CN108373512A - A method of the depth congruent melting solvent extraction Moringa seed polysaccharide made of choline chloride and urea - Google Patents
A method of the depth congruent melting solvent extraction Moringa seed polysaccharide made of choline chloride and urea Download PDFInfo
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- CN108373512A CN108373512A CN201810429835.7A CN201810429835A CN108373512A CN 108373512 A CN108373512 A CN 108373512A CN 201810429835 A CN201810429835 A CN 201810429835A CN 108373512 A CN108373512 A CN 108373512A
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- Prior art keywords
- moringa
- seed polysaccharide
- congruent melting
- moringa seeds
- melting solvent
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- C—CHEMISTRY; METALLURGY
- C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
- C08B—POLYSACCHARIDES; DERIVATIVES THEREOF
- C08B37/00—Preparation of polysaccharides not provided for in groups C08B1/00 - C08B35/00; Derivatives thereof
- C08B37/0003—General processes for their isolation or fractionation, e.g. purification or extraction from biomass
Abstract
The invention discloses the methods of the depth congruent melting solvent extraction Moringa seed polysaccharide made of choline chloride and urea, include the preparation of depth congruent melting solvent, the pretreatment of moringa seeds, the extraction of Moringa seed polysaccharide, the thick purifying of Moringa seed polysaccharide, the measurement of total starches content, the measurement of protein content, 52 column chromatographies of DEAE purify, and obtain polysaccharide sample.The advantages that present invention has operability strong, and simple process is at low cost, easily obtains, and the recovery rate of total starches is high.Meanwhile depth congruent melting solvent makes simply, initial substance is renewable, degradable, is widely present in nature.
Description
Technical field
The present invention relates to a kind of moringa seeds polyoses producing methods, and in particular to a kind of to be made by choline chloride and glycerine
The method of depth congruent melting solvent extraction Moringa seed polysaccharide, belongs to efficient biomass Resource Access technical field.
Background technology
Moringa (Moringa oleifera) also known as drumstick tree, foreign Ailanthus altissima, belong to all sorts of flowers dish mesh Moringaceae, because its rhizome have it is pungent
Pungent, therefore Moringa of gaining the name.It is a kind of perennial tropical trees, is distributed mainly on India, Africa, South America and Southeast Asia state
Family, the areas such as Taiwan, Guangdong, Yunnan are distributed mainly in China.Nutritional ingredient and active material there are many containing in moringa seeds, and
With a variety of physiological activity, in health products and pharmaceuticals industry tool has been widely used and development prospect.
Polysaccharide is a kind of high molecular polymer, is formed by glucosides key connection by thousands of a monosaccharide.Polysaccharide is extensive
It is distributed in nature, plant, animal have presence in microbial body.Natural polysaccharide can be used for clinical test, as anti-diabetic,
The treatment of the illnesss such as antitumor, reducing blood lipid or as anti-inflammatory, antiviral drug, polysaccharide is for maintaining human normal operation tool
There is important role.Moringa seed polysaccharide is natural products, nontoxic, has very high researching value.
Depth congruent melting solvent (Deep Eutectic Solvents, DES) is one kind by hydrogen bond receptor and hydrogen bond donor shape
At molecular complex, be a kind of salting liquid.Hydrogen bond receptor such as can cheaply obtain two kinds of compound generally use quaternary ammonium salts, easily
Degradation, nontoxic choline chloride, and urea, ethylene glycol, glycerine, oxalic acid etc. then may be selected in hydrogen bond receptor.By two kinds of chemical combination
Since the hydrogen bond formed between anion and hydrogen bond donor causes the delocalization of charge in object mixed process, lead to depth congruent melting solvent
Fusing point be less than each component compound fusing point.Such as under room temperature by choline chloride and urea with 1：2 molar ratio mixing, gained
The fusing point of depth congruent melting solvent be about 12 DEG C, and the fusing point of choline chloride is 302-305 DEG C, and the fusing point of urea is 132.7
DEG C, well below the fusing point of each component.Depth congruent melting solvent has cheap, easy acquisition compared to other ionic liquids
The advantages that.
With depth congruent melting solvent extraction Moringa seed polysaccharide than traditional extracting method extraction efficiency higher and easy to operate,
Moringa seed polysaccharide can be obtained to a greater extent, improve the utilization rate of moringa seeds.
