CN102640596B - 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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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 object of the invention is in order to provide one to utilize induced by magnetic field to process the method that improves buckwheat blastogenesis active substances-quercetin content.
Know-why of the present invention
By utilizing magnetic field physical means, in buckwheat seed germination process, excite the endogenous enzyme activity such as phenylalnine ammonialyase (PAL), enzyme, namely chalcone isomerase (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 bioactivator, mineral nutrient and the plant celluloses etc. such as protein, B family vitamin, Quercetin in buckwheat.The bioactivators such as Quercetin are a kind of very strong antioxidant, can effectively remove the oxygen radical in human body, can help human body to improve blood circulation, improve immunity, and prevention diabetes, high fat of blood, hypertension are had to important function.At present, the technology that improves the bioactive substance contents such as Quercetin mainly contains sprouts to buckwheat seed the technology of processing, there is no the complex art combining by technology such as induced by magnetic field processing before sprouting, thereby limited the content increase of the bioactivators such as Quercetin.
It is for further improving a kind of physical means that seed sprouting power adopts in seed process that magnetization is processed.The seed of paddy rice, wheat, soybean, corn, rape and many vegetable crops was carried out to magnetization research both at home and abroad, all confirm that magnetic field is conducive to promote seed sprouting, improve germination vigor and germination rate, but there is no correlative study for the content influence of the bioactivators such as Quercetin.
Summary of the invention
The object of this invention is to provide a kind of in improving buckwheat seed germinating capacity and vigor, effectively improve the ratio vigor of two kinds of enzymes (phenylalnine ammonialyase, enzyme, namely chalcone isomerase) important in the bioactivator forming processes such as Quercetin, and then improve the method for the bioactive substance contents such as Quercetin in buckwheat bud, to meet the requirement of buckwheat functional food processing.
Technical scheme of the present invention
One utilizes induced by magnetic field to process the method that improves buckwheat blastogenesis active substances-quercetin content, first adopt magnetic field to induce processing to buckwheat seed, to induce again buckwheat seed after treatment to sprout 7 days, obtain containing the buckwheat bud compared with high bioactivity material-quercetin content;
The magnetic field intensity in above-mentioned magnetic field is preferably 0.2T～0.6T, and the induction processing time is preferably 30min;
Above-mentioned buckwheat seed sprouting condition is preferably controlled 28 DEG C of temperature, humidity 85%.
Above-mentioned one utilizes induced by magnetic field to process the method that improves buckwheat blastogenesis active substances-quercetin content, specifically comprises the steps:
(1), the pretreatment of buckwheat seed
Adopt clear water to eluriate buckwheat seed to remove not plump seed and sick worm seed, then with 35 DEG C of water seed soaking 24h, drain away the water after then rinsing 2-3 time with clear water;
(2), step (1) being placed in to intensity through pretreatment the buckwheat seed that drains away the water is that 30min is processed in the magnetic field induction of 0.2T～0.6T, then under 28 DEG C of temperature, humidity 85% condition, sprout 7 days, obtain containing the buckwheat bud compared with high bioactivity material-quercetin content.
Beneficial effect of the present invention
One of the present invention utilizes induced by magnetic field to process the method that improves buckwheat blastogenesis active substances-quercetin content, in improving buckwheat seed germination rate and germination vigor, can significantly improve the activity of phenylalnine ammonialyase in buckwheat germination process (PAL), enzyme, namely chalcone isomerase (CHI), and then significantly improve the content of bioactivator-Quercetin in buckwheat bud, especially be that the buckwheat bud that 0.4T processed is not compared through the buckwheat bud of magnetic field processing through magnetic field intensity, its bioactivator-quercetin content has improved nearly 23%.
Embodiment
Below by embodiment, the present invention is set forth, but do not limit the present invention.
The method of testing of the various indexs in the embodiment of the present invention in buckwheat bud germination process is as follows:
method 1:the mensuration of germination vigor and germination rate (" international seed sprouting code " (ISTA))
In formula: M
1normal germination grain number in-germination vigor number of days;
M
2-all normal grain numbers that germinates;
Seed number is planted experimentally in M-confession
method 2:the extraction of PAL, CHI crude enzyme liquid (must string, Hu little Mei, Zhu Yanming etc. wild soybean phenylalnine ammonialyase (PAL) is Changing Pattern [J] under different inductive conditions. Soybean Science, 2011,30 (4): 703-705,709.)
