CN107582582A - A kind of technique of extraction purification Moringa flavones - Google Patents
A kind of technique of extraction purification Moringa flavones Download PDFInfo
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- CN107582582A CN107582582A CN201711072153.7A CN201711072153A CN107582582A CN 107582582 A CN107582582 A CN 107582582A CN 201711072153 A CN201711072153 A CN 201711072153A CN 107582582 A CN107582582 A CN 107582582A
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- moringa
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- flavones
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Abstract
The invention discloses a kind of technique of extraction purification Moringa flavones, comprise the following steps：Leaf of Moringa is taken, dry, pulverize as coarse powder, adds distillation watertight to swell 24h, filters moisture content, it is standby that coarse powder of swelling is made；60 80% ethanol of 15 20 times of amounts is added to coarse powder of swelling, while adds the alpha amylase of 0.1 0.15% times of amount, refluxing extraction 3 times, filtering, merges extract solution, Moringa flavone extractive is made；By Moringa flavones crude extract degreasing, decolourize, adjust pH, extracted repeatedly using absolute ethyl alcohol 3 times, then smart extract solution is obtained by D101 macroporous absorbent resin separating-purifyings；Smart extract solution is concentrated under reduced pressure, recrystallized, is dried, Moringa flavone powder is made.The technological operation of the present invention is simple, and production cost is low, can remove the glucide during Moringa extracting flavonoids, improves product purity；Meanwhile prepared Moringa flavones product stability is high, is advantageous to industrialized production.
Description
【Technical field】
The invention belongs to the extractive technique field of natural products effective ingredient, and in particular to a kind of extraction purification Moringa flavones
Technique.
【Background technology】
Moringa Moringaceae, Moringa, it is perennial woody plant, is a kind of tropical plants for having unique economic to be worth,
It is distributed mainly on the provinces and regions such as Yunnan, Guangdong, Guangxi, Hainan, Taiwan.Moringa is as vegetable food medicinal material in augment nutritional, dietotherapy
Health care, medical health etc. are used widely, and are described as " Tree of Life, the tree of mystery ".The polysaccharide contained in Moringa can
With activating cell strengthen immunity, contribute to excreting insulin and regulation blood glucose, there is abundant health care's effect, it is long-term use of
There is positive effect for antihypertensive, drop high fat of blood, reducing hyperglycaemia, cancer prevention tumour can also be prevented, strengthen immunity is protected
Shield heart is dirty, can be with balanced human's skin pigment, beautifying face and moistering lotion.Meanwhile the flavones contained in Moringa can effective against oxidation,
Green Tea Extract, eliminate human body active oxygen；Blood circulation can be improved, reduce cholesterol, improve the symptom of cardiovascular and cerebrovascular disease；
Stomach lining treatment gastric ulcer, pre- anti-osteoporosis, prevention fatty liver, alcoholic liver are protected, is refreshed the mind, treatment apoplexy, enhancing
Digest, keep fit taste, dispelling fatigue, to treat and prevent depression etc. effect notable." a kind of Moringa is total for Patent Application Publication
Extracting method of flavones and application thereof (patent No.：ZL201510157323.6) ", there is provided a kind of extraction side of Moringa general flavone
Method, it is made through stock, degreasing, extraction, purifying, extraction Moringa general flavone content is in 18-35%.But the Moringa general flavone extracted
Still suffer from that stability is not high enough, purity is low, be not easy to realize industrial applications.With the improvement of living standards, people are to Moringa
Food medicine effect is increasingly paid attention to, and the extraction process of the active ingredient such as pterigospermin, Moringa polysaccharide, Moringa flavones continues to develop how
The Moringa active ingredient of high-purity is extracted, while improves product properties, is allowed to realize industrialized production, has to production and living
Important promotion is important.
【The content of the invention】
A kind of technique of extraction purification Moringa flavones provided by the invention, to solve, Moringa extracting flavonoids speed is low to be caused to give birth to
The problems such as production cost is high, carbohydrate content causes Moringa flavones purity not high more.