Invention content
The object of the present invention is to provide a kind of depth congruent melting solvent extraction moringa seeds made of choline chloride and urea
The method of polysaccharide, the extracting method recovery rate is high, and at low cost, low energy consumption, environmental protection, easy to operate, and the polysaccharide of preparation passes through purifying,
Purity improves, and is a kind of excellent extracting method.Former technical solution is as follows：
A method of the depth congruent melting solvent extraction Moringa seed polysaccharide made of choline chloride and urea, including it is following
Step：
(1) preparation of depth congruent melting solvent：The choline chloride and urea being placed in vacuum drying chamber after drying are with 1:2
Molar ratio mixes, and grinding at room temperature is until obtain the liquid of stable homogeneous, i.e. depth congruent melting solvent；
(2) processing of raw material：Choose intact moringa seeds, decladding is crushed after dry 2-3d under the conditions of 50-70 DEG C
60 mesh sieve, and obtain moringa seeds fine powder；
(3) extraction of Moringa seed polysaccharide：It accurately weighs 10g steps (2) moringa seeds fine powder and 100mL steps (1) depth is total
Molten solvent mixing is placed in magneton stirring 20-40min under 50-70 DEG C of constant temperature waters pot, 240r/min rotating speeds, and mixture uses high
10-15min is centrifuged under fast refrigerated centrifuge 12000r/min and detaches solid particle, takes supernatant spare；
(4) the de- albumen of Moringa seed polysaccharide：It takes supernatant made from step (3) to be placed in ice water and 3%-5%TCA is added
4 DEG C of refrigerators stand 4-5h, NaOH tune pH to neutrality, the Moringa of 2 protein that is removed of repetitive operation after stirring 10-15min
Seed polysaccharide supernatant；
(5) moringa seeds polysaccharide alcohol precipitation：Take the Moringa seed polysaccharide supernatant of removing protein made from step (4) that 4 times of volumes are added
85% ethyl alcohol stands overnight at 4 DEG C, takes supernatant；
(6) it is concentrated and dried：Supernatant made from collection step (5) is concentrated in vacuo to original 1/5, concentrate freeze-drying
12-24h obtains moringa seeds Thick many candies；
(7) measurement of total starches content：Moringa seeds Thick many candies made from step (6) are measured using Coomassie Brilliant Blue；
(8) measurement of protein content：The moringa seeds Thick many candies of step (7) are measured using phend-sulphuric acid again；
(9) DEAE-52 column chromatographies purify：The solution that the Moringa Thick many candies of step (8) are configured to 10mg/mL crosses DEAE-
52 columns are eluted with 0-0.5mol/L NaCl solutions, are collected eluent and are concentrated and dried to get to Moringa seed polysaccharide.
The advantageous effect of the present invention compared with the existing technology is：
The present invention uses depth congruent melting solvent extraction Moringa polysaccharide, substantially increases the recovery rate of polysaccharide in moringa seeds；And
Extractant is easy to make, and the prices of raw materials are low, environmentally friendly, and production method is easy.The pure of polysaccharide is improved by column chromatography
Degree；The bioactivity of polysaccharide is greatly saved by being freeze-dried.
Specific implementation mode
The present invention is described in further details below by example, these examples are only used for illustrating the present invention, and unlimited
The scope of the present invention processed.
Embodiment 1
A method of the depth congruent melting solvent extraction Moringa seed polysaccharide made of choline chloride and urea, including it is following
Step：
(1) preparation of depth congruent melting solvent：The choline chloride and urea being placed in vacuum drying chamber after drying are with 1:2
Molar ratio mixes, and grinding at room temperature is until obtain the liquid of stable homogeneous, i.e. depth congruent melting solvent；
(2) processing of raw material：Choose intact moringa seeds, decladding crushed 60 mesh sieve under the conditions of 50 DEG C after dry 3d,
Obtain moringa seeds fine powder；
(3) extraction of Moringa seed polysaccharide：It accurately weighs 10g steps (2) moringa seeds fine powder and 100mL steps (1) depth is total
Molten solvent mixing is placed in magneton under 55 DEG C of thermostat water baths, 240r/min rotating speeds and stirs 30min, and mixture is freezed using high speed
15min is centrifuged under centrifuge 12000r/min and detaches solid particle, takes supernatant spare；
(4) the de- albumen of Moringa seed polysaccharide：It takes supernatant made from step (3) to be placed in ice water and 5%TCA stirrings is added
4 DEG C of refrigerators stand 4h, NaOH tune pH to neutrality, the Moringa seed polysaccharide supernatant of 2 protein that are removed of repetitive operation after 10min
Liquid；
(5) moringa seeds polysaccharide alcohol precipitation：Take the Moringa seed polysaccharide supernatant of removing protein made from step (4) that 4 times of volumes are added
85% ethyl alcohol stands overnight at 4 DEG C, takes supernatant；
(6) it is concentrated and dried：Supernatant made from collection step (5) is concentrated in vacuo to original 1/5, concentrate freeze-drying
20h obtains moringa seeds Thick many candies；
(7) measurement of total starches content：Moringa seeds Thick many candies made from step (6) are measured using Coomassie Brilliant Blue；
(8) measurement of protein content：The moringa seeds Thick many candies of step (7) are measured using phend-sulphuric acid again；
(9) DEAE-52 column chromatographies purify：The moringa seeds Thick many candies of step (8) are configured to the solution mistake of 10mg/mL
DEAE-52 columns are eluted with 0-0.5mol/L NaCl solutions, are collected eluent and are concentrated and dried to get more to moringa seeds
Sugar.