In sample thief 1 g liquid nitrogen, grind; The boric acid borate buffer solution (pH 8.8 includes the EDTA of 1 mmol/L, the mercaptoethanol of 5 mmol/L, 1% PVP, 1% glycerine) that adds 0.1 mol/L of 5 mL, grinds to form homogenate; 12 000 r/min, 4 DEG C of centrifugal 20 min; Get that supernatant (being crude enzyme liquid) uses immediately or-80 DEG C save backup.
method 3:mensuration (the Lister C E of PAL activity, 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-Phe (0.01molL
-1) 1.5mL, boric acid borate buffer solution 3mL mixing, as testing sample (cumulative volume is 5mL), with distilled water replacement crude enzyme liquid and L-Phe (0.01molL
-1), boric acid borate buffer solution mixes, as blank; The solution mixing is measured to OD with ultraviolet specrophotometer
290value, and record; Testing sample is incubated to 1h 0.2mL, 20% HCl cessation reaction at 30 DEG C; Again use the OD after ultraviolet specrophotometer (Shanghai essence instrument Science and Technology Ltd. 754 type ultraviolet-uisible spectrophotometers) assaying reaction
290value, and record.Changing 0.01 according to the every mL enzyme of every h liquid at 290nm place light absorption value is 1 Ge Meihuo unit, the activity of conversion PAL.
PAL activity=
method 4:the mensuration of CHI activity: (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 0.75mL crude enzyme liquid and add 2mL reactant liquor (pH7.4, the Tris-HCL solution of 0.05M, including final concentration is 7.5mg/mL bovine serum albumin(BSA)), then add 50 μ L, the hydroxylated chalcone solution of 1mg/mL, at 34 DEG C of isothermal reaction 30min, separately gets with the thick zyme extract of amount and adds 2mL reactant liquor, boiling water bath 10min in contrast, uses ultraviolet specrophotometer (Shanghai essence instrument Science and Technology Ltd. 754 type ultraviolet-uisible spectrophotometers) to measure OD
381place's light absorption value, changing 0.001 according to the every mL enzyme of every min liquid at 381nm place light absorption value is 1 Ge Meihuo unit, converses the activity of CHI.
CHI activity=
method 5:the mensuration of protein content: Coomassie brilliant blue (G-250) method (must string, Hu little Mei, Zhu Yanming etc. wild soybean phenylalnine ammonialyase (PAL) is Changing Pattern [J] under different inductive conditions. Soybean Science, 2011,30 (4): 703-705,709.)
The drafting of calibration curve: by the Coomassie brilliant blue G-250(5mL preparing) mix with the standard protein liquid in each pipe, after the standing 5min of room temperature, measure each pipe OD with visible spectrophotometer (Shanghai essence instrument Science and Technology Ltd. 754 type ultraviolet-uisible spectrophotometers)
595, and record.Taking light absorption value as ordinate, protein concentration is abscissa mapping, obtains calibration curve (y=6.2071x+0.0388, R
2=0.997).Determining the protein quantity in crude enzyme liquid: by 0.2mL crude enzyme liquid, 0.8mL distilled water and 5.0mL Coomassie brilliant blue G-250 solution fully mix, repeats 3 times after the standing 5min of room temperature and measures its OD
595value, obtains mean value and checks in protein content by calibration curve.
method 6:the calculating of specific activity of enzyme (must string, Hu little Mei, Zhu Yanming etc. wild soybean phenylalnine ammonialyase (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 are m).
Mobile phase: adopting acetonitrile and 0.05% phosphoric acid is eluant, eluent, wash-out ratio is in 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
Utilize the induced by magnetic field that magnetic field intensity is 0.2T to process a method that improves buckwheat blastogenesis active substances-quercetin content, comprise the steps:
(1), the pretreatment of buckwheat seed
Adopt clear water to eluriate buckwheat seed to remove not plump seed and sick worm seed, then with 35 DEG C of water seed soaking 24h, drain away the water after then rinsing 2-3 time with clear water;
(2), step (1) being placed in to magnetic field intensity through pretreatment the buckwheat seed that drains away the water is that the induction of 0.2T magnetic field is processed after 30min, put into the incubator of 28 DEG C of temperature, humidity 85% and sprout 7 days, must contain the buckwheat bud compared with high bioactivity material-quercetin content.
Employing method 1-7 measures containing compared with germination rate and the germination vigor of the buckwheat bud of high bioactivity material-quercetin content of above-mentioned gained, and phenylalnine ammonialyase (PAL) than vigor, enzyme, namely chalcone isomerase (CHI) than vigor and quercetin content.The results are shown in Table 2.
table 2 .0.2T magnetic field intensity is processed buckwheat germination test result
| Process 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: * represents P<0.05, i.e. significant difference; * represents P<0.01, and difference is extremely remarkable.
