material 10Kg is taken, 1Kg RMIM lipase and 900g water is added, inflated with nitrogen evacuation of air, is 50 in reaction temperature
DEG C, vacuum -0.05 ~ -0.1MPa, 3h is reacted under conditions of stirring frequency 180r/min.Filter after reaction, added into oil strain
800g partial glyceride lipase Lipase SMG1 and 650g water, inflated with nitrogen evacuation of air, it is 50 DEG C in reaction temperature, vacuum-
3h is reacted under conditions of 0.05 ~ -0.1MPa, stirring frequency 180r/min, is centrifuged after reaction, oil phase is filtered.Add into oil strain
Enter 1Kg calcium oxide, inflated with nitrogen evacuation of air, be 50 DEG C in reaction temperature, vacuum -0.05 ~ -0.1MPa, stirring frequency 150r/
3h is adsorbed under conditions of min, is filtered after absorption.The silicagel column for loading 1.1Kg glycol-based silica gel carries out column chromatography, is prepared into refined
Fish oil finished product 6.54Kg.Finished product indices are shown in Table 2.
Embodiment 7：
Fish oil raw material 20Kg is taken, 1.6Kg TLIM lipase and 1.4Kg water is added, inflated with nitrogen evacuation of air, is in reaction temperature
50 DEG C, vacuum -0.05 ~ -0.1MPa, 4h is reacted under conditions of stirring frequency 180r/min.Filter after reaction, add into oil strain
Enter 1.5Kg partial glyceride lipase Lipase G50 and 1.3Kg water, inflated with nitrogen evacuation of air, be 50 DEG C in reaction temperature, vacuum
- 0.05 ~ -0.1MPa is spent, 4h is reacted under conditions of stirring frequency 180r/min, is centrifuged after reaction, oil phase is filtered.Into oil strain
1.8Kg calcium oxide is added, inflated with nitrogen evacuation of air, is 50 DEG C in reaction temperature, vacuum -0.05 ~ -0.1MPa, stirring frequency
4h is adsorbed under conditions of 150r/min, is filtered after absorption.The silicagel column for loading 2Kg silica gel carries out column chromatography, is prepared into refined fish
Oily finished product 13.8Kg.Finished product indices are shown in Table 2.
Embodiment 8：
Fish oil raw material 50Kg is taken, 3Kg TLIM lipase and 2.8Kg water is added, inflated with nitrogen evacuation of air, is 50 in reaction temperature
DEG C, vacuum -0.05 ~ -0.1MPa, 5h is reacted under conditions of stirring frequency 180r/min.Filter after reaction, added into oil strain
2.5Kg partial glyceride lipase Lipase G50 and 2.2Kg water, inflated with nitrogen evacuation of air, it is 50 DEG C in reaction temperature, vacuum
- 0.05 ~ -0.1MPa is spent, 5h is reacted under conditions of stirring frequency 180r/min, is centrifuged after reaction, oil phase is filtered.Into oil strain
3.5Kg calcium hydroxides are added, inflated with nitrogen evacuation of air, are 50 DEG C in reaction temperature, vacuum -0.05 ~ -0.1MPa, stirring frequency
5h is adsorbed under conditions of rate 150r/min, is filtered after absorption.The silicagel column for loading 4.5Kg silica gel carries out column chromatography, is prepared into essence
Fish oil finished product 36.8Kg processed.Finished product indices are shown in Table 2.
Embodiment 9：
Fish oil raw material 100Kg is taken, 7Kg TLIM lipase and 6.5Kg water is added, inflated with nitrogen evacuation of air, is 50 in reaction temperature
DEG C, vacuum -0.05 ~ -0.1MPa, 6h is reacted under conditions of stirring frequency 160r/min.Filter after reaction, added into oil strain
6Kg partial glyceride lipase Lipase G50 and 5.5Kg water, inflated with nitrogen evacuation of air, it is 50 DEG C in reaction temperature, vacuum-
5h is reacted under conditions of 0.05 ~ -0.1MPa, stirring frequency 160r/min, is centrifuged after reaction, oil phase is filtered.Add into oil strain
Enter 8Kg calcium hydroxides, inflated with nitrogen evacuation of air, be 50 DEG C in reaction temperature, vacuum -0.05 ~ -0.1MPa, stirring frequency
5h is adsorbed under conditions of 160r/min, is filtered after absorption.Filtering, then extract fish oil crude product 3 times with 3 × 16L aqueous phases, take oil phase to add
Enter anhydrous sodium sulfate 5Kg to be dried, filter.Load 10Kg silica gel silicagel column carry out column chromatography, be prepared into refined fish oil into
Product 72.5Kg.Finished product indices are shown in Table 2.
