CN105949089A - Method for extracting natural taurine from mussels - Google Patents
Method for extracting natural taurine from mussels Download PDFInfo
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- CN105949089A CN105949089A CN201610556023.XA CN201610556023A CN105949089A CN 105949089 A CN105949089 A CN 105949089A CN 201610556023 A CN201610556023 A CN 201610556023A CN 105949089 A CN105949089 A CN 105949089A
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- XOAAWQZATWQOTB-UHFFFAOYSA-N taurine Chemical compound NCCS(O)(=O)=O XOAAWQZATWQOTB-UHFFFAOYSA-N 0.000 title claims abstract description 142
- 229960003080 taurine Drugs 0.000 title claims abstract description 71
- 241000237536 Mytilus edulis Species 0.000 title claims abstract description 35
- 235000020638 mussel Nutrition 0.000 title claims abstract description 33
- 238000000034 method Methods 0.000 title claims abstract description 26
- 239000007788 liquid Substances 0.000 claims abstract description 11
- 239000000047 product Substances 0.000 claims description 27
- 235000013372 meat Nutrition 0.000 claims description 11
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 11
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 claims description 9
- 239000006228 supernatant Substances 0.000 claims description 8
- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 claims description 6
- 238000002835 absorbance Methods 0.000 claims description 5
- 101800000263 Acidic protein Proteins 0.000 claims description 4
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 claims description 4
- 238000002360 preparation method Methods 0.000 claims description 4
- 230000008569 process Effects 0.000 claims description 4
- 102000004169 proteins and genes Human genes 0.000 claims description 4
- 108090000623 proteins and genes Proteins 0.000 claims description 4
- 238000003809 water extraction Methods 0.000 claims description 4
- 101710093543 Probable non-specific lipid-transfer protein Proteins 0.000 claims description 3
- 230000002378 acidificating effect Effects 0.000 claims description 3
- 239000012153 distilled water Substances 0.000 claims description 3
- 239000000706 filtrate Substances 0.000 claims description 3
- 239000011347 resin Substances 0.000 claims description 3
- 229920005989 resin Polymers 0.000 claims description 3
- 238000004042 decolorization Methods 0.000 claims description 2
- 238000005360 mashing Methods 0.000 claims description 2
- 238000000605 extraction Methods 0.000 abstract description 15
- 239000000126 substance Substances 0.000 abstract description 5
- 230000008901 benefit Effects 0.000 abstract description 4
- 238000003912 environmental pollution Methods 0.000 abstract description 3
- 239000002994 raw material Substances 0.000 abstract description 3
- 235000014102 seafood Nutrition 0.000 abstract 2
- 102000008186 Collagen Human genes 0.000 abstract 1
- 108010035532 Collagen Proteins 0.000 abstract 1
- 229920001436 collagen Polymers 0.000 abstract 1
- 239000003960 organic solvent Substances 0.000 abstract 1
- 238000002474 experimental method Methods 0.000 description 14
- YRKCREAYFQTBPV-UHFFFAOYSA-N acetylacetone Chemical compound CC(=O)CC(C)=O YRKCREAYFQTBPV-UHFFFAOYSA-N 0.000 description 12
- 239000000284 extract Substances 0.000 description 11
- 230000000694 effects Effects 0.000 description 9
- 239000000243 solution Substances 0.000 description 9
- 241000283690 Bos taurus Species 0.000 description 6
- WSFSSNUMVMOOMR-UHFFFAOYSA-N Formaldehyde Chemical compound O=C WSFSSNUMVMOOMR-UHFFFAOYSA-N 0.000 description 6
- 239000004365 Protease Substances 0.000 description 6
- 230000007062 hydrolysis Effects 0.000 description 6
