CN109370254A - A method of preparing High color values ant melanin from silky ant - Google Patents
A method of preparing High color values ant melanin from silky ant Download PDFInfo
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- CN109370254A CN109370254A CN201811639078.2A CN201811639078A CN109370254A CN 109370254 A CN109370254 A CN 109370254A CN 201811639078 A CN201811639078 A CN 201811639078A CN 109370254 A CN109370254 A CN 109370254A
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- C09—DYES; PAINTS; POLISHES; NATURAL RESINS; ADHESIVES; COMPOSITIONS NOT OTHERWISE PROVIDED FOR; APPLICATIONS OF MATERIALS NOT OTHERWISE PROVIDED FOR
- C09B—ORGANIC DYES OR CLOSELY-RELATED COMPOUNDS FOR PRODUCING DYES, e.g. PIGMENTS; MORDANTS; LAKES
- C09B61/00—Dyes of natural origin prepared from natural sources, e.g. vegetable sources
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- C—CHEMISTRY; METALLURGY
- C09—DYES; PAINTS; POLISHES; NATURAL RESINS; ADHESIVES; COMPOSITIONS NOT OTHERWISE PROVIDED FOR; APPLICATIONS OF MATERIALS NOT OTHERWISE PROVIDED FOR
- C09B—ORGANIC DYES OR CLOSELY-RELATED COMPOUNDS FOR PRODUCING DYES, e.g. PIGMENTS; MORDANTS; LAKES
- C09B67/00—Influencing the physical, e.g. the dyeing or printing properties of dyestuffs without chemical reactions, e.g. by treating with solvents grinding or grinding assistants, coating of pigments or dyes; Process features in the making of dyestuff preparations; Dyestuff preparations of a special physical nature, e.g. tablets, films
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Abstract
The method that the invention discloses a kind of to prepare High color values ant melanin from silky ant, includes the following steps: feedstock processing；High pressure microjet ultramicro crushing treatment；Separation: aqueous two-phase extraction is carried out to paste serous material is obtained, it obtains the upper phase of extraction and extracts lower phase, double-aqueous phase system used in aqueous two-phase extraction is made of polyethylene glycol, sodium chloride and water, mass percent of the polyethylene glycol in double-aqueous phase system is 10-25%, mass percent of the sodium chloride in double-aqueous phase system is 10-20%, surplus is water, and the weight average molecular weight of the polyethylene glycol is 1000-8000；Desalting processing；Column chromatography；It is dry: upper step eluent is dried to obtain ant melanin.
Description
Technical field
The present invention relates to the extracting methods of field of biotechnology, specifically a kind of that High color values ant is prepared from silky ant
The method of melanin.
Background technique
Silky ant (Polyrhachis vicina Roger), is called Polyhachis vicina Roger, polyrhachis vicina Roger, bidentate thorniness
Ant belongs to Arthropoda, Insecta, film wing day, Formicidae, Formicinae, Polyrhachis, for the social insect being gregariousness." herbal guiding principle
Mesh " in, to the habit of ant, toxicity, it is edible and it is medicinal all made detailed record, ant has " strengthening vital QI to eliminate pathogenic factors " and " different disease
With controlling " unique effects.The pharmacological evaluation of nineteen eighty-three Zhao's first-class prompts, and silky ant has anti-inflammatory, protect liver, calmness, relievings asthma, spasmolysis
The effects of, while carried out toxicity and subacute measurement prompt its is nontoxic.
Melanin (Melanin) is the nitrogenous of the complexity of one kind as made of indoles substance or polyphenols oxidation polymerization
High polymer, it is widely present in plant, animal and microorganism.Melanin is to be insoluble in generally in black, brown or brown
A kind of large biological molecule of water, acid and organic solvent.Melanin, which has, to be adjusted physiological function, improves immunity, anti-aging, salubrity
It is healthy and strong and waits health-care effects.In addition, melanin also has protection large biological molecule, has significant inhibition to AIDS virus in vitro
Effect.Currently, natural black pigment contained in Semen sesami nigrum, black rice, semen sojae atricolor, blank corn, black-bone chicken etc. has caused, people's is dense
Thick interest finds that they have effects that nourishing, health, anti-oxidant and anti-aging etc., but for silky ant melanin
Research is rarely reported.Silky ant is known as " microfauna high-grade nutrient treasure-house ", and 1993, health ministry determined it as medicine
Eat dual-purpose nontoxic excellent ant kind.The country has carried out the chemical component of silky ant, pharmacological action and clinical application more
Research.Result of study shows that silky ant has the effects that enhance immune function, anti-aging effects, antitumor, adjusting blood glucose.
