CN102640596A - Method for improving content of bioactive substance quercetin of buckwheat sprouts through magnetic field induction treatment - Google Patents
Method for improving content of bioactive substance quercetin of buckwheat sprouts through magnetic field induction treatment Download PDFInfo
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- CN102640596A CN102640596A CN2012101568639A CN201210156863A CN102640596A CN 102640596 A CN102640596 A CN 102640596A CN 2012101568639 A CN2012101568639 A CN 2012101568639A CN 201210156863 A CN201210156863 A CN 201210156863A CN 102640596 A CN102640596 A CN 102640596A
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Abstract
The invention discloses a method for improving the content of the bioactive substance quercetin of buckwheat sprouts through magnetic field induction treatment. The method comprises the following steps of: firstly performing induction treatment on buckwheat grains by means of a magnetic field; and then sprouting the induced buckwheat grains for 7 days, thereby obtaining the buckwheat sprouts containing high content of the bioactive substance quercetin. The method for improving the content of the bioactive substance quercetin of the buckwheat sprouts through the magnetic field induction treatment provided by the invention is capable of obviously improving the activity of phenylalnine ammonialyase (PAL) and chalcone isomerase (CHI) in the sprouting process of the buckwheat while improving the sprouting percentage and germinative force of the buckwheat grains, thereby obviously improving the content of the bioactive substance quercetin in the buckwheat.
Description
Technical field
The objective of the invention is in order to provide a kind of induced by magnetic field that utilizes to handle the method that improves buckwheat blastogenesis active substances-quercetin content.
Know-why of the present invention
Through utilizing the magnetic field physical means, in buckwheat seed germination process, excite PAL (PAL), enzyme, namely chalcone isomerase endogenous enzyme activities such as (CHI), thereby improve buckwheat blastogenesis active substances-quercetin content.
Background technology
Buckwheat is the dicotyledon of the distinctive medicine-food two-purpose of China, contains bioactivators such as protein, vitamin B group, Quercetin, mineral nutrient and plant cellulose etc. in the buckwheat.Bioactivators such as Quercetin are a kind of very strong antioxidant, can effectively remove the oxygen radical in the human body, can help human body to improve blood circulation, improve immunity, and prevent diabetes, high fat of blood, hypertension are had important function.At present; The technology that improves bioactive substance contents such as Quercetin mainly contains sprouts Treatment Technology to the buckwheat seed; Do not have the complex art that before sprouting, combines, thereby limited the content increase of bioactivators such as Quercetin through the induced by magnetic field process etc. techniques.
It is for further improving a kind of physical means that seed sprouting power adopts in the seed process that magnetization is handled.Both at home and abroad the seeds of paddy rice, wheat, soybean, corn, rape and many vegetable crops was carried out magnetization research; Confirm that all magnetic field helps promoting seed sprouting; Improve germination vigor and germination rate, but still do not have correlative study for the content influence of bioactivators such as Quercetin.
Summary of the invention
The purpose of this invention is to provide a kind of when improving buckwheat seed germinating capacity and vigor; The ratio vigor of important two kinds of enzymes (PAL, enzyme, namely chalcone isomerase) in the bioactivator forming processes such as effective raising Quercetin; And then improve the method for bioactive substance contents such as Quercetin in the buckwheat bud, to satisfy the requirement of buckwheat functional food processing.
Technical scheme of the present invention
A kind of induced by magnetic field that utilizes is handled the method that improves buckwheat blastogenesis active substances-quercetin content; Promptly at first adopt magnetic field that the buckwheat seed is induced processing; To induce the buckwheat seed after the processing to sprout again 7 days, promptly obtain containing the buckwheat bud of higher bioactivator-quercetin content;
The magnetic field intensity in above-mentioned magnetic field is preferably 0.2T～0.6T, induces the processing time to be preferably 30min;
Above-mentioned buckwheat seed sprouting condition is preferably controlled 28 ℃ of temperature, humidity 85%.