- 238000006460 hydrolysis reaction Methods 0.000 description 6
- 108090000790 Enzymes Proteins 0.000 description 5
- 102000004190 Enzymes Human genes 0.000 description 5
- 108090000526 Papain Proteins 0.000 description 5
- NINIDFKCEFEMDL-UHFFFAOYSA-N Sulfur Chemical compound [S] NINIDFKCEFEMDL-UHFFFAOYSA-N 0.000 description 5
- 229940088598 enzyme Drugs 0.000 description 5
- 229940055729 papain Drugs 0.000 description 5
- 235000019834 papain Nutrition 0.000 description 5
- 241000238557 Decapoda Species 0.000 description 4
- VMHLLURERBWHNL-UHFFFAOYSA-M Sodium acetate Chemical compound [Na+].CC([O-])=O VMHLLURERBWHNL-UHFFFAOYSA-M 0.000 description 4
- 239000005864 Sulphur Substances 0.000 description 4
- 238000010521 absorption reaction Methods 0.000 description 4
- 239000002253 acid Substances 0.000 description 4
- 239000013078 crystal Substances 0.000 description 4
- 238000001514 detection method Methods 0.000 description 4
- 238000001914 filtration Methods 0.000 description 4
- 239000001632 sodium acetate Substances 0.000 description 4
- 235000017281 sodium acetate Nutrition 0.000 description 4
- 239000003643 water by type Substances 0.000 description 4
- QTBSBXVTEAMEQO-UHFFFAOYSA-N Acetic acid Chemical compound CC(O)=O QTBSBXVTEAMEQO-UHFFFAOYSA-N 0.000 description 3
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 3
- LSNNMFCWUKXFEE-UHFFFAOYSA-M Bisulfite Chemical compound OS([O-])=O LSNNMFCWUKXFEE-UHFFFAOYSA-M 0.000 description 2
- UFHFLCQGNIYNRP-UHFFFAOYSA-N Hydrogen Chemical compound [H][H] UFHFLCQGNIYNRP-UHFFFAOYSA-N 0.000 description 2
- 238000005481 NMR spectroscopy Methods 0.000 description 2
- 238000006243 chemical reaction Methods 0.000 description 2
- 239000003153 chemical reaction reagent Substances 0.000 description 2
- HVYWMOMLDIMFJA-DPAQBDIFSA-N cholesterol Chemical compound C1C=C2C[C@@H](O)CC[C@]2(C)[C@@H]2[C@@H]1[C@@H]1CC[C@H]([C@H](C)CCCC(C)C)[C@@]1(C)CC2 HVYWMOMLDIMFJA-DPAQBDIFSA-N 0.000 description 2
- 150000001875 compounds Chemical class 0.000 description 2
- 238000013461 design Methods 0.000 description 2
- 238000010790 dilution Methods 0.000 description 2
- 239000012895 dilution Substances 0.000 description 2
- 230000002526 effect on cardiovascular system Effects 0.000 description 2
- 230000002708 enhancing effect Effects 0.000 description 2
- 229910052739 hydrogen Inorganic materials 0.000 description 2
- 239000001257 hydrogen Substances 0.000 description 2
- 150000002500 ions Chemical class 0.000 description 2
- 239000000463 material Substances 0.000 description 2
- 238000001953 recrystallisation Methods 0.000 description 2
- 238000002798 spectrophotometry method Methods 0.000 description 2
- 238000010189 synthetic method Methods 0.000 description 2
- 238000002604 ultrasonography Methods 0.000 description 2
- SLZAMKNXRDJWLA-UHFFFAOYSA-N 1-(5-acetyl-2,6-dimethyl-1,4-dihydropyridin-3-yl)ethanone Chemical compound CC(=O)C1=C(C)NC(C)=C(C(C)=O)C1 SLZAMKNXRDJWLA-UHFFFAOYSA-N 0.000 description 1
- 241000219112 Cucumis Species 0.000 description 1
- 235000015510 Cucumis melo subsp melo Nutrition 0.000 description 1
- DGAQECJNVWCQMB-PUAWFVPOSA-M Ilexoside XXIX Chemical compound C[C@@H]1CC[C@@]2(CC[C@@]3(C(=CC[C@H]4[C@]3(CC[C@@H]5[C@@]4(CC[C@@H](C5(C)C)OS(=O)(=O)[O-])C)C)[C@@H]2[C@]1(C)O)C)C(=O)O[C@H]6[C@@H]([C@H]([C@@H]([C@H](O6)CO)O)O)O.[Na+] DGAQECJNVWCQMB-PUAWFVPOSA-M 0.000 description 1
- 241000058338 Macrobrachium nipponense Species 0.000 description 1
- 241001465754 Metazoa Species 0.000 description 1
- 241000238413 Octopus Species 0.000 description 1
- 241000237509 Patinopecten sp. Species 0.000 description 1
- 108091005804 Peptidases Proteins 0.000 description 1
