CN102295667A - Method for extracting glucoraphanin compound from broccoli and cauliflower - Google Patents
Method for extracting glucoraphanin compound from broccoli and cauliflower Download PDFInfo
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- 238000000605 extraction Methods 0.000 claims abstract description 24
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Abstract
The invention discloses a method for extracting a glucoraphanin compound from broccoli and cauliflower, which belongs to the technical field of plant components extraction. The method comprises the following steps: (1) selecting and pretreating the plant materials of broccoli and cauliflower; (2) extracting a reagent, performing a crude extraction and concentrating; (3) extracting an organic solvent of the crude extract and removing impurity; (4) carrying out a preliminary purification of the extract; (5) purifying glucoraphanin by chromatographic column and other steps. The invention has the characteristics of simple extracting and purifying method, high extraction yield of glucoraphanin and single components and high purity. The products of the invention can be taken as standard sample for applying to the biochemical experiments and relative researches.
Description
Technical field
The present invention relates to plant component extractive technique field, relate in particular to a kind of broccoli and Cauliflower of utilizing and be material, extract the method for high purity cancer-resisting chemical protective agent precursor radish sulphur glycosides.
Technical background
In recent years, many epidemiological studies find that edible brassicaceous vegetable can reduce the generation of cancer, wherein, the bud seedling vegetable of edible broccoli and Cauliflower or heavy dose of bouquet (250 gram/sky), can reduce dna damage, the activity of raising stages 2 enzyme, thus prevent and reduce the generation of cancer.Studies show that a kind of sulforaphen that is produced by glucosinolate-radish sulphur glycosides (4-glucoraphanin glycosides) degraded is regulated carcinogenic metabolism as repressor specifically by double mechanism in the broccoli.Sulforaphen is the strongest phytochemicals of finding in the vegetables of antitumour activity, and multiple cancers such as lung cancer, intestinal cancer, prostate cancer and mammary cancer are all had very strong inhibition effect.Nearest studies show that, sulforaphen and some other glucosinolate meta-bolites may and promote that cancer cell-apoptosis hinders growth of tumor by the blocking-up cell cycle.But sulforaphen unstable chemcial property, easily decompose, and its precursor radish sulphur glycosides is relatively stable, and can be had anticancer by enteric microorganism degraded generation and the active different sulfo-nitrile hydrochlorate sulforaphen of anti-cancer in human body, so radish sulphur glycosides just can be used as the additive of food, healthcare products or medicine.In addition, the degraded product sulforaphen of radish sulphur glycosides is inhibited to pathogenic bacteria such as some Fusarium oxysporums, the false monospore bacillus etc. of various plants, its degraded product nitriles substance has also participated in the defence to various insects such as aphid, small cabbage moth etc., so radish sulphur glycosides is a kind of phytochemicals with various biological function.At present, the standard substance glucosinolate that can sell in the world has only two kinds of 2-propenyl mustard oil glycosides and phenmethyl glucosinolates, and produce by Sigma-Aldrich and AppliChem two companies respectively, and cost an arm and a leg, and have the strong anticancer and active precursor substance radish sulphur glycosides Shang Wu of anti-cancer company sale.Therefore, utilize own abundant broccoli or cauliflower germplasm resource, develop autonomous radish sulphur glycosides, meaningful work beyond doubt.
Summary of the invention
The present invention seeks to, less at present glucosinolate product category, and rely on external import, expensive defective, providing a kind of is material with broccoli Cauliflower plant, and the precursor that therefrom extracts sulforaphen also can participate in the method for the radish sulphur glycoside compound of the multiple defense response of cress.
The object of the invention is achieved by the following technical programs.