Embodiment 10：
Fish oil raw material 100Kg is taken, 12Kg RMIM lipase and 8Kg water is added, inflated with nitrogen evacuation of air, is 50 in reaction temperature
DEG C, vacuum -0.05 ~ -0.1MPa, 6h is reacted under conditions of stirring frequency 160r/min.Filter after reaction, added into oil strain
9Kg partial glyceride lipase Lipase SMG1 and 5.5Kg water, inflated with nitrogen evacuation of air, it is 50 DEG C in reaction temperature, vacuum-
5h is reacted under conditions of 0.05 ~ -0.1MPa, stirring frequency 160r/min, is centrifuged after reaction, oil phase is filtered.Add into oil strain
Enter 9Kg calcium hydroxides, inflated with nitrogen evacuation of air, be 50 DEG C in reaction temperature, vacuum -0.05 ~ -0.1MPa, stirring frequency
4h is adsorbed under conditions of 150r/min, is filtered after absorption.The silicagel column for loading 12Kg glycol-based silica gel carries out column chromatography, is prepared into
Refined fish oil finished product 64.6Kg.Finished product indices are shown in Table 2.
1. implement according to the methods described of Chinese patent CN101255380B embodiments 1, using food-grade fish oil as initiation material,
Prepare refined fish oil sample 1.
2. the Master's thesis delivered according to Song Shijun《The research of enzymatic clarification EPA, DHA triglycerides》Chapter 5, the side in
Method, using food-grade fish oil as initiation material, prepare refined fish oil sample 2.
3. implement according to the methods described of Chinese patent CN105821088A embodiments 1, using food-grade fish oil as initiation material,
Prepare refined fish oil sample 3.
The detection data of table 1, embodiment 1-5
| Testing index | Standard requirement | Embodiment 1 | Embodiment 2 | Embodiment 3 | Embodiment 4 | Embodiment 5 |
| Fatty-acid ethyl ester % | It must not detect | Do not detect | Do not detect | Do not detect | Do not detect | Do not detect |
| Diglyceride and monoglyceride % | Less than 0.5% | 0.31 | 0.15 | 0.01 | 0.11 | 0.05 |
| Triglycerides % | More than 95% | 99.6 | 99.7 | 99.9 | 99.8 | 99.9 |
| EPA% | More than 18% | 22.3 | 25.1 | 30.4 | 26.1 | 25.8 |
| DHA% | More than 15% | 19.2 | 22.3 | 27.8 | 23.4 | 23.1 |
| Acid number | Less than 0.2 | 0.15 | 0.05 | 0.04 | 0.10 | 0.05 |
| Peroxide value | Less than 1.0 | 0.91 | 0.41 | 0.12 | 0.39 | 0.34 |
| Anisidine value | Less than 5.0 | 4.0 | 2.1 | 0.32 | 1.9 | 1.4 |
The detection data of table 2, embodiment 6-10
| Testing index | Standard requirement | Embodiment 6 | Embodiment 7 | Embodiment 8 | Embodiment 9 | Embodiment 10 |
| Fatty-acid ethyl ester % | It must not detect | Do not detect | Do not detect | Do not detect | Do not detect | Do not detect |
| Diglyceride and monoglyceride % | Less than 0.5% | 0.13 | 0.14 | 0.15 | 0.10 | 0.08 |
| Triglycerides % | More than 95% | 99.8 | 99.8 | 99.8 | 99.8 | 99.9 |
| EPA% | More than 18% | 26.8 | 25.9 | 25.2 | 26.3 | 29.3 |
| DHA% | More than 15% | 24.2 | 23.3 | 22.8 | 24.4 | 27.6 |
| Acid number | Less than 0.2 | 0.08 | 0.05 | 0.11 | 0.10 | 0.05 |
| Peroxide value | Less than 1.0 | 0.34 | 0.37 | 0.35 | 0.32 | 0.18 |
| Anisidine value | Less than 5.0 | 1.8 | 2.1 | 1.9 | 1.5 | 0.53 |
The detection data of table 3, sample 1-3
| Testing index | Standard requirement | Sample 1 | Sample 2 | Sample 3 |