- 102100037486 Reverse transcriptase/ribonuclease H Human genes 0.000 description 1
- FJJCIZWZNKZHII-UHFFFAOYSA-N [4,6-bis(cyanoamino)-1,3,5-triazin-2-yl]cyanamide Chemical compound N#CNC1=NC(NC#N)=NC(NC#N)=N1 FJJCIZWZNKZHII-UHFFFAOYSA-N 0.000 description 1
- 230000009471 action Effects 0.000 description 1
- 238000013019 agitation Methods 0.000 description 1
- 239000003513 alkali Substances 0.000 description 1
- 238000004458 analytical method Methods 0.000 description 1
- 230000003064 anti-oxidating effect Effects 0.000 description 1
- 150000001576 beta-amino acids Chemical class 0.000 description 1
- 230000008827 biological function Effects 0.000 description 1
- 230000004641 brain development Effects 0.000 description 1
- 238000005119 centrifugation Methods 0.000 description 1
- 239000003610 charcoal Substances 0.000 description 1
- 235000012000 cholesterol Nutrition 0.000 description 1
- 238000007796 conventional method Methods 0.000 description 1
- 239000012043 crude product Substances 0.000 description 1
- 238000002425 crystallisation Methods 0.000 description 1
- 230000008025 crystallization Effects 0.000 description 1
- 238000007405 data analysis Methods 0.000 description 1
- 230000007812 deficiency Effects 0.000 description 1
- 238000002242 deionisation method Methods 0.000 description 1
- 230000004064 dysfunction Effects 0.000 description 1
- 238000010828 elution Methods 0.000 description 1
- 230000002124 endocrine Effects 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 230000007613 environmental effect Effects 0.000 description 1
- IDGUHHHQCWSQLU-UHFFFAOYSA-N ethanol;hydrate Chemical compound O.CCO IDGUHHHQCWSQLU-UHFFFAOYSA-N 0.000 description 1
- 235000013305 food Nutrition 0.000 description 1
- -1 formaldehyde 2,6-dimethyl 3,5-diacetyl 1,4 dihydropyridine Chemical compound 0.000 description 1
- AFVYYWDNIWVXOS-UHFFFAOYSA-N formaldehyde pentane-2,4-dione Chemical compound O=C.CC(=O)CC(C)=O AFVYYWDNIWVXOS-UHFFFAOYSA-N 0.000 description 1
- 210000000232 gallbladder Anatomy 0.000 description 1
- 238000010438 heat treatment Methods 0.000 description 1
- 230000036039 immunity Effects 0.000 description 1
- 238000002329 infrared spectrum Methods 0.000 description 1
- 238000002347 injection Methods 0.000 description 1
- 239000007924 injection Substances 0.000 description 1
- 238000011835 investigation Methods 0.000 description 1
- 238000005259 measurement Methods 0.000 description 1
- 230000010534 mechanism of action Effects 0.000 description 1
- 230000004060 metabolic process Effects 0.000 description 1
- 238000002156 mixing Methods 0.000 description 1
- 229930014626 natural product Natural products 0.000 description 1
- 230000007830 nerve conduction Effects 0.000 description 1
- 230000007935 neutral effect Effects 0.000 description 1
- 231100000252 nontoxic Toxicity 0.000 description 1
- 230000003000 nontoxic effect Effects 0.000 description 1
- 230000005311 nuclear magnetism Effects 0.000 description 1
- 235000015097 nutrients Nutrition 0.000 description 1
- 239000008188 pellet Substances 0.000 description 1
- 235000019419 proteases Nutrition 0.000 description 1
- 230000001105 regulatory effect Effects 0.000 description 1
- 235000020637 scallop Nutrition 0.000 description 1
- 239000011734 sodium Substances 0.000 description 1
- 229910052708 sodium Inorganic materials 0.000 description 1
- 239000007787 solid Substances 0.000 description 1
- 238000001228 spectrum Methods 0.000 description 1
- 239000012086 standard solution Substances 0.000 description 1
- 238000003756 stirring Methods 0.000 description 1
- 229910052717 sulfur Inorganic materials 0.000 description 1