In existing research, extraction for ant melanin is general to use: silky ant crushing-sour water solution-centrifugation-black
Precipitating-vacuum drying-melanin product technique is collected in the heavy operation of the heavy-centrifugation-precipitating-repetition alkali soluble acid of plain crude product-alkali soluble acid,
Such operation, one is complex process, and the time used in entire technique is long, the color value of another melanin for being
It is low.Research to ant melanin, there are also number of values to obtain improvements.
Summary of the invention
The method that the object of the present invention is to provide a kind of to prepare High color values ant melanin from silky ant, obtained black
The advantages such as plain color valence is high, and preparation method has simple process, is suitble to produce, and production cost is low.
The invention is realized by the following technical scheme:
A method of it preparing High color values ant melanin from silky ant, includes the following steps:
1) feedstock processing: the silky ant of drying is crushed, sieving；
2) high pressure microjet ultramicro crushing treatment: the powder that upper step is obtained, the pure water of the weight such as addition add phase
When handling 10-20min, revolving speed 8000- with high-speed shearing emulsion machine in the petroleum sodium sulfonate of powder weight 0.3-0.5%
Then 12000r/min is handled with high pressure microjet Ultra-Micro Grinding Equipment in 25 DEG C, processing pressure 80-120MPa, place
Managing number is 2-3 times, obtains paste serous material；
3) separation: carrying out aqueous two-phase extraction to paste serous material is obtained, and obtains the upper phase of extraction and extracts lower phase, aqueous two-phase extraction
Used double-aqueous phase system is made of polyethylene glycol, sodium chloride and water, quality of the polyethylene glycol in double-aqueous phase system
Percentage is 10-25%, and mass percent of the sodium chloride in double-aqueous phase system is 10-20%, and surplus is water, described poly-
The weight average molecular weight of ethylene glycol is 1000-8000；
4) desalting processing: phase in the extraction that step 3) is obtained, desalination, phase under the extraction that step 3) is obtained, desalination；
5) column chromatographs: merging the liquid obtained after desalination, filters, concentration is chromatographed by column, with D101 type macroporous absorption tree
Rouge is adsorbed, and cleaning resin to trickle with deionized water is colourless, then the ethanol solution for being 20-30% with volumetric concentration
Eluent is collected in elution；
6) dry: upper step eluent is dried to obtain ant melanin.
Sieving described in step 1) of the present invention is to sieve with 100 mesh sieve.
Desalination described in step 4) is using ultrafiltration or dialysis, and ultrafiltration uses molecular cut off for 1000-
100000 ultrafiltration membrane desalination, dialysis use molecular cut off for the bag filter desalination of 1000-100000.
Drying described in step 6), using spray drying or freeze-drying.
Compared with prior art, advantages of the present invention:
1, super using high pressure microjet by the present invention in that handling silky ant raw material with high pressure microjet superfine communication technique
Micro-crushing equipment generate extruding force, shearing force, impact force, by liquid-liquid, it is liquid-solid between high-speed impact, high shear, hole
Effect etc., makes the molecular structure of material change.High pressure microjet Ultra-Micro Grinding Equipment can accomplish nano-dispersed, refinement effect
For fruit up to 0.lum even hereinafter, pressure is up to 80-120MPa, obtained solution particles particle diameter distribution is uniform, has operation letter
It is single, pollution-free, can online cleaning-sterilizing, can high efficiency many advantages, such as carrying out breaking-wall cell.