As can be seen from Table 2, along with the increase of magnetic field intensity, when germination rate, germination vigor extremely significantly increase, phenylalnine ammonialyase (PAL) and enzyme, namely chalcone isomerase (CHI) also increase than vigor thereupon, and quercetin content also presents remarkable increase.
embodiment 2
Utilize the induced by magnetic field that magnetic field intensity is 0.4T to process a method that improves buckwheat blastogenesis active substances-quercetin content, comprise the steps:
(1), the pretreatment of buckwheat seed
Adopt clear water to eluriate buckwheat seed to remove not plump seed and sick worm seed, then with 35 DEG C of water seed soaking 24h, drain away the water after then rinsing 2-3 time with clear water;
(2), step (1) being placed in to intensity through pretreatment the buckwheat seed that drains away the water is that the induction of 0.4T magnetic field is processed after 30min, put into the incubator of 28 DEG C of temperature, humidity 85% and sprout 7 days, must contain the buckwheat bud compared with high bioactivity material-quercetin content.
Employing method 1-7 measures containing compared with germination rate and the germination vigor of the buckwheat bud of high bioactivity material-quercetin content of above-mentioned gained, and phenylalnine ammonialyase (PAL) than vigor, enzyme, namely chalcone isomerase (CHI) than vigor and quercetin content.The results are shown in Table 3.
table 3 .0.4T magnetic field intensity is processed buckwheat germination test result
| Process 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: * represents P<0.05, i.e. significant difference; * represents P<0.01, and difference is extremely remarkable.
As can be seen from Table 3, along with the increase of magnetic field intensity, when germination rate, germination vigor extremely significantly increase, phenylalnine ammonialyase (PAL) and enzyme, namely chalcone isomerase (CHI) also extremely significantly increase than vigor, and quercetin content also presents extremely significantly to be increased
embodiment 3
Utilize the induced by magnetic field that magnetic field intensity is 0.2T to process a method that improves buckwheat blastogenesis active substances-quercetin content, comprise the steps:
(1), the pretreatment of buckwheat seed
Adopt clear water to eluriate buckwheat seed to remove not plump seed and sick worm seed, then with 35 DEG C of water seed soaking 24h, drain away the water after then rinsing 2-3 time with clear water;
(2), step (1) being placed in to magnetic field intensity through pretreatment the buckwheat seed that drains away the water is that the induction of 0.6T magnetic field is processed after 30min, put into the incubator of 28 DEG C of temperature, humidity 85% and sprout 7 days, must contain the buckwheat bud compared with high bioactivity material-quercetin content.
Employing method 1-7 measures containing compared with germination rate and the germination vigor of the buckwheat bud of high bioactivity material-quercetin content of above-mentioned gained, and phenylalnine ammonialyase (PAL) than vigor, enzyme, namely chalcone isomerase (CHI) than vigor and quercetin content.The results are shown in Table 4.
table 4 .0.6T magnetic field intensity is processed buckwheat germination test result
| Process 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: * represents P<0.05, i.e. significant difference; * represents P<0.01, and difference is extremely remarkable.
As can be seen from Table 4, along with the increase of magnetic field intensity, when germination rate, germination vigor extremely significantly increase, phenylalnine ammonialyase (PAL) and enzyme, namely chalcone isomerase (CHI) also significantly increase than vigor, and quercetin content also presents remarkable increase.
in sum, can find out, the increase of magnetic field intensity when the present invention processes along with induced by magnetic field, in improving buckwheat seed germinating capacity and germination vigor, can effectively improve the ratio vigor of two kinds of enzymes (phenylalnine ammonialyase, enzyme, namely chalcone isomerase) important in Quercetin forming process, and then bioactivator-quercetin content in raising buckwheat bud, especially be that the buckwheat bud that 0.4T processed is not compared through the buckwheat bud of magnetic field processing through magnetic field intensity, its bioactivator-quercetin content has improved nearly 23%.
Foregoing is only 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 (2)
1. one kind is utilized induced by magnetic field to process the method that improves buckwheat blastogenesis active substances-quercetin content, first adopt magnetic field to induce processing to buckwheat seed, again buckwheat seed after treatment is sprouted 7 days, must contain the buckwheat bud compared with high bioactivity material-quercetin content, the magnetic field intensity that it is characterized in that described magnetic field is 0.6T, the described induced by magnetic field processing time is 30min, and described buckwheat seed sprouting condition is 28 DEG C of temperature, humidity 85%.
2. one as claimed in claim 1 utilizes induced by magnetic field to process the method that improves buckwheat blastogenesis active substances-quercetin content, it is characterized in that specifically comprising the steps:
(1), the pretreatment of buckwheat seed
Adopt clear water to eluriate buckwheat seed to remove not plump seed and sick worm seed, then with 35 DEG C of water seed soaking 24h, drain away the water after then rinsing 2-3 time with clear water;
(2), step (1) be placed in to magnetic field intensity through pretreatment the buckwheat seed that drains away the water induce processing, then sprout, must contain the buckwheat bud compared with high bioactivity material-quercetin content.
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| RU2239332C2 (en) * | 2002-07-22 | 2004-11-10 | Алтайский государственный технический университет им. И.И. Ползунова | Method for buckwheat flour production |
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