- 239000011593 sulfur Substances 0.000 description 1
- 238000012360 testing method Methods 0.000 description 1
- 230000001988 toxicity Effects 0.000 description 1
- 231100000419 toxicity Toxicity 0.000 description 1
- 238000002525 ultrasonication Methods 0.000 description 1
- 238000003828 vacuum filtration Methods 0.000 description 1
- 239000002023 wood Substances 0.000 description 1
Classifications
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
- C07C303/00—Preparation of esters or amides of sulfuric acids; Preparation of sulfonic acids or of their esters, halides, anhydrides or amides
- C07C303/42—Separation; Purification; Stabilisation; Use of additives
- C07C303/44—Separation; Purification
Landscapes
- Chemical & Material Sciences (AREA)
- Organic Chemistry (AREA)
- Coloring Foods And Improving Nutritive Qualities (AREA)
Abstract
The invention relates to a method for extracting natural taurine from mussels and belongs to the technical field of chemical extraction. The method for extracting the natural taurine from the mussels comprises the following steps that S1, the mussels are used for preparing clear liquid containing taurine; S2, collagen is removed from the clear liquid; S3, the product obtained by the step 2 is separated and purified. The method for extracting the natural taurine from the mussels has the advantages that the most common mussels in Dalian are taken as the raw material, the price is low, the mussels belong to low-end seafood, the extraction manner is mild, and no environmental pollution problem exits. The technological operation is simple, and the yield is large. The natural taurine is extracted from the seafood rich in taurine, no organic solvent residual exists after a series of operation, and natural extraction is realized.
Description
Technical field
The invention belongs to chemical extraction technical field, particularly one from mussel, extract natural cattle sulphur
The method of acid.
Background technology
Taurine is the beta amino acids of a kind of sulfur-bearing, has biological function widely, mainly has promotion
Brain development, vision enhancing, regulate nerve conduction, promote to absorb, digest fat, participate in gallbladder solid
The metabolism of alcohol, safeguards cardiovascular and cerebrovascular vessel, endocrine dysfunction, the effect of antioxidation etc..Thus have and carry
High memory, vision enhancing, fat reducing, reduces cholesterol, protects cardiovascular and cerebrovascular vessel, defying age, carry
High immunity etc. all have certain effect.Along with the further investigation of the mechanism of action to taurine, its
Application also will be more and more extensive.Taurine is as a kind of important nutrient of humans and animals, from sky
So product extracts attention always.
In common food, content of taurine is extremely low or need to find, but marine product is containing the highest
Content.Chinese Marine Biology Resources is enriched, such as content in the marine product such as Concha Ostreae, scallop, Octopus
Abundant, but the price comparison of product own is expensive, if extracting taurine from them, relatively costly.
This problem is investigated by early stage, finds in the leftover bits and pieces of mussel, marine rainbow and prawn head and shrimp same
Containing abundant taurine.These products are widely present in Dalian Area, and cheap.From sea
The technique extracting taurine in product is simple and pollution-free, and extracts cattle with the leftover bits and pieces of aquatic products
Sulfonic acid has again the meaning of environmental protection, and prospect is the best.