2, ant melanin is one kind biology for being insoluble in water, acid and organic solvent generally in black, brown or brown
Macromolecular.Existing extracting method is all after directly silky ant raw material is smashed or crushed by sour water solution, alkali soluble acid is heavy repeatedly
It extracts, or mentioned by water/modes such as alcohol extracting, enzymatic hydrolysis or Microwave Extraction directly extract, above processing, silky ant is thin
Grease in born of the same parents can be isolated simultaneously, and the ingredients such as group and melanin containing greasy mix, to influence subsequent effective
The separation of ingredient increases the processing difficulty of process.The present invention first uses high pressure microjet superfine communication technique to handle, and is using
When high pressure microjet superfine communication technique handles silky ant raw material, petroleum sodium sulfonate joined, have in molecular structure one strong
Hydrophilic sulfonic group is linked with alkyl, and surface-active is strong, so that the incompatible polar group in 2 in ant melanin kind
Group: hydrophilic group and lipophilic group separately adsorb, it is different with the affinity difference of water and grease specific gravity to oil using non-oil component and incite somebody to action
It is oily to be separated with non-oil component, conducive to the aqueous two-phase extraction of lower step, by different extraction conditions so that melanin and other at
Divide and is kept completely separate.
3, by means of the present invention, the separation of ant melanin is more complete, obtains the ant of High color values after making chromogenic reaction
Ant melanin, color value E214>345。
Detailed description of the invention
Fig. 1 is the UV-visible spectrum for the silky ant melanin that the embodiment of the present invention 3 obtains.
Specific embodiment
With embodiment, the invention will be further described below, but the invention is not limited to these embodiments.
Embodiment 1:
A method of it preparing High color values ant melanin from silky ant, includes the following steps:
1) feedstock processing: the silky ant of drying is crushed, sieving；
2) high pressure microjet ultramicro crushing treatment: the powder that upper step is obtained, the pure water of the weight such as addition add phase
When handling 20min, revolving speed 8000r/min with high-speed shearing emulsion machine, then in the petroleum sodium sulfonate of powder weight 0.3%
It is handled with high pressure microjet Ultra-Micro Grinding Equipment in 25 DEG C, processing pressure 120MPa, number of processes is 2 times, obtains cream
Shape slurries；
3) separation: carrying out aqueous two-phase extraction to paste serous material is obtained, and obtains the upper phase of extraction and extracts lower phase, aqueous two-phase extraction
Used double-aqueous phase system is made of polyethylene glycol, sodium chloride and water, quality of the polyethylene glycol in double-aqueous phase system
Percentage is 10%, and mass percent of the sodium chloride in double-aqueous phase system is 10%, and surplus is water, the polyethylene glycol
Weight average molecular weight be 1000-8000；
4) desalting processing: phase in the extraction that step 3) is obtained, desalination, phase under the extraction that step 3) is obtained, desalination；
5) column chromatographs: merging the liquid obtained after desalination, filters, concentration is chromatographed by column, with D101 type macroporous absorption tree
Rouge is adsorbed, and cleaning resin to trickle with deionized water is colourless, then the ethanol solution for being 20-30% with volumetric concentration
Gradient elution collects eluent；
6) dry: upper step eluent being freeze-dried to obtain ant melanin powder.
Embodiment 2:
A method of it preparing High color values ant melanin from silky ant, includes the following steps:
1) feedstock processing: the silky ant of drying is crushed, sieving；
2) high pressure microjet ultramicro crushing treatment: the powder that upper step is obtained, the pure water of the weight such as addition add phase
When handling 10min, revolving speed 12000r/min with high-speed shearing emulsion machine, then in the petroleum sodium sulfonate of powder weight 0.4%
It is handled with high pressure microjet Ultra-Micro Grinding Equipment in 25 DEG C, processing pressure 80MPa, number of processes is 3 times, obtains paste
Slurries；
3) separation: carrying out aqueous two-phase extraction to paste serous material is obtained, and obtains the upper phase of extraction and extracts lower phase, aqueous two-phase extraction
Used double-aqueous phase system is made of polyethylene glycol, sodium chloride and water, quality of the polyethylene glycol in double-aqueous phase system
Percentage is 15%, and mass percent of the sodium chloride in double-aqueous phase system is 20%, and surplus is water, the polyethylene glycol
Weight average molecular weight be 1000-8000；
4) desalting processing: phase in the extraction that step 3) is obtained, desalination, phase under the extraction that step 3) is obtained, desalination；
5) column chromatographs: merging the liquid obtained after desalination, filters, concentration is chromatographed by column, with D101 type macroporous absorption tree
Rouge is adsorbed, and cleaning resin to trickle with deionized water is colourless, then the ethanol solution for being 20-30% with volumetric concentration
Gradient elution collects eluent；
6) dry: upper step eluent being spray-dried to obtain ant melanin powder.
