CN103519170A - Preparation method of high-activity buckwheat leaf flavonoids - Google Patents

Preparation method of high-activity buckwheat leaf flavonoids Download PDF

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CN103519170A
CN103519170A CN201310478293.XA CN201310478293A CN103519170A CN 103519170 A CN103519170 A CN 103519170A CN 201310478293 A CN201310478293 A CN 201310478293A CN 103519170 A CN103519170 A CN 103519170A
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buckwheat leaf
preparation
buckwheat
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hydrolysis
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李艳琴
史建强
李彬春
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Shanxi University
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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
    • A61K36/18—Magnoliophyta (angiosperms)
    • A61K36/185—Magnoliopsida (dicotyledons)
    • A61K36/70—Polygonaceae (Buckwheat family), e.g. spineflower or dock
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K2236/00—Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
    • A61K2236/10—Preparation or pretreatment of starting material
    • A61K2236/15—Preparation or pretreatment of starting material involving mechanical treatment, e.g. chopping up, cutting or grinding
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K2236/00—Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
    • A61K2236/30—Extraction of the material
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K2236/00—Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
    • A61K2236/30—Extraction of the material
    • A61K2236/33—Extraction of the material involving extraction with hydrophilic solvents, e.g. lower alcohols, esters or ketones
    • A61K2236/333—Extraction of the material involving extraction with hydrophilic solvents, e.g. lower alcohols, esters or ketones using mixed solvents, e.g. 70% EtOH
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K2236/00—Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
    • A61K2236/50—Methods involving additional extraction steps
    • A61K2236/53—Liquid-solid separation, e.g. centrifugation, sedimentation or crystallization

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Abstract

The invention discloses a preparation method of high-activity buckwheat leaf flavonoids. The preparation method comprises the following steps: extracting in 65-75 DEG C water bath by using 75-85% edible ethyl alcohol as a solvent according to a ratio of material to liquid being 1: (30-50), concentrating suction filtration liquid to one eighth to one tenth of the original volume, putting in a high-pressure reaction kettle, hydrolyzing for 20-60 minutes under the vapor pressure of 0.6-0.8Mpa, centrifuging, collecting precipitates, and drying at 60 DEG C, thus obtaining the high-activity buckwheat leaf flavonoid product. The purity of the buckwheat leaf flavonoid product is increased to 90.6% from 73.5% before high-pressure hydrolysis, and the removal ratio of DPPH free radicals is increased by 9.7% compared with the removal ratio before hydrolysis. Due to the adoption of the preparation method, the content of isoquercetin and quercetin which have relatively high biological activity is increased, the content of the isoquercetin is increased to 3.27-20.6% from zero before hydrolysis, and the content of the quercetin is increased to 8.2-22.78% from 2.2% before hydrolysis. The preparation method has the advantages of waste utilization, low cost, no pollution, simple operation and the like.

Description

A kind of preparation method of high activity buckwheat leaf flavones
Technical field
The present invention relates to buckwheat leaf flavones, specifically belong to preparation method of a kind of high activity buckwheat leaf flavones and products thereof.
Background technology
Buckwheat is buckwheat, dicotyledon, buckwheat seed, root, stem, leaf and spend and all contain abundant flavone compound.In buckwheat leaf, the main component of Flavonoid substances is rutin, Quercetin trace.It is the rutinoside of Quercetin that chemical structure analysis discloses rutin, and isoquercitin is the glucoside of Quercetin, and rutin can be hydrolyzed to isoquercitin or Quercetin, and isoquercitin can be hydrolyzed to Quercetin.The structural formula of Quercetin, isoquercitin and rutin is as follows:
Figure BDA0000395244570000011
Their threes have different bioavailabilities and physiologically active, RESEARCH ON CELL-BIOLOGY shows that absorbing of chromocor compound is mainly to enter blood by small intestine, and bioavailability is determined by its molecular weight and hydrophily, isoquercitin is except part is by free diffusing, because its end contains glucose part, by the sodium ion dependent form GLUT (SGLT1) on cell membrane, be mainly that active transport enters blood plasma, bioavailability is higher; And Quercetin and rutin can only enter blood plasma by free diffusing, bioavailability is lower, Quercetin is because of its molecular weight and lipophilicity simultaneously, bioavailability than rutin high (referring to Wolffram et al.Quercetin-3-glucoside is transported by the glucose carrier SGLT1across the brush border membrane of rat small intestine[J] .J Nutr.2002,132 (4): 630-5; Gee JM et al.Intestinal transport of quercetin glycosides in rats involves both deglycosylation and interaction with the hexose transport pathway[J] .J Nutr.2000,130 (11): 2765-71.).In addition, Quercetin is except having the physiological function similar to rutin, also there is the atherosclerotic of control and reduce the effects such as cholesterol (
Figure BDA0000395244570000012
et al.Effect of quercetin on experimental hyperlipidemia and atherosclerosis in rabbits[J] .Pharmacol Rep.2005,57 (5): 604-9; Auger et al.Dietary wine phenolics catechin, quercetin, and resveratrol efficiently protect hypercholesterolemic hamsters against aortic fatty streak accumulation[J] .J Agric Food Chem.2005,53 (6): 2015-21.); Isoquercitin has better market prospects, and its market price is the more than 1000 times of rutin; Active determination in vitro shows that Quercetin and isoquercitin show higher antioxidation activity than rutin.Therefore, buckwheat leaf chromocor extract is hydrolyzed and changes isoquercitin and/or Quercetin into, can make its BA maximize and variation, thereby promote the exploitation of buckwheat flavone nutrient and healthcare products and medicine.
Common method for hydrolysis has chemical hydrolysis, enzymatic hydrolysis and physics highly pressured hydrolysis.Chemical hydrolysis is introduced chemical reagent in reaction system, and toxicity increases and security reduces; Though enzymatic hydrolysis green high-efficient, at present still without suitable enzyme source, enzyme preparation is more expensive, by contrast, and highly pressured hydrolysis a kind of preferred version of can yet be regarded as.In China, buckwheat cultivated area is extensive, but very low to the exploitation of buckwheat leaf, is substantially taken as firewood and has burnt.For improving buckwheat leaf utilization rate, turning waste into wealth, we extract chromocor compound from buckwheat leaf, and are changed into by highly pressured hydrolysis the flavones product that physiologically active is higher, to further developing into nutrient and healthcare products.
Summary of the invention
The object of the present invention is to provide a kind of preparation method of high activity buckwheat leaf flavones, in the buckwheat leaf flavones product that the method makes, the content of isoquercitin and Quercetin significantly increases, and physiologically active improves.
The preparation method of a kind of high activity buckwheat leaf flavones provided by the invention, comprises the steps:
1) buckwheat leaf clean is dried, ground, obtain buckwheat leaf powder, by solid-liquid ratio 1:30-50, to the edible ethanol that adds 75-85% in buckwheat leaf powder, mix, be heated to 65-75 ℃, stirring and leaching 4h;
2) after lixiviate finishes, suction filtration obtains filtrate while hot, and precipitation obtains filtrate, merging filtrate with 2-3 times of cleaning of 75-85% edible ethanol, suction filtration;
3) concentrated filtrate is to the 1/8-1/10 of original volume, put into autoclave, control still inner vapor and press 0.6-0.8MPa, hydrolysis 20-60min, after question response still is cooling, hydrolysate is centrifugal, collecting precipitation, use again distilled water washing precipitation 3-5 time, the sediment of gained is 60 ℃ of oven dry, obtain the buckwheat leaf flavones that physiologically active is higher, product general flavone content is 88.8-90.6%, wherein containing rutin 14.0-51.1%, isoquercitin 3.27-20.6%, Quercetin 8.2-22.78% and other micro-flavonoids composition.
In described step 1), solid-liquid ratio is preferably 1:40-50; Step 3) mesohigh hydrolysis steam pressure is preferably 0.7MPa; The concentration of the edible ethanol described in step is preferably 85%.
Compared with prior art, advantage of the present invention and effect:
First, in the buckwheat leaf flavones product that the present invention obtains, the content of isoquercitin and Quercetin significantly increases, and is significantly higher than like product, isoquercitin and Quercetin not only physiologically active higher than rutin, and molecular weight is little, fat-soluble good, absorption and bioavailability are also greatly improved.
Secondly, the purity of product significantly improves, and is conducive to further prepare the medicine of high-quality and different dosage form.
Finally, the present invention's ethanol used is edible ethanol and recyclable, safe; The highly pressured hydrolysis that the present invention uses belongs to Physical hydrolysis, does not introduce chemical reagent and heavy metal ion in system, green, safety, reduced simultaneously pollutant discharge, reduced the destruction to environment.The present invention selects the buckwheat leaf that is rich in flavone compound and is taken as firewood in buckwheat as extracting material, has not only improved the utilization rate of buckwheat, turns waste into wealth; And adopting buckwheat leaf to extract flavones, cost is low, yield is high.
Accompanying drawing explanation
The mark product chromatogram of Fig. 1 rutin, isoquercitin and Quercetin
Fig. 2 buckwheat leaf flavone gruff bring up substance of the present invention solution chromatogram
Fig. 3 high activity buckwheat leaf of the present invention flavonoids solution (0.7MPa, 20min) chromatogram
Fig. 4 high activity buckwheat leaf of the present invention flavonoids solution (0.7MPa, 60min) chromatogram
Fig. 5 high activity buckwheat leaf of the present invention flavones and the comparison of crude extract to DPPH free radical scavenging activity
The specific embodiment
Embodiment 1
The preparation of buckwheat leaf flavone gruff bring up substance: the buckwheat leaf that field is gathered in is cleaned dries, grind, cross 60 mesh sieves, obtains buckwheat leaf powder; Take buckwheat leaf powder 40g, add 85% edible ethanol 1600mL (solid-liquid ratio 1:40), mix, put into 70 ℃ of water-bath stirring and leaching 4h, after taking-up, suction filtration obtains filtrate while hot, by the 85% edible ethanol washing and precipitating of 2 times once, and merging filtrate; With Rotary Evaporators concentrated filtrate to 300mL, a large amount of Precipitations; Centrifugal concentrating liquid, sediment, in 60 ℃ of oven dry, pulverizing, obtains buckwheat leaf flavone gruff bring up substance 5.33g, product general flavone content >=73.5%.
High effective liquid chromatography for measuring flavonoid component and content: instrument: U.S. Agilent company 1100 series of high efficiency liquid chromatographs, are included in line vacuum degasser, quaternary gradient pump, automatic sampler, column oven, DAD PDAD and chem workstation.
Chromatographiccondition: Hypersil BDS C18 chromatographic column (4.6 * 250mm, 5 μ m), 30 ℃ of column temperatures, mobile phase A (pure water), C(acetonitrile), the acetic acid of D(2%, v/v); In detection, the change in concentration of A and C is 0～7min, A:56%, C:24%; 8～12min, A:20%, C:60%; 13～17min, C:100%.Flow velocity 1.0mL/min, detects wavelength 360nm, reference wavelength 450nm.
Mark product: rutin: isoquercitin: Quercetin=0.1:0.05:0.1 (mg/mL), according to mark product measurement result (see figure 1), the appearance time of rutin, isoquercitin and Quercetin is respectively 3.401,4.120 and 10.741min.
The buckwheat leaf flavone gruff bring up substance 0.2g that takes dry constant weight, is accurate to 0.0001g, and be placed in tool plug triangular flask and add 70% ethanol to extract 4h 70 ℃ of water-baths, suction filtration while hot, filtrate is placed in 25mL volumetric flask constant volume, then gets 1mL and be settled to 50mL as liquid to be measured.Accurately draw 5 μ L liquid to be measured, carry out high performance liquid chromatography and obtain chromatogram (see figure 2), in buckwheat leaf flavone gruff bring up substance, contain as calculated other flavonoids composition of rutin 60.4%, Quercetin 2.0% and trace.
Determination of Physiological Activity: the antioxidation activity of evaluating buckwheat leaf flavones with DPPH radical scavenging activity.With 70% ethanol preparation DPPH solution, concentration is 0.04mg/mL.In the colorimetric cylinder that adds in advance 2.0mL DPPH solution, adding respectively 2mL concentration is 4,6,8,12,16,20 μ g/mL liquid to be measured, shakes up, standing 30min, measures its light absorption value at 517nm place,
Figure BDA0000395244570000031
Wherein: A xlight absorption value for sample and DPPH reaction solution; A xoit is the absorbance that 70% ethanol replaces DPPH solution; A 0it is the light absorption value that 70% ethanol replaces sample solution.
Buckwheat leaf flavone gruff bring up substance the results are shown in Figure 5 to DPPH radical scavenging activity.
Embodiment 2
The preparation of high activity buckwheat leaf flavones: the buckwheat leaf that field is gathered in is cleaned dries, grind, cross 60 mesh sieves, obtains buckwheat leaf powder; Take buckwheat leaf powder 40g, add 85% edible ethanol 2000mL (solid-liquid ratio 1:50), mix, put into 70 ℃ of water-bath stirring and leaching 4h, after taking-up, suction filtration obtains filtrate while hot, by the 85% ethanol washing and precipitating of 2 times once, and merging filtrate; With Rotary Evaporators concentrated filtrate to 200mL, put into autoclave, at 0.7MPa hydrolysis 20min, after question response still is cooling, pour hydrolysate into Centrifuge Cup, 6000g centrifugal collecting precipitation, use again distilled water washing precipitation 4 times, 60 ℃ of oven dry of gained precipitation, obtain the higher buckwheat leaf flavones product of physiologically active, this product general flavone content >=88.8%.
The assay method of flavonoid component and content: with embodiment 1, contain other flavonoids composition (see figure 3) of rutin 51.1%, isoquercitin 20.6%, Quercetin 8.2% and trace in high activity buckwheat leaf flavones.
Determination of Physiological Activity method: with embodiment 1, high activity buckwheat leaf flavones the results are shown in Figure 5 to DPPH radical scavenging activity.
Embodiment 3
The preparation of high activity buckwheat leaf flavones: the buckwheat leaf that field is gathered in is cleaned dries, grind, cross 60 mesh sieves, obtains buckwheat leaf powder; Take buckwheat leaf powder 40g, add 85% edible ethanol 1600mL(solid-liquid ratio 1:40), mix, put into 70 ℃ of water-bath stirring and leaching 4h, after taking-up, suction filtration obtains filtrate while hot, by the 85% edible ethanol washing and precipitating of 2 times once, merging filtrate; With Rotary Evaporators concentrated filtrate to 160mL, put into autoclave, at 0.7MPa hydrolysis 60min, after question response still is cooling, pour hydrolysate into Centrifuge Cup, 6000g centrifugal collecting precipitation, use again distilled water washing precipitation 4 times, 60 ℃ of oven dry of gained precipitation, obtain the higher buckwheat leaf flavones product of physiologically active, this product general flavone content >=90.6%.
The assay method of flavonoid component and content: with embodiment 1, contain other flavonoids composition (see figure 4) of rutin 14.0%, isoquercitin 3.27%, Quercetin 22.78% and trace in high activity buckwheat leaf flavones.
Determination of Physiological Activity method: with embodiment 1, high activity buckwheat leaf flavones the results are shown in Figure 5 to DPPH radical scavenging activity.

Claims (4)

1. a preparation method for high activity buckwheat leaf flavones, is characterized in that, comprises the steps:
1) buckwheat leaf clean is dried, ground, obtain buckwheat leaf powder, by solid-liquid ratio 1:30-50, to the edible ethanol that adds 75-85% in buckwheat leaf powder, mix, be heated to 65-75 ℃, stirring and leaching 4h;
2) after lixiviate finishes, suction filtration obtains filtrate while hot, and precipitation obtains filtrate, merging filtrate with 2-3 times of cleaning of 75-85% edible ethanol, suction filtration;
3) concentrated filtrate is to the 1/8-1/10 of original volume, put into autoclave, control still inner vapor and press 0.6-0.8MPa, hydrolysis 20-60min is after question response still is cooling, centrifugal by hydrolysate, collecting precipitation, use distilled water washing precipitation 3-5 time, the sediment of gained, 60 ℃ of oven dry, obtains high activity buckwheat leaf flavones again.
2. the preparation method of a kind of high activity buckwheat leaf flavones as claimed in claim 1, is characterized in that, described step 3) mesohigh hydrolysis steam pressure is 0.7MPa.
3. the preparation method of a kind of high activity buckwheat leaf flavones as claimed in claim 1, is characterized in that, in described step 1), solid-liquid ratio is 1:40-50.
4. the preparation method of a kind of high activity buckwheat leaf flavones as claimed in claim 1, is characterized in that, the concentration of described edible ethanol is 85%.
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Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN106923171A (en) * 2017-01-19 2017-07-07 百色学院 A kind of Zhuang characteristic snack purple potato glycan glutinous rice cake and preparation method thereof
CN115974826A (en) * 2022-12-28 2023-04-18 西昌航飞苦荞科技发展有限公司 Method for preparing tartary buckwheat quercetin from tartary buckwheat flavone extract

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CN101045720A (en) * 2007-03-20 2007-10-03 陕西师范大学 Method for extracting buckwheat flavone from buckwheat shell
KR20090106310A (en) * 2008-04-04 2009-10-08 황보룡 Shochu production method using buckwheat
CN102948758A (en) * 2012-11-26 2013-03-06 陕西科技大学 Method for extracting buckwheat flavone from buckwheat bran

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Cited By (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN106923171A (en) * 2017-01-19 2017-07-07 百色学院 A kind of Zhuang characteristic snack purple potato glycan glutinous rice cake and preparation method thereof
CN106923171B (en) * 2017-01-19 2020-03-31 百色学院 Zhuang special snack purple sweet potato and purple yam glutinous rice cake and preparation method thereof
CN115974826A (en) * 2022-12-28 2023-04-18 西昌航飞苦荞科技发展有限公司 Method for preparing tartary buckwheat quercetin from tartary buckwheat flavone extract

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Application publication date: 20140122