Since at the beginning of nineteen fifty, start synthetic taurine in the world.At present, domestic and international city
The taurine used on field has taurine and the natural taurine of chemosynthesis, but mostly wherein is chemistry
Synthetic method produces.Although synthetic method taurine is cheap, but it is big to there is material toxicity, and technique is grasped
Make the problems such as complicated, environmental pollution.Along with raising and the progress of Green Chemistry of living standards of the people,
People increasingly advocate the taurine using natural extract.Therefore, from natural product, cattle sulphur is extracted
Acid becomes market urgent needs, has great application prospect.
Summary of the invention
It is an object of the invention to overcome the deficiencies in the prior art, taurine is passed through removing protein, takes off
Color, the method such as isolated and purified are extracted from shrimp.
The present invention solves it and technical problem is that and take techniques below scheme to realize: extract from mussel
The method of natural taurine, comprises the steps of S1 and utilizes the mussel preparation clear liquid containing taurine；
S2 is by described clear liquid removing protein；The product of described step S2 is carried out isolated and purified by S3.
Preferably, described step S1 comprises the steps of S1.1 and mussel is cleaned taking-up mussel of shelling
Meat；S1.2 is homogenized during just mussel meat puts into tissue mashing machine；Mussel meat after homogenate is claimed by S1.3
Weight also dilutes with the water of 3 times of quality；S1.4 regulates pH value；S1.5 puts into enter in constant incubator
Row water extraction；Water extraction product is filtered by S1.6, takes filtrate.
Preferably, described step S2 comprises the steps of S2.1 except acidic protein；S2.2 removes alkali
Property albumen.
Preferably, described step S2.1 is except acidic protein employing hydrochloric acid is by described step S1 product
PH is adjusted to 3-4, is then centrifuged 20 minutes under 4500r/min, and collects supernatant.
Preferably, described step S2.2 is except basic protein employing sodium hydroxide is by described step S2.1
The pH of product be adjusted to 9-10, under 4500r/min centrifugal 20 minutes, collect supernatant.
Preferably, the concretely comprising the following steps of described step S3: S3.1 regulates described step S2 product
PH to 4-5；The product of described step S3.1 is injected strong acidic ion resin exchange column by S3.2,
And use distilled water to carry out eluting；S3.3 collects the part of absorbance peak-peak about 0.55 and flows out
Liquid.
Preferably, exist between described step S2 and described step S3 described step S2 product is entered
The step of row decolouring.
Preferably, described decolorization process specifically comprises: add activity in the product of described step S2
Charcoal decolours.
Preferably, in the product of step S2 described in every 100mL, 2～3g activated carbons are added.
Advantages of the present invention and good effect be: advantages of nontoxic raw materials benefit obtains, and there is not environmental pollution
Problem.And technological operation is simple, yield is big.Use from the marine product the highest containing amount of taurine
Extract, take the most common mussel in Dalian Area as raw material.Gentle mode is extracted, and then passes through
After the operation of string, organic solvent-free remains, and accomplishes natural extract.
Accompanying drawing explanation
Fig. 1 is the canonical plotting of taurine；
Fig. 2 is that taurine is extracted influence curve figure by pH；
Fig. 3 is that taurine is extracted influence curve figure by enzymolysis time；
Fig. 4 is that taurine is extracted influence curve figure by hydrolysis temperature；
Fig. 5 is each factorial effect curve chart；
Fig. 6 is taurine standard substance infrared spectrogram；
Fig. 7 is the infrared spectrogram that taurine is extracted in experiment；
Fig. 8 is taurine standard substance hydrogen nuclear magnetic resonance spectrogram；
Fig. 9 is taurine sample product hydrogen nuclear magnetic resonance spectrogram.
Detailed description of the invention
Below in conjunction with the accompanying drawings, by specific embodiment, the invention will be further described.Following example
The most illustrative, it not determinate, it is impossible to limit protection scope of the present invention with this.Implement
Experimental technique described in example if no special instructions, is conventional method；If no special instructions, described
Reagent and equipment, the most commercially obtain.