Above-mentioned a kind of induced by magnetic field that utilizes is handled the method that improves buckwheat blastogenesis active substances-quercetin content, specifically comprises the steps:
(1), the preliminary treatment of buckwheat seed
Adopt clear water to eluriate the buckwheat seed,, drain away the water with after flushing with clean water 2-3 time then again with 35 ℃ of water seed soaking 24h to remove not plump seed and sick worm seed;
(2), step (1) being placed intensity through preliminary treatment and the buckwheat seed that drains away the water is that the magnetic field of 0.2T～0.6T is induced and handled 30min; Under 28 ℃ of temperature, humidity 85% condition, sprouted 7 days then, promptly obtain containing the buckwheat bud of higher bioactivator-quercetin content.
Beneficial effect of the present invention
A kind of induced by magnetic field that utilizes of the present invention is handled the method that improves buckwheat blastogenesis active substances-quercetin content; When improving buckwheat seed germination rate and germination vigor; Can significantly improve the activity of PAL in the buckwheat germination process (PAL), enzyme, namely chalcone isomerase (CHI); And then significantly improve the content of bioactivator-Quercetin in the buckwheat bud; Especially be that the buckwheat bud that 0.4T handled is not compared through the buckwheat bud of magnetic field treated through magnetic field intensity, its bioactivator-quercetin content has improved nearly 23%.
Embodiment
Through embodiment the present invention is set forth below, but do not limit the present invention.
The method of testing of the various indexs in the embodiment of the invention in the buckwheat bud germination process is following:
Method 1:The mensuration of germination vigor and germination rate (" international seed sprouting rules " (ISTA))
In the formula: M
1Normal germination grain number in the-germination vigor fate;
M
2-all normal grain the number that germinates;
The seed number is planted experimentally in M-confession
Method 2:The extraction of PAL, CHI crude enzyme liquid (opening must string, Hu Xiaomei, Zhu Yanming etc. and wild soybean PAL (PAL) is Changing Pattern [J] under different inductive conditions. soybean science, 2011,30 (4): 703-705,709.)
Grind in the sample thief 1 g liquid nitrogen; Add the boric acid borate buffer solution (pH 8.8, include the EDTA of 1 mmol/L, the mercaptoethanol of 5 mmol/L, 1% PVP, 1% glycerine) of 0.1 mol/L of 5 mL, grind to form homogenate; 12 000 r/min, 4 ℃ of centrifugal 20 min; Getting supernatant (being crude enzyme liquid) uses perhaps-80 ℃ preservation subsequent use immediately.
Method 3:Mensuration (the Lister C E that PAL is active; Lancaster J E. Developmental changes in enzymes of flavonoid biosynthesis in the skins of red and green apple cultivars [J]. Science of Food Agriculture; 1996,71:313-320.)
According to enzyme extract 0.5mL, L-phenyl alanine (0.01molL
-1) 1.5mL, boric acid borate buffer solution 3mL mix, and as testing sample (cumulative volume is 5mL), replaces crude enzyme liquid and L-phenyl alanine (0.01molL with distilled water
-1), the boric acid borate buffer solution mixes, as blank; The solution that mixes is measured OD with ultraviolet specrophotometer
290Value, and record; Testing sample is used 0.2mL, 20% HCl cessation reaction at 30 ℃ of insulation 1h; Use the OD behind ultraviolet specrophotometer (the Shanghai essence instrument Science and Technology Ltd. 754 type ultraviolet-uisible spectrophotometers) assaying reaction once more
290Value, and record.Light absorption value change 0.01 is 1 enzyme unit alive at the 290nm place according to the every mL enzyme of every h liquid, the activity of conversion PAL.
PAL activity=
Method 4:The mensuration that CHI is active: (Lister C E; Lancaster J E. Developmental changes in enzymes of flavonoid biosynthesis in the skins of red and green apple cultivars [J]. Science of Food Agriculture; 1996,71:313-320.)