Embodiment 2
A method of the depth congruent melting solvent extraction Moringa seed polysaccharide made of choline chloride and urea, including it is following
Step：
(1) preparation of depth congruent melting solvent：The choline chloride and urea being placed in vacuum drying chamber after drying are with 1:2
Molar ratio mixes, and grinding at room temperature is until obtain the liquid of stable homogeneous, i.e. depth congruent melting solvent；
(2) processing of raw material：Choose intact moringa seeds, decladding crushed 60 mesh sieve under the conditions of 60 DEG C after dry 3d,
Obtain moringa seeds fine powder；
(3) extraction of Moringa seed polysaccharide：It accurately weighs 10g steps (2) moringa seeds fine powder and 100mL steps (1) depth is total
Molten solvent mixing is placed in magneton under 60 DEG C of thermostat water baths, 240r/min rotating speeds and stirs 40min, and mixture is freezed using high speed
10min is centrifuged under centrifuge 12000r/min and detaches solid particle, takes supernatant spare；
(4) the de- albumen of Moringa seed polysaccharide：It takes supernatant made from step (3) to be placed in ice water and 5%TCA stirrings is added
4 DEG C of refrigerators stand 4h, NaOH tune pH to neutrality, the Moringa seed polysaccharide supernatant of 2 protein that are removed of repetitive operation after 10min
Liquid；
(5) moringa seeds polysaccharide alcohol precipitation：Take the Moringa seed polysaccharide supernatant of removing protein made from step (4) that 4 times of volumes are added
85% ethyl alcohol stands overnight at 4 DEG C, takes supernatant；
(6) it is concentrated and dried：Supernatant made from collection step (5) is concentrated in vacuo to original 1/5, concentrate freeze-drying
18h obtains moringa seeds Thick many candies；
(7) measurement of total starches content：Moringa seeds Thick many candies made from step (6) are measured using Coomassie Brilliant Blue；
(8) measurement of protein content：The moringa seeds Thick many candies of step (7) are measured using phend-sulphuric acid again；
(9) DEAE-52 column chromatographies purify：The moringa seeds Thick many candies of step (8) are configured to the solution mistake of 10mg/mL
DEAE-52 columns are eluted with 0-0.5mol/L NaCl solutions, are collected eluent and are concentrated and dried to get more to moringa seeds
Sugar.
Embodiment 3
A method of the depth congruent melting solvent extraction Moringa seed polysaccharide made of choline chloride and urea, including it is following
Step：
(1) preparation of depth congruent melting solvent：The choline chloride and urea being placed in vacuum drying chamber after drying are with 1:2
Molar ratio mixes, and grinding at room temperature is until obtain the liquid of stable homogeneous, i.e. depth congruent melting solvent；
(2) processing of raw material：Choose intact moringa seeds, decladding crushed 60 mesh sieve under the conditions of 70 DEG C after dry 2d,
Obtain moringa seeds fine powder；
(3) extraction of Moringa seed polysaccharide：It accurately weighs 10g steps (2) moringa seeds fine powder and 100mL steps (1) depth is total
Molten solvent mixing is placed in magneton under 70 DEG C of thermostat water baths, 240r/min rotating speeds and stirs 20min, and mixture is freezed using high speed
15min is centrifuged under centrifuge 12000r/min and detaches solid particle, takes supernatant spare；
(4) the de- albumen of Moringa seed polysaccharide：It takes supernatant made from step (3) to be placed in ice water and 5%TCA stirrings is added
4 DEG C of refrigerators stand 4h, NaOH tune pH to neutrality, the Moringa seed polysaccharide supernatant of 2 protein that are removed of repetitive operation after 12min
Liquid；
(5) moringa seeds polysaccharide alcohol precipitation：Take the Moringa seed polysaccharide supernatant of removing protein made from step (4) that 4 times of volumes are added
85% ethyl alcohol stands overnight at 4 DEG C, takes supernatant；
(6) it is concentrated and dried：Supernatant made from collection step (5) is concentrated in vacuo to original 1/5, concentrate freeze-drying
Moringa seeds Thick many candies are obtained for 24 hours；
(7) measurement of total starches content：Moringa seeds Thick many candies made from step (6) are measured using Coomassie Brilliant Blue；
(8) measurement of protein content：The moringa seeds Thick many candies of step (7) are measured using phend-sulphuric acid again；
(9) DEAE-52 column chromatographies purify：The moringa seeds Thick many candies of step (8) are configured to the solution mistake of 10mg/mL
DEAE-52 columns are eluted with 0-0.5mol/L NaCl solutions, are collected eluent and are concentrated and dried to get more to moringa seeds
Sugar.