Experiment reagent
Experimental apparatus
The determination of standard curve regression equation
Take 10mmol/L taurine titer 0mL, 1mL, 2mL, 4mL, 6mL, 8mL, 10mL respectively,
It is settled to 10mL, respectively takes one milliliter, add 1ml developer, 8ml 1mol/L sodium acetate, at 400nm
Lower measurement OD value, obtains standard curve (such as Fig. 1).
Developer is by 10ml 1mol/L sodium acetate, 0.4ml acetylacetone,2,4-pentanedione, 1ml formaldehyde deionization
Water is settled to 25ml preparation, need to face with now joining.Color reaction reason behind is, containing acetic acid
In the solution of sodium, taurine can produce N-substituent group under the high temperature conditions with acetylacetone,2,4-pentanedione and formaldehyde
2,6-dimethyl 3,5-diacetyl 1,4 dihydropyridine.This coordination compound can present significantly
Yellow, and content of taurine is the highest, then and color is the most obvious, maximum absorption wave a length of 390～400nm.
Taurine used in the present invention colour developing principle is: in the presence of sodium acetate, taurine and acetyl
Acetone and the heated reaction of formaldehyde generate N-substituent group 2,6-dimethyl 3,5-diacetyl 1,4 dihydro
Pyridine complex.This coordination compound displaing yellow, its absorbance is just becoming within the specific limits with content of taurine
Ratio, maximum absorption wave a length of 390～400nm.
Embodiment 1
Step 1: containing the preparation of taurine clear liquid
Mussel meat is pulverized, adds pure water, stir 10min, agitator speed 30r/min, will
PH value is regulated to neutral after the water dilution of the Macrobrachium nipponensis equivalent volumes of homogenate.
Step 2: ultrasonication
Preferably incorporate in homogenate for convenience of taurine, according to the difference of experimental group, use ultrasonic
Ripple crushes instrument and carries out the broken of 5min, 10min, 15min, 20min, 25min respectively, utilizes water
Homogenate is filtered by circulation sucking filtration machine, takes filtrate.
Step 3: enzyme process slightly carries taurine
Take the homogenate 30ml crushed, be added thereto to 0.5g neutrality papain, after mixing
Regulation pH puts in the pot of assigned temperature constant temperature waters to designated value, stands 20min, and middle agitation is several
Secondary.Homogenate extraction completed, puts into heating 15min in the water-bath of 100 degrees Celsius, makes wood
Melon protease loses activity under high temperature action.
During experiment of single factor, take three variablees, except the ultrasonic disruption time mentioned above,
Also having temperature and the pH of enzyme extraction, each variable respectively sets five levels.Through inquiry, experiment is used
The use pH of papain be 6～7, using temperature is 55～65 DEG C, and its pH scope is expanded by I
To 5.5～7.5, and it is that shelves carry out experiment of single factor often to differ 0.5；Its temperature range is expanded
To 45～65 DEG C, and it is shelves with every 5 DEG C.
In the middle of orthogonal experiment process, according to bell shaped curve, from five condition setting, select four bars
Part, carries out the orthogonal experiment of three factor four levels.
Step 4: removing protein
Take the Carnis Mactrae meat homogenate (extracting solution) after enzyme denaturing, use recirculated water Vacuum filtration device repeatedly
Sucking filtration 2～3 times to extracting solution in the clear solution slightly turned white.
Extract with 5%HCl (mass ratio, below-mentioned all percentage ratios are mass ratio) regulation
Liquid pH to 4～5, room temperature, 4500r/min are centrifuged 15min.Stay supernatant.
The supernatant concentration obtained in previous step is that 5%NaOH regulates pH to 9～10, and continues
Under normal temperature condition, with the centrifugation 15min of 4500r/min.Stay supernatant.
Step 5: decolouring
Adding appropriate activated carbon in the supernatant collected to decolour, preferred version is every 100mL
Adding 2～3g, the pH value of solution 5%HCl after filtering regulates to 4.5, stand-by.