Get the 0.75mL crude enzyme liquid and add 2mL reactant liquor (including final concentration is the 7.5mg/mL bovine serum albumin(BSA) for pH7.4, the Tris-HCL solution of 0.05M); Add 50 μ L then; The hydroxyl chalcone solution of 1mg/mL, at 34 ℃ of isothermal reaction 30min, the thick zyme extract that other gets with amount adds the 2mL reactant liquor; Boiling water bath 10min measures OD as contrast with ultraviolet specrophotometer (Shanghai essence instrument Science and Technology Ltd. 754 type ultraviolet-uisible spectrophotometers)
381Place's light absorption value, light absorption value variation 0.001 is 1 enzyme unit alive at the 381nm place according to the every mL enzyme of every min liquid, converses the activity of CHI.
CHI activity=
Method 5:The mensuration of protein content: Coomassie brilliant blue (G-250) method (opening must string, Hu Xiaomei, Zhu Yanming etc. and wild soybean PAL (PAL) is Changing Pattern [J] under different inductive conditions. soybean science, 2011,30 (4): 703-705,709.)
The drafting of calibration curve: with the standard protein liquid mixing in the Coomassie brilliant blue G-250 (5mL) for preparing and each pipe, room temperature is respectively managed OD with visible spectrophotometer (Shanghai essence instrument Science and Technology Ltd. 754 type ultraviolet-uisible spectrophotometers) mensuration after leaving standstill 5min
595, and record.With the light absorption value is ordinate, and protein concentration is the abscissa mapping, obtains calibration curve (y=6.2071x+0.0388, R
2=0.997).Determining the protein quantity in the crude enzyme liquid: with the 0.2mL crude enzyme liquid, 0.8mL distilled water and the abundant mixing of 5.0mL Coomassie brilliant blue G-250 solution, repetition was measured its OD 3 times after room temperature left standstill 5min
595Value obtains mean value and checks in protein content through calibration curve.
Method 6:The calculating of specific activity of enzyme (opening must string, Hu Xiaomei, Zhu Yanming etc. and wild soybean PAL (PAL) is Changing Pattern [J] under different inductive conditions. soybean science, 2011,30 (4): 703-705,709.)
Protein content in the vigor/crude enzyme liquid of the ratio vigor=enzyme of enzyme
Method 7:Quercetin content is measured (high performance liquid chromatography)
Instrument and equipment: Agilent 1100 high performance liquid chromatographs
Analytical column: anti-phase Zorbax Eclipse SB-C18 post (250 * 4.6 mm, 5 μ m).
Flowing phase: adopting acetonitrile and 0.05% phosphoric acid is eluant, eluent, and the wash-out ratio is seen table 1:
Table 1 elution requirement
| Time (min) | Acetonitrile (%) | 0.05% phosphoric acid (%) |
| 0.00 | 10 | 90 |
| 80.0 | 11 | 89 |
| 95.0 | 25 | 75 |
| 105.0 | 40 | 60 |
| 115.0 | 80 | 20 |
Detect wavelength: Quercetin is selected 370nm
Column temperature: room temperature
Flow velocity: 1.0mL/min
Sample size: 15 μ L
Analysis time: 120 min
Embodiment 1
A kind of magnetic field intensity of utilizing is handled the method that improves buckwheat blastogenesis active substances-quercetin content for the induced by magnetic field of 0.2T, comprises the steps:
(1), the preliminary treatment of buckwheat seed
Adopt clear water to eluriate the buckwheat seed,, drain away the water with after flushing with clean water 2-3 time then again with 35 ℃ of water seed soaking 24h to remove not plump seed and sick worm seed;
(2), with step (1) through preliminary treatment and the buckwheat seed that drains away the water place magnetic field intensity be 0.2T magnetic field induce handle 30min after; Put into the incubator of 28 ℃ of temperature, humidity 85% and sprouted 7 days, promptly get the buckwheat bud that contains higher bioactivator-quercetin content.
Employing method 1-7 measures the germination rate and the germination vigor of the buckwheat bud that contains higher bioactivator-quercetin content of above-mentioned gained, and PAL (PAL) than vigor, enzyme, namely chalcone isomerase (CHI) than vigor and quercetin content.The result sees table 2.