To solve above technical problem, the present invention uses following technical method：
A kind of technique of extraction purification Moringa flavones, comprises the following steps：
S1：Leaf of Moringa is taken, dry, pulverize as coarse powder, adds distillation watertight to swell 24h, filters moisture content, coarse powder of swelling is made
It is standby；
S2：To swell coarse powder add the 15-20 times of 60-80% measured ethanol, while add 0.1-0.15% times measure α-
Amylase, refluxing extraction 3 times, filtering, merge extract solution, Moringa flavones crude extract is made；
S3：By Moringa flavones crude extract degreasing, decolourize, add ammoniacal liquor regulation pH to 7.5-8, using absolute ethyl alcohol repeatedly
Extraction 3 times, chromatographic solution is abandoned, collect extract, using D101 macroporous absorbent resin separating-purifying extracts, condition is：Parsing
Agent is 42-55% ethanol solution, and its dosage is 3-4.5 times of macroporous absorbent resin volume, and water wash zone is deionized water, macropore
Polymeric adsorbent absorption regeneration solvent be 2-4% soda bath, adsorption zone flow velocity 8-12BV/h, desorption zone flow velocity 15-25BV/h,
Water wash zone flow velocity 25-35BV/h, renewing zone flow velocity 5-10BV/h, switching time 1-1.5h, temperature are 45-55 DEG C, and pressure is
0.5-0.8MPa, it is standby that smart extract solution is made；
S4：Smart extract solution is placed in 60-65 DEG C of rotary evaporator, while is decompressed to 0.01-0.03MPa, is concentrated into
The 8%-11% of flavones containing Moringa in every milliliter, recrystallize, dry, Moringa flavone powder is made.
Preferably, leaf of Moringa described in step S1 was advisable with growth period at 40-50 days.
Preferably, the addition of distilled water described in step S1 is 8-10 times of coarse powder.
Preferably, the effect of alpha-amylase described in step S2 is to hydrolyze the glucide in leaf of Moringa.
Preferably, the temperature control of refluxing extraction described in step S2 extracts 2-3h every time at 85-95 DEG C.
Preferably, alpha-amylase described in step S2 also includes following preparation method：
(1) tryptone, yeast extract, sodium chloride, potassium dihydrogen phosphate, starch, water are pressed 2:1:1.5:0.5:1:200 ratio
Example is well mixed, is put into sterile conical flask, adds ammoniacal liquor regulation pH to 7-7.5, is cultivated at 32-35 DEG C；
(2) when thalline enters exponential phase, the solid medium made of agar is transferred to 0.5% inoculum concentration
In, culture 48h is aerated at 35-37 DEG C, by allotment, centrifuges, drying, alpha-amylase is made.
Preferably, degreasing agent described in step S3 is petroleum ether.
Preferably, decolorization described in step S3 is activated carbon decolorizing, while can also go the removal of impurity.
Preferably, the temperature recrystallized described in step S4 is 8-12 DEG C.
Preferably, described in step S4 dry temperature be 65-70 DEG C, when a length of 1.5-3h.
The invention has the advantages that：
The extraction process of the present invention is simple to operate, and production cost is low, can remove caused by during Moringa extracting flavonoids
Glucide, effectively increase product purity；Meanwhile prepared Moringa flavones product stability is high, is advantageous to industrial metaplasia
Production.
【Embodiment】
It is specifically described with reference to specific embodiment.
Embodiment 1
A kind of technique of extraction purification Moringa flavones, comprises the following steps：
S1：Leaf of Moringa of the growth period at 40 days is taken, dry, pulverize as coarse powder, adds the distillation watertights of 8 times of amounts to swell 24h,
Moisture content is filtered, it is standby that coarse powder of swelling is made；
S2：60% ethanol of 15 times of amounts is added to coarse powder of swelling, while adds the alpha-amylase of 0.1% times of amount, it is described
The preparation method of alpha-amylase comprises the following steps：(1) by tryptone, yeast extract, sodium chloride, potassium dihydrogen phosphate, starch, water
By 2:1:1.5:0.5:1:200 ratio is well mixed, and is put into sterile conical flask, ammoniacal liquor regulation pH to 7 is added, at 32 DEG C
Under cultivated；(2) when thalline enters exponential phase, the solid culture made of agar is transferred to 0.5% inoculum concentration
In base, culture 48h is aerated at 35 DEG C, by allotment, centrifuges, drying, alpha-amylase is made.Alpha-amylase will be added again
Coarse powder and 60% ethanol refluxing extraction 3 times at 85 DEG C of swelling, extract 2h every time, filter, merge extract solution, be made Moringa
Flavones crude extract；
S3：Moringa flavones crude extract is used into petroleum ether degreasing, reuses decolorization and impurity removal by active carbon, adds ammoniacal liquor regulation
PH to 7.5, use absolute ethyl alcohol to extract 3 times repeatedly to separate polysaccharide component therein, abandon chromatographic solution, collect extract, adopt
With D101 macroporous absorbent resin separating-purifying extracts, condition is：The ethanol solution that agent is 42% is parsed, its dosage is that macropore is inhaled
3 times of attached resin volume, water wash zone are deionized water, and macroporous absorbent resin absorption regeneration solvent is 2% soda bath, is adsorbed
Area flow velocity 8BV/h, desorption zone flow velocity 15BV/h, water wash zone flow velocity 25BV/h, renewing zone flow velocity 5BV/h, switching time 1h, temperature
Spend for 45-55 DEG C, pressure 0.5MPa, it is standby that smart extract solution is made；
S4：Smart extract solution is placed in 60 DEG C of rotary evaporator, while is decompressed to 0.01MPa, be concentrated into every milliliter
Flavones containing Moringa 8%, is recrystallized at 8 DEG C, obtains yellow crystal body, and 1.5h is dried at 65 DEG C, and Moringa flavone powder is made.
