CN102295667B - 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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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, relating in particular to one, to utilize broccoli and Cauliflower be material, extracts the method for high purity cancer-resisting chemical protective agent precursor glucorphanin.
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 grams/day), can reduce DNA damage, the activity of raising stages 2 enzyme, thus prevent and reduce the generation of cancer.Research shows, in broccoli, a kind of sulforaphen being produced by glucosinolate-glucorphanin (Glucoraphanin) degraded, as repressor, regulates carcinogenic metabolism specifically by double mechanism.Sulforaphen is the strongest phytochemicals of antitumour activity of finding in vegetables, and the kinds cancers such as lung cancer, intestinal cancer, prostate cancer and mammary cancer are had to very strong inhibition effect.Nearest research shows, sulforaphen and some other glucosinolate meta-bolites may and promote cancer cell-apoptosis hinder the growth of tumour by blocking-up cell cycle.But sulforaphen unstable chemcial property, easily decompose, and its precursor glucorphanin is relatively stable, and can is generated and have different sulfo-nitrile hydrochlorate sulforaphen anticancer and that anti-cancer is active by enteric microorganism degraded in human body, therefore glucorphanin just can be used as the additive of food, healthcare products or medicine.In addition, the degraded product sulforaphen of glucorphanin is as inhibited in some Fusarium oxysporums, Pseudomonas etc. to the pathogenic bacteria of various plants, its degraded product nitriles substance has also participated in various insects as the defence of aphid, small cabbage moth etc., and therefore glucorphanin is a kind of phytochemicals with various biological function.At present, the standard substance glucosinolate that can sell in the world only has two kinds of 2-propenyl mustard oil glycosides and phenmethyl glucosinolates, and produced by Sigma-Aldrich and AppliChem two companies respectively, and expensive, there is no company's sale and there is the anticancer and active precursor substance glucorphanin of anti-cancer by force.Therefore, utilize own abundant broccoli or cauliflower germplasm resource, develop autonomous glucorphanin, a meaningful job beyond doubt.
Summary of the invention
The present invention seeks to, less for current glucosinolate product category, and rely on external import, expensive defect, provide a kind of taking broccoli Cauliflower plant as material, the precursor that therefrom extracts sulforaphen also can participate in the method for the glucorphanin compound of the multiple defense response of cress.
The object of the invention is achieved by the following technical programs.
The method of extracting glucorphanin compound from broccoli Cauliflower, the method is carried out as follows:
(1) selection of broccoli Cauliflower vegetable material and pre-treatment: selecting the seed, bud seedling, cauline leaf and the floral organ that account for total fats glucosinolate more than 90% and account for the more than 80% broccoli Cauliflower of total glucosinolate containing glucorphanin is material, avoiding as lancinated, extrude, wear and tear under crumbliness physical injury prerequisite, after water cleans, drains, for subsequent use;
(2) extract reagent, slightly mention concentrated: taking water, methyl alcohol or 80% methanol aqueous solution as extracting 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 DEG C, stir and soak the same process of 20-30min and repeat to extract 2-3 time, respectively after filtration, ultrafiltration removes merging filtrate after insolubles, is dissolved in after accounting in the water that extracts reagent 1-2% volume obtaining glucorphanin crude extract after rotary evaporation or lyophilize;
(3) organic solvent extraction of crude extract and impurity elimination: glucorphanin crude extract is extracted to colourless with organic solvent alternately repeatedly; 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 to preliminary purification in following program: 1) take after DEAE-Sephadex A-25 anionite-exchange resin according to the ratio of 1-2g resin extraction 100mg glucorphanin, with 80-100% methanol solution immersion 10-24h, pack the post height of bed in the glass column of 5cm; 2) be doubly that 0-10% methanol aqueous solution is washed post to the concentration of anionite-exchange resin volume with 3-5; 3) by the aqueous portion loading of removing after insolubles; 4) with after the water wash of two bed volumes, then with etc. 20% Virahol or 20% acetonitrile solution of column volume be that leacheate carries out drip washing; 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 rotary evaporation is dry, obtains the mixed solution that contains multiple glucosinolate component after 0.22 μ m water filter membrane ultrafiltration;
(5) chromatographic column purifying glucorphanin: taking preparative or semi-preparative chromatographic column as purification column, by preliminary purification mixed solution gradation sample introduction, taking containing the water of 0.1% trifluoroacetic acid and containing the acetonitrile of 0.1% trifluoroacetic acid as moving phase, acetonitrile concentration continues 5min by initial 0-5%, 5-20% continues 20min, and 20% maintains 5min arranges gradient, at flow velocity 2-3ml/min, detect under wavelength 210-230nm, collect the highest chromatographic peak and go out the moving phase flowing out during peak; Moving phase by above-mentioned gradation sample introduction, after collecting merges, through lyophilize, rotary evaporation or nitrogen dry up dry after, obtain purity higher than 95% glucorphanin product; The content of glucorphanin is with reference to Sun et al., Food Chemistry, and (2011) method is carried out quantitatively.
Described is methylene dichloride, ethyl acetate, acetone and normal hexane for the organic solvent extracting.
Described anionresin liquid is the acetonitrile solution of the aqueous isopropanol of the 5-20% that contains 0.5M Repone K, sodium-chlor, potassium sulfate or sodium sulfate or the 5-20% that contains 0.5M Repone K, sodium-chlor, potassium sulfate or sodium sulfate, wherein, step (4)-4) use the former as anionresin liquid using 20% Virahol as leacheate, if step (4)-4) use the latter as anionresin liquid using 20% acetonitrile as leacheate.
20% described aqueous isopropanol, 20% acetonitrile solution, 5-20% aqueous isopropanol, 5-20% acetonitrile solution is the aqueous solution of respective concentration Virahol or acetonitrile.
Described chromatographic column is preparation or semi-preparative, taking ODS BP as filler, particle diameter as 5-10 μ m, column length as 25-30cm, diameter is the reverse phase silica gel filled column of 10-20mm.
Described rotary evaporation or nitrogen dry up drying method, and its temperature is no more than 50 DEG C.
The content of described glucorphanin 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 to be rich in the broccoli of glucorphanin and Cauliflower plant as material, water or methanol solution extract glucosinolate complexes, carry out preliminary purification by extraction and anionite-exchange resin, after concentrated, recycle semi-preparative or preparative scale chromatography post can DNA purity up to more than 95% glucorphanin compound, it is simple that the method has method, with low cost, it is high that glucorphanin extracts yield, the single purity of component is high, can be used as standard substance and is applied in Biochemistry Experiment and correlative study.
The glucorphanin that the present invention extracts is to have cancer-resisting chemical protective agent---the precursor of sulforaphen; Also can participate in multiple cress and resist the important substance of various plants pathogenic bacteria and insect.
Brief description of the drawings
Fig. 1. from the extraction of broccoli Cauliflower, purifying glucorphanin process flow sheet.
Fig. 2. select for the preparation of glucosinolate high-efficient liquid phase chromatogram in the broccoli kind of glucorphanin.
Standard substance are 2-oil of mirbane-β-D-galactoside (120uM), detection method is with reference to Sun et al., Food Chemistry, (2011), in material, glucorphanin accounts for 98% of total glucosinolate, accounts for 98.4% of fats glucosinolate.
Fig. 3. the color atlas that glucorphanin enriching and purifying is forward and backward.
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 glucorphanin result;
Figure B is desulfurization glucorphanin detection by quantitative result after semipreparative column purifying.
Embodiment
By following examples, the present invention is described in further detail, but content of the present invention is not limited to this.
Instrument and material involved in the present invention are as follows:
Liquid phase systems is Waters company of the U.S., comprises pump Waters1525, automatic sample handling system 717plus Autosampler, detector Waters2487;
Institute's water is ultrapure water, and acetonitrile and trifluoroacetic acid are that chromatographically pure is produced by TEDIA company of the U.S.;
Chromatographic column used is that Dalian produces according to Lyntech Corporation (US) 10177 South 77th East Avenue Tulsa, Oklahoma 74133 U.S., wherein semi-preparative chromatographic column filler is SinoChrom ODS-BP C18, particle diameter 5 μ m, column length 25cm, diameter 10mm, preparative scale chromatography column packing is SinoChrom ODS-BP C18, particle diameter 10 μ m, column length 30cm, diameter 20mm;
Glucorphanin is the one belonging in fats glucosinolate; In different broccolis and Cauliflower material there is poor very large difference in the component of contained glucosinolate and content; Therefore in the time selecting vegetable material, first needing to select containing glucorphanin content and the high plant of ratio is material, to improve output and the purity of purifying products; Can be with reference to Sun et al. in the time of concrete selection, Food Chemistry, component and the content of (2011) method to glucosinolate in vegetable material detects; The selected material of the present invention is that glucorphanin accounts for total fats glucosinolate more than 90%, and accounts for more than 80% broccoli or the Cauliflower material of total glucosinolate.
Embodiment 1:(utilizes broccoli seeds to extract the method for glucorphanin)
(1) selection of broccoli seeds material and pre-treatment: with reference to Sun et al., Food Chemistry, (2011) method, selecting to account for total fats glucosinolate more than 90% and account for more than 80% broccoli seeds of total glucosinolate containing glucorphanin is material (seeing Fig. 2), under avoiding as physical injury prerequisites such as lancinating, extrude, wear and tear and be broken, water cleans and removes after surface contaminant and impurity for 2 times, drains, for subsequent use;
(2) extract reagent, slightly mention concentrated: the extraction reagent using methyl alcohol as glucorphanin, by extract reagent and broccoli seeds by volume the ratio of weight 2.5L: 1kg and 85 DEG C, stir the same process that soaks 20min and repeat to extract after 2 times, respectively through Whatman2 filter paper filtering, then after insolubles is removed in 0.45 μ m and 0.22 μ m filter membrane ultrafiltration merging filtrate; Be no more than the rotary evaporation of 50 DEG C through temperature and be dissolved in after dry and account in the pure water that extracts reagent volume 2%, obtain the crude extract of glucorphanin;
(3) organic solvent extraction of crude extract and impurity elimination: by thick glucorphanin water lift for solution organic solvent dichloromethane, ethyl acetate, acetone and normal hexane be repeatedly extracted to alternately colourless after, collect water, centrifugal 10min under 4 DEG C, 13200g, removes insolubles, collects supernatant liquor;
(4) preliminary purification of extraction liquid: the supernatant liquor of extraction liquid is carried out to preliminary purification in following program: 1) take after DEAE-Sephadex A-25 anionite-exchange resin according to the ratio of 1-2g resin extraction 100mg glucorphanin, with 80% methanol solution immersion 10h, pack the post height of bed in the glass column of 5cm; 2) wash post with 3 times of pure water to anionite-exchange resin volume; 3) supernatant liquor of extraction liquid is added to loading in purification column; 4) with after the pure water drip washing of two bed volumes, then with etc. 20% Virahol of column volume be that leacheate carries out drip washing; 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 no more than the rotary evaporation of 50 DEG C through temperature and is dissolved in pure water after dry, obtains the mixing solutions that contains multiple glucosinolate component after 0.22 μ m water filter membrane ultrafiltration;
(5) chromatographic column purifying glucorphanin: taking semi-preparative chromatographic column as purification column, preliminary purification mixed solution is pressed to 100 μ l gradation sample introductions, taking containing the water of 0.1% trifluoroacetic acid and containing the acetonitrile of 0.1% trifluoroacetic acid as moving phase, acetonitrile concentration continues 5min by 0%, 0-20% continues 20min, and 20% maintains 5min arranges gradient, at flow velocity 2ml/min, detect under wavelength 229nm, collect the highest chromatographic peak and go out the moving phase flowing out during peak; The moving phase of above-mentioned gradation sample introduction, collection is merged, after lyophilize, obtain purity higher than 95% glucorphanin product; The glucorphanin of enrichment is with reference to Sun et al., Food Chemistry, and (2011) method is carried out quantitative and qualitative analysis detection (seeing Fig. 3); The glucorphanin 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 glucorphanin in plant.
Embodiment 2:(utilizes broccoli bud seedling to extract the method for glucorphanin)
In this example, step (1) material is the broccoli bud seedling of germination 3-4 days, the extraction solvent adopting in step (2) is water, extract 100 DEG C of temperature, extraction time 25min, the ratio of extracting reagent and material is 1L: 1kg, repeats to extract 3 times, and the sample after lyophilize is dissolved in the water of 1.5% extraction reagent volume; In step (3), impurity-removing method is the centrifugal 10min of 13200g at 4 DEG C; In step (4), soak solution used is 85% methyl alcohol, 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 waiting 20% acetonitrile solution of column volume; Elutriant is 5% acetonitrile solution containing 0.5M sodium-chlor of 3 times of column volumes; Step 5 adopts preparative scale chromatography post, and applied sample amount is 200 μ l, and adopting moving phase starting point concentration is 2.5% acetonitrile, and the flow velocity of moving phase is 3ml/min, and detection wavelength is 210nm, and other step, technique are with embodiment 1.
Embodiment 3:(utilizes Cauliflower bud seedling to extract the method for glucorphanin)
In this example, step (1) material therefor is broccoli cauline leaf, the extraction solvent adopting in step (2) is 80% methyl alcohol, extract 90 DEG C of temperature, extraction time 25min, the ratio of extracting reagent and material is 1L: 2kg, repeats to extract 3 times, and the sample after lyophilize is dissolved in the water of 1.0% extraction reagent volume; In step (3), impurity-removing method is with 0.45 μ m and 0.22 μ m ultrafiltration membrance filter; In step (4), soak solution used is 90% methyl alcohol, 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 waiting 20% Virahol of column volume; Elutriant is 20% aqueous isopropanol containing 0.5M Repone K of 2 times of column volumes; In step (5), adopt preparative scale chromatography post, applied sample amount is 200 μ l, and adopting moving phase starting point concentration is 2% acetonitrile, and the flow velocity of moving phase is 3ml/min, and detection wavelength is 230nm, and other step, technique are with embodiment 1.
Embodiment 4:(utilizes cauliflower seed to extract the method for glucorphanin)
In this example, step (1) material therefor is cauliflower seed, the extraction solvent adopting in step (2) is water, extract 100 DEG C of temperature, extraction time 30min, the ratio of extracting reagent and material is 3L: 1kg, repeats to extract 2 times, and the sample after lyophilize is dissolved in the water of 2.0% extraction reagent volume; In step (3), impurity-removing method is 0.45 μ m and 0.22 μ m ultrafiltration membrance filter; In step (4), soak solution used is 100% methyl alcohol, 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 waiting 20% acetonitrile solution of column volume; Elutriant is 10% acetonitrile solution containing 0.5M sodium sulfate of 3 times of column volumes; Step (5) adopts preparative scale chromatography column purification, and applied sample amount is 150 μ l, and adopting moving phase starting point concentration is 2% acetonitrile, and the flow velocity of moving phase is 3ml/min, and detection wavelength is 227nm, and other step, technique are with embodiment 1.
Embodiment 5:(utilizes Cauliflower bud seedling to extract the method for glucorphanin)
In this example, step (1) material therefor is the Cauliflower bud seedling of germination 3-4 days, the extraction solvent adopting in step (2) is methyl alcohol, extract 85 DEG C of temperature, extraction time 20min, the ratio of extracting reagent and material is 3L: 2kg, repeats to extract 3 times, and the sample after lyophilize is dissolved in the water of 1.5% extraction reagent volume; In step (3), impurity-removing method is the centrifugal 10min of 13200g at 4 DEG C; In step (4), soak solution used is 80% methyl alcohol, 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 waiting 20% aqueous isopropanol of column volume; Elutriant is 10% aqueous isopropanol containing 0.5M potassium sulfate of 2 times of column volumes; Step (5) adopts semi-preparative chromatographic column purifying, and applied sample amount is 100 μ l, and adopting moving phase starting point concentration is 1% acetonitrile, and the flow velocity of moving phase is 2ml/min, and detection wavelength is 226nm, and other step, technique are with embodiment 1.
Embodiment 6:(utilizes broccoli curd to extract the method for glucorphanin)
In this example, step (1) material therefor is ripe broccoli curd, the extraction solvent adopting in step (2) is 80% methyl alcohol, extract 90 DEG C of temperature, extraction time 20min, the ratio of extracting reagent and material is 2L: 1kg, repeats to extract 2 times, and the sample after lyophilize is dissolved in the water of 1.0% extraction reagent volume; In step (3), impurity-removing method is the centrifugal 10min of 13200g at 4 DEG C; In step (4), soak solution used is 90% methyl alcohol, 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 waiting 20% acetonitrile solution of column volume; Elutriant is 15% acetonitrile solution containing 0.5M sodium sulfate of 3 times of column volumes; Step (5) adopts semi-preparative chromatographic column purifying, and applied sample amount is 150 μ l, and adopting moving phase starting point concentration is 1.5% acetonitrile, and the flow velocity of moving phase is 2ml/min, and detection wavelength is 220nm, and other step, technique are with embodiment 1.
Claims (7)
1. the method for extracting glucorphanin compound from broccoli Cauliflower, is characterized in that carrying out as follows:
(1) selection of broccoli Cauliflower vegetable material and pre-treatment: selecting the seed, bud seedling, cauline leaf and the floral organ that account for total fats glucosinolate more than 90% and account for the more than 80% broccoli Cauliflower of total glucosinolate containing glucorphanin is material, avoiding lancinating, under the prerequisite of extruding, after water cleans, drains, for subsequent use;
(2) extract reagent, slightly mention concentrated: taking water, methyl alcohol or 80% methanol aqueous solution as extracting 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 DEG C, stir and soak the same process of 20-30min and repeat to extract 2-3 time, respectively after filtration, ultrafiltration removes merging filtrate after insolubles, is dissolved in after accounting in the water that extracts reagent 1-2% volume obtaining glucorphanin crude extract after rotary evaporation or lyophilize;
(3) organic solvent extraction of crude extract and impurity elimination: glucorphanin crude extract is extracted to colourless with organic solvent alternately repeatedly; 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 to preliminary purification in following program: 1) take after DEAE-Sephadex A-25 anionite-exchange resin according to the ratio of 1-2g resin extraction 100mg glucorphanin, with 80-100% methanol solution immersion 10-24h, pack the post height of bed in the glass column of 5cm; 2) be doubly that 0-10% methanol aqueous solution is washed post to the concentration of anionite-exchange resin volume with 3-5; 3) by the aqueous portion loading of removing after insolubles; 4) with after the water wash of two bed volumes, then with etc. 20% Virahol or 20% acetonitrile solution of column volume be that leacheate carries out drip washing; 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 rotary evaporation is dry, obtains the mixed solution that contains multiple glucosinolate component after 0.22 μ m water filter membrane ultrafiltration;
(5) chromatographic column purifying glucorphanin: taking preparative or semi-preparative chromatographic column as purification column, by preliminary purification mixed solution gradation sample introduction, taking containing the water of 0.1% trifluoroacetic acid and containing the acetonitrile of 0.1% trifluoroacetic acid as moving phase, acetonitrile concentration continues 5min by initial 0-5%, 5-20% continues 20min, and 20% maintains 5min arranges gradient, at flow velocity 2-3ml/min, detect under wavelength 210-230nm, collect the highest chromatographic peak and go out the moving phase flowing out during peak; Moving phase by above-mentioned gradation sample introduction, after collecting merges, through lyophilize, rotary evaporation or nitrogen dry up dry after, obtain purity higher than 95% glucorphanin product.
2. by method claimed in claim 1, it is characterized in that described is methylene dichloride, ethyl acetate, acetone and normal hexane for the organic solvent extracting.
3. by method claimed in claim 1, it is characterized in that described anionresin liquid is the acetonitrile solution of the aqueous isopropanol of the 5-20% that contains 0.5M Repone K, sodium-chlor, potassium sulfate or sodium sulfate or the 5-20% that contains 0.5M Repone K, sodium-chlor, potassium sulfate or sodium sulfate, wherein, step (4)-4) use the former as anionresin liquid using 20% Virahol as leacheate, if step (4)-4) use the latter as anionresin liquid using 20% acetonitrile as leacheate.
4. by method claimed in claim 1, it is characterized in that 20% described aqueous isopropanol, 20% acetonitrile solution is the aqueous solution of respective concentration Virahol or acetonitrile.
5. by method claimed in claim 3, it is characterized in that described 5-20% aqueous isopropanol, 5-20% acetonitrile solution is the aqueous solution of respective concentration Virahol or acetonitrile.
6. by method claimed in claim 1, it is characterized in that described chromatographic column is for preparation or semi-preparative, taking ODS BP as filler, particle diameter as 5-10 μ m, column length as 25-30cm, diameter is the reverse phase silica gel filled column of 10-20mm.
7. by method claimed in claim 1, it is characterized in that described rotary evaporation or nitrogen dry up drying method, its temperature is no more than 50 DEG C.
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| CN107501354B (en) * | 2017-08-18 | 2019-11-26 | 赣州华汉生物科技有限公司 | A method of extracting high-purity glucorphanin |
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| CN108690103B (en) * | 2018-04-03 | 2022-02-25 | 山东中医药大学 | Method for preparing high-purity glucoraphanin extract by taking radish seeds as raw materials |
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| Bo Sun等.Variation of glucosinolates in three edible parts of Chinese kale (Brassica alboglabra Bailey) varieties.《Food Chemistry》.2011,第124卷(第3期),941-947. |
| Extraction and Purification of Glucoraphanin by Preparative High-Performance Liquid Chromatography (HPLC);Iris Lee等;《Journal of chemical education》;20110331;第88卷;第833页左栏最后一段 * |
| Iris Lee等.Extraction and Purification of Glucoraphanin by Preparative High-Performance Liquid Chromatography (HPLC).《Journal of chemical education》.2011,第88卷832-834. |
| Variation of glucosinolates in three edible parts of Chinese kale (Brassica alboglabra Bailey) varieties;Bo Sun等;《Food Chemistry》;20110201;第124卷(第3期);全文 * |
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| Publication number | Publication date |
|---|---|
| CN102295667A (en) | 2011-12-28 |
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