CN103860620A - Antioxidant active extract of snow chrysanthemum and preparation method thereof - Google Patents
Antioxidant active extract of snow chrysanthemum and preparation method thereof Download PDFInfo
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Abstract
The invention relates to the technical field of snow chrysanthemum extraction, and discloses an antioxidant active extract of snow chrysanthemum and a preparation method thereof. The antioxidant active extract of the snow chrysanthemum is prepared by the following preparation method: firstly, extracting by using an ethanol water solution after crushing the snow chrysanthemum and then carrying out vacuum concentration to obtain an ethanol extract; adsorbing by using macroporous resin, eluting by using an ethanol water solution, carrying out vacuum common peony and drying to obtain the antioxidant active extract of the snow chrysanthemum after extracting by petroleum ether, trichloromethane and ethyl acetate sequentially. The result of an experiment of the antioxidant active extract of the snow chrysanthemum obtained by the method in the ability of removing and restoring DPPH and ABTS free radicals proves that the antioxidant active extract of the snow chrysanthemum disclosed by the embodiment of the invention has good antioxidant action; the process disclosed by the invention is simple and convenient to operate, safe and reliable in extraction process, and is a technique capable of effectively extracting the antioxidant active extract of the snow chrysanthemum.
Description
Technical field
The present invention relates to avenge chrysanthemum extractive technique field, is a kind of snow chrysanthemum antioxidative active extractive and preparation method thereof.
Background technology
Along with the raising of people's living standard, the various diseases serious harm that reactive oxygen species and free radicals causes the mankind's health, caused the extensive concern of various circles of society.In recent years, evaluate and natural resources that screening has a strong anti-oxidation activity have become the new trend of biology, medical science and food scientific research.
Snow chrysanthemum, have another name called " blood chrysanthemum ", formal name used at school is dichromatism Coreopsis basalis (Coreopsis tinetoria), is Compositae (Compositae) golden pheasant Chrysanthemum annual herb plant, is mainly distributed in the Kunlun Mountains of the about 3000m of hotan area height above sea level.For a long time, the Kunlun snow chrysanthemum is drunk when scented tea by local resident, is also a kind of Xinjiang medicinal herbs most in use.It has the effects such as heat-clearing and toxic substances removing, blood circulation promoting and blood stasis dispelling, stomach function regulating spleen invigorating, blood pressure lowering, blood fat reducing and blood sugar lowering.In the Kunlun snow chrysanthemum, containing abundant nutritional labeling, and the oxidation-resistant active ingredient such as polyphenol, flavone, anthocyanidin, in control some diseases relevant to radical damage of the mankind and defying age process, may play a very important role.But there is no the technological means of efficiently producing snow chrysanthemum antioxidative active extractive at present both at home and abroad, therefore, yet there are no the research report of the snow chrysanthemum oxidation-resistant active ingredient using antioxidant activity as index separation and Extraction, this exploitation to snow chrysanthemum resource has great importance.
Summary of the invention
The invention provides a kind of snow chrysanthemum antioxidative active extractive and preparation method thereof, overcome the deficiency of above-mentioned prior art, it can effectively solve the problem that there is no the technological means of efficiently producing snow chrysanthemum antioxidative active extractive at present both at home and abroad.
One of technical scheme of the present invention realizes by following measures: a kind of snow chrysanthemum antioxidative active extractive obtains as follows: the first step, broken snow inulin and mistake 10 mesh sieve to 50 mesh sieves are obtained avenging inulin to mince, adding volume fraction in mincing to snow inulin is that 70% to 95% ethanol water extracts 1 time to 3 times, each time of extracting is 0.5 hour to 2 hours, temperature is 76 DEG C to 80 DEG C, it is each that to extract to adding the mince volume fraction of 6 times of weight portion to 20 times weight portions of snow inulin during snow inulin minces be 70% to 95% ethanol water, after each extraction, filter and obtain filtrate, after being merged, filtrate carries out concentrating under reduced pressure, obtain ethanol extraction,
Second step, after adding the water suspendible of 1 times of weight portion to 4 times weight portion of ethanol extraction in ethanol extraction, obtain suspension at normal temperatures, after adding petroleum ether to extract 3 times to 6 times in suspension, obtain petroleum ether raffinate again, each extraction adds the petroleum ether of 1 times of weight portion to 4 times weight portion of ethanol extraction;
The 3rd step obtains chloroform raffinate at normal temperatures after adding chloroform to extract 3 times to 6 times in petroleum ether raffinate, and each extraction adds the chloroform of 1 times of weight portion to 4 times weight portion of ethanol extraction;
The 4th step, after adding ethyl acetate to extract 3 times to 5 times in chloroform raffinate, obtain acetic acid ethyl acetate extract at normal temperatures, each extraction adds the ethyl acetate of 1 times of weight portion to 4 times weight portion of ethanol extraction, and acetic acid ethyl acetate extract is carried out after concentrating under reduced pressure is also dried obtaining acetic acid ethyl ester extract;
The 5th step, by acetic acid ethyl ester extract macroporous resin adsorption, then carries out eluting with the ethanol water that volumetric concentration is 10% to 70%, collects stream part, carry out concentrating under reduced pressure and dry after obtain avenging chrysanthemum antioxidative active extractive.