| Fatty-acid ethyl ester % | It must not detect | 15.3 | 2.81 | 2.92 |
| Diglyceride and monoglyceride % | Less than 0.5% | 29.4 | 0.91 | 5.43 |
| Triglycerides % | More than 95% | 55.2 | 96.2 | 91.5 |
| EPA% | More than 18% | 32.1 | 38.1 | 25.6 |
| DHA% | More than 15% | 26.1 | 25.1 | 17.4 |
| Acid number | Less than 0.2 | 3.4 | 0.63 | 2.1 |
| Peroxide value | Less than 1.0 | 5.2 | 3.5 | 4.6 |
| Anisidine value | Less than 5.0 | 20.6 | 15.7 | 17.3 |
Table 1-3 result is shown, using the technical method of existing patent report, can not be prepared without fatty-acid ethyl ester, sweet
The impurity contents such as oily diester and monoglyceride are low, the oxidation index such as peroxide value, anisidine value it is low possess natural characteristic
The ester type refined fish oil of glycerol for injection three.The present invention from fatty electrodes method hydrolysis, partial glyceride lipase specific for hydrolysis,
The technical methods such as alkali-refining deacidification, column chromatography removal of impurities, are used enzyme dosage, enzyme hydrolysis condition, solid base amount, depickling condition, silica gel
The factors such as amount have carried out many experiments exploration, by the preferred compositions of above-mentioned technology, have obtained a kind of holding fish oil triglycerides
Native configurations and characteristic, low impurity（No fatty acids ethyl ester, diglyceride and monoglyceride are less than 0.5%）, suboxides index
（Acid number is less than 5.0 less than 0.2, peroxide value less than 1.0, anisidine value）, high EPA and DHA content（33%~60%）Note
Penetrate and use refined fish oil, reach the inaccessiable effect of existing patented technology institute.
Claims (10)
1. a kind of preparation method of injection refined fish oil, it is characterized in that using food grade fish oil as raw material, using lipase as urging
Reaction is hydrolyzed in agent, is hydrolyzed using partial glyceride lipase and produces diglyceride and monoglyceride, alkali-refining deacidification, column chromatography
The ester type refined fish oil of glycerol for injection three is prepared in removal of impurities.
2. the preparation method of injection refined fish oil according to claim 1, it is characterized in that：The lipase is Sn-1,3
Position specific lipase.
3. the preparation method of injection refined fish oil according to claim 2, it is characterized in that：The Sn-1,3 positions specificity
Lipase is TLIM or RMIM lipase.
4. the preparation method of injection refined fish oil according to claim 1, it is characterized in that：Lipase catalyzed hydrolysis reacts
Middle weight proportion is fish oil 100, lipase 1 ~ 15, water 1 ~ 10.
5. the preparation method of injection refined fish oil according to claim 1, it is characterized in that：Lipase catalyzed hydrolysis reacts
Condition be：Stirring, vacuum -0.05 ~ -0.1MPa, 30 ~ 70 DEG C of temperature, time 2h ~ 6h.
6. the preparation method of injection refined fish oil according to claim 1, it is characterized in that：The partial glyceride lipase
For Lipase G50 or Lipase SMG1 lipase.
7. the preparation method of injection refined fish oil according to claim 1, it is characterized in that：Partial glyceride lipase hydrolyzes
Weight proportion is fish oil 100 in step, partial glyceride lipase 0.1 ~ 10, water 0.5 ~ 10.