From the method for broccoli Cauliflower extraction radish sulphur glycoside compound, this method is carried out as follows:
(1) selection of broccoli Cauliflower vegetable material and pre-treatment: select to contain radish sulphur glycosides account for total fats glucosinolate more than 90% and the seed, bud seedling, cauline leaf and the floral organ that account for the broccoli Cauliflower of total glucosinolate more than 80% be material, under avoiding as the crumbliness physical injury prerequisite of lancinating, extrude, wear and tear, after water cleans, drains, standby;
(2) extract reagent, slightly mention concentrated: with water, methyl alcohol or 80% methanol aqueous solution serves as to extract reagent; To extract reagent and the pretreated vegetable material of broccoli Cauliflower, by volume the ratio of weight 1-3L: 1-2kg and 85-100 ℃, the same process that stir to soak 20-30min repeat to extract 2-3 time, respectively after filtration, ultrafiltration removes merging filtrate behind the insolubles, after rotary evaporation or lyophilize, be dissolved in after accounting in the water that extracts reagent 1-2% volume radish sulphur glycosides crude extract;
(3) organic solvent extraction of crude extract and impurity elimination: alternately repeatedly be extracted to colourless with organic solvent radish sulphur glycosides crude extract; Collect water, after filtration, ultrafiltration or the centrifugal insolubles of removing, collect supernatant liquor;
(4) preliminary purification of extraction liquid: the supernatant liquor of extraction liquid is carried out preliminary purification in following program: after 1) ratio of extracting 100mg radish sulphur glycosides according to the 1-2g resin takes by weighing DEAE-Sephadex A-25 anionite-exchange resin, soak 10-24h with the 80-100% methanol solution, pack the post height of bed in the glass column of 5cm; 2) doubly wash post for the 0-10% methanol aqueous solution with 3-5 to the concentration of anionite-exchange resin volume; 3) will remove sample on the aqueous portion behind the insolubles; 4), be that leacheate carries out drip washing with 20% Virahol or 20% acetonitrile solution that wait column volume again with behind the water wash of two bed volumes; 5) carry out wash-out with anionresin liquid 2-3 times of column volume and that match with above-mentioned leacheate, and collect with the isopyknic dehydrated alcohol of anionresin liquid in; 6) elutriant is water-soluble after the rotary evaporation drying, obtains containing the mixed solution of multiple glucosinolate component after 0.22 μ m water filter membrane ultrafiltration;
(5) chromatographic column purifying radish sulphur glycosides: with preparation type or half preparative scale chromatography post is purification column, with preliminary purification mixed solution gradation sample introduction, with water that contains 0.1% trifluoroacetic acid and the acetonitrile that contains 0.1% trifluoroacetic acid is moving phase, acetonitrile concentration continues 5min by initial 0-5%, 5-20% continues 20min, and 20% keeps 5min is provided with gradient, at flow velocity 2-3ml/min, detect under the wavelength 210-230nm, collect the highest chromatographic peak and go out effusive moving phase during the peak; Moving phase after above-mentioned gradation sample introduction, the collection is merged, after lyophilize, rotary evaporation or nitrogen dry up drying, promptly obtain purity and be higher than 95% radish sulphur glycoside product; The content of radish sulphur glycosides is with reference to Sun et al., Food Chemistry, and (2011) method is carried out quantitatively.
The described organic solvent that is used to extract is methylene dichloride, ethyl acetate, acetone and normal hexane.
Described anionresin liquid be contain 0.5M Repone K, sodium-chlor, vitriolate of tartar or sodium sulfate 5-20% aqueous isopropanol or contain the acetonitrile solution of the 5-20% of 0.5M Repone K, sodium-chlor, vitriolate of tartar or sodium sulfate, wherein, step (4)-4) be that leacheate then uses the former as anionresin liquid with 20% Virahol, if step (4)-4) be that leacheate then uses the latter as anionresin liquid with 20% acetonitrile.
Described 20% aqueous isopropanol, 20% acetonitrile solution, the 5-20% aqueous isopropanol, the 5-20% acetonitrile solution is the aqueous solution of respective concentration Virahol or acetonitrile.
Described chromatographic column is that filler, particle diameter are that 5-10 μ m, column length are that 25-30cm, diameter are the reverse phase silica gel filled column of 10-20mm for preparation or half preparation type with ODS BP.
Described rotary evaporation or nitrogen dry up drying method, and its temperature is no more than 50 ℃.
The content of described radish sulphur glycosides adopts Sun et al., Food Chemistry, and (2011) method is carried out quantitative assay.
The invention has the beneficial effects as follows:
The present invention is a material with the broccoli and the Cauliflower plant of being rich in radish sulphur glycosides, water or methanol solution extract the glucosinolate mixture, carry out preliminary purification by extraction and anionite-exchange resin, after concentrating, utilize half preparation type or preparative scale chromatography post to get final product dna purity again up to the radish sulphur glycoside compound more than 95%, it is simple that this method has method, with low cost, radish sulphur glycosides extracts the yield height, the single purity height of component can be used as standard substance and is applied in Biochemistry Experiment and the correlative study.
The radish sulphur glycosides that the present invention extracts is to have the cancer-resisting chemical protective agent---the precursor of sulforaphen; Also can participate in the important substance that multiple cress is resisted various plants pathogenic bacteria and insect.
