CN102512501A - Preparation method of compound Chinese magnoliavine fruit preparation and antioxidant activity thereof - Google Patents
Preparation method of compound Chinese magnoliavine fruit preparation and antioxidant activity thereof Download PDFInfo
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- CN102512501A CN102512501A CN2012100099516A CN201210009951A CN102512501A CN 102512501 A CN102512501 A CN 102512501A CN 2012100099516 A CN2012100099516 A CN 2012100099516A CN 201210009951 A CN201210009951 A CN 201210009951A CN 102512501 A CN102512501 A CN 102512501A
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- schisandrae chinensis
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Abstract
The invention belongs to the technical field of research and development of medical products, and relates to a preparation method for a compound Chinese magnoliavine fruit preparation and antioxidant activity thereof. The method comprises the following steps of: adding Chinese magnoliavine fruit and astragalus root into 70 percent ethanol to lixiviate at room temperature; performing ultrasonic processing for 30 minutes at the temperature of 30 DEG C and at the power of 80 W; filtering; repeatedly extracting filter residue by using 70 percent ethanol in an amount which is 6 times the amount of the filter residue and in an amount which is 4 times the amount of the filter residue respectively; filtering; combining filtrates; recovering the ethanol to obtain Chinese magnoliavine fruit and astragalus root extract; centrifuging porphyridium cruentum by using a centrifuge of 5,000r/min for 10 minutes; suspending in 50mmol/L phosphoric acid buffer solution with pH of 7.4; performing ultrasonic crushing; centrifuging by using a high-speed freezing centrifuge to obtain supernatant; freeze-drying to obtain a dry product; and uniformly mixing the dry product with the Chinese magnoliavine fruit and astragalus root extract to obtain the compound Chinese magnoliavine fruit preparation. The product has an antioxidant effect.
Description
Technical field
Medical product research and development technical field under the present invention.The present invention relates to a kind of method for preparing and antioxidant activity thereof of compound recipe Fructus Schisandrae Chinensis preparation.
Background technology
Fructus Schisandrae Chinensis is perennial fallen leaves liana; For magnoliaceae schisandra (Schisandra chinensis (Turcz.) Baill.) is practised title " Radix Schisandrae Bicoloris ", or schisandra chinensis (Schisandra sphenantheraRehd.etWils.) is practised the dry mature fruit of claiming " Fructus Schisandrae Sphenantherae ".Treasure once called: " there is the branch in north and south five tastes the present, and southern product person color is red, and northern product person darkish complexion, people's tonic must use the northern product person good ".It is two kinds that Chinese Pharmacopoeia version in 2000 is recorded Radix Schisandrae Bicoloris and Fructus Schisandrae Sphenantherae respectively.Fructus Schisandrae Chinensis only refers in particular to the dry mature fruit of Radix Schisandrae Bicoloris.And Fructus Schisandrae Sphenantherae refers in particular to the dry mature fruit of schisandra chinensis.Temperature is sour, sweet; Return lung, the heart, kidney channel.Mainly contain schizandrin (schizandrin), deoxyschisandrin (de-oxyschizandrin), newly plain (neoschizandrin), schisandrol (schizan-drol) of son, schizantherin (schisantherin simply; Gomisin), A, B, C, D, E, F, G, H, J, K1, K2, K3, L1, L2, M1, M2, N2; O, R etc.Effect is astringent or styptic treatment for spontaneous sweating for convergence, supplementing QI for promoting the production of body fluid, kidney calming.Be used for the chronic cough dyspnea due to deficiency, emission, enuresis frequent micturition, incessant chronic diarrhea, spontaneous perspiration, night sweat, Tianjin wound is thirsty, the deficient pulse of losing heart, interior-heat is quenched one's thirst, palpitation and insomnia.
The Radix Astragali is the root of the Radix Astagali Astragalus membranaceus of leguminous herbaceous plant (Fisch.) Bge.var.mongholicus (Bge.) Hsiao, Radix Astragali Astragalus membranaceus (Fisch.), has effects such as invigorating QI to consolidate the body surface resistance, diuretic detumescent, expelling pus and toxin by strengthening QI, granulation promoting.The Radix Astragali contains various trace elements such as saponin, sucrose, polysaccharide, several amino acids, folic acid and selenium, zinc, copper.The enhancing human body immunity function arranged, protect the liver, diuresis, defying age, anti-stress, blood pressure lowering and antibacterial action more widely.