Step 6: isolated and purified
Draw solution injection 7cm × 60cm strong acidic ion resin exchange column that 8.0mL collects,
And using distilled water to carry out eluting, elution speed, at 2mL/min, is collected about absorbance peak-peak
Part effluent.
Step 7: the mensuration of extracted amount
Formaldehyde-acetylacetone method is used to measure the extracted amount of taurine.Take 1mL effluent or its dilution
Liquid, adds 1mol/L sodium acetate solution 8mL, and addition 1mL formaldehyde-acetylacetone,2,4-pentanedione, as developer, is joined
10mL solution processed, is incubated 15 minutes in 100 DEG C of water-baths, is cooled to room temperature, at 400nm ripple
Strong point measures absorbance, by taurine standard solution absorption curve (as shown in Figure 1), determines cattle
The extracted amount of sulfonic acid.
Step 8: detection crystallization
Use the ultraviolet spectrophotometry detection component containing taurine, put it to 4 DEG C of refrigerators are put
Put 2h, obtain taurine crude crystalline.Crude product is redissolved in water, after filtration, adds nothing
Water-ethanol, is positioned over 4 DEG C of recrystallization, is separated by solid-liquid, so crystallize, then recrystallization repeatedly,
I.e. can get pure taurine crystal.
Experiment of single factor
Take 50 grams of mussel meats, add 150 grams of deionized waters, put into tissue smashing machine and make homogenate；
Homogenate is divided into 5 parts of equalization, and successively by homogenate pH value furnishing 5,5.5,6,6.5,
7, temperature controls at 55 DEG C, and adding concentration is the papain of 0.6%, at ultrasonic cell-break
Carry out cell breakage under instrument effect and extract 15min.Show that what taurine extracted by pH affects result such as
Shown in Fig. 2.
When obtaining pH=5.5 according to Fig. 2 and data analysis taurine extract yield the highest, pH5.5 with
After, the yield of taurine is declining successively.Therefore determine that Optimal pH condition is 5.5.
Take 50 grams of mussel meats, add 150 grams of deionized waters, put into tissue smashing machine and make homogenate；
Homogenate is divided into 5 parts of equalization, and sets gradually enzymolysis time when ultrasound wave _ enzymolysis crushes and be
5min, 10min, 15min, 20min, 25min, pH controls 5.5, and hydrolysis temperature is 55 DEG C.
Adding concentration is the papain of 0.6%, carries out cell and break under ultrasonic cell disruption instrument effect
Broken extraction.
According to enzymolysis time knowable to Fig. 3 when 20min, the extraction rate reached of taurine to peak value, enzyme
After solving 20min, taurine yield is on a declining curve, therefore determines that the peak enzymolysis-ability time is 20min.
Take 50 grams of mussel meats, add 150 grams of deionized waters, put into tissue smashing machine and make homogenate；
Homogenate is divided into 5 parts of equalization, and sets gradually when ultrasound wave enzymolysis crushes hydrolysis temperature successively
Being 45 DEG C, 50 DEG C, 55 DEG C, 60 DEG C, 65 DEG C, pH controls 5.5, and adding concentration is 0.6%
Papain, carries out cell breakage under ultrasonic cell disruption instrument effect and extracts 15min.Obtain enzyme
What taurine was extracted by solution temperature affects result as shown in Figure 4.
According to Fig. 4, initial extraction time gradually continues, the extraction yield of taurine also with
The continuity pole of time significantly improves, and before 60 DEG C, taurine extraction yield increase trend is fairly obvious,
Between upon extracting when 60 DEG C, the extraction yield of taurine is maximum, and hereafter the time continues again, cattle sulphur
The extraction yield of acid is below yield when 60 DEG C, therefore selects 60 DEG C for the optimum extraction time.
Orthogonal experiment
Table 1 empirical factor table
Orthogonal design table according to design operates, and obtains orthogonal result table 2.
