CN102293868B - Application of total flavonoids of lilac daphne root in preparation of medicament for reducing blood sugar - Google Patents
Application of total flavonoids of lilac daphne root in preparation of medicament for reducing blood sugar Download PDFInfo
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Abstract
The invention relates to application of total flavonoids of lilac daphne root extracted from a lilac daphne plant to preparation of a medicament for reducing blood sugar. The total flavonoids of lilac daphne root, which is disclosed by the invention, respectively represents remarkable activity of reducing blood sugar at each dosage; and the total flavonoids of lilac daphne root can be used for preparing a medicament or health-card food for preventing or treating hyperglycemia symptom.
Description
Technical field
The present invention relates to from the Radix Brassicae Rapae plant, extract the purposes that the total flavonoids from radix Daphne genkwa that obtains prepares hypoglycemic drug, belong to medicinal natural product field.
Background technology
A large amount of epidemiological studies show that it is the pathogenetic risk factor of glycosuria that pathoglycemia raises, and diabetes are a kind of common endocrine metabolism diseases, are the third-largest illness of harm humans health after cardiovascular disease and cancer.Because diabetes have prevalence, disability rate and complication height, therefore become one of disease that medical circle is rather attracted attention and extraordinarily pay attention to.
Present hypoglycemic drug mostly is the big compound recipe of Western medicine or Chinese medicine, and Western medicine (comprising injection and oral antidiabetic drug) effect is strong and rapidly, the excessive patient's of causing hypoglycemia and be forced to drug withdrawal, and the result causes the withdrawal symptom knock-on again; The big compound recipe of Chinese medicine all is to be concocted and extract and make through simple by natural Chinese medicinal herb basically, and not only composition is quite complicated, preparation inconvenience, and is difficult to carry out more deep structure/drug efficacy study, also is unfavorable for drug quality control and the modernization of Chinese medicine.
There is abundant Chinese herbal medicine resource in China, these natural plant resources of reasonable development how, utilize Modern Pharmaceutical Chemistry analytical technology and biological experiment means, seek a class safely and effectively the natural plants hypoglycemic drug become the focus of medical science, field of biology extensive concern, also be the key that promotes China's hypoglycemic drug industrial competitiveness.
Radix Brassicae Rapae (Brassica rapa L.) has another name called turnip, and biennial herb is now extensively planted in the Qinghai-Tibet Platean.Radix Brassicae Rapae is grown in height above sea level, the temperature difference is big, sunshine the is strong altitude environment, and adaptive capacity is strong, the output height, nutritious, usually as feed applications in animal husbandry, its fleshy tap root can also be as vegetable or fruit human consumption, have the effect of heat-clearing and toxic substances removing, nourishing oxygen supplement.At present in the disclosed method, Shang Weijian is about the report of total flavonoids from radix Daphne genkwa effect of lowering blood sugar aspect for the development and use Radix Brassicae Rapae.Glucosides or other forms of derivant that flavone compound refers to belong on the chemical constitution chemical compound such as flavonoid, flavonols, flavanone, flavanone alcohols, anthocyan, flavanol compound, two benzene pyrrones, osajin and forms with monosaccharide or oligosaccharide.They extensively are present in nature, in higher plant and the pteridophyte plant, are the important natural organic-compounds of a class especially, and its physiologically active is varied.Studies show that effects such as flavone compound has the hypertension of control, treatment coronary heart disease, suppresses thrombosis, anti-liver poison, antiinflammatory, rushes down down, blood fat reducing, estrogen-like effects in a large number.Therefore, the development and use plant flavone has huge biology, medical significance.The present invention is by extracting total flavonoids from radix Daphne genkwa and it being carried out hypoglycemic activity detect, widened the approach of Radix Brassicae Rapae comprehensive exploitation, natural for exploring, effectively hypoglycemic drug provides theoretical foundation, meet the needs of the utilization of plateau plant resources effective and Tibetan medicine new drug development, be conducive to promote the ethnic mimority area expanding economy.
Summary of the invention
It is the total flavonoids from radix Daphne genkwa extract of main component that one of purpose of the present invention provides with the flavone compound;
The present invention also provides above-mentioned total flavonoids from radix Daphne genkwa preparation method of extract;
The present invention also provide above-mentioned total flavonoids from radix Daphne genkwa extract treatment and/purposes in the medication preparation of prevention pathoglycemia rising disease.
For achieving the above object, the technical solution used in the present invention is:
Extract total flavones from Radix Brassicae Rapae root piece, yield is 0.2～1.0 weight %, is solid-state faint yellow, is soluble in ethanol, and is soluble in water.Utilize the diabetic mice model due to alloxan and the epinephrine to carry out the experiment of yuenkanin total saponins hypoglycemic activity effect, except as otherwise noted, the % among the present invention is weight percentage.
Total flavonoids from radix Daphne genkwa preparation method of extract of the present invention is specially:
Method A:
(1) distilled water of Radix Brassicae Rapae raw material with 5～15 times of volumes boiled;
Slowly add lime cream or NaOH alkaline solution when (2) stirring, transfer pH to 8～10, continue to boil 20～60min, use 4 layers of gauze sucking filtration while hot, filtrate is filtered again with middling speed filter paper;
(3) filtering residue decocts with 5～15 times of volume distilled water again, and sucking filtration repeats to extract 1～3 time while hot,
(4) merge twice filtrate, under 60～80 ℃ of conditions, transfer pH to 4～6 with concentrated hydrochloric acid, leave standstill 24h after stirring evenly, sucking filtration;
(5) with distilled water precipitate is washed till neutrality, obtains rough total flavones after the drying,
(6) use the boiling water recrystallization, obtain refined total flavonoids after the drying, yield is 0.2～0.7 weight %, and purity is 37%～75%.
Method B:
(1) with 50～95% ethanol of 5～15 times of volumes or methanol as solvent, in 50～80 ℃ the Radix Brassicae Rapae raw material is carried out reflux, extract, 1～4h, carry out repeating for 1～4 time extracting;
(2) with middling speed filter paper filtering extracting solution, merging filtrate, being evaporated to filtrate does not have the alcohol flavor;
(3) in filtrate, add petroleum ether extraction 3～5 times by 1: 1～3 volumes, fling to [60 ℃ of water-baths vapor away the petroleum ether in the solution] petroleum ether for 60 ℃, adding ethyl acetate by 1: 1～3 volumes again extracts 3～5 times concentrated solution, the solution decompression of ethyl acetate part is concentrated, dry, obtain thick total flavonoids extract;
(4) thick total flavonoids extract is dissolved in a small amount of 50～95% ethanol, obtain total flavones upper prop liquid, utilize absorption with macroporous adsorbent resin, after colourless to effluent with distilled water flushing, be 1～2.5ml/min with flow velocity, it is extremely colourless to carry out gradient elution with 20～95% ethanol of 2～8 times of column volumes, collect eluent, with the total flavones that eluent 50～80 ℃ of concentrating under reduced pressure, dryings obtain making with extra care, yield is 0.5～0.7 weight %, and purity is 75%～95%.
Method C
(1) with 60～95% soak with ethanol 1～3h of Radix Brassicae Rapae raw material with 10～30 times of volumes;
(2) under normal pressure, with microwave power 500～900w, microwave radiation 20～60s, intermittent radiation 2～5 times carries out the microwave-assisted leaching;
(3) filter collection filtrate, filtrate is condensed into the extractum that does not have the alcohol flavor;
(4) extractum is dissolved in a small amount of hot water, successively with petroleum ether and ethyl acetate extractum liquid is extracted by 1: 1～3 volume ratios, collect the ethyl acetate part, decompression and solvent recovery obtains thick total flavonoids extract after the drying;
(5) thick total flavonoids extract is dissolved in a small amount of 50～95% ethanol, obtains total flavones upper prop liquid.Utilize absorption with macroporous adsorbent resin, colourless to effluent with distilled water flushing after, be 1～2.5ml/min with flow velocity, carry out gradient elution to colourless, the collection eluent with 20～95% ethanol of 2～8 times of column volumes.The total flavonoids from radix Daphne genkwa that eluent 50～80 ℃ of concentrating under reduced pressure, dryings are obtained making with extra care.Yield is 0.3～0.9 weight %.Purity is 43%～75%.
Radix Brassicae Rapae raw material in the described preparation method is the tuber of Radix Brassicae Rapae, and natural air drying after cutting into slices is pulverized then and obtained powder, and smashing fineness reaches 40～200 orders;
Used macroporous adsorbent resin is D101 or AB-8 among said method B, the method C;
Described drying means is reduced vacuum drying, lyophilization or spray drying.
Total flavonoids from radix Daphne genkwa is prepared into various, for example powder, tablet, capsule, granule, slow releasing tablet, slow releasing capsule, drop pill or liquid preparation.Described preparation is applied to the clinical treatment hyperglycemia and makes contain the total flavonoids from radix Daphne genkwa health-caring product capable of reducing blood sugar.
The present invention is with the total flavonoids from radix Daphne genkwa that hydrochloric acid-the magnesium powder reaction detection is carried, and the foam place presents aubergine; After 60% alcoholic solution of sample thief drops on the filter paper, with 1%AlCl
3Alcoholic solution is reflected under the visible light and presents lark, presents the yellow fluorescence speckle under ultraviolet light; 60% drips of solution of sample thief ethanol on filter paper, smoked with strong aqua ammonia after, under ultraviolet light, present yellowish-brown fluorescence speckle, illustrate that extract is flavone really.The quantitative assay of total flavonoids from radix Daphne genkwa is to utilize sodium nitrite-aluminum nitrate-sodium hydroxide colorimetry, is standard substance drawing standard curve with the rutin, utilizes determined by ultraviolet spectrophotometry total flavonoids from radix Daphne genkwa content.
Total flavonoids from radix Daphne genkwa by method for preparing is having significant effect aspect the blood sugar lowering.Utilize the diabetic mice model due to alloxan and the epinephrine to carry out the experiment of total flavonoids from radix Daphne genkwa hypoglycemic activity effect, the result shows, total flavonoids from radix Daphne genkwa causes diabetic mice to alloxan and epinephrine and has remarkable hypoglycemic activity.
The total flavonoids from radix Daphne genkwa raw material that the inventive method obtains is easy to get, and method is simple, and production cost is low, and general flavone content is higher in the product, steady quality, be easy to control.
The specific embodiment:
Embodiment 1
Employing method A gets and dries the Radix Brassicae Rapae tuber and be crushed to 100 orders, boils with the distilled water of 5 times of volumes, when not stopping stirring after boiling, slowly adds NaOH, and solution PH is transferred to 10.Continue to keep boiling 20min.Carry out sucking filtration with four layers of gauze pad at buchner funnel while hot, sucking filtration finishes and with middling speed filter paper filtrate is filtered again.Collect filtrate.Filtering residue is decocted 20min again with the distilled water of 5 times of volumes, and sucking filtration, filtration repeat to extract 3 times merging filtrate like this to filtering residue.Filtrate is heated to 60 ℃, slowly adds concentrated hydrochloric acid, solution PH is transferred to 4, after stirring, leave standstill 24h after, sucking filtration, collecting precipitation thing.With a large amount of distilled water flushing precipitate, be 7,60 ℃ of reduced vacuum dry sediments until PH, obtain thick total flavones.With the thick total flavones recrystallization that boiling water will obtain, 60 ℃ of reduced vacuum drying crystallines obtain flaxen refining total flavonoids from radix Daphne genkwa, and yield is 0.32%, and purity is 57%.
Embodiment 2
Employing method A gets and dries the Radix Brassicae Rapae tuber and be crushed to 40 orders, boils with the distilled water of 15 times of volumes, when not stopping stirring after boiling, slowly adds lime cream, and solution PH is transferred to 9.Continue to keep boiling 60min.Carry out sucking filtration with four layers of gauze pad at buchner funnel while hot, sucking filtration finishes and with middling speed filter paper filtrate is filtered again.Collect filtrate.Filtering residue is decocted 60min again with the distilled water of 15 times of volumes, and sucking filtration, filtration repeat to extract 1 time merging filtrate like this to filtering residue.Filtrate is heated to 80 ℃, slowly adds concentrated hydrochloric acid, solution PH is transferred to 4, after stirring, leave standstill 24h after, sucking filtration, collecting precipitation thing.With a large amount of distilled water flushing precipitate, be 7,80 ℃ of dry sediments until PH, obtain thick total flavones.With the thick total flavones recrystallization that boiling water will obtain, the crystallization of lyophilization gained obtains flaxen refining total flavonoids from radix Daphne genkwa, and yield is 0.62%, and purity is 39%.
Embodiment 3
Employing method A gets and dries the Radix Brassicae Rapae tuber and be crushed to 200 orders, boils with the distilled water of 10 times of volumes, when not stopping stirring after boiling, slowly adds lime cream, and solution PH is transferred to 8.Continue to keep boiling 40min.Carry out sucking filtration with four layers of gauze pad at buchner funnel while hot, sucking filtration finishes and with middling speed filter paper filtrate is filtered again.Collect filtrate.Filtering residue is decocted 40min again with the distilled water of 5 times of volumes, and sucking filtration, filtration repeat to extract 2 times merging filtrate like this to filtering residue.Filtrate is heated to 70 ℃, slowly adds concentrated hydrochloric acid, solution PH is transferred to 4, after stirring, leave standstill 24h after, sucking filtration, collecting precipitation thing.With a large amount of distilled water flushing precipitate, be 7,70 ℃ of reduced vacuum dry sediments until PH, obtain thick total flavones.With the thick total flavones recrystallization that boiling water will obtain, 70 ℃ of reduced vacuum drying crystallines obtain flaxen refining total flavonoids from radix Daphne genkwa, and yield is 0.24%, and purity is 73%.
Embodiment 4
Employing method B as solvent, carries out reflux, extract, 1h in 50 ℃ to the Radix Brassicae Rapae raw material with 95% ethanol of 5 times of volumes, carries out repeating for 4 times extracting.With middling speed filter paper filtering extracting solution, merging filtrate, being evaporated to filtrate does not have the alcohol flavor.In filtrate, add petroleum ether extraction 3 times by 1: 1 volume, fling to petroleum ether for 60 ℃, add ethyl acetate by 1: 2 volume again concentrated solution is extracted 3 times, the solution decompression of ethyl acetate part is concentrated, dry, obtain thick total flavonoids extract.Thick total flavonoids extract is dissolved in a small amount of 95% ethanol, obtains total flavones upper prop liquid.Utilize the D101 absorption with macroporous adsorbent resin, colourless to effluent with distilled water flushing after, be 2.5ml/min with the flow velocity, carry out gradient elution to colourless, the collection eluent with 20%, 60%, 95% ethanol of 8 times of column volumes.50 ℃ of concentrating under reduced pressure of eluent, reduced vacuum drying are obtained flaxen refined total flavonoids, and yield is that 0.33% purity is 85%.
Embodiment 5
Employing method B as solvent, carries out reflux, extract, 1h in 60 ℃ to the Radix Brassicae Rapae raw material with 50% ethanol of 10 times of volumes, carries out repeating for 5 times extracting.With middling speed filter paper filtering extracting solution, merging filtrate, being evaporated to filtrate does not have the alcohol flavor.In filtrate, add petroleum ether extraction 4 times by 1: 1 volume, fling to petroleum ether for 60 ℃, add ethyl acetate by 1: 1 volume again concentrated solution is extracted 4 times, the solution decompression of ethyl acetate part is concentrated, dry, obtain thick total flavonoids extract.Thick total flavonoids extract is dissolved in a small amount of 60% ethanol, obtains total flavones upper prop liquid.Utilize the AB-8 absorption with macroporous adsorbent resin, colourless to effluent with distilled water flushing after, be 2ml/min with the flow velocity, carry out gradient elution to colourless, the collection eluent with 30%, 50%, 80%, 95% ethanol of 4 times of column volumes.Eluent 60 ℃ of concentrating under reduced pressure, lyophilizations are obtained flaxen refined total flavonoids, and yield is 0.63%, and purity is 77%.
Embodiment 6
Employing method B as solvent, carries out reflux, extract, 1h in 70 ℃ to the Radix Brassicae Rapae raw material with 70% ethanol of 15 times of volumes, carries out repeating for 3 times extracting.With middling speed filter paper filtering extracting solution, merging filtrate, being evaporated to filtrate does not have the alcohol flavor.In filtrate, add petroleum ether extraction 4 times by 1: 1 volume, fling to petroleum ether for 60 ℃, add ethyl acetate by 1: 3 volume again concentrated solution is extracted 4 times, the solution decompression of ethyl acetate part is concentrated, dry, obtain thick total flavonoids extract.Thick total flavonoids extract is dissolved in a small amount of 70% ethanol, obtains total flavones upper prop liquid.Utilize the AB-8 absorption with macroporous adsorbent resin, colourless to effluent with distilled water flushing after, be 1ml/min with the flow velocity, carry out gradient elution to colourless, the collection eluent with 30%, 60%, 95% ethanol of 2 times of column volumes.Eluent 60 ℃ of concentrating under reduced pressure, lyophilizations are obtained flaxen refined total flavonoids, and yield is 0.43%, and purity is 91%.
Embodiment 7
Employing method C is with the 60% soak with ethanol 1h of Radix Brassicae Rapae raw material with 20 times of volumes.Under normal pressure, with microwave power 700w, microwave radiation 40s, intermittent radiation 3 times carries out the microwave-assisted leaching.Filter and collect filtrate, filtrate is condensed into the extractum that does not have the alcohol flavor.Extractum is dissolved in a small amount of hot water, successively with petroleum ether and ethyl acetate extractum liquid is extracted by 1: 3 volume ratio, collect the ethyl acetate part, decompression and solvent recovery obtains thick total flavonoids extract after the drying.Thick total flavonoids extract is dissolved in a small amount of 60% ethanol, obtains total flavones upper prop liquid.Utilize the AB-8 absorption with macroporous adsorbent resin, colourless to effluent with distilled water flushing after, be 2ml/min with the flow velocity, carry out gradient elution to colourless, the collection eluent with 20%, 50%, 80%, 95% ethanol of 5 times of column volumes.Eluent 60 ℃ of concentrating under reduced pressure, dryings are obtained flaxen refining total flavonoids from radix Daphne genkwa, and yield is 0.83%, and purity is 45%.
Embodiment 8
Employing method C is with the 75% soak with ethanol 3h of Radix Brassicae Rapae raw material with 10 times of volumes.Under normal pressure, with microwave power 500w, microwave radiation 60s, intermittent radiation 2 times carries out the microwave-assisted leaching.Filter and collect filtrate, filtrate is condensed into the extractum that does not have the alcohol flavor.Extractum is dissolved in a small amount of hot water, successively with petroleum ether and ethyl acetate extractum liquid is extracted by 1: 1 volume ratio, collect the ethyl acetate part, decompression and solvent recovery obtains thick total flavonoids extract after the drying.Thick total flavonoids extract is dissolved in a small amount of 75% ethanol, obtains total flavones upper prop liquid.Utilize the AB-8 absorption with macroporous adsorbent resin, colourless to effluent with distilled water flushing after, be 1ml/min with the flow velocity, carry out gradient elution to colourless, the collection eluent with 30%, 60%, 90% ethanol of 3 times of column volumes.Eluent 60 ℃ of concentrating under reduced pressure, dryings are obtained flaxen refining total flavonoids from radix Daphne genkwa, and yield is 0.46%, and purity is 60%.
Embodiment 9
Employing method C, with the Radix Brassicae Rapae raw material with 30 times of volumes, 95% soak with ethanol 2h.Under normal pressure, with microwave power 900w, microwave radiation 20s, intermittent radiation 5 times carries out the microwave-assisted leaching.Filter and collect filtrate, filtrate is condensed into the extractum that does not have the alcohol flavor.Extractum is dissolved in a small amount of hot water, successively with petroleum ether and ethyl acetate extractum liquid is extracted by 1: 2 volume ratio, collect the ethyl acetate part, decompression and solvent recovery obtains thick total flavonoids extract after the drying.Thick total flavonoids extract is dissolved in a small amount of 95% ethanol, obtains total flavones upper prop liquid.Utilize the D101 absorption with macroporous adsorbent resin, colourless to effluent with distilled water flushing after, be 2.5ml/min with the flow velocity, carry out gradient elution to colourless, the collection eluent with 30%, 60%, 90% ethanol of 8 times of column volumes.Eluent 50～80 ℃ of concentrating under reduced pressure, dryings are obtained flaxen refining total flavonoids from radix Daphne genkwa, and yield is 0.59%, and purity is 72%.
Embodiment 10
Tablet: get total flavonoids from radix Daphne genkwa 50g, add 50g and can press starch and 40g carboxymethyl starch sodium, mix homogeneously, the ethanol with 70% are wetting agent, soft material processed, 18 mesh sieves are granulated, oven dry, 16 mesh sieve granulate, adding the 3g magnesium stearate is lubricant, mix homogeneously, tabletting, coating are made 1000 coating tablets sheets.
Embodiment 11
Capsule: get total flavonoids from radix Daphne genkwa 50g, add 80g and can press starch and 20g sodium carboxymethyl cellulose, mix homogeneously, the ethanol with 70% are wetting agent, soft material processed, 24 mesh sieves are granulated, oven dry, 20 mesh sieve granulate, add the 3g magnesium stearate, mix homogeneously, encapsulated, make 1000 capsules.
Embodiment 12
Granule: get total flavonoids from radix Daphne genkwa 100g, add 450g cane sugar powder, 100g sodium carboxymethylcellulose, 100g microcrystalline Cellulose and 50g citric acid, mix homogeneously, be wetting agent soft material processed with 3%PVP, 20 mesh sieves are granulated, oven dry, 18 mesh sieve granulate, packing, making particle diameter is 1000 bags of granules of 830～880 μ m.Every bag heavy 0.72～0.78g.
Embodiment 13
Injectable powder: get total flavonoids from radix Daphne genkwa 12g, hydroxypropyl-p-cyclodextrin 135g, add injection water 3L, stir into pasty state, add 70 ℃～85 ℃ water for injection 3L again, temperature control with mannitol 450g dissolving, is used 1moL/L hydrochloric acid solution adjust pH to 5.6 at 70 ℃～85 ℃ again during dissolving, add pin with charcoal after stirring at room, filtering decarbonization, fine straining. filtrate is carried out packing by every 2ml, adopts the quick freezing drying, goods are taken out, seal, namely obtaining character is white loose block or powder, namely gets injectable powder.
Embodiment 14
Drop pill: with the 150g Macrogol 4000 water-bath heat fused solution, add total flavonoids from radix Daphne genkwa 50g while hot, stir, 65 ℃ are incubated 1 hour, the dropping-pill machine system of using, drip 120 droplets/minute of speed, drip 65～75 ℃ of temperature processed, cool off in the liquid paraffin about 15 ℃, make the heavily about 60mg of ball, drying, coating namely gets drop pill.
Embodiment 15
Oral liquid: it is an amount of to get distilled water, adds 20g total flavonoids from radix Daphne genkwa, 180g cane sugar powder, 20g citric acid, and stirring and dissolving is filtered, and filtrate is replenished distilled water to 10000ml, it is an amount of to add essence, stirs packing, every 10ml, gland was sterilized 1 hour for 100 ℃, and passed examination namely gets oral liquid.
Embodiment 16
Powder: get total flavonoids from radix Daphne genkwa 30g, being ground into particle diameter is 20 μ m powders, adds 500g cane sugar powder, 100g sodium carboxymethylcellulose, 100g microcrystalline Cellulose mix homogeneously, and 1000 bags of powder are made in packing.Every bag heavy 0.68～0.73g.Addition is 0.01～1% of food or feedstuff.
The total flavonoids from radix Daphne genkwa of above-mentioned preparation embodiment 2 gained is carried out following pharmacodynamic experiment:
With the hypoglycemic activity experiment of embodiment 2 prepared total flavonoids from radix Daphne genkwa to the alloxan diabetes mouse model
1. experiment material and method:
Get 60 of male mice in kunming, 28-32g, adaptability raise back random packet, fasting 12 hours; The alloxan of every mouse tail vein injection 85mg/kg, 72h posterior orbit measuring blood sugar of blood extracting, blood glucose reach the value of defining (〉=16.9mmol/L) then be defined as diabetic mice, press the height random packet of blood glucose, be respectively model group, positive group and total flavones I (0.5g/kg), total flavones II (1g/kg) and total flavones III (2g/kg) group; Give 0.5% every day to model group
Sodium carboxymethyl cellulose(CMC) solution 0.4ml, positive group gives 50mg/kg glibenclamide, and each group of total flavones gives the extractive of general flavone of corresponding dosage, and administration continues 7 days, and eye socket was got blood in the 8th day, with determination of glucose oxidase serum blood glucose.Put to death after the 8th day eye socket of mice got blood, dissect and get liver, kidney, thymus and spleen respectively, weigh, calculate organ index.Experimental data is carried out statistical procedures with the SPSS10.0 statistical software, and experimental data represents with mean ± standard deviation that all employing t check is compared between organizing.
2. experimental result:
Table 1 total flavonoids from radix Daphne genkwa is to the influence of experimental mouse blood glucose
| Group and dosage | Initial blood glucose (d1) | Administration is blood glucose (d7) after 7 days |
| Model group- | 23.1±2.8 | 22.3±2.1 |
| Positive group (50mg/kg) | 23.0±2.6 | 18.9±3.3* |
| Total flavones I (0.5g/kg) | 23.0±2.2 | 18.7±2.0* |
| Total flavones II (1g/kg) | 22.9±2.4 | 16.7±3.2** |
| Total flavones III (2g/kg) | 23.2±2.9 | 17.5±3.1** |
Annotate: compare " * " P＜0.5, " * * " P＜0.01 with model group
Table 2 total flavonoids from radix Daphne genkwa is to the influence of experimental mouse organ index
Annotate: 1. with normal group relatively, a significance＞0.05, a+ significance＞0.01;
2. with model group relatively, b significance＞0.05, b+ significance＞0.01
Interpretation: the blood glucose of positive group, total flavones I, total flavones II and total flavones III group mice is compared model group significant decline, and wherein total flavones II and total flavones III group is lower than model group extremely significantly.Experimental result shows that total flavonoids from radix Daphne genkwa causes diabetic mice to alloxan and has remarkable hypoglycemic activity.
Organ index is that reflection environmental stimuli commonly used is to the index of laboratory animal toxicity.Thymus is the mice important immune organs, though this index of all improper mices all is markedly inferior to normal group, positive group and total flavones group mice still are significantly improved.The liver of model group is obviously than normal group enlargement, and the liver index of total flavones III group is almost identical with normal group.In addition, the kidney of model group is serious enlargement also, and total flavones III group also is significantly improved to the pathological changes of kidney.
In sum, total flavonoids from radix Daphne genkwa not only can reduce the blood glucose that alloxan causes diabetic mice, can also improve the pathological changes of internal organs simultaneously.The relevant test of pesticide effectiveness has also been carried out to the animal hyperglycemia due to the epinephrine simultaneously in this chamber, and the result shows also significant hypoglycemic activity.
Above-mentioned experimental result shows that total flavonoids from radix Daphne genkwa of the present invention all demonstrates significant hypoglycemic activity at each dosage.Can utilize total flavonoids from radix Daphne genkwa to prepare medicine or health food and be used for prevention or treatment hyperglycemia disease.
The preparation method of total flavonoids from radix Daphne genkwa of the present invention and purposes are described by concrete example, those skilled in the art can use for reference content of the present invention, links such as appropriate change raw material, process conditions realize corresponding other purpose, its relevant change does not all break away from content of the present invention, all similar replacements and change will become apparent to those skilled in the art that and all be regarded as comprising within the scope of the present invention.
Claims (6)
1. a total flavonoids from radix Daphne genkwa prepares the purposes of hypoglycemic drug, and it is characterized in that: the total flavonoids from radix Daphne genkwa extract is prepared into hypoglycemic medicine, and described total flavonoids from radix Daphne genkwa prepares as follows:
(1) distilled water of Radix Brassicae Rapae raw material with 5～15 times of volumes boiled;
Slowly add lime cream or NaOH alkaline solution when (2) stirring, transfer pH to 8～10, continue to boil 20～60min, use 4 layers of gauze sucking filtration while hot, filtrate is filtered again with middling speed filter paper;
(3) filtering residue decocts with 5～15 times of volume distilled water again, and sucking filtration repeats to extract 1～3 time while hot,
(4) merge twice filtrate, under 60～80 ℃ of conditions, transfer pH to 4～6 with concentrated hydrochloric acid, leave standstill 24h after stirring evenly, sucking filtration;
(5) with distilled water precipitate is washed till neutrality, obtains rough total flavones after the drying,
(6) use the boiling water recrystallization, obtain refined total flavonoids after the drying, yield is 0.2～0.7 weight %, and purity is 37%～75%;
The root piece that described Radix Brassicae Rapae raw material is Radix Brassicae Rapae.
2. a total flavonoids from radix Daphne genkwa prepares the purposes of hypoglycemic drug, and it is characterized in that: the total flavonoids from radix Daphne genkwa extract is prepared into hypoglycemic medicine, and described total flavonoids from radix Daphne genkwa prepares as follows:
(1) with 50～95% ethanol of 5～15 times of volumes or methanol as solvent, in 50～80 ℃ the Radix Brassicae Rapae raw material is carried out reflux, extract, 1～4h, carry out repeating for 1～4 time extracting;
(2) with middling speed filter paper filtering extracting solution, merging filtrate, being evaporated to filtrate does not have the alcohol flavor;
(3) in filtrate, add petroleum ether extraction 3～5 times by 1: 1～3 volumes, fling to petroleum ether for 60 ℃, add ethyl acetate by 1: 1～3 volumes again concentrated solution is extracted 3～5 times, the solution decompression of ethyl acetate part is concentrated, dry, obtain thick total flavonoids extract;
(4) thick total flavonoids extract is dissolved in a small amount of 50～95% ethanol, obtain total flavones upper prop liquid, utilize absorption with macroporous adsorbent resin, after colourless to effluent with distilled water flushing, be 1～2.5ml/min with flow velocity, it is extremely colourless to carry out gradient elution with 20～95% ethanol of 2～8 times of column volumes, collect eluent, with the total flavones that eluent 50～80 ℃ of concentrating under reduced pressure, dryings obtain making with extra care, yield is 0.5～0.7 weight %, and purity is 75%～95%;
The root piece that described Radix Brassicae Rapae raw material is Radix Brassicae Rapae.
3. a total flavonoids from radix Daphne genkwa prepares the purposes of hypoglycemic drug, and it is characterized in that: the total flavonoids from radix Daphne genkwa extract is prepared into hypoglycemic medicine, and described total flavonoids from radix Daphne genkwa prepares as follows:
(1) with 60～95% soak with ethanol 1～3h of Radix Brassicae Rapae raw material with 10～30 times of volumes;
(2) under normal pressure, with microwave power 500～900w, microwave radiation 20～60s, intermittent radiation 2～5 times carries out the microwave-assisted leaching;
(3) filter collection filtrate, filtrate is condensed into the extractum that does not have the alcohol flavor;
(4) extractum is dissolved in a small amount of hot water, successively with petroleum ether and ethyl acetate extractum liquid is extracted by 1: 1～3 volume ratios, collect the ethyl acetate part, decompression and solvent recovery obtains thick total flavonoids extract after the drying;
(5) thick total flavonoids extract is dissolved in a small amount of 50～95% ethanol, obtains total flavones upper prop liquid; Utilize absorption with macroporous adsorbent resin, colourless to effluent with distilled water flushing after, be 1～2.5ml/min with flow velocity, carry out gradient elution to colourless, the collection eluent with 20～95% ethanol of 2～8 times of column volumes; The total flavonoids from radix Daphne genkwa that eluent 50～80 ℃ of concentrating under reduced pressure, dryings are obtained making with extra care; Yield is 0.3～0.9 weight %; Purity is 43%～75%;
The root piece that described Radix Brassicae Rapae raw material is Radix Brassicae Rapae.
4. according to the arbitrary described purposes of claim 1-3, wherein said Radix Brassicae Rapae raw material is the tuber of Radix Brassicae Rapae, and natural air drying after cutting into slices is pulverized then and obtained powder, and smashing fineness reaches 40～200 orders.
5. according to the arbitrary described purposes of claim 1-3, wherein said drying means is reduced vacuum drying, lyophilization or spray drying.
6. according to the arbitrary described purposes of claim 1-3, wherein the total flavonoids from radix Daphne genkwa extract is prepared into powder, tablet, capsule, granule, slow releasing tablet, slow releasing capsule, drop pill or liquid preparation.
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| CN105748569A (en) * | 2016-03-31 | 2016-07-13 | 淮阴师范学院 | Extracting and separating method of total flavones of wisteria flowers |
| CN111214588A (en) * | 2020-01-21 | 2020-06-02 | 沈阳药科大学 | Total flavone in lilac daphne root and preparation and application thereof |
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