Table 2 .0.2T magnetic field intensity is handled the buckwheat germination and is measured the result
| Handle intensity | Germination rate (%) | Germination vigor (%) | PAL is than vigor (U/mg) | CIH is than vigor (U/mg) | Quercetin (%) |
| 0T | 53%±0.016 | 57%±0.012 | 212.41±1.593 | 79.81±1.247 | 3.53±0.104 |
| 0.2T | 65%±0.034** | 68%±0.029** | 217.27±2.011** | 86.20±1.841** | 3.82±0.086* |
Significant difference: * represent P 0.05, i.e. significant difference; * representes P, and < 0.01, promptly difference is extremely remarkable.
Can find out that from table 2 along with the increase of magnetic field intensity, when germination rate, germination vigor extremely significantly increased, PAL (PAL) and enzyme, namely chalcone isomerase (CHI) also increased than vigor thereupon, quercetin content also presents remarkable increase.
Embodiment 2
A kind of magnetic field intensity of utilizing is handled the method that improves buckwheat blastogenesis active substances-quercetin content for the induced by magnetic field of 0.4T, comprises the steps:
(1), the preliminary treatment of buckwheat seed
Adopt clear water to eluriate the buckwheat seed,, drain away the water with after flushing with clean water 2-3 time then again with 35 ℃ of water seed soaking 24h to remove not plump seed and sick worm seed;
(2), with step (1) through preliminary treatment and the buckwheat seed that drains away the water place intensity be 0.4T magnetic field induce handle 30min after; Put into the incubator of 28 ℃ of temperature, humidity 85% and sprouted 7 days, promptly get the buckwheat bud that contains higher bioactivator-quercetin content.
Employing method 1-7 measures the germination rate and the germination vigor of the buckwheat bud that contains higher bioactivator-quercetin content of above-mentioned gained, and PAL (PAL) than vigor, enzyme, namely chalcone isomerase (CHI) than vigor and quercetin content.The result sees table 3.
Table 3 .0.4T magnetic field intensity is handled the buckwheat germination and is measured the result
| Handle intensity | Germination rate (%) | Germination vigor (%) | PAL is than vigor (U/mg) | CIH is than vigor (U/mg) | Quercetin (%) |
| 0T | 53%±0.016 | 57%±0.012 | 212.41±1.593 | 79.81±1.247 | 3.53±0.104 |
| 0.4T | 70%±0.009** | 73%±0.008** | 223.58±1.894** | 88.74±0.769** | 4.34±0.165** |
Significant difference: * represent P 0.05, i.e. significant difference; * representes P, and < 0.01, promptly difference is extremely remarkable.
Can find out that from table 3 along with the increase of magnetic field intensity, when germination rate, germination vigor extremely significantly increased, PAL (PAL) and enzyme, namely chalcone isomerase (CHI) also extremely significantly increased than vigor, quercetin content also appears extremely significantly to be increased
Embodiment 3
A kind of magnetic field intensity of utilizing is handled the method that improves buckwheat blastogenesis active substances-quercetin content for the induced by magnetic field of 0.2T, comprises the steps:
(1), the preliminary treatment of buckwheat seed
Adopt clear water to eluriate the buckwheat seed,, drain away the water with after flushing with clean water 2-3 time then again with 35 ℃ of water seed soaking 24h to remove not plump seed and sick worm seed;
(2), with step (1) through preliminary treatment and the buckwheat seed that drains away the water place magnetic field intensity be 0.6T magnetic field induce handle 30min after; Put into the incubator of 28 ℃ of temperature, humidity 85% and sprouted 7 days, promptly get the buckwheat bud that contains higher bioactivator-quercetin content.
Employing method 1-7 measures the germination rate and the germination vigor of the buckwheat bud that contains higher bioactivator-quercetin content of above-mentioned gained, and PAL (PAL) than vigor, enzyme, namely chalcone isomerase (CHI) than vigor and quercetin content.The result sees table 4.