Claims (1)
1. a kind of method of the depth congruent melting solvent extraction Moringa seed polysaccharide made of choline chloride and urea, feature include
Following steps：
(1) preparation of depth congruent melting solvent：The choline chloride and urea being placed in vacuum drying chamber after drying are with 1:2 mole
Than mixing, grinding at room temperature is until obtain the liquid of stable homogeneous, i.e. depth congruent melting solvent；
(2) processing of raw material：Choose intact moringa seeds, decladding crushed 60 mesh under the conditions of 50-70 DEG C after dry 2-3d
Sieve, obtains moringa seeds fine powder；
(3) extraction of Moringa seed polysaccharide：It accurately weighs 10g steps (2) moringa seeds fine powder and 100mL steps (1) depth congruent melting is molten
Agent mixes, and is placed in magneton stirring 20-40min, mixture under 50-70 DEG C of constant temperature waters pot, 240r/min rotating speeds and uses high speed cold
Freeze centrifugation 10-15min under centrifuge 12000r/min and detach solid particle, takes supernatant spare；
(4) the de- albumen of Moringa seed polysaccharide：It takes supernatant made from step (3) to be placed in ice water and 3%-5%TCA stirrings is added
4 DEG C of refrigerators stand 4-5h after 10-15min, and the moringa seeds of NaOH tune pH to neutrality, 2 protein that are removed of repetitive operation are more
Sugared supernatant；
(5) moringa seeds polysaccharide alcohol precipitation：Take the Moringa seed polysaccharide supernatant of removing protein made from step (4) that 4 times of volumes 85% are added
Ethyl alcohol, stand overnight at 4 DEG C, take supernatant；
(6) it is concentrated and dried：Supernatant made from collection step (5) is concentrated in vacuo to original 1/5, and concentrate is freeze-dried 12-
Moringa seeds Thick many candies are obtained for 24 hours；
(7) measurement of total starches content：Moringa seeds Thick many candies made from step (6) are measured using Coomassie Brilliant Blue；
(8) measurement of protein content：The moringa seeds Thick many candies of step (7) are measured using phend-sulphuric acid again；
(9) DEAE-52 column chromatographies purify：The solution that the Moringa Thick many candies of step (8) are configured to 10mg/mL crosses DEAE-52 columns,
It is eluted with 0-0.5mol/L NaCl solutions, collects eluent and be concentrated and dried to get to Moringa seed polysaccharide.
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| CN110856518A (en) * | 2018-08-22 | 2020-03-03 | 丁晓 | Technological method for comprehensively treating moringa seeds, and product and application thereof |
| CN111345392A (en) * | 2020-02-24 | 2020-06-30 | 天津科技大学 | Eutectic solvent, preparation method thereof and application of eutectic solvent in extraction of plant protein |
| CN112062868A (en) * | 2020-06-12 | 2020-12-11 | 湖南农业大学 | Method for extracting Chinese yam polysaccharide |
| CN113134053A (en) * | 2021-03-30 | 2021-07-20 | 广东丸美生物技术股份有限公司 | Aloe extract and preparation method and application thereof |
| CN113336867A (en) * | 2021-05-28 | 2021-09-03 | 中国计量大学 | Auricularia auricula polysaccharide, and total wall-broken extraction method and application thereof |
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Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110856518A (en) * | 2018-08-22 | 2020-03-03 | 丁晓 | Technological method for comprehensively treating moringa seeds, and product and application thereof |
| CN111345392A (en) * | 2020-02-24 | 2020-06-30 | 天津科技大学 | Eutectic solvent, preparation method thereof and application of eutectic solvent in extraction of plant protein |
| CN112062868A (en) * | 2020-06-12 | 2020-12-11 | 湖南农业大学 | Method for extracting Chinese yam polysaccharide |
| CN113134053A (en) * | 2021-03-30 | 2021-07-20 | 广东丸美生物技术股份有限公司 | Aloe extract and preparation method and application thereof |
| CN113336867A (en) * | 2021-05-28 | 2021-09-03 | 中国计量大学 | Auricularia auricula polysaccharide, and total wall-broken extraction method and application thereof |
| CN113336867B (en) * | 2021-05-28 | 2022-08-09 | 中国计量大学 | Auricularia auricula polysaccharide, and total wall-broken extraction method and application thereof |
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Application publication date: 20180807 |