The further optimization and/or improvements to one of foregoing invention technical scheme below:
In the above-mentioned first step, the mass water content being evaporated in ethanol extraction after filtrate is merged is 40% to 50%; Or/and in the first step, to be 50 DEG C to 70 DEG C, pressure in temperature carry out for-0.06Mpa concentrating under reduced pressure to-0.1Mpa condition.
In above-mentioned the 4th step, acetic acid ethyl acetate extract is carried out to concentrating under reduced pressure and the dry mass water content that obtains is 5.5% to 6.9% acetic acid ethyl ester extract; Or/and concentrating under reduced pressure carries out to-0.1Mpa at 50 DEG C to 70 DEG C ,-0.06Mpa; Or/and in the 4th step, dry is in temperature is-60 DEG C to-40 DEG C, the pressure vacuum freeze drier that is 0Mpa to 0.01Mpa, or carry out under the spraying of 130 DEG C to 190 DEG C of inlet temperature, 30 DEG C to 90 DEG C of leaving air temps is dried.
In above-mentioned the 5th step, by acetic acid ethyl ester extract AB-8 macroporous resin adsorption, during by AB-8 macroporous resin adsorption, upper prop flow velocity is 1.4 bv/h to 1.7 bv/h, then is that 40% to 50% ethanol water carries out eluting with the flow velocity of 1.4 bv/h to 1.7 bv/h by volumetric concentration; Or/and, in the 5th step, stream part of collection is carried out to concentrating under reduced pressure and the dry mass water content that obtains is 5.5% to 6.9% snow chrysanthemum antioxidative active extractive; Or/and, in the 5th step, concentrating under reduced pressure temperature be 50 DEG C to 70 DEG C, pressure be-0.06Mpa carries out to-0.1Mpa condition; Or/and, in the 5th step, dry be in temperature is-60 DEG C to-40 DEG C, the pressure vacuum freeze drier that is 0Mpa to 0.01Mpa or carry out under the spraying of 130 DEG C to 190 DEG C of inlet temperature, 30 DEG C to 90 DEG C of leaving air temps is dried; Or/and in the 5th step, while carrying out eluting with ethanol water, the volumetric concentration of ethanol water is respectively 10% or 30% or 50% or 70%.
Two of technical scheme of the present invention realizes by following measures: a kind of preparation method of avenging chrysanthemum antioxidative active extractive, carry out in the steps below: the first step, broken snow inulin and mistake 10 mesh sieve to 50 mesh sieves are obtained avenging inulin to mince, adding volume fraction in mincing to snow inulin is that 70% to 95% ethanol water extracts 1 time to 3 times, each time of extracting is 0.5 hour to 2 hours, temperature is 76 DEG C to 80 DEG C, it is each that to extract to adding the mince volume fraction of 6 times of weight portion to 20 times weight portions of snow inulin during snow inulin minces be 70% to 95% ethanol water, after each extraction, filter and obtain filtrate, after being merged, filtrate carries out concentrating under reduced pressure, obtain ethanol extraction,
Second step, after adding the water suspendible of 1 times of weight portion to 4 times weight portion of ethanol extraction in ethanol extraction, obtain suspension at normal temperatures, after adding petroleum ether to extract 3 times to 6 times in suspension, obtain petroleum ether raffinate again, each extraction adds the petroleum ether of 1 times of weight portion to 4 times weight portion of ethanol extraction;
The 3rd step obtains chloroform raffinate at normal temperatures after adding chloroform to extract 3 times to 6 times in petroleum ether raffinate, and each extraction adds the chloroform of 1 times of weight portion to 4 times weight portion of ethanol extraction;
The 4th step, after adding ethyl acetate to extract 3 times to 5 times in chloroform raffinate, obtain acetic acid ethyl acetate extract at normal temperatures, each extraction adds the ethyl acetate of 1 times of weight portion to 4 times weight portion of ethanol extraction, and acetic acid ethyl acetate extract is carried out after concentrating under reduced pressure is also dried obtaining acetic acid ethyl ester extract;
The 5th step, by acetic acid ethyl ester extract macroporous resin adsorption, then carries out eluting with the ethanol water that volumetric concentration is 10% to 70%, collects stream part, carry out concentrating under reduced pressure and dry after obtain avenging chrysanthemum antioxidative active extractive.
Two the further optimization and/or improvements to foregoing invention technical scheme below:
In the above-mentioned first step, the mass water content being evaporated in ethanol extraction after filtrate is merged is 40% to 50%; Or/and in the first step, to be 50 DEG C to 70 DEG C, pressure in temperature carry out for-0.06Mpa concentrating under reduced pressure to-0.1Mpa condition.
In above-mentioned the 4th step, acetic acid ethyl acetate extract is carried out to concentrating under reduced pressure and the dry mass water content that obtains is 5.5% to 6.9% acetic acid ethyl ester extract; Or/and concentrating under reduced pressure carries out to-0.1Mpa at 50 DEG C to 70 DEG C ,-0.06Mpa; Or/and in the 4th step, dry is in temperature is-60 DEG C to-40 DEG C, the pressure vacuum freeze drier that is 0Mpa to 0.01Mpa, or carry out under the spraying of 130 DEG C to 190 DEG C of inlet temperature, 30 DEG C to 90 DEG C of leaving air temps is dried.