Embodiment 2
A kind of technique of extraction purification Moringa flavones, comprises the following steps：
S1：Leaf of Moringa of the growth period at 45 days is taken, dry, pulverize as coarse powder, adds the distillation watertights of 9 times of amounts to swell 24h,
Moisture content is filtered, it is standby that coarse powder of swelling is made；
S2：65% ethanol of 16 times of amounts is added to coarse powder of swelling, while adds the alpha-amylase of 0.11% times of amount, it is described
The preparation method of alpha-amylase comprises the following steps：(1) by tryptone, yeast extract, sodium chloride, potassium dihydrogen phosphate, starch, water
By 2:1:1.5:0.5:1:200 ratio is well mixed, and is put into sterile conical flask, ammoniacal liquor regulation pH to 7.2 is added, 33
Cultivated at DEG C；(2) when thalline enters exponential phase, the solid made of agar is transferred to 0.5% inoculum concentration and trained
Support in base, culture 48h is aerated at 36 DEG C, by allotment, centrifuge, drying, alpha-amylase is made.Alphalise starch will be added again
Coarse powder and 65% ethanol refluxing extraction 3 times at 88 DEG C of swelling of enzyme, extract 2.5h every time, filtering, merge extract solution, are made
Moringa flavones crude extract；
S3：Moringa flavones crude extract is used into petroleum ether degreasing, reuses decolorization and impurity removal by active carbon, adds ammoniacal liquor regulation
PH to 7.8, use absolute ethyl alcohol to extract 3 times repeatedly to separate polysaccharide component therein, abandon chromatographic solution, collect extract, adopt
With D101 macroporous absorbent resin separating-purifying extracts, condition is：The ethanol solution that agent is 50% is parsed, its dosage is that macropore is inhaled
4 times of attached resin volume, water wash zone are deionized water, and macroporous absorbent resin absorption regeneration solvent is 2.5% soda bath, is inhaled
Attached area's flow velocity 10BV/h, desorption zone flow velocity 18BV/h, water wash zone flow velocity 30BV/h, renewing zone flow velocity 8BV/h, switching time be
1.2h, temperature are 50 DEG C, pressure 0.6MPa, and it is standby that smart extract solution is made；
S4：Smart extract solution is placed in 65 DEG C of rotary evaporator, while is decompressed to 0.03MPa, be concentrated into every milliliter
Flavones containing Moringa 11%, is recrystallized at 10 DEG C, obtains yellow crystal body, and 2h is dried at 66 DEG C, and Moringa flavone powder is made.
Embodiment 3
A kind of technique of extraction purification Moringa flavones, comprises the following steps：
S1：Leaf of Moringa of the growth period at 50 days is taken, dry, pulverize as coarse powder, adds the distillation watertights of 10 times of amounts to swell
24h, moisture content is filtered, it is standby that coarse powder of swelling is made；
S2：80% ethanol of 20 times of amounts is added to coarse powder of swelling, while adds the alpha-amylase of 0.15% times of amount, it is described
The preparation method of alpha-amylase comprises the following steps：(1) by tryptone, yeast extract, sodium chloride, potassium dihydrogen phosphate, starch, water
By 2:1:1.5:0.5:1:200 ratio is well mixed, and is put into sterile conical flask, ammoniacal liquor regulation pH to 7.5 is added, 35
Cultivated at DEG C；(2) when thalline enters exponential phase, the solid made of agar is transferred to 0.5% inoculum concentration and trained
Support in base, culture 48h is aerated at 37 DEG C, by allotment, centrifuge, drying, alpha-amylase is made.Alphalise starch will be added again
Coarse powder and 80% ethanol refluxing extraction 3 times at 95 DEG C of swelling of enzyme, extract 3h every time, filtering, merge extract solution, are made peppery
Genitein crude extract；
S3：Moringa flavones crude extract is used into petroleum ether degreasing, reuses decolorization and impurity removal by active carbon, adds ammoniacal liquor regulation
PH to 8, use absolute ethyl alcohol to extract 3 times repeatedly to separate polysaccharide component therein, abandon chromatographic solution, collect extract, use
D101 macroporous absorbent resin separating-purifying extracts, condition are：The ethanol solution that agent is 55% is parsed, its dosage is macroporous absorption
4.5 times of resin volume, water wash zone are deionized water, and macroporous absorbent resin absorption regeneration solvent is 4% soda bath, is adsorbed
Area flow velocity 12BV/h, desorption zone flow velocity 25BV/h, water wash zone flow velocity 35BV/h, renewing zone flow velocity 10BV/h, switching time be
1.5h, temperature are 55 DEG C, pressure 0.8MPa, and it is standby that smart extract solution is made；
S4：Smart extract solution is placed in 62 DEG C of rotary evaporator, while is decompressed to 0.02MPa, be concentrated into every milliliter
Flavones containing Moringa 9%, is recrystallized at 12 DEG C, obtains yellow crystal body, and 3h is dried at 70 DEG C, and Moringa flavone powder is made.
