CN102793754A - Gordon euryale seed extract with antioxidation and anti-ageing effects as well as preparation method and application thereof - Google Patents
Gordon euryale seed extract with antioxidation and anti-ageing effects as well as preparation method and application thereof Download PDFInfo
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Abstract
The invention discloses a gordon euryale seed extract with antioxidation and anti-ageing effects as well as a preparation method and application thereof. Through system optimization of an extraction solvent, an extracting method, an extraction method and a macroporous resin separation method, an optimal preparation process is obtained to prepare the gordon euryale seed extract. An experimental result shows that the gordon euryale seed extract provided by the invention is capable of obviously improving the motion capability and the exploratory behavior of a subacute ageing mouse in an open field, the academic record and memory ability of the mouse in water maze tests and the spleen and kidney indexes of the mouse, increasing the activity and protein contents of GSH-Px (Glutathione Peroxidase) and NOS (Nitric Oxide Synthase) in brain tissues, reducing the activity of AChE (acetylcholinesterase) in the brain tissues, improving the pathological change of the brain tissues, and having excellent antioxidation and anti-ageing effects.
Description
Technical field
The present invention relates to a kind of Chinese medicine extraction method, be specifically related to a kind of Semen euryales extract with antioxidation, delaying senility function.
Background technology
Semen Euryales beginning is stated from Shennong's Herbal, classifies as top gradely, has another name called Caput Gallus domesticus; " Chinese pharmacopoeia is all on the books, and the dry mature kernal for the annual aquatic herbaceous plant of Nymphaeceae Gorgon euryale has supplementing the kidney to control the nocturnal to go through version; Spleen reinforcing antidiarrheal, the effect of damp eliminating leukorrhagia stopping cures mainly nocturnal emission, spermatorrhea; The enuresis, frequent micturition, insufficiency of the spleen chronic diarrhea, nebulousurine, leukorrhagia have the good reputation of " Radix Ginseng in the water " and " Arillus Longan in the water ", are traditional Chinese medicines material and precious natural tonic.
China is one of country with the fastest developing speed of aging in the world, also is one of maximum country of the aged.1% population sampling in the whole nation will show that over-65s population accounts for 7.7% of total population number, predicts the year two thousand twenty over-65s aging population and will reach 11.92% in the end of the year 2005.
Aging is a kind of biosis of complicacy, and each item function of body all is tending towards descending when old and feeble, shows as the aging and dystropy of a series of bodies usually, as alopecia, cutis laxa, senile plaque pile up, the thinking and judgment ability weakens even dementia etc.Therefore, the aging and behavioristics of body investigates and is used as the successful sign of aging model preparation usually.
Relevant at present old and feeble theory is a lot, comprises free radical theory, genetic program theory, lipofuscin accumulation theory, hypoendocrinism theory etc., and wherein radical damage occupies crucial status in old and feeble Study on Mechanism.GSH-Px is a kind of antioxidative important enzyme of body, it specifically catalytic reduction type glutathion (GSH) reduction reaction of peroxide its role is to block the infringement of the radical pair body that is caused by peroxide, the protection cell membrane is avoided peroxide injury.Nitric oxide synthetase (NOS) can generate by catalyzing N O in vivo, and NO is a kind of neurotransmitter, also is a kind of extremely unsettled strong oxidizer, but cell-cytotoxic reaction can take place too much NO deposition in the brain, and this possibly express relevant with the mistake that induces type NOS.Acetylcholinesterase (AChE) is that the variation of AChE content can embody the functional activity of cholinergic neuron to a certain extent at the enzyme of the synapse place hydrolysis neurotransmitter acetylcholine (ACh) of cholinergic neuron.
Chinese medicine is in the advantage that has independent body aspect antioxidation, the activity of fighting against senium, therefore from Chinese herbal medicine, seeks pure natural, antioxidation, material that activity of fighting against senium is strong has great importance.
Summary of the invention:
Goal of the invention: the objective of the invention is in order to solve the deficiency of prior art; Extraction separation obtains antioxidation from the Chinese medicine Semen Euryales, activity of fighting against senium is strong; The extract that untoward reaction is low, another object of the present invention provide the method for preparing and its application of Semen euryales extract.