In above-mentioned the 5th step, by acetic acid ethyl ester extract AB-8 macroporous resin adsorption, during by AB-8 macroporous resin adsorption, upper prop flow velocity is 1.4 bv/h to 1.7 bv/h, then is that 40% to 50% ethanol water carries out eluting with the flow velocity of 1.4 bv/h to 1.7 bv/h by volumetric concentration; Or/and, in the 5th step, stream part of collection is carried out to concentrating under reduced pressure and the dry mass water content that obtains is 5.5% to 6.9% snow chrysanthemum antioxidative active extractive; Or/and, in the 5th step, concentrating under reduced pressure temperature be 50 DEG C to 70 DEG C, pressure be-0.06Mpa carries out to-0.1Mpa condition; Or/and, in the 5th step, dry be in temperature is-60 DEG C to-40 DEG C, the pressure vacuum freeze drier that is 0Mpa to 0.01Mpa or carry out under the spraying of 130 DEG C to 190 DEG C of inlet temperature, 30 DEG C to 90 DEG C of leaving air temps is dried; Or/and in the 5th step, while carrying out eluting with ethanol water, the volumetric concentration of ethanol water is respectively 10% or 30% or 50% or 70%.
The snow chrysanthemum antioxidative active extractive that the present invention obtains shows in the experimental result of DPPH, ABTS free radical scavenging and reducing power, the snow chrysanthemum antioxidative active extractive obtaining according to the embodiment of the present invention has good antioxidation, and the simple and convenient operation of technique of the present invention, leaching process is safe and reliable, is a kind of process that can effectively extract snow chrysanthemum antioxidative active extractive.
Brief description of the drawings
Accompanying drawing 1 is for 10% extract, 30% extract, 50% extract, 70% extract, BHT and Vc in the present invention are to reducing power measurement result curve chart.
Detailed description of the invention
The present invention is not subject to the restriction of following embodiment, can determine concrete embodiment according to technical scheme of the present invention and practical situation.
Below in conjunction with embodiment, the invention will be further described:
Embodiment 1, a kind of snow chrysanthemum antioxidative active extractive obtains by following preparation method: the first step, to avenge inulin broken and cross 10 mesh sieve to 50 mesh sieves, the volume fraction that adds snow chrysanthemum 6 times of weight portion to 20 times weight portions is 70% to 95% ethanol water, is dipped to the heart, extract 1 time to 3 times, each extraction 0.5 hour to 2 hours, each temperature of extracting is 76 DEG C to 80 DEG C, after each extraction, filters and obtains filtrate, after filtrate is merged, carry out concentrating under reduced pressure, obtain ethanol extraction; Second step, after adding the water suspendible of 1 times of weight portion to 4 times weight portion of ethanol extraction in ethanol extraction, obtain suspension at normal temperatures, after adding petroleum ether to extract 3 times to 6 times in suspension, obtain petroleum ether raffinate again, each extraction adds the petroleum ether of 1 times of weight portion to 4 times weight portion of ethanol extraction, and water used herein is pure water; The 3rd step obtains chloroform raffinate at normal temperatures after adding chloroform to extract 3 times to 6 times in petroleum ether raffinate, and each extraction adds the chloroform of 1 times of weight portion to 4 times weight portion of ethanol extraction; The 4th step, after adding ethyl acetate to extract 3 times to 5 times in chloroform raffinate, obtain acetic acid ethyl acetate extract at normal temperatures, each extraction adds the ethyl acetate of 1 times of weight portion to 4 times weight portion of ethanol extraction, and acetic acid ethyl acetate extract is carried out after concentrating under reduced pressure is also dried obtaining acetic acid ethyl ester extract; The 5th step, by acetic acid ethyl ester extract macroporous resin adsorption, then carries out eluting with the ethanol water that volumetric concentration is 10% to 70%, collects stream part, carry out concentrating under reduced pressure and dry after obtain avenging chrysanthemum antioxidative active extractive.
Embodiment 2, a kind of snow chrysanthemum antioxidative active extractive obtains by following preparation method: the first step, by broken snow inulin and cross 10 mesh sieves or 50 mesh sieves and obtain avenging inulin and mince, adding volume fraction in mincing to snow inulin is that 70% or 95% ethanol water extracts 1 time or 3 times, each time of extracting is 0.5 hour or 2 hours, temperature is 76 DEG C or 80 DEG C, it is each that to extract to adding the mince volume fraction of 6 times of weight portions or 20 times of weight portions of snow inulin during snow inulin minces be 70% or 95% ethanol water, after each extraction, filter and obtain filtrate, after being merged, filtrate carries out concentrating under reduced pressure, obtain ethanol extraction, second step, after adding the water suspendible of 1 times of weight portion of ethanol extraction or 4 times of weight portions in ethanol extraction, obtain suspension at normal temperatures, after adding petroleum ether to extract 3 times or 6 times in suspension, obtain petroleum ether raffinate again, each extraction adds the petroleum ether of 1 times of weight portion of ethanol extraction or 4 times of weight portions, the 3rd step obtains chloroform raffinate at normal temperatures after adding chloroform to extract 3 times or 6 times in petroleum ether raffinate, and each extraction adds the chloroform of 1 times of weight portion of ethanol extraction or 4 times of weight portions, the 4th step, after adding ethyl acetate to extract 3 times or 5 times in chloroform raffinate, obtain acetic acid ethyl acetate extract at normal temperatures, each extraction adds the ethyl acetate of 1 times of weight portion of ethanol extraction or 4 times of weight portions, and acetic acid ethyl acetate extract is carried out after concentrating under reduced pressure is also dried obtaining acetic acid ethyl ester extract, the 5th step, by acetic acid ethyl ester extract macroporous resin adsorption, then carries out eluting with the ethanol water that volumetric concentration is 10% or 70%, collects stream part, carry out concentrating under reduced pressure and dry after obtain avenging chrysanthemum antioxidative active extractive.
Embodiment 3, as above-described embodiment preferably, in the first step, the mass water content being evaporated in ethanol extraction after filtrate is merged is 40% to 50%; Or/and in the first step, to be 50 DEG C to 70 DEG C, pressure in temperature carry out for-0.06Mpa concentrating under reduced pressure to-0.1Mpa condition.
Embodiment 4, as above-described embodiment preferably, in the 4th step, acetic acid ethyl acetate extract is carried out to concentrating under reduced pressure and the dry mass water content that obtains is 5.5% to 6.9% acetic acid ethyl ester extract; Or/and concentrating under reduced pressure carries out to-0.1Mpa at 50 DEG C to 70 DEG C ,-0.06Mpa; Or/and in the 4th step, dry is in temperature is-60 DEG C to-40 DEG C, the pressure vacuum freeze drier that is 0Mpa to 0.01Mpa, or carry out under the spraying of 130 DEG C to 190 DEG C of inlet temperature, 30 DEG C to 90 DEG C of leaving air temps is dried.
Embodiment 5, as above-described embodiment preferably, in the 5th step, by acetic acid ethyl ester extract AB-8 macroporous resin adsorption, during by AB-8 macroporous resin adsorption, upper prop flow velocity is 1.4 bv/h to 1.7 bv/h, then is that 40% to 50% ethanol water carries out eluting with the flow velocity of 1.4 bv/h to 1.7 bv/h by volumetric concentration; Or/and, in the 5th step, stream part of collection is carried out to concentrating under reduced pressure and the dry mass water content that obtains is 5.5% to 6.9% snow chrysanthemum antioxidative active extractive; Or/and, in the 5th step, concentrating under reduced pressure temperature be 50 DEG C to 70 DEG C, pressure be-0.06Mpa carries out to-0.1Mpa condition; Or/and, in the 5th step, dry be in temperature is-60 DEG C to-40 DEG C, the pressure vacuum freeze drier that is 0Mpa to 0.01Mpa or carry out under the spraying of 130 DEG C to 190 DEG C of inlet temperature, 30 DEG C to 90 DEG C of leaving air temps is dried; Or/and in the 5th step, while carrying out eluting with ethanol water, the volumetric concentration of ethanol water is respectively 10% or 30% or 50% or 70%.
Measure the oxidation resistance of the snow chrysanthemum antioxidative active extractive obtaining according to the above embodiment of the present invention method:
According to the method for the above embodiment of the present invention, by acetic acid ethyl ester extract with being 10% by volumetric concentration respectively after macroporous resin adsorption, 30%, 50% and 70% ethanol water carries out the oxidation resistance of the snow chrysanthemum antioxidative active extractive obtaining after eluting and measures, and compare with positive reference substance BHT and Vc respectively, in following experimentation, volumetric concentration is that 10% ethanol water carries out the snow chrysanthemum antioxidative active extractive that eluting obtains and is called for short 10% extract, volumetric concentration is that 30% ethanol water carries out the snow chrysanthemum antioxidative active extractive that eluting obtains and is called for short 30% extract, volumetric concentration is that 50% ethanol water carries out the snow chrysanthemum antioxidative active extractive that eluting obtains and is called for short 50% extract, volumetric concentration is that 70% ethanol water carries out the snow chrysanthemum antioxidative active extractive that eluting obtains and is called for short 70% extract.
, DPPH free radical scavenging activity measure:
10% extract is configured to 0.002mg/ml with absolute methanol, 0.004mg/ml, 0.006mg/ml, 0.008mg/ml, 0.01mg/ml, the solution of 0.02mg/ml variable concentrations, is configured to 0.001mg/ml by 30% extract with absolute methanol, 0.002mg/ml, 0.004mg/ml, 0.006mg/ml, 0.008mg/ml, the solution of 0.01mg/ml variable concentrations, is configured to 0.002mg/ml by 50% extract with absolute methanol, 0.004mg/ml, 0.006mg/ml, 0.008mg/ml, 0.01mg/ml, the solution of 0.02mg/ml variable concentrations, is configured to 0.002mg/ml by 70% extract with absolute methanol, 0.004mg/ml, 0.006mg/ml, 0.008mg/ml, 0.01mg/ml, 0.015mg/ml, the solution of 0.02mg/ml variable concentrations, is configured to 0.002mg/ml by BHT with absolute methanol, 0.004mg/ml, 0.006mg/ml, 0.008mg/ml, 0.01mg/ml, 0.015mg/ml, the solution of 0.02mg/ml variable concentrations, is configured to 0.002mg/ml by Vc with absolute methanol, 0.004mg/ml, 0.006mg/ml, 0.008mg/ml, 0.01mg/ml, 0.015mg/ml, the solution of 0.02mg/ml variable concentrations, gets respectively 10% extract of 2mL variable concentrations, 30% extract, 50% extract, 70% extract, BHT and Vc add respectively 2mL DPPH methanol solution, mix, and under room temperature, lucifuge is reacted 30 min, measure absorbance, 10% extract of every kind of variable concentrations at 517nm place, 30% extract, 50% extract, 70% extract, BHT survey parallel with Vc three times, averages, and BHT and Vc, as with reference to product, calculate clearance rate.
Clearance rate %=(1-A
sample/ A
control) × 100%
In formula, A
samplethe absorbance of 10% extract, 30% extract, 50% extract, 70% extract, BHT and the Vc of variable concentrations, A
controlbe the absorbance of DPPH background, experimental result is in table 1.
From table 1,10% extract, 30% extract, 50% extract, 70% extract all have certain scavenging action to DPPH free radical, 30% ethanol elution thing and 50% ethanol elution thing all have good removing ability to DPPH free radical, wherein, the ability that 10% extract, 30% extract, 50% extract, 70% extract are removed DPPH free radical is better than positive reference substance BHT, approaches with Vc, and 50% extract to DPPH radical scavenging activity much larger than positive reference substance BHT and Vc, its IC
50for 3.21ug/ml.
, ABTS free radical scavenging activity measure:
10% extract is configured to 0.03mg/ml with absolute methanol, 0.05mg/ml, 0.07mg/ml, 0.1mg/ml, 0.15mg/ml, the solution of 0.2mg/ml variable concentrations, is configured to 0.02mg/ml by 30% extract with absolute methanol, 0.04mg/ml, 0.06mg/ml, 0. 08mg/ml, 0.1mg/ml, 0.2mg/ml, the solution of 0.3mg/ml variable concentrations, is configured to 0.02mg/ml by 50% extract with absolute methanol, 0.04mg/ml, 0.06mg/ml, 0.08 mg/ml, the solution of 0.1mg/ml variable concentrations, is configured to 0.02mg/ml by 70% extract with absolute methanol, 0.04mg/ml, 0.06mg/ml, 0.08mg/ml, the solution of 0.1mg/ml variable concentrations, is configured to 0.02mg/ml by BHT with absolute methanol, 0.04mg/ml, 0.06mg/ml, 0.08mg/ml, the solution of 0.1mg/ml variable concentrations, is configured to 0.02mg/ml by Vc with absolute methanol, 0.04mg/ml, 0.06mg/ml, 0.08mg/ml, the solution of 0.1mg/ml variable concentrations, gets respectively 10% extract of 2mL variable concentrations, 30% extract, 50% extract, 70% extract, BHT and Vc add respectively 2mL DPPH methanol solution, mix, and under room temperature, lucifuge is reacted 30 min, measure absorbance, 10% extract of every kind of variable concentrations at 517 nm places, 30% extract, 50% extract, 70% extract, BHT survey parallel with Vc three times, averages, and BHT and Vc, as with reference to product, calculate clearance rate.
Clearance rate %=(1-A
sample/ A
control) × 100%
In formula, A
samplethe absorbance of 10% extract, 30% extract, 50% extract, 70% extract, BHT and the Vc of variable concentrations, A
controlbe the absorbance of ABTS background, experimental result is in table 1.
From table 1,10% extract, 30% extract, 50% extract, 70% extract all have certain scavenging action to ABTS free radical.Wherein, 50% extract, 70% extract are better than positive reference substance Vc, approach with BHT, and 50% extract is greater than positive reference substance Vc and BHT to the removing ability of ABTS free radical, its IC
50be 0.06288 mg/ml.
, reducing power measure:
With Prussian blue [Fe
4(Fe (CN)
6)
3] growing amount as index, by Hexacyanoferrate potassium [K
3fe (CN)
6] be reduced into K
4fe (CN)
6, recycling Fe
3+form Fe
4(Fe (CN)
6)
3, by 700 nm place light absorption value change-detection reducing power sizes, the reducing power of the higher expression sample of light absorption value is stronger.
10% extract is configured to 0.002mg/ml with absolute methanol, 0.004mg/ml, 0.006mg/ml, 0.008mg/ml, 0.01mg/ml, 0.015mg/ml, the solution of 0.02mg/ml variable concentrations, is configured to 0.002mg/ml by 30% extract with absolute methanol, 0.004mg/ml, 0.006mg/ml, 0.008mg/ml, 0.01mg/ml, 0.015mg/ml, the solution of 0.02mg/ml variable concentrations, is configured to 0.002mg/ml by 50% extract with absolute methanol, 0.004mg/ml, 0.006mg/ml, 0.008mg/ml, 0.01mg/ml, 0.015mg/ml, the solution of 0.02mg/ml variable concentrations, is configured to 0.002mg/ml by 70% extract with absolute methanol, 0.004mg/ml, 0.006mg/ml, 0.008mg/ml, 0.01mg/ml, 0.015mg/ml, the solution of 0.02mg/ml variable concentrations, is configured to 0.002mg/ml by BHT with absolute methanol, 0.004mg/ml, 0.006mg/ml, 0.008mg/ml, 0.01mg/ml, 0.015mg/ml, the solution of 0.02mg/ml variable concentrations, is configured to 0.002mg/ml by Vc with absolute methanol, 0.004mg/ml, 0.006mg/ml, 0.008mg/ml, 0.01mg/ml, 0.015mg/ml, the solution of 0.02mg/ml variable concentrations, respectively gets 10% extract of 2.5mL variable concentrations, 30% extract, 50% extract, 70% extract, BHT and Vc, add respectively 2.5mL PBS (0.2mol/ml, pH=6.6), 2.5mL mass concentration is 1% K
3fe (CN)
6solution, constant temperature 20 min in 50 DEG C of water-baths, cooling fast, add 2.5ml mass concentration and be 10% trichloroacetic acid solution, centrifugal 10 min of 3000r/min, get supernatant 2.5mL, add the liquor ferri trichloridi that 2.5mL distilled water, 0.5ml mass concentration are 0.1%, mix rear standing l0 min, under 700 nm, survey absorbance.All mensuration is parallel carries out three times, averages.
Experimental result: generally, the reducing power of sample and antioxidant activity have obvious dependency, reducing power is stronger, show that oxidation resistance is stronger, measuring in concentration range, 10% extract, 30% extract, 50% extract, 70% extract all demonstrate certain reducing power, and reducing power and extract concentrations are obvious dose-effect relationship, when 50% extract concentrations is 0.1 mg/mL, absorbance and positive control Vc approach, and are 1.453.
Accompanying drawing 1 is for 10% extract, 30% extract, 50% extract, 70% extract, BHT and Vc in the present invention are to reducing power measurement result curve chart.
In sum, the present invention has mainly verified the antioxidant activity effect of snow chrysanthemum antioxidative active extractive from three aspects, result shows that the snow chrysanthemum antioxidative active extractive obtaining according to the above embodiment of the present invention has good antioxidant activity effect, especially be that 50% ethanol water carries out the snow chrysanthemum antioxidative active extractive that eluting obtains by volumetric concentration, compare with BHT with reference substance Vc, be that the antioxidant effect that 50% ethanol water carries out the snow chrysanthemum antioxidative active extractive that eluting obtains is all better than Vc and BHT by volumetric concentration, therefore, the snow chrysanthemum antioxidative active extractive obtaining according to the above embodiment of the present invention has good antioxidant activity effect and has good antioxidation, can be for the preparation of antioxidant health-care product.
The snow chrysanthemum antioxidative active extractive that the present invention obtains shows in the experimental result of DPPH, ABTS free radical scavenging and reducing power, the snow chrysanthemum antioxidative active extractive obtaining according to the embodiment of the present invention has good antioxidation, and the simple and convenient operation of technique of the present invention, leaching process is safe and reliable, is a kind of process that can effectively extract snow chrysanthemum antioxidative active extractive.
Table 1. is removed the IC of DPPH, ABTS free radical
50value
| Sample | Remove DPPH free radical IC 50Value (mg/ml) | Remove ABTS free radical IC 50Value (mg/ml) |
| 10% ethanol elution thing | 0.0121 | 0.1296 |
| 30% ethanol elution thing | 0.0082 | 0.1139 |
| 50% ethanol elution thing | 0.00321 | 0.06288 |
| 70% ethanol elution thing | 0.0196 | 0.0665 |
| Vc | 0.0072 | 0.1008 |
| BHT | 0.0283 | 0.0655 |
Above technical characterictic has formed embodiments of the invention, and it has stronger adaptability and implementation result, can increase and decrease according to actual needs non-essential technical characterictic, meets the demand of different situations.
Claims (10)
1. a snow chrysanthemum antioxidative active extractive, it is characterized in that obtaining as follows: the first step, broken snow inulin and mistake 10 mesh sieve to 50 mesh sieves are obtained avenging inulin to mince, adding volume fraction in mincing to snow inulin is that 70% to 95% ethanol water extracts 1 time to 3 times, each time of extracting is 0.5 hour to 2 hours, temperature is 76 DEG C to 80 DEG C, it is each that to extract to adding the mince volume fraction of 6 times of weight portion to 20 times weight portions of snow inulin during snow inulin minces be 70% to 95% ethanol water, after each extraction, filter and obtain filtrate, after being merged, filtrate carries out concentrating under reduced pressure, obtain ethanol extraction,
Second step, after adding the water suspendible of 1 times of weight portion to 4 times weight portion of ethanol extraction in ethanol extraction, obtain suspension at normal temperatures, after adding petroleum ether to extract 3 times to 6 times in suspension, obtain petroleum ether raffinate again, each extraction adds the petroleum ether of 1 times of weight portion to 4 times weight portion of ethanol extraction;
The 3rd step obtains chloroform raffinate at normal temperatures after adding chloroform to extract 3 times to 6 times in petroleum ether raffinate, and each extraction adds the chloroform of 1 times of weight portion to 4 times weight portion of ethanol extraction;
The 4th step, after adding ethyl acetate to extract 3 times to 5 times in chloroform raffinate, obtain acetic acid ethyl acetate extract at normal temperatures, each extraction adds the ethyl acetate of 1 times of weight portion to 4 times weight portion of ethanol extraction, and acetic acid ethyl acetate extract is carried out after concentrating under reduced pressure is also dried obtaining acetic acid ethyl ester extract;
The 5th step, by acetic acid ethyl ester extract macroporous resin adsorption, then carries out eluting with the ethanol water that volumetric concentration is 10% to 70%, collects stream part, carry out concentrating under reduced pressure and dry after obtain avenging chrysanthemum antioxidative active extractive.
2. snow chrysanthemum antioxidative active extractive according to claim 1, is characterized in that in the first step, and the mass water content being evaporated in ethanol extraction after filtrate is merged is 40% to 50%; Or/and in the first step, to be 50 DEG C to 70 DEG C, pressure in temperature carry out for-0.06Mpa concentrating under reduced pressure to-0.1Mpa condition.
3. snow chrysanthemum antioxidative active extractive according to claim 2, is characterized in that in the 4th step, acetic acid ethyl acetate extract is carried out to concentrating under reduced pressure and the dry mass water content that obtains is 5.5% to 6.9% acetic acid ethyl ester extract; Or/and concentrating under reduced pressure carries out to-0.1Mpa at 50 DEG C to 70 DEG C ,-0.06Mpa; Or/and in the 4th step, dry is in temperature is-60 DEG C to-40 DEG C, the pressure vacuum freeze drier that is 0Mpa to 0.01Mpa, or carry out under the spraying of 130 DEG C to 190 DEG C of inlet temperature, 30 DEG C to 90 DEG C of leaving air temps is dried.
4. snow chrysanthemum antioxidative active extractive according to claim 1 and 2, it is characterized in that in the 5th step, by acetic acid ethyl ester extract AB-8 macroporous resin adsorption, during by AB-8 macroporous resin adsorption, upper prop flow velocity is 1.4 bv/h to 1.7 bv/h, then is that 40% to 50% ethanol water carries out eluting with the flow velocity of 1.4 bv/h to 1.7 bv/h by volumetric concentration; Or/and, in the 5th step, stream part of collection is carried out to concentrating under reduced pressure and the dry mass water content that obtains is 5.5% to 6.9% snow chrysanthemum antioxidative active extractive; Or/and, in the 5th step, concentrating under reduced pressure temperature be 50 DEG C to 70 DEG C, pressure be-0.06Mpa carries out to-0.1Mpa condition; Or/and, in the 5th step, dry be in temperature is-60 DEG C to-40 DEG C, the pressure vacuum freeze drier that is 0Mpa to 0.01Mpa or carry out under the spraying of 130 DEG C to 190 DEG C of inlet temperature, 30 DEG C to 90 DEG C of leaving air temps is dried; Or/and in the 5th step, while carrying out eluting with ethanol water, the volumetric concentration of ethanol water is respectively 10% or 30% or 50% or 70%.
5. snow chrysanthemum antioxidative active extractive according to claim 3, it is characterized in that in the 5th step, by acetic acid ethyl ester extract AB-8 macroporous resin adsorption, during by AB-8 macroporous resin adsorption, upper prop flow velocity is 1.4 bv/h to 1.7 bv/h, then is that 40% to 50% ethanol water carries out eluting with the flow velocity of 1.4 bv/h to 1.7 bv/h by volumetric concentration; Or/and, in the 5th step, stream part of collection is carried out to concentrating under reduced pressure and the dry mass water content that obtains is 5.5% to 6.9% snow chrysanthemum antioxidative active extractive; Or/and, in the 5th step, concentrating under reduced pressure temperature be 50 DEG C to 70 DEG C, pressure be-0.06Mpa carries out to-0.1Mpa condition; Or/and, in the 5th step, dry be in temperature is-60 DEG C to-40 DEG C, the pressure vacuum freeze drier that is 0Mpa to 0.01Mpa or carry out under the spraying of 130 DEG C to 190 DEG C of inlet temperature, 30 DEG C to 90 DEG C of leaving air temps is dried; Or/and in the 5th step, while carrying out eluting with ethanol water, the volumetric concentration of ethanol water is respectively 10% or 30% or 50% or 70%.
6. avenge the preparation method of chrysanthemum antioxidative active extractive for one kind, it is characterized in that carrying out in the steps below: the first step, broken snow inulin and mistake 10 mesh sieve to 50 mesh sieves are obtained avenging inulin to mince, adding volume fraction in mincing to snow inulin is that 70% to 95% ethanol water extracts 1 time to 3 times, each time of extracting is 0.5 hour to 2 hours, temperature is 76 DEG C to 80 DEG C, it is each that to extract to adding the mince volume fraction of 6 times of weight portion to 20 times weight portions of snow inulin during snow inulin minces be 70% to 95% ethanol water, after each extraction, filter and obtain filtrate, after being merged, filtrate carries out concentrating under reduced pressure, obtain ethanol extraction,
Second step, after adding the water suspendible of 1 times of weight portion to 4 times weight portion of ethanol extraction in ethanol extraction, obtain suspension at normal temperatures, after adding petroleum ether to extract 3 times to 6 times in suspension, obtain petroleum ether raffinate again, each extraction adds the petroleum ether of 1 times of weight portion to 4 times weight portion of ethanol extraction; Water used herein is pure water;
The 3rd step obtains chloroform raffinate at normal temperatures after adding chloroform to extract 3 times to 6 times in petroleum ether raffinate, and each extraction adds the chloroform of 1 times of weight portion to 4 times weight portion of ethanol extraction;
The 4th step, after adding ethyl acetate to extract 3 times to 5 times in chloroform raffinate, obtain acetic acid ethyl acetate extract at normal temperatures, each extraction adds the ethyl acetate of 1 times of weight portion to 4 times weight portion of ethanol extraction, and acetic acid ethyl acetate extract is carried out after concentrating under reduced pressure is also dried obtaining acetic acid ethyl ester extract;
The 5th step, by acetic acid ethyl ester extract macroporous resin adsorption, then carries out eluting with the ethanol water that volumetric concentration is 10% to 70%, collects stream part, carry out concentrating under reduced pressure and dry after obtain avenging chrysanthemum antioxidative active extractive.
