CN107174605A - A kind of proso millet prolamin n-butanol extract and its preparation method and application - Google Patents

A kind of proso millet prolamin n-butanol extract and its preparation method and application Download PDF

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CN107174605A
CN107174605A CN201710357078.2A CN201710357078A CN107174605A CN 107174605 A CN107174605 A CN 107174605A CN 201710357078 A CN201710357078 A CN 201710357078A CN 107174605 A CN107174605 A CN 107174605A
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proso millet
millet prolamin
prolamin
butanol extract
extract
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高良才
林泽杰
陆诗韵
周梦璇
陈清
林青楠
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East China Normal University
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    • AHUMAN NECESSITIES
    • A23FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23LFOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
    • A23L33/00Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
    • A23L33/10Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
    • A23L33/105Plant extracts, their artificial duplicates or their derivatives
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61KPREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K36/00Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
    • A61K36/18Magnoliophyta (angiosperms)
    • A61K36/185Magnoliopsida (dicotyledons)
    • A61K36/73Rosaceae (Rose family), e.g. strawberry, chokeberry, blackberry, pear or firethorn
    • AHUMAN NECESSITIES
    • A23FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23VINDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
    • A23V2200/00Function of food ingredients
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61KPREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K2236/00Isolation 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
    • A61K2236/30Extraction of the material
    • A61K2236/33Extraction of the material involving extraction with hydrophilic solvents, e.g. lower alcohols, esters or ketones
    • A61K2236/333Extraction of the material involving extraction with hydrophilic solvents, e.g. lower alcohols, esters or ketones using mixed solvents, e.g. 70% EtOH
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61KPREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K2236/00Isolation 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
    • A61K2236/30Extraction of the material
    • A61K2236/35Extraction with lipophilic solvents, e.g. Hexane or petrol ether
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61KPREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K2236/00Isolation 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
    • A61K2236/30Extraction of the material
    • A61K2236/39Complex extraction schemes, e.g. fractionation or repeated extraction steps
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61KPREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K2236/00Isolation 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
    • A61K2236/50Methods involving additional extraction steps
    • A61K2236/51Concentration or drying of the extract, e.g. Lyophilisation, freeze-drying or spray-drying

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  • Life Sciences & Earth Sciences (AREA)
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Abstract

The invention belongs to active constituents of medicine extraction and separation technology field, one kind is disclosed using proso millet prolamin as raw material, using ethanol extracting n-butyl alcohol, the method to prepare proso millet prolamin n-butanol extract.One of proso millet prolamin extract active ingredient that the present invention is prepared is flavone compound, and this compound structure is similar with wild scutellaria glycosides structure.The proso millet prolamin n-butanol extract prepared can effectively reduce the fasting blood-glucose of diabetes, put on weight, and alleviate many food diuresis of many drinks.Available for treatment/alleviation of the hyperglycaemia as caused by diabetes, weight loss and " more than three " symptom, available for reparation mouse hearing impairment.The present invention provides new approaches for hypoglycemic and protection hearing, the exploitation for the medicine for repairing hearing impairment.

Description

A kind of proso millet prolamin n-butanol extract and its preparation method and application
Technical field
The invention belongs to medicinal active ingredient extraction and separation technology and food, pharmaceutical product use technical field, it is related to Herba pogonatheri criniti The extracting and developing preparation method of certain kind of berries Flavonoid substances.The proso millet prolamin n-butanol extract prepared can reduce blood glucose, change Become thin caused by kind diabetes, improve diabetes " more than three " symptom, repair hearing impairment.