Description of drawings
Fig. 1. from the extraction of broccoli Cauliflower, purifying radish sulphur glycosides process flow sheet.
Fig. 2. select to be used for preparing the broccoli kind glucosinolate high-efficient liquid phase chromatogram of radish sulphur glycosides.
Standard substance are 2-oil of mirbane-β-D-galactoside (120uM), and detection method is with reference to Sun et al., Food Chemistry, (2011), radish sulphur glycosides accounts for 98% of total glucosinolate in the material, accounts for 98.4% of fats glucosinolate.
Fig. 3. the forward and backward color atlas of radish sulphur glycosides enriching and purifying.
Wherein, concentration is 5mM, and 2-propenyl mustard oil glycosides (sinigrin) is as interior mark, and concentration is 3mM
Figure A is that semipreparative column purifying front evaporator light has detected type radish sulphur glycosides result;
Figure B is desulfurization radish sulphur glycosides detection by quantitative result behind the semipreparative column purifying.
Embodiment
The present invention is described in further detail by following examples, but content of the present invention is not limited thereto.
Instrument and material involved in the present invention are as follows:
Liquid phase systems is a U.S. Waters company, comprises pump Waters1525, automatic sample handling system 717plus Autosampler, detector Waters2487;
Institute's water is a ultrapure water, and acetonitrile and trifluoroacetic acid are that chromatographically pure is produced by U.S. TEDIA company;
Used chromatographic column is that Dalian produces according to Lyntech Corporation (US) 10177 South 77th East Avenue Tulsa, Oklahoma 74133 U.S., wherein half preparative scale chromatography column packing is SinoChrom ODS-BP C18, particle diameter 5 μ m, column length 25cm, diameter 10mm, preparative scale chromatography column packing are SinoChrom ODS-BP C18, particle diameter 10 μ m, column length 30cm, diameter 20mm;
Radish sulphur glycosides is to belong to a kind of in the fats glucosinolate; There are the very big difference of difference in the component of contained glucosinolate and content in different broccolis and the Cauliflower material; So when selecting vegetable material, at first need to select to contain radish sulphur glycosides content and the high plant of ratio is a material, to improve the output and the purity of purifying products; Can be when concrete the selection with reference to Sun et al., Food Chemistry, (2011) method detects the component and the content of glucosinolate in the vegetable material; The selected material of the present invention is that radish sulphur glycosides accounts for total fats glucosinolate more than 90%, and accounts for broccoli or the Cauliflower material of total glucosinolate more than 80%.
Embodiment 1:(utilizes the broccoli seed to extract the method for radish sulphur glycosides)
(1) selection of broccoli seed material and pre-treatment: with reference to Sun et al., Food Chemistry, (2011) method, selection contains radish sulphur glycosides and accounts for total fats glucosinolate more than 90% and to account for the broccoli seed of total glucosinolate more than 80% be the material (see figure 2), avoid as lancinate, extrude, wear and tear and physical injury prerequisite such as fragmentation under, water drains after cleaning and removing surface contaminant and impurity 2 times, and is standby;
(2) extract reagent, slightly mention concentrated: with the extraction reagent of methyl alcohol as radish sulphur glycosides, to extract reagent and broccoli seed by volume the ratio of weight 2.5L: 1kg and 85 ℃, stir the same process that soaks 20min and repeat to extract 2 times after, respectively through the Whatman2 filter paper filtering, merging filtrate after insolubles is removed in 0.45 μ m and 0.22 μ m filter membrane ultrafiltration again; After temperature is no more than 50 ℃ rotary evaporation drying, be dissolved in and account in the pure water that extracts reagent volume 2% crude extract of acquisition radish sulphur glycosides;
(3) organic solvent extraction of crude extract and impurity elimination: with the thick water lift solution of radish sulphur glycosides with organic solvent dichloromethane, ethyl acetate, acetone and normal hexane be extracted to repeatedly alternately colourless after, collect water, centrifugal 10min under 4 ℃, 13200g removes insolubles, collects supernatant liquor;