Table 2 Orthogonal experiment results analytical table directly perceived
According to the extracted amount of spectrophotometry taurine, determine that standard curve equation is
Y=2.2257x-0.01857, R2=0.99118, it may be determined that the single factor test bar selected in material Carnis Mactrae
Part meets linear rule, can determine that information is as shown in Figure 5 as the level conditions of orthogonal experiment.
According to data in analytical table directly perceived, hydrolysis temperature, pH, the extreme difference of broken time are successively
Reduce, illustrate that taurine is extracted the impact order of experiment successively by hydrolysis temperature, pH, broken time
For hydrolysis temperature > pH > broken time.According to table determine optimum extraction condition be pH be 5.5, ultrasonic
It is 20min that ripple-enzymolysis crushes the time, and breaking temperature is 60 DEG C.
Infrared detection
In the infrared detection of taurine is tested, have employed pellet technique, the reality of specific experiment
Execute and carry out with reference to standard GB/T/T 6040-2002 " infrared spectrum analysis general rule ".
As Fig. 6,7, the infrared absorption peak picture of comparison sample and taurine standard substance, would know that through anhydrous
The white needle-like crystals that Ethanol Treatment separates out is exactly high-purity natural taurine.
Nuclear-magnetism detects
As Fig. 8,9, shown, in nmr spectrum occur three signals, illustrated three kinds the most of the same race
The H of class, non-be not δ 4.646, δ 3.296 and δ 3.132 [27] through comparison, these data and mark
Quasi-condition is similar to, and illustrates that the white needle-like crystals obtained from shrimp homogenate is high-purity natural cattle sulphur
Acid crystal.
Claims (9)
1. the method extracting natural taurine from mussel, it is characterised in that comprise the steps of
S1 utilizes the mussel preparation clear liquid containing taurine；
S2 is by described clear liquid removing protein；
The product of described step S2 is carried out isolated and purified by S3.
The method extracting natural taurine from mussel the most according to claim 1, its feature exists
In, described step S1 comprises the steps of
Mussel is cleaned the taking-up mussel meat that shells by S1.1；
Mussel meat is put in tissue mashing machine and is homogenized by S1.2；
Mussel meat after homogenate is weighed and uses the water of 3 times of quality to dilute by S1.3；
S1.4 regulates pH value；
S1.5 puts into and carries out water extraction in constant incubator；
Water extraction product is filtered by S1.6, takes filtrate.
The method extracting natural taurine from mussel the most according to claim 1, its feature exists
In, described step S2 comprises the steps of
S2.1 removes acidic protein；
S2.2 removes basic protein.
The method extracting natural taurine from mussel the most according to claim 3, its feature exists
In, described step S2.1 uses hydrochloric acid that the pH of described step S1 product is adjusted to 3-4 except acidic protein,
Then it is centrifuged 20 minutes under 4500r/min, and collects supernatant.
The method extracting natural taurine from mussel the most according to claim 3, its feature exists
In, described step S2.2 uses sodium hydroxide by the product of described step S2.1 except basic protein
PH is adjusted to 9-10, is centrifuged 20 minutes under 4500r/min, collects supernatant.
The method extracting natural taurine from mussel the most according to claim 1, its feature exists
In, concretely comprising the following steps of described step S3:
S3.1 regulates the pH to 4.5 of described step S2 product；
The product of described step S3.1 is injected strong acidic ion resin exchange column by S3.2, and uses
Distilled water carries out eluting；
S3.3 collects the part effluent of absorbance peak-peak about 0.55.
7. according to extracting natural taurine from mussel described in claim 1-6 any claim
Method, it is characterised in that exist described step S2 between described step S2 and described step S3
Product carries out the step decoloured.
The method extracting natural taurine from mussel the most according to claim 7, its feature exists
In, described decolorization process specifically comprises: adds activated carbon in the product of described step S2 and takes off
Color.
The method extracting natural taurine from mussel the most according to claim 8, its feature exists
In, in the product of step S2 described in every 100mL, add 2～3g activated carbons.
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| CN113200889A (en) * | 2020-12-18 | 2021-08-03 | 浙江经贸职业技术学院 | Method for preparing taurine from mussels |
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