Porphyridium cruentum (Porphyridium cruentum) is a kind of unicellular red algae that belongs to Rhodophyta, in growth course, can synthesize bioactive substances such as polysaccharide sulfate, phycobniliprotein and polyunsaturated fatty acid outside the born of the same parents in a large number.The Porphyridium cruentum phycoerythrin accounts for phycobniliprotein 84%, and is wherein maximum with B-phycoerythrin content.Phycoerythrin is of many uses; It both can be used as natural pigment and had been applied to industry such as food, cosmetics, dyestuff; Can be made into fluorometric reagent again and be used for clinical diagnose and fields such as immunochemistry and biological engineering; Phycoerythrin still is a kind of photosensitizer with potentiality to be exploited, is used for the optical dynamic therapy of tumor.The outer sulfated polysaccharides ntiviral characteristic of the born of the same parents of Porphyridium cruentum is outstanding, has functions such as antitumor, antiallergic, antioxidation, blood fat reducing, accent blood fat.Therefore, comprehensive exploitation Porphyridium cruentum phycoerythrin, polysaccharide and algae powder product series, economic results in society are obvious.
Human body can constantly produce various free radicals in the oxidative metabolism process of vital movement; Too much radical pair biomacromolecule causes damage in the body; Cause the 26S Proteasome Structure and Function of cell and tissue to go down, and the aging course of body is exerted an influence, finally cause the generation of disease.And the antioxidant of synthetic has certain toxicity and carcinogenesis.So estimate and screen and have the new trend that the active natural plants of strong anti-oxidation has become medical science, biology and food scientific research.Adopting typical antioxidation in vitro system, is contrast with the ascorbic acid, estimates the antioxidant activity of compound recipe Fructus Schisandrae Chinensis preparation, is the antioxidant activity product for better developing compound recipe Fructus Schisandrae Chinensis resource, and theoretical foundation is provided.
DPPH method: DPPH is 1, and 1-diphenyl-2-trinitrophenyl-hydrazine (1,1-Diphenyl-2-picrylhydrazyl radical2; 2-Diphenyl-1-(2; 4,6-trinitrophenyl) hydrazyl) the DPPH method was suggested in 1958, was widely used in the oxidation resistance of measuring food.This method is according to the DPPH free radical single electron to be arranged, and has the last one to absorb at the 517nm place, and its alcoholic solution is the characteristic of purple.When free radical scavenger existed, owing to its single electron pairing its absorption being faded away, its fading extent became quantitative relationship with the electron amount of its acceptance, thereby available spectrophotometer carries out fast quantitative analysis.
Summary of the invention
1. method for preparing with compound recipe Fructus Schisandrae Chinensis preparation of antioxidation is characterized in that comprising following process:
(1) weight proportion of crude drug is: Fructus Schisandrae Chinensis 9g-12g, Radix Astragali 6g-9g, Porphyridium cruentum 10g-20g;
(2) Fructus Schisandrae Chinensis and Radix Astragali adding 8-10 are doubly measured lixiviate 8-10h under the 70% ethanol room temperature, seal ultrasonic 30min again, temperature is 30 ℃; Power is 80W, uses filtered through gauze then, and filtering residue repeats to extract each once with the ethanol of 6 times of amounts and 4 times of amounts 70% respectively again; Filter, must filtrate;
(3) filtrating in (2) is merged, reclaim ethanol, obtain Fructus Schisandrae Chinensis, Radix Astragali extract;
(4) with Porphyridium cruentum with the centrifugal 10min of 5000r/min centrifuge, be suspended in pH7.4 then, in the phosphate buffer of 50mmol/L, carry out ultrasonication; Broken 5s, interval 5s, 20 times, with the centrifugal 15min of 10000r/min High speed refrigerated centrifuge; Get supernatant, lyophilization gets dry thing;
(5), get compound recipe Fructus Schisandrae Chinensis preparation with Fructus Schisandrae Chinensis, Radix Astragali extract mix homogeneously in the dry thing in (4) and (3).
2. compound recipe Fructus Schisandrae Chinensis preparation is made an experiment, said preparation has antioxidation
According to the present invention, " % " among the present invention is percentage by weight.
The specific embodiment
Method for preparing and the antioxidant activity thereof that relates to a kind of compound recipe Fructus Schisandrae Chinensis preparation of the present invention comprises following examples, and following embodiment can further specify the present invention, but does not limit the present invention in any way.
Embodiment 1:
1. method for preparing with compound recipe Fructus Schisandrae Chinensis preparation of antioxidation is characterized in that comprising following process:
(1) weight proportion of crude drug is: Fructus Schisandrae Chinensis 9g, Radix Astragali 6g, Porphyridium cruentum 10g;
(2) Fructus Schisandrae Chinensis and the Radix Astragali are added lixiviate 8 under 8 times of amount 70% ethanol room temperatures, seal ultrasonic 30min again, temperature is 30 ℃; Power is 80W, uses filtered through gauze then, and filtering residue repeats to extract each once with the ethanol of 6 times of amounts and 4 times of amounts 70% respectively again; Filter, must filtrate;
(3) filtrating in (2) is merged, reclaim ethanol, obtain Fructus Schisandrae Chinensis, Radix Astragali extract;
(4) with Porphyridium cruentum with the centrifugal 10min of 5000r/min centrifuge, be suspended in pH7.4 then, in the phosphate buffer of 50mmol/L, carry out ultrasonication; Broken 5s, interval 5s, 20 times, with the centrifugal 15min of 10000r/min High speed refrigerated centrifuge; Get supernatant, lyophilization gets dry thing;
(5), get compound recipe Fructus Schisandrae Chinensis preparation with Fructus Schisandrae Chinensis, Radix Astragali extract mix homogeneously in the dry thing in (4) and (3).
2. compound recipe Fructus Schisandrae Chinensis preparation is made an experiment, said preparation has antioxidation
Embodiment 2:
1. method for preparing with compound recipe Fructus Schisandrae Chinensis preparation of antioxidation is characterized in that comprising following process:
(1) weight proportion of crude drug is: Fructus Schisandrae Chinensis 12g, Radix Astragali 9g, Porphyridium cruentum 20g;
(2) Fructus Schisandrae Chinensis and the Radix Astragali are added lixiviate 10h under 10 times of amount 70% ethanol room temperatures, seal ultrasonic 30min again, temperature is 30 ℃; Power is 80W, uses filtered through gauze then, and filtering residue repeats to extract each once with the ethanol of 6 times of amounts and 4 times of amounts 70% respectively again; Filter, must filtrate;
(3) filtrating in (2) is merged, reclaim ethanol, obtain Fructus Schisandrae Chinensis, Radix Astragali extract;
(4) with Porphyridium cruentum with the centrifugal 10min of 5000r/min centrifuge, be suspended in pH7.4 then, in the phosphate buffer of 50mmol/L, carry out ultrasonication; Broken 5s, interval 5s, 20 times, with the centrifugal 15min of 10000r/min High speed refrigerated centrifuge; Get supernatant, lyophilization gets dry thing;
(5), get compound recipe Fructus Schisandrae Chinensis preparation with Fructus Schisandrae Chinensis, Radix Astragali extract mix homogeneously in the dry thing in (4) and (3).
2. compound recipe Fructus Schisandrae Chinensis preparation is made an experiment, said preparation has antioxidation
A kind of compound recipe Fructus Schisandrae Chinensis preparation antioxidation test
1 compound recipe Fructus Schisandrae Chinensis preparation is to the mensuration of DPPH clearance rate
1.1 the preparation of DPPH solution
DPPH sample 0.01g decided in accurate title, adds dehydrated alcohol and be settled in the 250ml volumetric flask, obtains DPPH solution, and concentration is 0.04mg/ml.
1.2 the preparation of sample solution
Precision takes by weighing compound recipe Fructus Schisandrae Chinensis preparation 0.4g and places 50ml tool plug volumetric flask, adds an amount of distilled water, seals ultrasonic 30min, treat its dissolving fully after, add distilled water again and be settled to 50ml, obtain the sample mother solution, concentration is 8mg/ml, and is subsequent use.In the sample mother solution, using the liquid-transfering gun precision to pipette volume is 0.003125,0.00625,0.0125,0.025; 0.05,0.1,0.2, the sample mother solution of 0.4ml; Using the distilled water standardize solution in the 10ml volumetric flask, to obtain concentration is 0.0025,0.005,0.01,0.02; 0.04,0.08,0.16, the sample solution of 0.32mg/ml.
1.3 sample is to the scavenging action of DPPH
The sample solution that accurately pipettes variable concentrations is mixed in the different test tubes with DPPH solution and ethanol solution equal-volume respectively, shakes up, and places 30min in the dark, under 515nm, measures its absorbance and is respectively A
iAnd A
jOther gets distilled water and DPPH solution equal-volume is mixed in the test tube, and recording absorbance is A.Wherein be mixed into the blank test with distilled water and dehydrated alcohol.And by following formula calculating clearance rate.
Clearance rate=(A-A
i+ A
j)/A * 100%
2 compound recipe Fructus Schisandrae Chinensis preparations are to the mensuration of hydroxy radical clearance rate
2.1 the preparation of sample solution
Precision takes by weighing compound recipe Fructus Schisandrae Chinensis preparation 0.25g and places 50ml tool plug volumetric flask, adds an amount of distilled water, seals ultrasonic 30min, treat its dissolving fully after, be settled to scale, obtain the sample mother solution, subsequent use.In the sample mother solution, using the liquid-transfering gun precision to pipette volume is 5 μ l, 10 μ l, 20 μ l, 40 μ l; 80 μ l, 160 μ l, 320 μ l, the sample mother solution of 640 μ l; Using the distilled water standardize solution in the 10ml volumetric flask, to obtain concentration is 0.0025,0.005,0.01,0.02; 0.04,0.08,0.16, the sample solution of 0.32mg/ml.
2.2 the preparation of reagent
Take by weighing 0.2032g crystal violet with the deionized water standardize solution in the 100ml volumetric flask, subsequent use.Use hydrochloric acid to transfer Tris buffer solution PH to be 5, subsequent use.Get 30% H
2O
2Solution 1.0mL, in the 100mL volumetric flask, subsequent use with the deionized water standardize solution.Take by weighing 0.0695gFeSO
47H
2O water standardize solution in the 50mL volumetric flask, subsequent use (existing with join at present).
2.3 the generation of hydroxy radical OH and mensuration
Get the crystal violet solution 2mL for preparing, Tris buffer 1mL puts into the graded tube of 10mL in order.And be settled to scale with deionized water, and shake up and leave standstill 30 minutes, be designated as A
578
Get the crystal violet solution 2mL for preparing, Tris buffer 1mL, Fe
2+Solution 1mL, H
2O
2Solution 1mL puts into the graded tube of 10mL in order.And be settled to scale with deionized water, and shake up and leave standstill 30 minutes, be designated as A
0
Get the crystal violet solution 2mL for preparing, Tris buffer 1mL, the liquid 2mL to be measured of variable concentrations gradient, Fe
2+Solution 1mL, H
2O
2Solution 1mL puts into the graded tube of 10mL in order.And be settled to scale with deionized water, shake up and leave standstill 30 minutes.Parallel three times, be designated as As.
Do blank experiment with deionized water, under 578 wavelength, measure its absorbance with 721 ultraviolet spectrophotometers.
2.4 the calculating of hydroxy radical OH clearance rate
Utilize formula: clearance rate %=(As-A
0)/(A
578-A
0) * 100% draws the clearance rate of sample extracting solution to hydroxy radical.
The antioxidation result of 3 compound recipe Fructus Schisandrae Chinensis preparations
3.1 compound recipe Fructus Schisandrae Chinensis preparation is to the clearance rate of DPPH and hydroxy radical
See table 1.
Table 1 compound recipe Fructus Schisandrae Chinensis preparation is to the clearance rate result of DPPH and hydroxy radical
3.2 vitamin C (VC) is to the clearance rate of DPPH and hydroxy radical
See table 2.
Table 2 VC is to the clearance rate result of DPPH and hydroxy radical
Claims (2)
1. method for preparing with compound recipe Fructus Schisandrae Chinensis preparation of antioxidation is characterized in that comprising following process:
(1) weight proportion of crude drug is: Fructus Schisandrae Chinensis 9g-12g, Radix Astragali 6g-9g, Porphyridium cruentum 10g-20g;
(2) Fructus Schisandrae Chinensis and Radix Astragali adding 8-10 are doubly measured lixiviate 8-10h under the 70% ethanol room temperature, seal ultrasonic 30min again, temperature is 30 ℃; Power is 80W, uses filtered through gauze then, and filtering residue repeats to extract each once with the ethanol of 6 times of amounts and 4 times of amounts 70% respectively again; Filter, must filtrate;
(3) filtrating in (2) is merged, reclaim ethanol, obtain Fructus Schisandrae Chinensis, Radix Astragali extract;
(4) with Porphyridium cruentum with the centrifugal 10min of 5000r/min centrifuge, be suspended in pH7.4 then, in the phosphate buffer of 50mmol/L, carry out ultrasonication; Broken 5s, interval 5s, 20 times, with the centrifugal 15min of 10000r/min High speed refrigerated centrifuge; Get supernatant, lyophilization gets dry thing;
(5), get compound recipe Fructus Schisandrae Chinensis preparation with Fructus Schisandrae Chinensis, Radix Astragali extract mix homogeneously in the dry thing in (4) and (3).
2. the purposes of a kind of compound recipe Fructus Schisandrae Chinensis preparation described in claim 1 is characterized in that preparing the application in the antioxidation medicine.
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105412391A (en) * | 2015-12-03 | 2016-03-23 | 哈尔滨医科大学 | Health care food formula having auxiliary antioxidant function and preparation method of health care food formula |
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1405294A (en) * | 2002-11-03 | 2003-03-26 | 陈克拉 | Wine for modulating immune from gene of ancient organism at deep sea and its production method |
| CN1430982A (en) * | 2003-01-13 | 2003-07-23 | 徐春华 | Capsule of astragalus and honey as well as its preparing technique |
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Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1405294A (en) * | 2002-11-03 | 2003-03-26 | 陈克拉 | Wine for modulating immune from gene of ancient organism at deep sea and its production method |
| CN1430982A (en) * | 2003-01-13 | 2003-07-23 | 徐春华 | Capsule of astragalus and honey as well as its preparing technique |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105412391A (en) * | 2015-12-03 | 2016-03-23 | 哈尔滨医科大学 | Health care food formula having auxiliary antioxidant function and preparation method of health care food formula |
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