(4) preliminary purification of extraction liquid: the supernatant liquor of extraction liquid is carried out preliminary purification in following program: after 1) ratio of extracting 100mg radish sulphur glycosides according to the 1-2g resin takes by weighing DEAE-Sephadex A-25 anionite-exchange resin, soak 10h with 80% methanol solution, pack the post height of bed in the glass column of 5cm; 2) wash post with 3 times of pure water to the anionite-exchange resin volume; 3) go up sample in the supernatant liquor adding purification column with extraction liquid; 4), be that leacheate carries out drip washing with 20% Virahol that waits column volume again with after the pure water drip washing of two bed volumes; 5) contain 0.5M K with 2 times of column volumes
2SO
45% aqueous isopropanol carry out wash-out, and collect with the isopyknic dehydrated alcohol of above-mentioned anionresin liquid in; 6) elutriant is dissolved in pure water be no more than 50 ℃ rotary evaporation drying through temperature after, obtains containing the mixing solutions of multiple glucosinolate component after 0.22 μ m water filter membrane ultrafiltration;
(5) chromatographic column purifying radish sulphur glycosides: with half preparative scale chromatography post is purification column, the preliminary purification mixed solution is pressed 100 μ l gradation sample introductions, with water that contains 0.1% trifluoroacetic acid and the acetonitrile that contains 0.1% trifluoroacetic acid is moving phase, acetonitrile concentration continues 5min by 0%, 0-20% continues 20min, and 20% keeps 5min is provided with gradient, at flow velocity 2ml/min, detect under the wavelength 229nm, collect the highest chromatographic peak and go out effusive moving phase during the peak; The moving phase of above-mentioned gradation sample introduction, collection is merged, after lyophilize, promptly obtain purity and be higher than 95% radish sulphur glycoside product; The radish sulphur glycosides of enrichment is with reference to Sun et al., Food Chemistry, and (2011) method is carried out qualitative and the detection by quantitative (see figure 3); The radish sulphur glycosides that purifying obtains can be used for its anticancer mechanism research in animal model, human body and animal isolated cells, also can be used for studying the biological function research of radish sulphur glycosides in plant.
Embodiment 2:(utilizes broccoli bud seedling to extract the method for radish sulphur glycosides)
In this example, step (1) material is germination 3-4 days a broccoli bud seedling, the extraction solvent that adopts in the step (2) is a water, extract 100 ℃ of temperature, extraction time 25min, the ratio of extracting reagent and material is 1L: 1kg, repeats to extract 3 times, and the sample after the lyophilize is dissolved in 1.5% and extracts in the water of reagent volume; Impurity-removing method is 4 ℃ of centrifugal 10min of following 13200g in the step (3); Used soak solution is 85% methyl alcohol in the step (4), and soak time is 15h, and scavenging solution is 5% methyl alcohol of 5 times of column volumes, is that leacheate carries out drip washing with 20% acetonitrile solution that waits column volume; Elutriant is 5% acetonitrile solution that contains 0.5M sodium-chlor of 3 times of column volumes; Step 5 adopts the preparative scale chromatography post, and applied sample amount is 200 μ l, and adopting the moving phase starting point concentration is 2.5% acetonitrile, and the flow velocity of moving phase is 3ml/min, and the detection wavelength is 210nm, and other step, technology are with embodiment 1.
Embodiment 3:(utilizes Cauliflower bud seedling to extract the method for radish sulphur glycosides)
In this example, step (1) material therefor is the broccoli cauline leaf, the extraction solvent that adopts in the step (2) is 80% methyl alcohol, extract 90 ℃ of temperature, extraction time 25min, the ratio of extracting reagent and material is 1L: 2kg, repeats to extract 3 times, and the sample after the lyophilize is dissolved in 1.0% and extracts in the water of reagent volume; Impurity-removing method is with 0.45 μ m and 0.22 μ m ultrafiltration membrance filter in the step (3); Used soak solution is 90% methyl alcohol in the step (4), and soak time is 20h, and scavenging solution is 10% methyl alcohol of 4 times of column volumes, is that leacheate carries out drip washing with 20% Virahol that waits column volume; Elutriant is 20% aqueous isopropanol that contains 0.5M Repone K of 2 times of column volumes; Adopt the preparative scale chromatography post in the step (5), applied sample amount is 200 μ l, and adopting the moving phase starting point concentration is 2% acetonitrile, and the flow velocity of moving phase is 3ml/min, and the detection wavelength is 230nm, and other step, technology are with embodiment 1.