8. the preparation method of injection refined fish oil according to claim 1, it is characterized in that：Partial glyceride lipase hydrolyzes
The condition of step is：Stirring, vacuum -0.05 ~ -0.1MPa, 30 ~ 70 DEG C of temperature, time 2h ~ 6h.
9. the preparation method of injection refined fish oil according to claim 1, it is characterized in that：Used in the alkali-refining deacidification
Alkali is solid base, is calcium hydroxide or calcium oxide；The step of alkali-refining deacidification is：It is fish oil 100 to add solid base proportioning, solid base
6 ~ 12, in vacuum -0.05 ~ -0.1MPa, 40 ~ 70 DEG C of temperature, stir depickling under conditions of filtering after 2h ~ 6h.
10. the preparation method of injection refined fish oil according to claim 1, it is characterized in that：The column chromatography removal of impurities institute
Filler is silica gel or glycol-based silica gel, and the consumption proportion of column chromatography removal step is fish oil 100, silica gel 5 ~ 40.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201711336867.4A CN107823137B (en) | 2017-12-14 | 2017-12-14 | Preparation method of refined fish oil for injection |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201711336867.4A CN107823137B (en) | 2017-12-14 | 2017-12-14 | Preparation method of refined fish oil for injection |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN107823137A true CN107823137A (en) | 2018-03-23 |
| CN107823137B CN107823137B (en) | 2020-11-13 |
Family
ID=61644521
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201711336867.4A Active CN107823137B (en) | 2017-12-14 | 2017-12-14 | Preparation method of refined fish oil for injection |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN107823137B (en) |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109593606A (en) * | 2019-01-31 | 2019-04-09 | 福建师范大学 | A kind of deep sea fish oil and its processing technology of high DHA content |
| CN112625786A (en) * | 2020-12-31 | 2021-04-09 | 浙江工业大学 | Preparation method of tuna oil |
| CN113584092A (en) * | 2021-07-27 | 2021-11-02 | 华南理工大学 | Method for enriching EPA and DHA in fish oil through enzymatic hydrolysis |
Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2005071053A1 (en) * | 2004-01-26 | 2005-08-04 | Uniliver N.V | Enzymatic modification of triglyceride fats |
| CN101348807A (en) * | 2008-08-26 | 2009-01-21 | 江南大学 | Method for enrichment of n-3 polyunsaturated fatty acid glyceride from fish oil |
| CN101824364A (en) * | 2010-04-09 | 2010-09-08 | 浙江兴业集团有限公司 | Method for deacidifying and refining high-acid value fish oil by using enzyme method |
| CN102994580A (en) * | 2012-11-30 | 2013-03-27 | 华南理工大学 | Preparation method of high-purity triglyceride type PUFA (Polyunsaturated Fatty Acid) |
| CN103757065A (en) * | 2014-01-13 | 2014-04-30 | 青岛澳海生物有限公司 | Preparation method of high-purity conjugated linoleic triglyceride |
| CN105821088A (en) * | 2016-05-26 | 2016-08-03 | 暨南大学 | Method for preparing glyceride rich in EPA and DHA through enzyme catalysis |
-
2017
- 2017-12-14 CN CN201711336867.4A patent/CN107823137B/en active Active
Patent Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2005071053A1 (en) * | 2004-01-26 | 2005-08-04 | Uniliver N.V | Enzymatic modification of triglyceride fats |
| CN101348807A (en) * | 2008-08-26 | 2009-01-21 | 江南大学 | Method for enrichment of n-3 polyunsaturated fatty acid glyceride from fish oil |
| CN101824364A (en) * | 2010-04-09 | 2010-09-08 | 浙江兴业集团有限公司 | Method for deacidifying and refining high-acid value fish oil by using enzyme method |
| CN102994580A (en) * | 2012-11-30 | 2013-03-27 | 华南理工大学 | Preparation method of high-purity triglyceride type PUFA (Polyunsaturated Fatty Acid) |