Background technology
Proso millet prolamin is rose family strawberry plants, and alias rust hair strawberry, yellowhairy strawberry herb are grown in height above sea level 700-3000m's Patana or limes marginis sylvan life, aboundresources, are distributed in the ground such as Yunnan Province of China, Guangxi, Sichuan, Guizhou, there is clearing heat and detoxicating, promoting blood circulation Effect of dissolving stasis, with preferable medical value.Diabetes be it is a kind of as caused by heredity or day after tomorrow defect of insulin secretion with The metabolic disease that carbohydrate metabolism disturbance and hyperglycaemia are characterized.Diabetic persistently raises along with blood glucose, and body weight drastically subtracts Gently, liver, kidney, pancreas dysfunction.In addition, the disorderly metabolism of diabetic often produces a large amount of oxygen radicals, cause other The generation of complication.Yet there are no proso millet prolamin has reduction blood glucose, improves and become thin caused by diabetes, improve diabetes " more than three " symptom, the report for repairing hearing impairment.
The content of the invention
Present invention firstly provides the extracting method of proso millet prolamin n-butanol extract, using proso millet prolamin as raw material, second is used Alcohol-extracting n-butyl alcohol, to prepare proso millet prolamin n-butanol extract.The method of the present invention is simple to operate, the yellow hair prepared Fragaia ananassa Duchesne extract has the effect of substantially reduction blood glucose.
The preparation method for the proso millet prolamin n-butanol extract that the present invention is provided, the described method comprises the following steps:
(1) shredded after proso millet prolamin is dried, using alcohol steep, obtain proso millet prolamin alcohol steep thing;
(2) by after the proso millet prolamin extract evaporation in step (1), supernatant is filtered to take, proso millet prolamin ethanol is obtained and slightly carries Thing;
(3) extracted after the proso millet prolamin alcohol extracts in step (2) are mixed with enough n-butanols, obtain proso millet prolamin N-butanol extract;
(4) the proso millet prolamin n-butanol extract in step (3) is evaporated, after n-butanol volatilization is clean, obtains yellow hair Used after strawberry n-butanol extract, dilution.
In step (1), the temperature of the drying is 25-37 DEG C;Preferably, it is 32 DEG C.
In step (1), the concentration of alcohol is 50%-95% (volume fraction);Preferably, it is 75%.
In step (1), the temperature of the extraction is 18-32 DEG C;Preferably, it is 25 DEG C (room temperatures).
In step (1), the time of the extraction is 2-4h;Preferably, it is 3h.
In step (1), the amount ratio of the proso millet prolamin and ethanol is (6-20) g:(300-600)ml;Preferably, it is 6g:400ml.
In step (1), the proso millet prolamin includes root, stem, leaf or its mixture of yellow hair proso millet prolamin plant.
In step (2), the evaporation can be using Rotary Evaporators evaporation.
In step (2), the time of the evaporation is 1-2 hours;Preferably, it is 1.5 hours.
In step (2), the evaporating temperature is 35 DEG C -45 DEG C;Preferably, it is 40 DEG C.
In step (2), the crude drug concentration of the yellow hair proso millet prolamin alcohol extracts thing can be 0.5g/ml;Wherein, institute State crude drug concentration and refer to evaporation gained original liquid concentration.
In step (2), preferably under vacuo under the conditions of carry out.
In step (3), the consumption of n-butanol is 15~20 times (V/V) of proso millet prolamin alcohol extracts.
In step (3), the temperature of the extraction is 18-32 DEG C;Preferably, it is 25 DEG C (room temperatures).
In step (3), the time of the extraction is 2-4h;Preferably, it is 3h.
In step (3), the n-butanol can be substituted by ethyl acetate.
In step (4), the evaporation can be using Rotary Evaporators evaporation.
In step (4), preferably under vacuo under the conditions of carry out.
In step (4), the temperature of the evaporation is 50 DEG C -65 DEG C;Preferably, it is 60 DEG C.
In step (4), the time of the evaporation is 1-2 hours;Preferably, it is 1.5 hours.
In step (4), it is generally the case that when the volume of remaining proso millet prolamin n-butanol extract in Rotary Evaporators and Huang When hair strawberry alcohol extracts volume is identical, n-butanol is that volatilization is complete.