Embodiment 4
A kind of technique of extraction purification Moringa flavones, comprises the following steps：
S1：Leaf of Moringa of the growth period at 40 days is taken, dry, pulverize as coarse powder, add the 8-10 times of distillation measured watertight to swell
24h, moisture content is filtered, it is standby that coarse powder of swelling is made；
S2：80% ethanol of 20 times of amounts is added to coarse powder of swelling, while adds the alpha-amylase of 0.1% times of amount, it is described
The preparation method of alpha-amylase comprises the following steps：(1) by tryptone, yeast extract, sodium chloride, potassium dihydrogen phosphate, starch, water
By 2:1:1.5:0.5:1:200 ratio is well mixed, and is put into sterile conical flask, ammoniacal liquor regulation pH to 7 is added, at 35 DEG C
Under cultivated；(2) when thalline enters exponential phase, the solid culture made of agar is transferred to 0.5% inoculum concentration
In base, culture 48h is aerated at 35 DEG C, by allotment, centrifuges, drying, alpha-amylase is made.Alpha-amylase will be added again
Coarse powder and 80% ethanol refluxing extraction 3 times at 85 DEG C of swelling, extract 2h every time, filter, merge extract solution, be made Moringa
Flavones crude extract；
S3：Moringa flavones crude extract is used into petroleum ether degreasing, reuses decolorization and impurity removal by active carbon, adds ammoniacal liquor regulation
PH to 8, use absolute ethyl alcohol to extract 3 times repeatedly to separate polysaccharide component therein, abandon chromatographic solution, collect extract, use
D101 macroporous absorbent resin separating-purifying extracts, condition are：The ethanol solution that agent is 55% is parsed, its dosage is macroporous absorption
3 times of resin volume, water wash zone are deionized water, and macroporous absorbent resin absorption regeneration solvent is 2%-4% soda bath, is inhaled
Attached area's flow velocity 11BV/h, desorption zone flow velocity 22BV/h, water wash zone flow velocity 35BV/h, renewing zone flow velocity 10BV/h, switching time be
1.5h, temperature are 55 DEG C, pressure 0.7MPa, and it is standby that smart extract solution is made；
S4：Smart extract solution is placed in 63 DEG C of rotary evaporator, while is decompressed to 0.01MPa, be concentrated into every milliliter
Flavones containing Moringa 9%, is recrystallized at 9 DEG C, obtains yellow crystal body, and 3h is dried at 70 DEG C, and Moringa flavone powder is made.
Experiment 1：
Comparative example 1, using a kind of " extracting method of Moringa general flavone and application thereof (patent No.：
ZL201510157323.6 the method for embodiment 1-3) " prepares Moringa flavones.According to a kind of " extracting method of Moringa general flavone
And application thereof (the patent No.：ZL201510157323.6 the method that absorbance is known in the inspection under 510nm wavelength employed in) " is right
The carry out assay of Moringa flavones, as a result as shown in the table made from embodiment 1-4 and comparative example 1.
Experiment 2：
Comparative example 1, using a kind of " extracting method of Moringa general flavone and application thereof (patent No.：
ZL201510157323.6 the method for embodiment 1-3) " prepares Moringa flavones.Using the stable duration of detection absorbance
Method, it is under equal conditions, stable to the carry out absorbance of Moringa flavones made from embodiment 1-4 and comparative example 1 to continue
Duration determines, as a result as shown in the table.
Learn that the content of Moringa flavones exists made from method of the invention by the experimental data of experiment 1 and experiment 2
More than 40.01%, it is above the component content of Moringa flavones made from prior art；Moringa made from the method for the present invention simultaneously
Flavones stability is higher than Moringa flavones made from prior art.