Technical scheme: in order to realize above purpose, the Semen euryales extract with antioxidation, delaying senility function provided by the invention, it is to prepare through following method:
(1) gets the Semen Euryales medical material, clean, remove impurity, put in the multi-function extractor, add 4 to 8 times of amount concentration and be 80% to 90% ethanol; Extract 1 to 3 time, extracted 6 to 8 hours at every turn, merge extractive liquid, filters, and must filtrate; Concentrating under reduced pressure, evaporate to dryness gets the Semen Euryales ethanol extraction, and is subsequent use.
(2) get the Semen Euryales ethanol extraction that above-mentioned steps (1) prepares, add the water suspendible after, with ethyl acetate or n-butanol extraction 3 to 6 times, ethyl acetate or butanol extraction liquid, concentrating under reduced pressure, evaporate to dryness, the ethyl acetate or the n-butanol extract of Semen Euryales, subsequent use;
(3) get macroporous resin column on ethyl acetate that step (2) prepares or the n-butanol extract; Water and concentration are 10% ethanol elution respectively earlier; Reuse concentration is 50%～60% ethanol elution; Thin layer chromatography inspection is known, is collected 50%～60% ethanol elution, concentrates, drying under reduced pressure obtains containing 5,7,4 '-Semen euryales extract of trihydroxy flavanone.
As preferred version, in the above-described Semen euryales extract with antioxidation, delaying senility function 5,7,4 '-percentage composition of trihydroxy flavanone is 10% to 20%.
As preferred version, above-described Semen euryales extract with antioxidation, delaying senility function, the described method for distilling of its step (1) is a reflux extraction; The concentration of the described extraction etoh solvent of step (1) is 80%.The present invention through a large amount of experiment screening method for distilling with extract solvent, experimental result shows, is that concentration is 80% ethanol when extracting solvent, at the reflux, extract, FAXIA, antioxidation in the Semen Euryales, activity of fighting against senium composition extraction efficiency are the highest.
Step of the present invention (1) obtains ethanol extraction; For further purification active component, the present invention extracts through preferred solvent, shows through a large amount of experiment screenings; Adopt ethyl acetate or n-butyl alcohol to extract and can the antioxidation in the Semen Euryales ethanol extraction, activity of fighting against senium composition be isolated to greatest extent; Impurity is removed, thereby guarantee drug effect, at utmost reduce untoward reaction.
Method for preparing with Semen euryales extract of antioxidation, delaying senility function provided by the invention, it may further comprise the steps:
(1) gets the Semen Euryales medical material, clean, remove impurity, put in the multi-function extractor, add 4 to 8 times of amount concentration and be 80% to 90% ethanol; Extract 1 to 3 time, extracted 6 to 8 hours at every turn, merge extractive liquid, filters, and must filtrate; Concentrating under reduced pressure, evaporate to dryness gets the Semen Euryales ethanol extraction, and is subsequent use.
(2) get the Semen Euryales ethanol extraction that above-mentioned steps (1) prepares, add the water suspendible after, with ethyl acetate or n-butanol extraction 3 to 6 times, ethyl acetate or butanol extraction liquid, concentrating under reduced pressure, evaporate to dryness, the ethyl acetate or the n-butanol extract of Semen Euryales, subsequent use;
(3) get macroporous resin column on ethyl acetate that step (2) prepares or the n-butanol extract; Water and concentration are 10% ethanol elution respectively earlier; Reuse concentration is 50%～60% ethanol elution; Thin layer chromatography inspection is known, is collected 50%～60% ethanol elution, concentrates, drying under reduced pressure obtains containing 5,7,4 '-Semen euryales extract of trihydroxy flavanone.
As preferred version, above-described method for preparing with Semen euryales extract of antioxidation, delaying senility function, the described method for distilling of step (1) is that reflux extraction, the concentration of extracting etoh solvent are 80%.
As preferred version, above-described method for preparing with Semen euryales extract of antioxidation, delaying senility function, the described macroporous adsorbent resin of step (3) is a D101 type macroporous adsorbent resin.The present invention is through the macroporous resin of orthogonal experiment screening opposed polarity different model, the ethanol elution agent of variable concentrations (10%, 20%, 30%, 40%, 50%, 60%, 70%); With antioxidation, defying age is the evaluating drug effect index; Experimental result shows with nonpolar macroporous adsorption resin D101 type macroporous adsorbent resin impurity-eliminating effect relatively good; And experimental result shows that concentration is that 10% ethanol can the big slightly impurity of the most of polarity of eluting; And that concentration is 50% to 60% ethanol is best to the eluting power of antioxidation, activity of fighting against senium composition, thus the present invention to determine best adsorbing material through the orthogonal experiment The selection result be that D101 type macroporous adsorbent resin is that concentration is 50% to 60% ethanol with best eluant.
As preferred version, the method for preparing of above-described Semen euryales extract with antioxidation, delaying senility function is separated the Semen euryales extract that obtains, and detection architecture shows in the extract 5,7,4 '-percentage composition of trihydroxy flavanone is 10% to 20%.
Semen euryales extract The pharmacological results with antioxidation, delaying senility function provided by the invention shows the effect with good antioxidation, slow down aging, can be used to prepare antioxidation, antisenescence health product or be used to prepare ischemia resisting property damage disease medicine.
Beneficial effect: the Semen euryales extract with antioxidation, delaying senility function provided by the invention is compared with prior art and is had the following advantages:
1, the present invention is an evaluation index with antioxidation, defying age, optimizes best method for preparing through a large amount of experiments, separates the Semen euryales extract that obtains; Active component content is high; Impurity is few, and drug effect is obvious, in the experimental result confirmation of spacious field, Morris water maze; Semen euryales extract with antioxidation, delaying senility function provided by the invention has good improvement effect to the ability of learning and memory of subacute mouse aging; To spleen, the effect of being significantly improved of kidney device index of subacute mouse aging, and can improve subacute mouse aging cerebral tissue NOS, GSH-Px vigor, reduce the AChE vigor.
2, Semen euryales extract provided by the invention can improve the vigor of subacute mouse aging cerebral tissue GSH-Px, and with the increasing of dosage, enzymatic activity has the trend of increase, has demonstrated good antioxidation, removes free radical and anti-aging effects.
3, experimental result shows that Semen euryales extract provided by the invention can reduce the vigor of AChE in the subacute mouse aging cerebral tissue, thereby can improve the content of ACh in the mouse brain, has to improve the effect that the old and feeble learning and memory function that causes descends.
4, the D-galactose causes the aging model experimental result and shows; Semen euryales extract provided by the invention can obviously suppress the apoptosis of hippocampal neurons; Even meet or exceed normal level, further can improve the D-galactose and cause the mouse aging ability of learning and memory from pathologic angle proof Semen euryales extract.
Semen euryales extract provided by the invention, active strong, drug effect is comprehensive, and untoward reaction is low, compares with prior art and has obtained good technical effect.
The specific embodiment
According to following embodiment, can understand the present invention better.Yet, those skilled in the art will readily understand that the described concrete material proportion of embodiment, process conditions and result thereof only are used to explain the present invention, and the present invention that should also can not limit in claims to be described in detail.
Embodiment 1
A kind of Semen euryales extract with antioxidation, delaying senility function, it prepares through following method:
(1) get the Semen Euryales medical material, clean, remove impurity, put in the multi-function extractor, add 8 times of amount concentration and be 80% ethanol, reflux, extract, 3 times was extracted 8 hours at every turn, and merge extractive liquid, filters, must filtrate, concentrating under reduced pressure, evaporate to dryness, the Semen Euryales ethanol extraction, subsequent use.
(2) get the Semen Euryales ethanol extraction that above-mentioned steps (1) prepares, add the water suspendible after, with the ethyl acetate extraction of equal volume 6 times, acetic acid ethyl acetate extract, concentrating under reduced pressure, evaporate to dryness, the ethyl acetate extract of Semen Euryales, subsequent use;
(3) get D101 type macroporous adsorbent resin on the ethyl acetate extract that step (2) prepares; Water and concentration are 10% ethanol elution respectively earlier; Reuse concentration is 50%～60% ethanol elution; Thin layer chromatography inspection is known, is collected 50%～60% ethanol elution, concentrates, drying under reduced pressure obtains containing 10%5,7,4 '-Semen euryales extract of trihydroxy flavanone.
Embodiment 2
A kind of Semen euryales extract with antioxidation, delaying senility function, it prepares through following method:
(1) get the Semen Euryales medical material, clean, remove impurity, put in the multi-function extractor, add 6 times of amount concentration and be 80% ethanol, reflux, extract, 3 times was extracted 8 hours at every turn, and merge extractive liquid, filters, must filtrate, concentrating under reduced pressure, evaporate to dryness, the Semen Euryales ethanol extraction, subsequent use.
(2) get the Semen Euryales ethanol extraction that above-mentioned steps (1) prepares, add the water suspendible after, with the n-butanol extraction of equal volume 6 times, butanol extraction liquid, concentrating under reduced pressure, evaporate to dryness, the n-butanol extract of Semen Euryales, subsequent use;
(3) get D101 type macroporous adsorbent resin on the n-butanol extract that step (2) prepares; Water and concentration are 10% ethanol elution respectively earlier; Reuse concentration is 50%～60% ethanol elution; Thin layer chromatography inspection is known, is collected 50%～60% ethanol elution, concentrates, drying under reduced pressure obtains containing 20%5,7,4 '-Semen euryales extract of trihydroxy flavanone.
Embodiment 3 antioxidation, defying age experiment
1, experiment material
1.1 trial drug
Semen Euryales is purchased the green first Science and Technology Ltd. of Pueraria lobota in Beijing, lot number: 110901; The extract that the embodiment of the invention 1 and enforcement 2 prepare.
The D-galactose, the graceful biological company limited of last Hypon, lot number: 110523.
GSH-Px, NOS, AChE, Coomassie brilliant blue protein determination kit, Nanjing build up bio-engineering research institute, lot number: 20120416.
1.2 experimental animal
A cleaning level KM mice is provided by Jiangsu University's Experimental Animal Center, the animal quality certification number: SCXK (Soviet Union) 2009-0002:.
1.3 experimental apparatus
The spacious field device of mice, the soft grand development in science and technology company limited in Shanghai.
SLY-WMS type Morris water maze, the large hunting park in Beijing Science and Technology Ltd..
FSH-2A is adjustable high speed electric refiner, the Jin Nan of Jintan City instrument plant.
LXJ-II B low speed high capacity multitube centrifuge, Nanjing instrument and equipment company limited dawn dawn.
The long ELIASA of Powerwave X340 all-wave, U.S. Bio-Tek company.
2. experimental technique
2.1 extract is to the effect of the subacute aging model of D-galactose induced mice among the present invention
Get 104 of the male KM mices of body weight 18～22g; Be divided into 8 groups at random; Be normal control group, D-galactose model control group, Semen Euryales of the present invention 80% ethanol extraction high and low dose group, the embodiment of the invention 1 extract high and low dose group, the embodiment of the invention 2 extract high and low dose groups, every group of 15 mices are except that the normal control group; All the other respectively organize mice lumbar injection every day 0.5%D-galactose; 0.1ml/10g, injected for 6 weeks continuously, the physiological saline solution of normal control group lumbar injection equivalent; Begin administration simultaneously, the high and low dose group mice gastric infusion every day dosage of Semen Euryales 80% ethanol extraction, embodiment 1 extract and embodiment 2 extracts is respectively 0.5g/kg and 0.25g/kg; Normal control group and model control group are irritated the normal saline of stomach with volume every day.Successive administration is after 6 weeks, measures the autonomic activities situation of respectively organizing mice, the average running speed of 90 seconds inherent spacious center court districts of record mice and surrounding zone and stride the lattice number of times; Adopt water maze training learning and memory abilities in aging mice then, write down its school grade, after finding the incubation period of fixed platform in promptly 2 minutes and withdrawing platform, pass through the number of times of original platform place quadrant; The comparable group differences also carries out statistical procedures; Experiment is last, takes off cervical vertebra and puts to death mice, and the body weight of record mice is got spleen, kidney is weighed, and calculates organ index.After other gets the brain tissue homogenate of mice, the activity of the GSH-Px in the mensuration cerebral tissue, NOS, AChE, the comparable group differences also carries out statistical procedures.Comment the action effect of each extract among the present invention in conjunction with above-mentioned each item index.Separate mouse brain and organize hippocampus, paraffin section after HE dyeing, observation of cell dyeing situation under the optical microscope.
3. experimental result
3.1 extract is to the effect of the subacute aging model of D-galactose induced mice among the present invention
Continuously after 6 weeks of lumbar injection D-galactose, except that the normal control group, each organizes phenomenons such as the visible fur of mice naked eyes is matt and sparse, action is comparatively slow, lethargy is depressed, and table 1 is seen in the influence of the autonomic activities of mice; Table 2 is seen in influence to the mice water maze laboratory; Table 3 is seen in exponential influence to the mice organ; Assay to each item biochemical indicator in the mouse brain tissue is seen table 4.
Table 1 extract of the present invention is to the influence
of spacious experiment of subacute mouse aging
Compare with matched group,
*P＜0.05,
*P＜0.01; Compare with model group,
▲P＜0.05,
▲ ▲P＜0.01.
Experimental result by table 1 shows; Compare with matched group; The movement velocity of model group mice and total time number average of crossing over obviously reduce (P＜0.01); Compare with model group; The present invention implements the 1 Semen euryales extract high and low dose group group that provides with embodiment 2 all can make the movement velocity of mouse aging and total number of times of crossing over obviously increase (P＜0.01), and the result shows that the present invention implements 1 Semen euryales extract that provides with embodiment 2 has better raising mouse aging than ethanol extraction movement velocity and total effect of crossing over number of times.
Table 2 extract of the present invention is to the influence
of the water maze laboratory of subacute aging model mice
Compare with matched group,
*P＜0.05,
*P＜0.01; Compare with model group,
▲P＜0.05,
▲ ▲P＜0.01.
Experimental result by table 2 shows, there is some difference with passing through number of times incubation period for each experimental group, and the model group mice is compared with matched group, obviously increases (P＜0.01) incubation period, passes through number of times and reduces.The extract of the embodiment of the invention 1 and embodiment 2 is compared with model group, and all obviously reduce incubation period, and passes through number of times and obviously increase (P＜0.05; P＜0.01); And the extract of the embodiment of the invention 1 and embodiment 2 is compared in the Semen Euryales ethanol extraction, has better minimizing incubation period, increases the effect of passing through number of times; Further show to have better antidotal effect than ethanol extraction after extraction of Semen euryales extract process and the macroporous resin separation and purification.
Table 3 extract of the present invention is to subacute aging model mice spleen, the exponential influence of kidney device
Compare with matched group,
*P＜0.05,
*P＜0.01; Compare with model group,
▲P＜0.05,
▲ ▲P＜0.01.
Experimental result by table 3 can be known, long term injections D-galactose, and atrophy all takes place in spleen of mice and kidney; Compare with the normal control group and to have significant difference (P＜0.01); Point out subacute aging model to prepare successfully, the extract that the embodiment of the invention 1 and embodiment 2 prepare is along with the increase of dosage, and atrophy degree is eased; Compare in model group and to have significant difference (P＜0.01); And show that through contrast and experiment the extract effect that the embodiment of the invention 2 prepares is owing to embodiment 1 extract, embodiment 1 extract is because the Semen Euryales ethanol extraction; Further show the Semen euryales extract for preparing through the preferred purifying process of the present invention has better raising mouse aging than ethanol extraction spleen, the exponential effect of kidney device.
Table 4 extract of the present invention is organized the influence
of biochemical indicator to subacute aging model mouse brain
Annotate: compare with blank control group,
*P＜0.05,
*P＜0.01; Compare with model group,
▲P＜0.05,
▲ ▲P＜0.01.Experimental result by table 4 shows, compares with matched group, and the model group mouse brain organizes NOS, GSH-Px significance to descend; AChE obviously raises (P＜0.05), compares with model group, the Semen euryales extract NOS that enforcement 1 provided by the invention and enforcement 2 prepare, active raise (P＜0.05) of GSH-Px; The active significance of AChE reduces (P＜0.01); And can find out that by the experimental result contrast Semen euryales extract that the embodiment of the invention 2 provides is superior to embodiment 1, be superior to the Semen Euryales ethanol extraction.Experimental result shows that further the Semen euryales extract for preparing by method provided by the invention has more superior antioxidation, antidotal effect.
3.2 extract of the present invention is to the general pathological observation of the subacute aging model cerebral tissue of D-galactose induced mice
Get brain fast, blot with filter paper, 10% neutral formalin is fixed, and carries out routine paraffin wax embedded section (5 μ m), tissues observed section pathology variation and taking pictures under optical microscope after HE dyeing and the resin glue mounting.
The HE colouring method: paraffin section, thickness 5 μ m mount on clean slide, and roasting sheet is 2 hours on 60 ℃ of roasting sheet machines; Xylene dewaxing 2 times each 5 minutes, is handled to transparent Lignum Sappan uniformly dyeing nuclear born of the same parents 6 minutes successively with 100% ethanol, 95% ethanol, 80% ethanol, distilled water; 1% hydrochloride alcohol breaks up a moment, and tap water dashed 15 minutes, and 0.5% Yihong ethanol was contaminated 2 minutes; Tap water dashed 1 minute, through dewater, transparent after, use the resin glue mounting.
Tissue paraffin section de is after HE dyeing, and to be homogeneous light blue or blue for normal cell nuclear under the optical microscope, and endochylema is pale red, and after birth is complete continuously.During tissue abnormalities, cell rounding diminishes, and local engrain appears in karyon pyknosis, cracked, and is inhomogeneous, and perhaps nuclear disperse to disappear and loses nuclear staining, only see endochylema be homogenizing red dye the non-structure characteristic.
Normal group: hippocampus cell marshalling, level is tight, complete form, cell edges is clear, even dyeing, kernel engrain.
D-galactose model group: cell quantity is less, and intercellular substance increases, and arranges loose uneven; Most of entoblast understain, chromatin is inhomogeneous; The circular karyon volume of a small amount of neurocyte reduces, pyknosis, have in addition disappearance, nuclear membrane shrinkage; Cell space diminishes, endochylema concentrates red dying.
Semen Euryales 80% ethanol extraction high and low dose group: with model group and compared with normal, cell quantity increases, and most of cellular morphology is complete, and cell edges is more clear, and chromatin is more even, the kernel engrain.
The embodiment of the invention 1 and 2 Semen euryales extract high and low dose group: compare with model group, cell quantity increases, the cell marshalling, and level is tight, the kernel engrain.
The presentation of results embodiment of the invention 1 and 2 Semen euryales extract high and low dose group cause to the D-galactose to some extent all that nerve cell damage and apoptosis have good protective effect in the mouse aging cerebral tissue.
Above experimental result shows; The Semen euryales extract that the present invention prepares; Can obviously improve mobility and exploratory behavior and mice water maze in spacious the experiment of subacute mouse aging school grade and the memory ability in testing, and spleen, the kidney device index of mice obviously improved; And activity and the protein content of GSH-Px, NOS in the cerebral tissue that can raise; Reduce the activity of cerebral tissue AChE; Can also obviously improve brain tissue pathology change, have good antioxidation, antidotal effect.
Claims (9)
1. the Semen euryales extract with antioxidation, delaying senility function is characterized in that, it prepares through following method:
(1) gets the Semen Euryales medical material, clean, remove impurity, put in the multi-function extractor, add 4 to 8 times of amount concentration and be 80% to 90% ethanol; Extract 1 to 3 time, extracted 6 to 8 hours at every turn, merge extractive liquid, filters, and must filtrate; Concentrating under reduced pressure, evaporate to dryness gets the Semen Euryales ethanol extraction, and is subsequent use.
(2) get the Semen Euryales ethanol extraction that above-mentioned steps (1) prepares, add the water suspendible after, with ethyl acetate or n-butanol extraction 3 to 6 times, ethyl acetate or butanol extraction liquid, concentrating under reduced pressure, evaporate to dryness, the ethyl acetate or the n-butanol extract of Semen Euryales, subsequent use;
(3) get macroporous resin column on ethyl acetate that step (2) prepares or the n-butanol extract; Water and concentration are 10% ethanol elution respectively earlier; Reuse concentration is 50%～60% ethanol elution; Thin layer chromatography inspection is known, is collected 50%～60% ethanol elution, concentrates, drying under reduced pressure obtains containing 5,7,4 '-Semen euryales extract of trihydroxy flavanone.
2. the Semen euryales extract with antioxidation, delaying senility function according to claim 1 is characterized in that, the described method for distilling of step (1) is a reflux extraction.
3. the Semen euryales extract with antioxidation, delaying senility function according to claim 1 is characterized in that, the concentration of the described extraction etoh solvent of step (1) is 80%.
4. the described method for preparing with Semen euryales extract of antioxidation, delaying senility function of claim 1 is characterized in that, may further comprise the steps:
(1) gets the Semen Euryales medical material, clean, remove impurity, put in the multi-function extractor, add 4 to 8 times of amount concentration and be 80% to 90% ethanol; Extract 1 to 3 time, extracted 6 to 8 hours at every turn, merge extractive liquid, filters, and must filtrate; Concentrating under reduced pressure, evaporate to dryness gets the Semen Euryales ethanol extraction, and is subsequent use.
(2) get the Semen Euryales ethanol extraction that above-mentioned steps (1) prepares, add the water suspendible after, with ethyl acetate or n-butanol extraction 3 to 6 times, ethyl acetate or butanol extraction liquid, concentrating under reduced pressure, evaporate to dryness, the ethyl acetate or the n-butanol extract of Semen Euryales, subsequent use;
(3) get macroporous resin column on ethyl acetate that step (2) prepares or the n-butanol extract; Water and concentration are 10% ethanol elution respectively earlier; Reuse concentration is 50%～60% ethanol elution; Thin layer chromatography inspection is known, is collected 50%～60% ethanol elution, concentrates, drying under reduced pressure obtains containing 5,7,4 '-Semen euryales extract of trihydroxy flavanone.
5. the method for preparing with Semen euryales extract of antioxidation, delaying senility function according to claim 4 is characterized in that, the described method for distilling of step (1) is that reflux extraction, the concentration of extracting etoh solvent are 80%.
6. the method for preparing with Semen euryales extract of antioxidation, delaying senility function according to claim 4 is characterized in that, the described macroporous adsorbent resin of step (3) is a D101 type macroporous adsorbent resin.
7. the method for preparing with Semen euryales extract of antioxidation, delaying senility function according to claim 4 is characterized in that, in the described Semen euryales extract of step (3) 5,7,4 '-percentage composition of trihydroxy flavanone is 10 to 20%.
8. each described Semen euryales extract application in preparation antioxidation, antisenescence health product of claim 1 to 3 with antioxidation, delaying senility function.
9. each described Semen euryales extract application in preparation control ischemia injury disease medicament of claim 1 to 3 with antioxidation, delaying senility function.
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| CN113652434A (en) * | 2020-05-12 | 2021-11-16 | 南京中医药大学 | Gorgon fruit DNA molecule with effect of promoting rice grain enlargement and application thereof |
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| CN113652434A (en) * | 2020-05-12 | 2021-11-16 | 南京中医药大学 | Gorgon fruit DNA molecule with effect of promoting rice grain enlargement and application thereof |
| CN113652434B (en) * | 2020-05-12 | 2023-04-28 | 南京中医药大学 | Gorgon fruit DNA molecule with function of promoting rice grain enlargement and application thereof |
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