Embodiment 3:
A method of it preparing High color values ant melanin from silky ant, includes the following steps:
1) feedstock processing: the silky ant of drying is crushed, sieving；
2) high pressure microjet ultramicro crushing treatment: the powder that upper step is obtained, the pure water of the weight such as addition add phase
When handling 15min, revolving speed 10000r/min with high-speed shearing emulsion machine, then in the petroleum sodium sulfonate of powder weight 0.5%
It is handled with high pressure microjet Ultra-Micro Grinding Equipment in 25 DEG C, processing pressure 100MPa, number of processes is 3 times, obtains cream
Shape slurries；
3) separation: carrying out aqueous two-phase extraction to paste serous material is obtained, and obtains the upper phase of extraction and extracts lower phase, aqueous two-phase extraction
Used double-aqueous phase system is made of polyethylene glycol, sodium chloride and water, quality of the polyethylene glycol in double-aqueous phase system
Percentage is 25%, and mass percent of the sodium chloride in double-aqueous phase system is 15%, and surplus is water, the polyethylene glycol
Weight average molecular weight be 1000-8000；
4) desalting processing: phase in the extraction that step 3) is obtained, desalination, phase under the extraction that step 3) is obtained, desalination；
5) column chromatographs: merging the liquid obtained after desalination, filters, concentration is chromatographed by column, with D101 type macroporous absorption tree
Rouge is adsorbed, and cleaning resin to trickle with deionized water is colourless, then the ethanol solution for being 20-30% with volumetric concentration
Gradient elution collects eluent；
6) dry: upper step eluent being freeze-dried to obtain ant melanin powder.
Comparative example 1:
1) feedstock processing: the silky ant of drying is crushed, sieving；
2) high pressure microjet ultramicro crushing treatment: the powder that upper step is obtained the pure water of weight such as is added, with cutting at a high speed
Mulser processing 15min, revolving speed 10000r/min are cut, then with high pressure microjet Ultra-Micro Grinding Equipment at 25 DEG C
Reason, processing pressure 100MPa, number of processes are 3 times, obtain paste serous material；
3) separation: carrying out aqueous two-phase extraction to paste serous material is obtained, and obtains the upper phase of extraction and extracts lower phase, aqueous two-phase extraction
Used double-aqueous phase system is made of polyethylene glycol, sodium chloride and water, quality of the polyethylene glycol in double-aqueous phase system
Percentage is 25%, and mass percent of the sodium chloride in double-aqueous phase system is 15%, and surplus is water, the polyethylene glycol
Weight average molecular weight be 1000-8000；
4) desalting processing: phase in the extraction that step 3) is obtained, desalination, phase under the extraction that step 3) is obtained, desalination；
5) column chromatographs: merging the liquid obtained after desalination, filters, concentration is chromatographed by column, with D101 type macroporous absorption tree
Rouge is adsorbed, and cleaning resin to trickle with deionized water is colourless, then the ethanol solution for being 20-30% with volumetric concentration
Gradient elution collects eluent；
6) dry: upper step eluent being freeze-dried to obtain ant melanin powder.
Comparative example 2:
The heavy operation of the heavy-centrifugation-precipitating-repetition alkali soluble acid of silky ant crushing-sour water solution-centrifugation-melanin crude product-alkali soluble acid
Collect precipitating-vacuum drying-melanin product.
Experimental example 1: the UV-visible spectrum for the silky ant melanin that embodiment 3 obtains
Melanin polymer generates spectral absorption characteristics because of the conjugation of its height, silky ant melanin it is ultraviolet-
Visible spectrum, a length of 214nm of its maximum absorption wave as shown in Figure 1:, without characteristic absorption peak, absorption value in visible region
It is reduced with the increase of wavelength.This with it is existing studies have reported that separate sources melanin maximum absorption wavelength in 210nm or so
It is consistent.
Experimental example 2: the measurement of silky ant melanin color value
0.1000g sample is accurately weighed, 0.0002g is accurate to, is dissolved in the phosphate buffer solution that pH is 8.0,40 DEG C
Water-bath 1h is settled in 100mL volumetric flask after dissolving melanin.A certain amount of weak solution dilution is drawn, accurately again with absorbance
It is advisable in 0.3-0.7.At melanin maximum absorbance 214nm, make reference liquid with phosphate buffer, surveys its absorbance.