Above content is to combine specific preferred embodiment further description made for the present invention, it is impossible to is assert
The specific implementation of the present invention is confined to these explanations, for person of an ordinary skill in the technical field, is not departing from
On the premise of present inventive concept, some simple deduction or replace can also be made, should all be considered as belonging to the present invention by being submitted
Claims determine scope of patent protection.
Claims (10)
1. a kind of technique of extraction purification Moringa flavones, it is characterised in that comprise the following steps：
S1：Leaf of Moringa is taken, dry, pulverize as coarse powder, adds distillation watertight to swell 24h, filters moisture content, it is standby that coarse powder of swelling is made
With；
S2：To swelling, coarse powder adds the 15-20 times of 60-80% measured ethanol, while adds the 0.1-0.15% times of alphalise starch measured
Enzyme, refluxing extraction 3 times, filtering, merge extract solution, Moringa flavones crude extract is made；
S3：By Moringa flavones crude extract degreasing, decolourize, add ammoniacal liquor regulation pH to 7.5-8, extracted repeatedly using absolute ethyl alcohol
3 times, chromatographic solution is abandoned, collects extract, using D101 macroporous absorbent resin separating-purifying extracts, condition is：Parsing agent is
42-55% ethanol solution, its dosage are 3-4.5 times of macroporous absorbent resin volume, and water wash zone is deionized water, macroporous absorption
Resin adsorption regenerated solvent is 2-4% soda bath, adsorption zone flow velocity 8-12BV/h, desorption zone flow velocity 15-25BV/h, is washed
Area flow velocity 25-35BV/h, renewing zone flow velocity 5-10BV/h, switching time 1-1.5h, temperature are 45-55 DEG C, pressure 0.5-
0.8MPa, it is standby that smart extract solution is made；
S4：Smart extract solution is placed in 60-65 DEG C of rotary evaporator, while is decompressed to 0.01-0.03MPa, is concentrated into every milli
The 8%-11% of flavones containing Moringa in liter, recrystallize, dry, Moringa flavone powder is made.
2. the technique of extraction purification Moringa flavones according to claim 1, it is characterised in that：Leaf of Moringa described in step S1
It was advisable with growth period at 40-50 days.
3. the technique of extraction purification Moringa flavones according to claim 1, it is characterised in that：Distilled water described in step S1
Addition be 8-10 times of coarse powder.
4. the technique of extraction purification Moringa flavones according to claim 1, it is characterised in that：Alphalise starch described in step S2
The effect of enzyme is to hydrolyze the glucide in leaf of Moringa.
5. the technique of extraction purification Moringa flavones according to claim 1, it is characterised in that：Backflow carries described in step S2
The temperature control taken extracts 2-3h every time at 85-95 DEG C.
6. the technique of extraction purification Moringa flavones according to claim 1, it is characterised in that：Alphalise starch described in step S2
Enzyme also includes following preparation method：
(1) tryptone, yeast extract, sodium chloride, potassium dihydrogen phosphate, starch, water are pressed 2:1:1.5:0.5:1:200 ratio is mixed
Close uniformly, be put into sterile conical flask, add ammoniacal liquor regulation pH to 7-7.5, cultivated at 32-35 DEG C；
(2) when thalline enters exponential phase, it is transferred to 0.5% inoculum concentration by solid medium made of agar,
Culture 48h is aerated at 35-37 DEG C, by allotment, centrifuges, drying, alpha-amylase is made.
7. the technique of extraction purification Moringa flavones according to claim 1, it is characterised in that：Degreasing agent described in step S3
For petroleum ether.
8. the technique of extraction purification Moringa flavones according to claim 1, it is characterised in that：Decolourized described in step S3
Journey is activated carbon decolorizing, while can also go the removal of impurity.
9. the technique of extraction purification Moringa flavones according to claim 1, it is characterised in that：Recrystallized described in step S4
Temperature be 8-12 DEG C.
10. the technique of extraction purification Moringa flavones according to claim 1, it is characterised in that：Dried described in step S4
Temperature be 65-70 DEG C, when a length of 1.5-3h.
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| CN116622002A (en) * | 2023-06-12 | 2023-08-22 | 湖南绿蔓生物科技股份有限公司 | Preparation method of moringa oleifera leaf extract |
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| CN103980242B (en) * | 2014-04-29 | 2016-01-27 | 天津大学 | The method of Moringa flavones is extracted from leaf of Moringa |
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| CN116622002A (en) * | 2023-06-12 | 2023-08-22 | 湖南绿蔓生物科技股份有限公司 | Preparation method of moringa oleifera leaf extract |
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Application publication date: 20180116 |