In step (4), the concentration of the dilution can be:0.4g/ml and 0.8g/ml etc..
In step (4), the active ingredient of the yellow hair proso millet prolamin n-butanol extract is Flavonoid substances, structure and open country Scutellaria glycosides is similar.
After step (4) terminates, step (5) can also be included:N-butanol extracting liquid in step (4) is subjected to freezing dry It is dry, obtain proso millet prolamin extract dry powder.
In the present invention, Flavonoid substances are soluble in ethanol and n-butanol, are dissolved in after ethanol, and ethanol evaporation obtains Huang Letones and it is other dissolve in second alcohols material, further extracted using n-butanol, can will be partially soluble in ethanol but not dissolve in N-butanol debris is removed.
Present invention also offers a kind of proso millet prolamin n-butanol extract or its dry powder, by being carried to proso millet prolamin n-butanol Thing or its dry powder is taken to carry out Active Components, the main component of the proso millet prolamin extract is similar with wild scutellaria glycosides structure Flavone compound.
Present invention also offers a kind of proso millet prolamin n-butanol extract or its dry powder prepared by the above method, institute The main component for stating proso millet prolamin n-butanol extract or its dry powder is flavone compound.Wherein, wild scutellaria glycosides structure is as follows Shown in formula (I):
Present invention also offers a kind of identification of proso millet prolamin n-butanol extract active ingredient and separation method, including with Lower step:
(a) the proso millet prolamin extract dry powder for preparing above method step (5) is dissolved in alcohol, adds corresponding reagent and determines Absorbance is surveyed after appearance, reference standard determines dry powder general flavone content.
(b) proso millet prolamin flavones semifinished product is dissolved in methanol in step (a), and its species is identified using thin layer.
(c) proso millet prolamin flavones semifinished product HPLC is analyzed and is collected active ingredient in step (5)
In step (a), the dry powder is dissolved in 60% ethanol, mixes that (volume ratio is 500 with NaNO2 (5%):1), After 6min, 1ml 10%AlCl3 and 10mLNaOH (1M) are added, mixture is finally settled to 25ml with 60% ethanol, shaken up, Place 15min.
In step (a), the absorbance is determined at 500nm wavelength.
In step (a), the standard items are rutin.
In step (c), the HPLC chromatogram post is:Eclipse XDB-C18 (4.6 × 250mm, 5mm), mobile phase:First The phosphoric acid solution of alcohol -0.06% (50:50), flow velocity:1ml/min, Detection wavelength:360nm, column temperature:25 DEG C, sample size:10ml.
In step (c), the standard items are wild scutellaria glycosides.
In step (c), the sample and standard items after filtering with microporous membrane 2 times with using.
Hypoglycemic related food/health products and medicine are being prepared the invention provides the proso millet prolamin n-butanol extract Application in thing.
Cause the food/guarantor become thin in preparation improvement diabetes the invention provides the proso millet prolamin n-butanol extract Application in strong product and medicine.
The invention provides the proso millet prolamin n-butanol extract prepare improve diabetes " more than three " symptom food/ Application in health products and medicine.Referring to many drinks, many foods, diuresis described three more.
The invention provides the proso millet prolamin n-butanol extract in the food for preparing protection hearing, repairing hearing impairment Application in product/health products and medicine.
The beneficial effects of the present invention are the method that the present invention prepares proso millet prolamin n-butanol extract is simple to operate, obtains To the important component of proso millet prolamin extract be flavone compound, its structure is similar to wild scutellaria glycosides.The inventive method system The blood glucose in diabetic mice rise tool that standby proso millet prolamin extract is induced STZ is significantly reduced effect, and its concentration is higher, Reduction effect is more obvious.The inventive method prepare proso millet prolamin extract STZ induce diabetic mice body weight mitigation, it is many " more than three " symptom of many drink diuresis of food has significant mitigation, and its concentration is higher, and reduction effect is more obvious.The inventive method system Standby proso millet prolamin extract has significant restitution to hearing impairment mouse hearing ability, and its concentration is higher, and reduction is made With more obvious.
Brief description of the drawings
Fig. 1 is influence of the proso millet prolamin n-butanol extract of embodiment 3 to STZ mouse fasting blood sugars.
Fig. 2 be the proso millet prolamin n-butanol extract of embodiment 4 to STZ mouse weights, drinking-water, food ration influence;A is Huang Influence of the hair strawberry n-butanol extract to STZ mouse amounts of drinking water;B is proso millet prolamin n-butanol extract to STZ mouse weights Influence;C is influence of the proso millet prolamin n-butanol extract to STZ mouse weight food rations.
Fig. 3 is the rutin standard curve of embodiment 5.
Fig. 4 is thin-layer chromatogram;Wherein, 1. Quercetin, 2. wild scutellaria glycosides, 3. Nobiletins, 4. sample liquids.
Fig. 5 is the HPLC result figures of wild scutellaria glycosides and proso millet prolamin n-butanol extract;A is wild scutellaria glycosides, and B is Herba pogonatheri criniti Certain kind of berries n-butanol extract.
Embodiment
With reference to specific examples below and accompanying drawing, the present invention is described in further detail.The process of the implementation present invention, Condition, experimental method etc., are the universal knowledege and common knowledge of this area, this hair in addition to the following content specially referred to It is bright that content is not particularly limited.
The preparation of the proso millet prolamin n-butanol extract of embodiment 1
Proso millet prolamin is dried into complete stool (including root, stem, leaf) chopping, 75% ethanol 400ml was extracted after 3 hours at room temperature, It is fully transferred in Rotary Evaporators, boiling 1.5h under vacuum filters to take supernatant, the Huang that crude drug concentration is 0.5g/ml is made Hair strawberry alcohol extracts.
Room temperature extracts 3h after proso millet prolamin alcohol extracts are mixed with enough n-butanols, is transferred in Rotary Evaporators, Boiling under 60 DEG C of vacuum, n-butanol is volatilized net, obtains proso millet prolamin n-butanol extract.
Freeze drier is transferred to after proso millet prolamin n-butanol extract is freezed, proso millet prolamin flavones dry powder is obtained.
Diabetic mice (STZ mouse) model construction of embodiment 2STZ inductions
Mice by intraperitoneal injection Streptozotocin (STZ) sets up type 1 diabetes model.Press 10mlkg-1Volume injection, Normal group injects pH4.6 sodium citrate cushioning liquid, and model group presses 40mgkg-1Dosage injects 0.4%STZ- citric acids Sodium solution, 1 time/d, continuously injects 5d, model stability after STZ is injected 14 days, continuous fasting measures fasting blood sugar > in 6 hours 12mmol·L-1It is considered as modeling success, is then discarded less than this value.
Influence of the proso millet prolamin n-butanol extract of embodiment 3 to STZ mouse fasting blood-glucoses
Each group mouse presses 10mLkg-1Volume gavage, normal group and model group gavage distilled water, low dose group, high agent Amount group presses 0.4gmL respectively-1With 0.8g/ml gavage proso millet prolamin n-butanol extracts, 1 time/d, totally 4 weeks.Mouse is determined weekly Tail vein blood after blood glucose, fasting 6h, determines fasting blood-glucose.
Experimental result as shown in figure 1, proso millet prolamin n-butanol extract can significantly reduce diabetic mice fasting blood sugar, And concentration more high effect is more obvious.As a result show that proso millet prolamin n-butanol extract has the effect of reduction fasting blood-glucose.
The proso millet prolamin n-butanol extract of embodiment 4 to diabetic mice body weight, ingest, the influence of amount of drinking water
As described in Example 3 handle after, respectively determine mouse body weight, ingest, amount of drinking water;Experimental result such as Fig. 2 institutes Show, proso millet prolamin n-butanol crude extract can effectively improve the symptom of diabetic mice body weight mitigation, hence it is evident that improve its " more than three " disease Shape.As a result illustrate that proso millet prolamin extract has and increase diabetic mice body weight, improve the effect of its many many food of drink
The constituent analysis of the proso millet prolamin n-butanol extract dry powder of embodiment 5
The proso millet prolamin extract dry powder that precision weighs the preparation of embodiment 1 is dissolved in 60% alcohol, takes 500mL and 1mL NaNO2 (5%) is mixed, after 6min, 1mL 10%AlCl3 and 10mLNaOH (1M) is added, finally by 60% ethanol of mixture 25ml is settled to, is shaken up, 15min is placed, absorbance, using 60% ethanol as blank control group, rutin are measured at 500nm wavelength For reference standard, dry powder general flavone content is demarcated with rutin standard curve.Fig. 3 is rutin standard curve, y=3.9064x+ 0.0021, R2=0.9991.
Proso millet prolamin n-butanol extract dry powder is mixed with vanillic aldehyde glacial acetic acid (5%, w/v, 0.5mL), 1mL perchloric acid, 60 DEG C are incubated after 10min, ice bath 15min, then add 5mL glacial acetic acid and mix, extinction is read under 538nm wavelength after 6min Degree.Using ursolic acid as standard items, dry powder total triterpene contentses are determined according to its standard curve.
As shown in table 1, the main component of proso millet prolamin n-butanol extract dry powder is flavone compound to experimental result.
The effective sugar-lowering components detection of the proso millet prolamin n-butanol extract dry powder of table 1.
Flavones Identification of Species in the proso millet prolamin n-butanol extract of embodiment 6
1) flavones species is determined using chromogenic reaction.
The chromogenic reaction of sample liquid:Proso millet prolamin n-butanol extract prepared by 2ml embodiments 1 is taken in test tube, in ripple Observed under long 254nm uviol lamps, record color.A little sample liquid is dipped with filter paper, further determining that color with glass bar again And record.
Ammoniacal liquor reacts:2ml sample liquids and 2ml ammoniacal liquor are taken in test tube, is mixed, is observed under 254nm uviol lamps.Walk below The rapid chromogenic reaction with sample liquid.
Sodium hydroxide reacts:2ml sample liquids and 2ml NaOH solutions are taken in test tube, is mixed, is observed under 254nm lamps. Chromogenic reaction of the following steps with sample liquid.
Alchlor reacts:The alchlor ethanol of 2ml sample liquids and 2ml 1% is taken, is mixed, is observed at 254nm. Chromogenic reaction of the following steps with sample liquid.
2) thin layer chromatography analysis flavones species is used
2mg/ml proso millet prolamin n-butanol extract, wild scutellaria glycosides, Nobiletin, rutin, Quercetin is prepared with methanol Reference substance solution is standby.From different exhibition coating systems, such as table 2.Distinguish pipette samples and reference substance solution with capillary, be added drop-wise to On silica gel plate, hair-dryer drying, repeatedly point sample three times.Silica gel plate is positioned in the exhibition layer cylinder containing developing agent after point sample, Yu Zi Observe, record under External Observation instrument, calculate Rf values.
Rf=X/L
Formula (II)
In formula (II), X is the flavone compound displacement (cm) isolated;L is the distance (cm) that solvent is moved.
Thin-layer chromatography development system is as shown in table 2;Thin-layer chromatography result is as shown in figure 4, wherein, 1. Quercetins, 2. open countries are yellow Cen glycosides, 3. Nobiletins, 4. sample liquids;As seen from Figure 4, proso millet prolamin n-butanol extract of the invention and wild scutellaria glycosides Displacement is the most close, can be learnt by calculating its Rf value, proso millet prolamin n-butanol extract and wild scutellaria glycosides structure phase Seemingly, table 4 further illustrates that both structures are similar.
As shown in table 3, flavonoid species may in proso millet prolamin n-butanol extract for chromogenic reaction experimental result It is flavones, flavonols or isoflavones.
Thin-layer chromatography experimental result is as shown in table 4, identical with wild scutellaria glycosides when sample liquid Rf values are 0.35, and is shone ultraviolet Penetrate lower presentation yellow-green fluorescence, therefore contain flavones and flavonoid drugs in sample, and structure is identical with open country scutellaria glycosides or phase Seemingly.
The thin-layer chromatography development system of table 2.
The chromogenic reaction of the flavone compound of table 3.
The Rf values of the flavone compound of table 4.
Flavones species in the high-efficient liquid phase chromatogram technique analysis proso millet prolamin n-butanol extract of embodiment 7
High performance liquid chromatography parameter is:Chromatographic column:Eclipse XDB-C18 (4.6 × 250mm, 5 μm), mobile phase:First The phosphoric acid solution of alcohol -0.06% (50:50), flow velocity:1ml/min, Detection wavelength:360nm, column temperature:25 DEG C, sample size:10μl.Will Sample and standard items after filtering with microporous membrane 2 times with using.
Using wild scutellaria glycosides as standard items, proso millet prolamin n-butanol extract prepared by embodiment 1 is analyzed.
Experimental result is as shown in figure 5, wherein A is wild scutellaria glycosides result figure, and B is proso millet prolamin n-butanol extract result Figure.Baseline is steady at 360 nm, and wild scutellaria glycosides retention time 5.296min, sample liquid has two peaks in 5.258,5.727min, Close with wild scutellaria glycosides retention time, both total peak area ratios are 64.5%, account for crude extract main component.Show Herba pogonatheri criniti Contain the material similar with wild scutellaria glycosides structure in certain kind of berries n-butanol extract, collection 5-6min is washed under the conditions of can collecting this separation Go out liquid, be the effective sugar-lowering components of proso millet prolamin.Open country scutellaria glycosides is hypoglycemic at present and protection liver effect does not have been reported that also.
The structure of the hearing impairment mouse model of embodiment 8
After 50 kunming mice adaptability are raised one week, random selection 10 is only used as normal group, remaining mouse exposed to 4~ In 45kHZ, 110dBSPL white noise, 9h is exposed daily, 10d is exposed altogether, auricle reflex, i.e. auricle hold up to adapt to glouglou After the phenomenon of sound is eliminated, model group, flunarizine hydrochloride group, low dose group, high dose group are randomly divided into.
Influence of the proso millet prolamin n-butanol extract of embodiment 9 to mouse hearing ability
Each group mouse presses 10mLkg-1Volume gavage, normal group and model group gavage distilled water, flunarizine hydrochloride group Gavage 1.5X 10-3g·kg-1, low dose group and high dose group gavage 0.4gkg-1And 0.8gkg-1Huang prepared by embodiment 1 Hair strawberry 1 time/d of n-butanol extract, totally 4 weeks.After gavage terminates, each group Cochlea of Mouse electrograph (ECochG), listening property brain stem are surveyed React (ABR).
Mouse peritoneal injection 4mg/kg sodium phenobarbital liquid is anaesthetized, using the multi-functional evoked potentuial measuring systems of YSSD-1000, Stimulation sound is shortwave, and sound pressure level 90dB, sensitivity 6.2uv, band logical 100~1000HZ of frequency carry out ABR and ECochG experiments. Record phase and complex action potential (CAP) amplitude variations between mouse ears each ripple incubation period, ripple.
Influence result to CAP amplitudes is as shown in table 5, compared with model group, (low dose of proso millet prolamin n-butanol extract group Amount group and high dose group) CAP amplitudes substantially rise.As a result illustrate that proso millet prolamin n-butanol extract can significantly recover mouse hearing Damage, and concentration more high effect is more obvious.
Influence result to the phase between incubation period and ripple is as shown in table 6, compared with model group, proso millet prolamin n-butanol extract Group (low dose group and high dose group) between I, III ripple incubation period and I-IV, I-V ripple the phase have notable recovery.Illustrate that embodiment 1 is made Standby proso millet prolamin n-butanol extract can be obviously improved auditory center function.As a result proso millet prolamin n-butanol extract pair is shown Noise hearing loss has therapeutic action.
The proso millet prolamin n-butanol extract of 5. embodiment of table 1 to CAP amplitudes influence (N=10)
Note:Compared with model group,*P < 0.05,*P < 0.01
The proso millet prolamin n-butanol extract of 6. embodiment of table 1 to the phase between incubation period and ripple influence (N=10)
Note:Compared with model group,*P < 0.05,*P < 0.01
The protection content of the present invention is not limited to above example.Under the spirit and scope without departing substantially from inventive concept, this Art personnel it is conceivable that change and advantage be all included in the present invention, and using appended claims as protect Protect scope.

Claims (10)

1. a kind of preparation method of proso millet prolamin n-butanol extract, it is characterised in that the described method comprises the following steps:
(1) shredded after the whole strain of proso millet prolamin is dried, using alcohol steep, obtain proso millet prolamin alcohol steep thing;
(2) by after the proso millet prolamin extract evaporation in step (1), supernatant is filtered to take, proso millet prolamin alcohol extracts are obtained;
(3) extracted after the proso millet prolamin alcohol extracts in step (2) are mixed with enough n-butanols, obtain the positive fourth of proso millet prolamin Alcoholic extract;
(4) the proso millet prolamin n-butanol extract in step (3) is evaporated, obtains proso millet prolamin n-butanol extracting liquid.
2. method as claimed in claim 1, it is characterised in that in step (1), the concentration of the ethanol is 50%-95%;Institute Dry temperature is stated for 25-37 DEG C;The temperature of the extraction is 18-32 DEG C, and the time of the extraction is 2-4h.
3. method as claimed in claim 1, it is characterised in that in step (1), the amount ratio of the proso millet prolamin and ethanol is (6-20)g:(300-600)ml.
4. method as claimed in claim 1, it is characterised in that in step (2), the temperature of the evaporation is 35 DEG C -45 DEG C;Institute The time for stating evaporation is 1-2 hours;In step (3), the consumption of n-butanol is 15~20 times of (V/ of proso millet prolamin alcohol extracts V)。
5. method as claimed in claim 1, it is characterised in that in step (3), the temperature of the extraction is 18-32 DEG C;It is described The time of extraction is 2-4h;In step (4), the temperature of the evaporation is 50 DEG C -65 DEG C;The time of the evaporation is 1-2 hours.
6. the proso millet prolamin n-butanol extract that a kind of any one methods described such as claim 1-5 is prepared.
7. proso millet prolamin n-butanol extract as claimed in claim 6 is in hypoglycemic food/health products and medicine are prepared Application.
8. proso millet prolamin n-butanol extract as claimed in claim 6 causes the food/guarantor become thin in preparation improvement diabetes Application in strong product and medicine.
9. proso millet prolamin n-butanol extract as claimed in claim 6 prepare improve diabetes " more than three " symptom food/ Application in health products and medicine.
10. proso millet prolamin n-butanol extract as claimed in claim 6 is in the food for preparing protection hearing, repairing hearing impairment Application in product/health products and medicine.
CN201710357078.2A 2017-05-19 2017-05-19 A kind of proso millet prolamin n-butanol extract and its preparation method and application Pending CN107174605A (en)

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Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN111803546A (en) * 2017-05-19 2020-10-23 华东师范大学 Application of composition of strawberries with centella asiatica

Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN111803546A (en) * 2017-05-19 2020-10-23 华东师范大学 Application of composition of strawberries with centella asiatica

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Application publication date: 20170919