The color value of melanin solution is calculated according to the following formula: (in formula: A is actual measurement light absorption value to E=Axf/m；F is dilution times
Number；M is sample mass)
Claims (4)
1. a kind of method for preparing High color values ant melanin from silky ant, which comprises the steps of:
1) feedstock processing: the silky ant of drying is crushed, sieving；
2) high pressure microjet ultramicro crushing treatment: the powder that upper step is obtained, the pure water of the weight such as addition are added and are equivalent to
The petroleum sodium sulfonate of powder weight 0.3-0.5% handles 10-20min, revolving speed 8000-12000r/ with high-speed shearing emulsion machine
Min, is then handled with high pressure microjet Ultra-Micro Grinding Equipment in 25 DEG C, processing pressure 80-120MPa, and number of processes is
2-3 times, obtain paste serous material；
3) it separates: carrying out aqueous two-phase extraction to paste serous material is obtained, acquisition extraction is upper mutually to be made with the lower phase of extraction, aqueous two-phase extraction
Double-aqueous phase system is made of polyethylene glycol, sodium chloride and water, quality percentage of the polyethylene glycol in double-aqueous phase system
Than for 10-25%, mass percent of the sodium chloride in double-aqueous phase system is 10-20%, and surplus is water, the poly- second two
The weight average molecular weight of alcohol is 1000-8000；
4) desalting processing: phase in the extraction that step 3) is obtained, desalination, phase under the extraction that step 3) is obtained, desalination；
5) column chromatographs: merge obtained liquid after desalination, filter, concentration is chromatographed by column, with D101 type macroporous absorbent resin into
Row absorption, it is colourless for cleaning resin to trickle with deionized water, then is washed with the ethanol solution that volumetric concentration is 20-30%
It is de-, collect eluent；
6) dry: upper step eluent is dried to obtain ant melanin.
2. a kind of method for preparing High color values ant melanin from silky ant according to claim 1, it is characterised in that:
Sieving described in step 1) is to sieve with 100 mesh sieve.
3. a kind of method for preparing High color values ant melanin from silky ant according to claim 1, it is characterised in that:
Desalination described in step 4) is using ultrafiltration or dialysis, and ultrafiltration uses molecular cut off for the ultrafiltration of 1000-100000
Film desalination, dialysis use molecular cut off for the bag filter desalination of 1000-100000.
4. a kind of method for preparing High color values ant melanin from silky ant according to claim 1, it is characterised in that:
Drying described in step 6), using spray drying or freeze-drying.
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN113861720A (en) * | 2021-10-23 | 2021-12-31 | 哈尔滨工业大学 | Preparation method of insect-source water-soluble protein-free melanin |
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| CN104672345A (en) * | 2015-03-31 | 2015-06-03 | 广西还珠海洋生物科技有限公司 | Method for preparing crude polysaccharide from steindachner soft-shelled turtle carapace |
| CN104693834A (en) * | 2015-03-31 | 2015-06-10 | 桂林得坤生物科技股份有限公司 | Method for preparing persimmon fruit pigments |
| CN107365284A (en) * | 2017-08-16 | 2017-11-21 | 桂林茗兴生物科技有限公司 | A kind of method that taxol is extracted from Chinese yew |
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2018
- 2018-12-29 CN CN201811639078.2A patent/CN109370254A/en active Pending
Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102276758A (en) * | 2011-08-10 | 2011-12-14 | 绵阳师范学院 | Method for comprehensively utilizing insect Chinese medicine dregs |
| CN103497535A (en) * | 2013-10-10 | 2014-01-08 | 西南科技大学 | Animal melanin extraction method and application of extraction product thereof |
| CN104672345A (en) * | 2015-03-31 | 2015-06-03 | 广西还珠海洋生物科技有限公司 | Method for preparing crude polysaccharide from steindachner soft-shelled turtle carapace |
| CN104693834A (en) * | 2015-03-31 | 2015-06-10 | 桂林得坤生物科技股份有限公司 | Method for preparing persimmon fruit pigments |
| CN107365284A (en) * | 2017-08-16 | 2017-11-21 | 桂林茗兴生物科技有限公司 | A kind of method that taxol is extracted from Chinese yew |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN113861720A (en) * | 2021-10-23 | 2021-12-31 | 哈尔滨工业大学 | Preparation method of insect-source water-soluble protein-free melanin |
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