Table 4 .0.6T magnetic field intensity is handled the buckwheat germination and is measured the result
| Handle intensity | Germination rate (%) | Germination vigor (%) | PAL is than vigor (U/mg) | CIH is than vigor (U/mg) | Quercetin (%) |
| 0T | 53%±0.016 | 57%±0.012 | 212.41±1.593 | 79.81±1.247 | 3.53±0.104 |
| 0.6T | 69%±0.029** | 73%±0.024** | 220.58±2.018** | 82.97±1.09* | 4.11±0.073** |
Significant difference: * represent P 0.05, i.e. significant difference; * representes P, and < 0.01, promptly difference is extremely remarkable.
Can find out that from table 4 along with the increase of magnetic field intensity, when germination rate, germination vigor extremely significantly increased, PAL (PAL) and enzyme, namely chalcone isomerase (CHI) also significantly increased than vigor, quercetin content also presents remarkable increase.
In sum; Can find out; The increase of magnetic field intensity when the present invention handles along with induced by magnetic field; When improving buckwheat seed germinating capacity and germination vigor, can effectively improve the ratio vigor of two kinds of enzymes (PAL, enzyme, namely chalcone isomerase) important in the Quercetin forming process, and then improve bioactivator-quercetin content in the buckwheat bud; Especially be that the buckwheat bud that 0.4T handled is not compared through the buckwheat bud of magnetic field treated through magnetic field intensity, its bioactivator-quercetin content has improved nearly 23%.
Foregoing is merely the basic explanation of the present invention under conceiving, and according to any equivalent transformation that technical scheme of the present invention is done, all should belong to protection scope of the present invention.
Claims (5)
1. one kind is utilized induced by magnetic field to handle the method that improves buckwheat blastogenesis active substances-quercetin content; It is characterized in that at first adopting magnetic field that the buckwheat seed is induced processing; Buckwheat seed after will handling was again sprouted 7 days, promptly got the buckwheat bud that contains higher bioactivator-quercetin content.
2. a kind of induced by magnetic field that utilizes as claimed in claim 1 is handled the method that improves buckwheat blastogenesis active substances-quercetin content; The magnetic field intensity that it is characterized in that described magnetic field is 0.2T～0.6T, and described buckwheat seed sprouting condition is 28 ℃ of temperature, humidity 85%.
3. a kind of induced by magnetic field that utilizes as claimed in claim 2 is handled the method that improves buckwheat blastogenesis active substances-quercetin content, it is characterized in that the described induced by magnetic field processing time is 30min.
4. a kind of induced by magnetic field that utilizes as claimed in claim 3 is handled the method that improves buckwheat blastogenesis active substances-quercetin content, and the magnetic field intensity that it is characterized in that described magnetic field is 0.4T.
5. handle the method that improves buckwheat blastogenesis active substances-quercetin content like claim 1,2,3 or 4 described a kind of induced by magnetic field that utilize, it is characterized in that specifically comprising the steps:
(1), the preliminary treatment of buckwheat seed
Adopt clear water to eluriate the buckwheat seed,, drain away the water with after flushing with clean water 2-3 time then again with 35 ℃ of water seed soaking 24h to remove not plump seed and sick worm seed;
(2), place magnetic field intensity to induce processing through preliminary treatment and the buckwheat seed that drains away the water step (1), sprout then, promptly get the buckwheat bud that contains higher bioactivator-quercetin content.
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Cited By (2)
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| CN111557424A (en) * | 2020-05-25 | 2020-08-21 | 江南大学 | Method for improving antioxidant capacity of germinated quinoa by magnetic field treatment |
| CN113853871A (en) * | 2021-09-10 | 2021-12-31 | 吉林农业大学 | Application of low-frequency static magnetic field in improving grain seed bud polyphenol content and activity |
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| CN111557424A (en) * | 2020-05-25 | 2020-08-21 | 江南大学 | Method for improving antioxidant capacity of germinated quinoa by magnetic field treatment |
| CN111557424B (en) * | 2020-05-25 | 2022-12-27 | 江南大学 | Method for improving antioxidant capacity of germinated quinoa by magnetic field treatment |
| CN113853871A (en) * | 2021-09-10 | 2021-12-31 | 吉林农业大学 | Application of low-frequency static magnetic field in improving grain seed bud polyphenol content and activity |
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