| CN103757065A (en) * | 2014-01-13 | 2014-04-30 | 青岛澳海生物有限公司 | Preparation method of high-purity conjugated linoleic triglyceride |
| CN105821088A (en) * | 2016-05-26 | 2016-08-03 | 暨南大学 | Method for preparing glyceride rich in EPA and DHA through enzyme catalysis |
Non-Patent Citations (5)
| Title |
|---|
| GERALD P. MCNEILL，等: "Lipase-Catalyzed Enrichment of Long-Chain Polyunsaturated Fatty Acids", 《JAOCS》 * |
| 刘书成，等: "酶法制备n-3多不饱和脂肪酸甘油酯的研究进展", 《海洋水产研究》 * |
| 孙兆敏，等: "酶法制备n-3多不饱和脂肪酸甘油三酯的工艺", 《食品工业科技》 * |
| 朱东奇，等: "重组米根霉脂肪酶的酶学性质及其催化水解鳀鱼油富集DHA的研究", 《粮油食品科技》 * |
| 高巍，等: "TLIM脂肪酶对油脂水解特性的研究", 《粮油加工》 * |
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109593606A (en) * | 2019-01-31 | 2019-04-09 | 福建师范大学 | A kind of deep sea fish oil and its processing technology of high DHA content |
| CN109593606B (en) * | 2019-01-31 | 2022-07-08 | 福建师范大学 | Deep sea fish oil with high DHA content and processing technology thereof |
| CN112625786A (en) * | 2020-12-31 | 2021-04-09 | 浙江工业大学 | Preparation method of tuna oil |
| CN113584092A (en) * | 2021-07-27 | 2021-11-02 | 华南理工大学 | Method for enriching EPA and DHA in fish oil through enzymatic hydrolysis |
Also Published As
| Publication number | Publication date |
|---|---|
| CN107823137B (en) | 2020-11-13 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN111088296B (en) | Method for enriching n-3 polyunsaturated fatty acid glyceride in grease | |
| CN104186705B (en) | Method based on enzymatic acidolysis palmitic acid three Lipase absobed structured lipid | |
| CN101940241B (en) | Pangasianodon gigas lipid, application and method for preparing breast milk lipid substitutes thereof | |
| CN107823137A (en) | A kind of preparation method of injection refined fish oil | |
| CN105821089A (en) | Method for enzymatically synthesizing medium-long chain structure triglyceride | |
| EP3208261B1 (en) | Method for preparing glyceride type polyunsaturated fatty acids | |
| CN103952448B (en) | A kind of method utilizing enzyme-chemically method directional preparation OPO | |
| JP7242769B2 (en) | Free polyunsaturated fatty acid-containing composition and method for producing the same | |
| CN103305559A (en) | Preparation method for natural flavor fatty acid ester | |
| CN105779140A (en) | Preparation method of ethyl ester type fish oil with high EPA content | |
| CN102994580A (en) | Preparation method of high-purity triglyceride type PUFA (Polyunsaturated Fatty Acid) | |
| CN101161819A (en) | Method for preparing n-3PUFA ocean glycerin ester by enzymatical process | |
| WO2015024055A1 (en) | Separation of omega-3 fatty acids | |
| CN107698447B (en) | Method for preparing fatty glyceride | |
| CN113789354A (en) | Process for preparing edible oil diglyceride | |
| CN105400837A (en) | Method for preparing diglyceride through enzyme catalysis | |
| US11396667B2 (en) | Enzymatic method for preparation of lecithin polyunsaturated fatty acids (PUFAs) | |
| US20220119852A1 (en) | METHOD FOR IMPROVING n-3 POLYUNSATURATED FATTY ACIDS ENRICHMENT EFFICIENCY IN GLYCERIDE | |
| CN108911981B (en) | Preparation method of omega-3-fatty acid triglyceride for injection | |
| CN105132153B (en) | A method of preparing squid liver oil | |
| JP4310387B2 (en) | Omega-3 highly unsaturated fatty acid-containing partial glyceride composition and method for producing the same | |
| CN109880858B (en) | Method for reducing content of free fatty acid in marine phospholipid | |
| CN114574280A (en) | Preparation method of high-content polyunsaturated fatty acid grease | |
| CN106135902A (en) | A kind of diglyceride fluid composition rich in catalpic acid and its preparation method and application | |
| KR101115283B1 (en) | Process for preparing a complex of omega-3 unsaturated fatty acid and ascorbic acid |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant |