CN102586352A - Method for enriching and extracting sulforaphane from brassica vegetable seeds - Google Patents
Method for enriching and extracting sulforaphane from brassica vegetable seeds Download PDFInfo
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- CN102586352A CN102586352A CN2012100115519A CN201210011551A CN102586352A CN 102586352 A CN102586352 A CN 102586352A CN 2012100115519 A CN2012100115519 A CN 2012100115519A CN 201210011551 A CN201210011551 A CN 201210011551A CN 102586352 A CN102586352 A CN 102586352A
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Abstract
The invention relates to a method for enriching and extracting sulforaphane from brassica vegetable seeds and belongs to the technical field of food biological technology. The method is characterized by comprising the following steps of: taking the sulforaphane brassica vegetable seeds with germinating ability as raw materials; crushing the raw materials and adding a Tris-HCl buffer solution according to the ratio of the materials to liquid being (w/v) 1:(3-5); adjusting the pH value to 6.0-8.0 to form an incubation buffer; adding vitamin C and Zn<2+>, activating endogenous myrosinase in the vegetable seeds and hydrolyzing glucosinolate to form the sulforaphane with an anti-cancer effect; and extracting the sulforaphane from the incubation buffer with ethyl acetate and carrying out filtering, vacuum concentration, ultrasonic extraction and vacuum condensation to obtain a sulforaphane product. The method disclosed by the invention has the characteristics of wide application range of the raw material, low production cost, high enrichment quantity of the sulforaphane and the like. The method is suitable for health-care food sulforaphane products.
Description
One, technical field
The present invention relates to a kind of rape and belong in the vegetable seed sulforaphen enrichment and process for extracting in sulforaphen enrichment and process for extracting, the especially vegetable seed such as Caulis et Folium Brassicae capitatae, wild cabbage, belong to technical field of food biotechnology.
Two, background technology
Epidemiological study shows; Edible Cruciferae rape belongs to vegetables; But like the generation of multiple cancers such as prevention of pancreatic cancer such as Caulis et Folium Brassicae capitatae, kale, brussels sprouts, Cauliflower, Chinese cabbage, lung cancer, the rectum cancer, mammary cancer and prostate cancer, this is mainly relevant with the hydrolysate-sulforaphen of the rape genus intravital glucosinolates of vegetables (sulphur glycosides).Sulforaphen is the isothiocyanate that is formed through the myrosin catalytic hydrolysis by 4-methylsulfinyl butyl sulphur glycosides (Glucoraphanin, GR belong to the analiphatic sulphur glycosides).The plant of being rich in the sulphur glycosides digests the sulforaphen that the back produces biologically active in human body, play the anticancer health-care effect.Along with people's deepens constantly cancer understanding, and increasing medical circle and the scientific worker of nutrition educational circles recognize, in the prevention and treatment problem of cancer, should classify prevention work as first place, and the adjustment dietary structure is the anti-cancer measure of accomplishing the most easily.Have a lot of natural cancer-resisting materials in the world, wherein majority is the vegetable food of integration of drinking and medicinal herbs, and it is one of focuses of studying over nearly 10 years that rape belongs to vegetables.
Occurring in nature; Sulforaphen (1-isothiocyano-4-methylsulfinyl butane; L-isothiocyanato-4-methyl sulfiny butane is that a kind of that plant produces when receiving extraneous injury can avoid the further sulfocompound of injury by protective plant sulfbraphane).Ubiquity a kind of " sulphur glycosides-myrosin " system in the brassica plant tissues such as Caulis et Folium Brassicae capitatae, cabbage, kale, Chinese cabbage, Cauliflower, leaf mustard, kohlrabi, brussels sprouts, turnip, horseradish; In the long-term evolution process, plant materials has produced a series of Mechanism and FunctionsDNAs that conform, protect self.
Though glucosinolate has certain research to the conversion of sulforaphen; But at present less with the extraction and purification Study on Technology report of control condition, sulforaphen for the glucosinolate enzymolysis process, and utilize plant endogenous myrosin with self glucosinolate be hydrolyzed into sulforaphen also in addition the technology of enrichment lack research especially.
In recent years; Chinese scholar has been carried out some researchs about sulforaphen and precursor substance thereof; What at present research was more is horseradish and sulforaphen in the mustard and the precursor substance of using as seasonings thereof, lacks research for enrichment, the raising sulforaphen extraction yield of sulforaphen.The research to sulforaphen abroad rests on laboratory stage at present, lacks practical beneficiation technologies.Glucosinolate is complicated to the mechanism that sulforaphen transforms, and how to regulate and control conversion condition and still lacks further investigation at present.The enrichment of sulforaphen can promote the product development relevant with preventing cancer with the realization of extraction and separation technology, has vast market prospect.
At present, reporting more both at home and abroad is about from natural matter, extracting the method for sulforaphen, but these methods mainly are through adding the external source myrosin glucosinolate to be changed into sulforaphen.For example; Patent (publication number CN1149986A; Open day on May 21st, 1997) " brassica vegetable supplement and preparation method thereof ", patent (publication number CN101544995A are disclosed; Open day on September 30th, 2009) " a kind of process for extracting of sulforaphen " and patent (publication number CN 102102113A are disclosed; Open day on June 22nd, 2011) disclose in the extraction process of " a kind of method of from the Cauliflower blade, extracting sulforaphen " must the expensive external source myrosin preparation of interpolation, cause the sulforaphen production cost too high, thereby this method is limited in production application.
In the disclosed in the past technology; Also there are many drawbacks in the process aspect that extracts for the organic solvent extraction sulforaphen; For example; Patent (publication number CN 101143842A; Open day on March 19th, 2008) disclose " a kind of method of extracting sulforaphen ", patent (publication number CN 101229211A, open day on July 30th, 2008) disclose " a kind of broccoli seed extractive and preparation method thereof " and patent (publication number CN 101514174A, open day on August 26th, 2009) and discloses the method for the multi-functional sulforaphane of extraction " a kind of from the Caulis et Folium Brassicae capitatae bud seedling vegetable " middle to extract the organic solvent that is adopted be toxic reagents such as acetone, methylene dichloride and sherwood oil; Be difficult to avoid residuing in the finished product, thus the use of limits product.
Three, summary of the invention
Technical problem:
The object of the present invention is to provide a kind of rape to belong to the enrichment and the extractive technique of sulforaphen in the vegetable seed, solve in the past in the sulforaphen process for extracting the high and technical barrier such as the product yield is low, toxicity is big of production cost.
Technical scheme:
Technical scheme of the present invention is summarized as follows: belonging to vegetable seed with rape is raw material, pulverizes back adding Tris-HCl damping fluid and also regulates pH, processes Incubating Solution; Add vitamins C and Zn
2+, activate endogenous myrosin in the vegetable seed, the glucosinolate of hydrolysis self; With ethyl acetate extraction, filtration, vacuum concentration,, make sulforaphen product with certain purity again through ultrasonic lixiviate, vacuum concentration.
Above-mentioned rape belongs to the enrichment and the extractive technique of sulforaphen in the vegetable seed, comprising:
The rapes such as broccoli seed or cabbage, kale, brussels sprouts, Chinese cabbage, Cauliflower, leaf mustard, kohlrabi, turnip and horseradish that (1) will have a germinating power belong to a certain seeds of vegetables pulverize after; Press solid-liquid ratio (w/v) 1: 3～5 and add 0.2mol/L Tris-HCl damping fluid; Regulate pH to 6.0～8.0, process Incubating Solution;
(2) in Incubating Solution, add 0.2～0.8mmol/L vitamins C and 0.4～0.6mmol/L Zn
2+, hatch 2～6h in 30～50 ℃;
(3) hatch end after, add 1～2mmol/L sodium-chlor, and with equal volume of ethyl acetate 1～3 time, combining extraction liquid filters also to collect and filtrates;
(4) by known concentration technology, vacuum concentration to the butter shape liquid (or evaporate to dryness) under 25～45 ℃ of conditions of will filtrating.
(5) in liquid concentrator, add ethyl acetate solution, in 20～40 ℃ of ultrasonic lixiviate 5～10min, ultrasonic frequency is 90khz, and power is 150～190W, makes the sulforaphen crude extract;
(6) the sulforaphen crude extract is crossed 0.45 μ m filter membrane after, by known concentration technology,, make through 20～40 ℃ of vacuum concentration.
Beneficial effect:
Compared with prior art, sulforaphen enrichment and process for extracting have the following advantages in a kind of rape genus vegetable seed of the present invention:
(1) raw material range that is suitable for of the present invention is wide, utilizes raw material self myrosin hydrolysis glucosinolate, has reduced production cost, has simplified production technique.
(2) the present invention is through adding proper vitamin C and Zn
2+, activate myrosin, accelerate the glucosinolate hydrolysis, improved the sulforaphen yield.
(3) the present invention adopts nontoxic ethyl acetate extraction sulforaphen, and adds proper amount of sodium chloride and help the layering extraction, improves percentage extraction, and obtains to be applicable to the sulforaphen product of protective foods level.
Four, specific embodiments
Embodiment 1:
After will having the broccoli seed pulverizing of germinating power, press solid-liquid ratio (w/v) and add 0.2mol/L Tris-HCl damping fluid at 1: 5, regulate pH to 6.0, process Incubating Solution; Add 0.2mmol/L vitamins C and 0.6mmol/L Zn
2+After, hatch 2h in 50 ℃; In Incubating Solution, add 2.0mmol/L sodium-chlor, with equal-volume ethyl acetate extraction 1 time, combining extraction liquid.Extraction liquid is vacuum concentration to butter shape liquid (or evaporate to dryness) under 45 ℃ of conditions after filtering.Add ethyl acetate solution in the liquid behind vacuum concentration,, make the sulforaphen crude extract, cross 0.45 μ m filter membrane,, make through 30 ℃ of vacuum concentration in 20 ℃ of ultrasonic lixiviate 10min (ultrasonic frequency is 90kHz, and power is 140W).
Embodiment 2:
After will having the cabbage seed pulverizing of germinating power, press solid-liquid ratio (w/v) and add 0.2mol/L Tri-HCls damping fluid at 1: 4, regulate pH to 6.5, process Incubating Solution; Add 0.4mmol/L vitamins C and 1.5mmol/L Zn
2+After, hatch 4h in 40 ℃, in Incubating Solution, add 2mmol/L sodium-chlor, with equal-volume ethyl acetate extraction 2 times, combining extraction liquid.Extraction liquid is vacuum concentration to butter shape liquid (or evaporate to dryness) under 40 ℃ of conditions after filtering.Add ethyl acetate solution in the liquid behind vacuum concentration,, make the sulforaphen crude extract, cross 0.45 μ m filter membrane,, make through 40 ℃ of vacuum concentration in 40 ℃ of ultrasonic lixiviate 5min (ultrasonic frequency is 90khz, and power is 190W).
Embodiment 3:
After will having the cauliflower seed pulverizing of germinating power, press solid-liquid ratio (w/v) and add 0.2mol/L Tris-HCl damping fluid at 1: 3, regulate pH to 7.0, process Incubating Solution; Add 0.6mmol/L vitamins C and 0.4mmol/L Zn
2+6h is hatched in 45 ℃ in the back, in Incubating Solution, adds 1.5mmol/L sodium-chlor, with equal-volume ethyl acetate extraction 3 times, and combining extraction liquid.Extraction liquid is vacuum concentration to butter shape liquid (or evaporate to dryness) under 30 ℃ of conditions after filtering.Add ethyl acetate solution in the liquid behind vacuum concentration,, make the sulforaphen crude extract, cross 0.45 μ m filter membrane,, make through 35 ℃ of vacuum concentration in 30 ℃ of ultrasonic lixiviate 8min (ultrasonic frequency is 90kHz, and power is 170W).
Embodiment 4:
After the horseradish seed that will have a germinating power is fully pulverized, press solid-liquid ratio (w/v) and add 0.2mol/L Tris-HCl damping fluid at 1: 4, regulate pH to 8.0, process Incubating Solution; Add 0.8mmol/L vitamins C and 0.3mmol/L Zn
2+6h is hatched in 30 ℃ in the back, in Incubating Solution, adds 1.0mmol/L sodium-chlor, with equal-volume ethyl acetate extraction 3 times, and combining extraction liquid.Extraction liquid is vacuum concentration to butter shape liquid (or evaporate to dryness) under 25 ℃ of conditions after filtering.Behind vacuum concentration, add ethyl acetate solution in the liquid,, make the sulforaphen crude extract, cross 0.45 μ m filter membrane,, make through 40 ℃ of vacuum concentration in 35 ℃ of ultrasonic lixiviate 7min (ultrasonic frequency is 90kHz, and power is 180W).
More than specified embodiment of the present invention, but this instance of just lifting for the ease of understanding should not be considered to be limitation of the scope of the invention.Equally; Any person of ordinary skill in the field all can be according to the description of technical scheme of the present invention and preferred embodiment thereof; Make various possible being equal to and change or replacement, but all these changes or replacement all should belong to the protection domain of claim of the present invention.
Claims (4)
1. a rape belongs to the enrichment and the process for extracting of sulforaphen in the vegetable seed, it is characterized in that: belonging to vegetable seed with the rape with germinating power is raw material, pulverizes the back and adds the Tris-HCl damping fluid and regulate pH by solid-liquid ratio, processes Incubating Solution; Add vitamins C and Zn
2+, activate endogenous myrosin in the vegetable seed, the glucosinolate of hydrolysis self; With the sulforaphen that forms in the ethyl acetate extraction Incubating Solution, through filtration, vacuum concentration, ultrasonic lixiviate, vacuum concentration makes sulforaphen again.
2. according to claim 1, a kind of rape belongs to the enrichment and the process for extracting of sulforaphen in the vegetable seed, may further comprise the steps:
The rape that (1) will have a germinating power belongs to vegetable seed pulverize after, add zero(ppm) water by solid-liquid ratio (w/v) 1: 3～5 and make Incubating Solution, with 0.2mol/L Tris-HCl damping fluid adjusting pH to 6.0～8.0;
(2) in Incubating Solution, add vitamins C and Zn
2+, hatch 2～6h in 30～50 ℃;
(3) hatch end after, add 1～2mmol/L sodium-chlor, and with equal volume of ethyl acetate 1～3 time, combining extraction liquid filters also to collect and filtrates;
(4) by known concentration technology, vacuum concentration to the butter shape liquid (or evaporate to dryness) under 25～45 ℃ of conditions of will filtrating;
(5) in liquid concentrator, add ethyl acetate solution, in 20～40 ℃ of ultrasonic lixiviate 5～10min, ultrasonic frequency is 90khz, and power is 150～190W, makes the sulforaphen crude extract;
(6) the sulforaphen crude extract is crossed 0.45 μ m filter membrane after, by known concentration technology,, make through 20～40 ℃ of vacuum concentration.
3. belong to sulforaphen enriching method in the vegetable seed like the said a kind of rape of claim 1-2, it is characterized in that the vitamins C and the Zn that add in the Incubating Solution
2+Amount is respectively 0.2～0.8mmol/L and 0.3～0.6mmol/L.
4. belong to sulforaphen enrichment and process for extracting in the vegetable seed like the said a kind of rape of claim 1-3; It is characterized in that it can be broccoli seed that rape belongs to vegetable seed, or seeds such as cabbage, kale, Chinese cabbage, Cauliflower, leaf mustard, kohlrabi, brussels sprouts, turnip, horseradish.
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Cited By (9)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102898341A (en) * | 2012-10-26 | 2013-01-30 | 贵州大学 | Extracting and purifying method of high-purity sulforaphane |
| CN103436565A (en) * | 2013-09-10 | 2013-12-11 | 甘肃农业大学 | Method for extracting sulforaphane from broccoli |
| CN104086469A (en) * | 2014-05-22 | 2014-10-08 | 苏州泽达兴邦医药科技有限公司 | Method for extracting and purifying sulforaphane from broccoli seeds |
| CN104131047A (en) * | 2014-01-14 | 2014-11-05 | 南京农业大学 | High-efficiency enrichment technology for isothiocyanate in brassica bud seedlings |
| CN104673768A (en) * | 2015-03-20 | 2015-06-03 | 北京农学院 | Process for extracting myrosinase from horseradish |
| CN105851942A (en) * | 2016-03-30 | 2016-08-17 | 扬州大学 | Flavoring type high-isothiocyanate-content brassica juncea sprouts and production technology thereof |
| CN106256250A (en) * | 2015-06-15 | 2016-12-28 | 浙江海正药业股份有限公司 | A kind of Caulis et Folium Brassicae capitatae hatches the preparation method of powder |
| CN107630051A (en) * | 2017-10-12 | 2018-01-26 | 临沂大学 | It is a kind of that the method for preparing reduced form sulforaphen extract with extracting coupling is digested using two-phase system |
| CN116135038A (en) * | 2021-11-17 | 2023-05-19 | 南京农业大学 | Method for hydrolyzing rapeseed meal thioglucoside by using bud-source myrosinase |
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Cited By (11)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102898341A (en) * | 2012-10-26 | 2013-01-30 | 贵州大学 | Extracting and purifying method of high-purity sulforaphane |
| CN103436565A (en) * | 2013-09-10 | 2013-12-11 | 甘肃农业大学 | Method for extracting sulforaphane from broccoli |
| CN103436565B (en) * | 2013-09-10 | 2015-08-19 | 甘肃农业大学 | A kind of method extracting sulforaphen from Caulis et Folium Brassicae capitatae |
| CN104131047A (en) * | 2014-01-14 | 2014-11-05 | 南京农业大学 | High-efficiency enrichment technology for isothiocyanate in brassica bud seedlings |
| CN104086469A (en) * | 2014-05-22 | 2014-10-08 | 苏州泽达兴邦医药科技有限公司 | Method for extracting and purifying sulforaphane from broccoli seeds |
| CN104673768A (en) * | 2015-03-20 | 2015-06-03 | 北京农学院 | Process for extracting myrosinase from horseradish |
| CN106256250A (en) * | 2015-06-15 | 2016-12-28 | 浙江海正药业股份有限公司 | A kind of Caulis et Folium Brassicae capitatae hatches the preparation method of powder |
| CN105851942A (en) * | 2016-03-30 | 2016-08-17 | 扬州大学 | Flavoring type high-isothiocyanate-content brassica juncea sprouts and production technology thereof |
| CN107630051A (en) * | 2017-10-12 | 2018-01-26 | 临沂大学 | It is a kind of that the method for preparing reduced form sulforaphen extract with extracting coupling is digested using two-phase system |
| CN107630051B (en) * | 2017-10-12 | 2021-03-26 | 临沂大学 | Method for preparing reduced sulforaphane extract by utilizing two-phase system enzymolysis and extraction coupling |
| CN116135038A (en) * | 2021-11-17 | 2023-05-19 | 南京农业大学 | Method for hydrolyzing rapeseed meal thioglucoside by using bud-source myrosinase |
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| CN102586352B (en) | 2014-02-26 |
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