7. the preparation method of snow chrysanthemum antioxidative active extractive according to claim 6, is characterized in that in the first step, and the mass water content being evaporated in ethanol extraction after filtrate is merged is 40% to 50%; Or/and in the first step, to be 50 DEG C to 70 DEG C, pressure in temperature carry out for-0.06Mpa concentrating under reduced pressure to-0.1Mpa condition.
8. according to the preparation method of the snow chrysanthemum antioxidative active extractive described in claim 6 or 7, it is characterized in that in the 4th step, acetic acid ethyl acetate extract is carried out to concentrating under reduced pressure and the dry mass water content that obtains is 5.5% to 6.9% acetic acid ethyl ester extract; Or/and concentrating under reduced pressure carries out to-0.1Mpa at 50 DEG C to 70 DEG C ,-0.06Mpa; Or/and in the 4th step, dry is in temperature is-60 DEG C to-40 DEG C, the pressure vacuum freeze drier that is 0Mpa to 0.01Mpa, or carry out under the spraying of 130 DEG C to 190 DEG C of inlet temperature, 30 DEG C to 90 DEG C of leaving air temps is dried.
9. according to the preparation method of the snow chrysanthemum antioxidative active extractive described in claim 6 or 7, it is characterized in that in the 5th step, by acetic acid ethyl ester extract AB-8 macroporous resin adsorption, during by AB-8 macroporous resin adsorption, upper prop flow velocity is 1.4 bv/h to 1.7 bv/h, then is that 40% to 50% ethanol water carries out eluting with the flow velocity of 1.4 bv/h to 1.7 bv/h by volumetric concentration; Or/and, in the 5th step, stream part of collection is carried out to concentrating under reduced pressure and the dry mass water content that obtains is 5.5% to 6.9% snow chrysanthemum antioxidative active extractive; Or/and, in the 5th step, concentrating under reduced pressure temperature be 50 DEG C to 70 DEG C, pressure be-0.06Mpa carries out to-0.1Mpa condition; Or/and, in the 5th step, dry be in temperature is-60 DEG C to-40 DEG C, the pressure vacuum freeze drier that is 0Mpa to 0.01Mpa or carry out under the spraying of 130 DEG C to 190 DEG C of inlet temperature, 30 DEG C to 90 DEG C of leaving air temps is dried; Or/and in the 5th step, while carrying out eluting with ethanol water, the volumetric concentration of ethanol water is respectively 10% or 30% or 50% or 70%.
10. the preparation method of snow chrysanthemum antioxidative active extractive according to claim 8, it is characterized in that in the 5th step, by acetic acid ethyl ester extract AB-8 macroporous resin adsorption, during by AB-8 macroporous resin adsorption, upper prop flow velocity is 1.4 bv/h to 1.7 bv/h, then is that 40% to 50% ethanol water carries out eluting with the flow velocity of 1.4 bv/h to 1.7 bv/h by volumetric concentration; Or/and, in the 5th step, stream part of collection is carried out to concentrating under reduced pressure and the dry mass water content that obtains is 5.5% to 6.9% snow chrysanthemum antioxidative active extractive; Or/and, in the 5th step, concentrating under reduced pressure temperature be 50 DEG C to 70 DEG C, pressure be-0.06Mpa carries out to-0.1Mpa condition; Or/and, in the 5th step, dry be in temperature is-60 DEG C to-40 DEG C, the pressure vacuum freeze drier that is 0Mpa to 0.01Mpa or carry out under the spraying of 130 DEG C to 190 DEG C of inlet temperature, 30 DEG C to 90 DEG C of leaving air temps is dried; Or/and in the 5th step, while carrying out eluting with ethanol water, the volumetric concentration of ethanol water is respectively 10% or 30% or 50% or 70%.
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| CN113651687A (en) * | 2021-09-26 | 2021-11-16 | 右江民族医学院 | Method for extracting antioxidant active substance from tsaoko amomum fruit |
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Inventor after: Zhou Juxing Inventor after: Wang Qiang Inventor after: Liu Yaya Inventor after: Yang Jie Inventor after: Li Qiang Inventor before: Zhou Juxing Inventor before: Wang Qiang Inventor before: Liu Yaya Inventor before: Yang Jie Inventor before: Cao Baoyong Inventor before: Li Qiang Inventor before: Zhang Zhiqiang |
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Application publication date: 20140618 |