Embodiment 4:(utilizes cauliflower seed to extract the method for radish sulphur glycosides)
In this example, step (1) material therefor is a cauliflower seed, the extraction solvent that adopts in the step (2) is a water, extract 100 ℃ of temperature, extraction time 30min, the ratio of extracting reagent and material is 3L: 1kg, repeats to extract 2 times, and the sample after the lyophilize is dissolved in 2.0% and extracts in the water of reagent volume; Impurity-removing method is 0.45 μ m and 0.22 μ m ultrafiltration membrance filter in the step (3); Used soak solution is 100% methyl alcohol in the step (4), and soak time is 24h, and scavenging solution is 5% methyl alcohol of 5 times of column volumes, is that leacheate carries out drip washing with 20% acetonitrile solution that waits column volume; Elutriant is 10% acetonitrile solution that contains 0.5M sodium sulfate of 3 times of column volumes; Step (5) adopts the preparative scale chromatography column purification, and applied sample amount is 150 μ l, and adopting the moving phase starting point concentration is 2% acetonitrile, and the flow velocity of moving phase is 3ml/min, and the detection wavelength is 227nm, and other step, technology are with embodiment 1.
Embodiment 5:(utilizes Cauliflower bud seedling to extract the method for radish sulphur glycosides)
In this example, step (1) material therefor is germination 3-4 days a Cauliflower bud seedling, the extraction solvent that adopts in the step (2) is a methyl alcohol, extract 85 ℃ of temperature, extraction time 20min, the ratio of extracting reagent and material is 3L: 2kg, repeats to extract 3 times, and the sample after the lyophilize is dissolved in 1.5% and extracts in the water of reagent volume; Impurity-removing method is 4 ℃ of centrifugal 10min of following 13200g in the step (3); Used soak solution is 80% methyl alcohol in the step (4), and soak time is 16h, and scavenging solution is 10% methyl alcohol of 4 times of column volumes, is that leacheate carries out drip washing with 20% aqueous isopropanol that waits column volume; Elutriant is 10% aqueous isopropanol that contains the 0.5M vitriolate of tartar of 2 times of column volumes; Step (5) adopts half preparative scale chromatography column purification, and applied sample amount is 100 μ l, and adopting the moving phase starting point concentration is 1% acetonitrile, and the flow velocity of moving phase is 2ml/min, and the detection wavelength is 226nm, and other step, technology are with embodiment 1.
Embodiment 6:(utilizes broccoli curd to extract the method for radish sulphur glycosides)
In this example, step (1) material therefor is sophisticated broccoli curd, the extraction solvent that adopts in the step (2) is 80% methyl alcohol, extract 90 ℃ of temperature, extraction time 20min, the ratio of extracting reagent and material is 2L: 1kg, repeats to extract 2 times, and the sample after the lyophilize is dissolved in 1.0% and extracts in the water of reagent volume; Impurity-removing method is 4 ℃ of centrifugal 10min of following 13200g in the step (3); Used soak solution is 90% methyl alcohol in the step (4), and soak time is 20h, and scavenging solution is 0% methyl alcohol of 3 times of column volumes, is that leacheate carries out drip washing with 20% acetonitrile solution that waits column volume; Elutriant is 15% acetonitrile solution that contains 0.5M sodium sulfate of 3 times of column volumes; Step (5) adopts half preparative scale chromatography column purification, and applied sample amount is 150 μ l, and adopting the moving phase starting point concentration is 1.5% acetonitrile, and the flow velocity of moving phase is 2ml/min, and the detection wavelength is 220nm, and other step, technology are with embodiment 1.
Claims (7)
1. extract the method for radish sulphur glycoside compound from the broccoli Cauliflower, it is characterized in that carrying out as follows:
(1) selection of broccoli Cauliflower vegetable material and pre-treatment: select to contain radish sulphur glycosides account for total fats glucosinolate more than 90% and the seed, bud seedling, cauline leaf and the floral organ that account for the broccoli Cauliflower of total glucosinolate more than 80% be material, under avoiding as the crumbliness physical injury prerequisite of lancinating, extrude, wear and tear, after water cleans, drains, standby;
(2) extract reagent, slightly mention concentrated: with water, methyl alcohol or 80% methanol aqueous solution serves as to extract reagent; To extract reagent and the pretreated vegetable material of broccoli Cauliflower, by volume the ratio of weight 1-3L: 1-2kg and 85-100 ℃, the same process that stir to soak 20-30min repeat to extract 2-3 time, respectively after filtration, ultrafiltration removes merging filtrate behind the insolubles, after rotary evaporation or lyophilize, be dissolved in after accounting in the water that extracts reagent 1-2% volume radish sulphur glycosides crude extract;
(3) organic solvent extraction of crude extract and impurity elimination: alternately repeatedly be extracted to colourless with organic solvent radish sulphur glycosides crude extract; Collect water, after filtration, ultrafiltration or the centrifugal insolubles of removing, collect supernatant liquor;
(4) preliminary purification of extraction liquid: the supernatant liquor of extraction liquid is carried out preliminary purification in following program: after 1) ratio of extracting 100mg radish sulphur glycosides according to the 1-2g resin takes by weighing DEAE-Sephadex A-25 anionite-exchange resin, soak 10-24h with the 80-100% methanol solution, pack the post height of bed in the glass column of 5cm; 2) doubly wash post for the 0-10% methanol aqueous solution with 3-5 to the concentration of anionite-exchange resin volume; 3) will remove sample on the aqueous portion behind the insolubles; 4), be that leacheate carries out drip washing with 20% Virahol or 20% acetonitrile solution that wait column volume again with behind the water wash of two bed volumes; 5) carry out wash-out with anionresin liquid 2-3 times of column volume and that match with above-mentioned leacheate, and collect with the isopyknic dehydrated alcohol of anionresin liquid in; 6) elutriant is water-soluble after the rotary evaporation drying, obtains containing the mixed solution of multiple glucosinolate component after 0.22 μ m water filter membrane ultrafiltration;
(5) chromatographic column purifying radish sulphur glycosides: with preparation type or half preparative scale chromatography post is purification column, with preliminary purification mixed solution gradation sample introduction, with water that contains 0.1% trifluoroacetic acid and the acetonitrile that contains 0.1% trifluoroacetic acid is moving phase, acetonitrile concentration continues 5min by initial 0-5%, 5-20% continues 20min, and 20% keeps 5min is provided with gradient, at flow velocity 2-3ml/min, detect under the wavelength 210-230nm, collect the highest chromatographic peak and go out effusive moving phase during the peak; Moving phase after above-mentioned gradation sample introduction, the collection is merged, after lyophilize, rotary evaporation or nitrogen dry up drying, promptly obtain purity and be higher than 95% radish sulphur glycoside product; The content of radish sulphur glycosides is with reference to Sun et al., Food Chemistry, and (2011) method is carried out quantitatively.
2. by the described method of claim 1, it is characterized in that the described organic solvent that is used to extract is methylene dichloride, ethyl acetate, acetone and normal hexane.
3. by the described method of claim 1, it is characterized in that described anionresin liquid be contain 0.5M Repone K, sodium-chlor, vitriolate of tartar or sodium sulfate 5-20% aqueous isopropanol or contain the acetonitrile solution of the 5-20% of 0.5M Repone K, sodium-chlor, vitriolate of tartar or sodium sulfate, wherein, step (4)-4) be that leacheate then uses the former as anionresin liquid with 20% Virahol, if step (4)-4) be that leacheate then uses the latter as anionresin liquid with 20% acetonitrile.
4. by the described method of claim 1, it is characterized in that described 20% aqueous isopropanol, 20% acetonitrile solution, the 5-20% aqueous isopropanol, the 5-20% acetonitrile solution is the aqueous solution of respective concentration Virahol or acetonitrile.
5. by the described method of claim 1, it is characterized in that described chromatographic column for preparation or half preparation type, is that filler, particle diameter are that 5-10 μ m, column length are that 25-30cm, diameter are the reverse phase silica gel filled column of 10-20mm with ODS BP.
6. by the described method of claim 1, it is characterized in that described rotary evaporation or nitrogen dry up drying method, its temperature is no more than 50 ℃.
7. by the described method of claim 1, it is characterized in that the content of described radish sulphur glycosides adopts Sun et al. in 2011, the method for Food Chemistry is carried out quantitative assay.
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| CN103416733A (en) * | 2012-05-24 | 2013-12-04 | 宁波海逸生物科技有限公司 | Glucosinolate extract from fresh broccoli and preparation method thereof |
| CN103709211A (en) * | 2013-12-13 | 2014-04-09 | 大兴安岭嘉迪欧营养原料有限公司 | Method for preparing glucoraphanin from broccoli |
| CN104880526A (en) * | 2015-05-25 | 2015-09-02 | 宁波立华植物提取技术有限公司 | Method for determining content of benzyl glucosinolate in lepidium meyenii walp |
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| CN107501354A (en) * | 2017-08-18 | 2017-12-22 | 赣州华汉生物科技有限公司 | A kind of method for extracting high-purity glucorphanin |
| CN107998167A (en) * | 2017-12-14 | 2018-05-08 | 武汉北度生物科技有限公司 | A kind of method that oxidation-resistant active ingredient is extracted from cauliflower |
| CN108690103A (en) * | 2018-04-03 | 2018-10-23 | 山东中医药大学 | A method of preparing high-purity radish glucoside extract by raw material of radish seed |
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