CN1594246A - Rhizoma Anemarrhenae total phenol extract, its preparation method and application - Google Patents
Rhizoma Anemarrhenae total phenol extract, its preparation method and application Download PDFInfo
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Abstract
The invention provides a method for preparing wind-weed rhizome extract which comprises the steps of, (1) backflow extracting with water 6-14 times of wind-weed rhizome weight, (2) filtering to remove solid body, concentrating the filtrate to obtain soup, (3) adjusting ethanol concentration to 75%+-15%, depositing, separation, filtering to obtain filtrate, (4) reclaiming alcohol, charging water to dilute, (5) adjusting pH=2-8, (6) passing the soup to resin adsorbing column, (7) washing column with water, (8) elution with pH=10-14 alkaline solution, (9) adjusting pH=2-8, and filtering.
Description
Technical field
The present invention relates to the extraction process field of effective ingredient in Chinese, relate to the extraction and the preparation method of total phenol extraction thing of the wind-weed particularly.The invention still further relates to the composition that contains the total phenol extraction thing of the wind-weed.
Background technology
The wind-weed is the liliaceous plant wind-weed (Anemarrhena asphodeloides) rhizome, and bitter but sweet flavor is cold in nature, goes into lung, stomach, kidney channel, has heat-clearing and fire-purging, promotes the production of body fluid effect such as to moisturize.Be China's traditional treatment diabetes Chinese medicine.
Existing report shows, the wind-weed has effect (chemical ingredients of the wind-weed and the Advance on Pharmacological Activities of resisting pathogenic microbes, analgesic, platelet aggregation-against, hypoglycemic, phosphoesterase inhibition, anti-inflammatory and reduction transaminase, Yang Lirong etc., 2002 the 24th the 4th phases of volume of foreign medical science traditional Chinese medicine fascicle, pp.207-210).The document has mainly been summarized the effect of saponin in the wind-weed, and Mangiferin and glucosides thereof are given the credit in the main position of hypoglycemic.
Other people has also studied the hypoglycemic activity of the wind-weed, and it is given the credit to (herbal medicine, 1996 the 27th the 10th phases of volume such as terpene in the wind-weed, peptide class, amino acid, flavones, glycan, polysaccharide, saponin, pp.605-606, China's biochemical drug magazine, 2003 the 24th the 2nd phases of volume, pp.81-82).The method that is used to extract zhimusaponin is also disclosed, and usefulness ethanol and macroporous resin backflow washing (Chinese experimental pharmacology of traditional Chinese medical formulae magazine, February calendar year 2001, Vol.7, No.1, pp.9-11).
Though for the existing multiple different reports of the composition of various blood sugar reducing functions in the wind-weed, wherein any composition is that main hypoglycemic position does not obtain confirming actually.Do not test yet a plurality of extract parts are compared with the blood sugar reducing function of decocting liquid, obtain the report of the main efficient part of wind-weed hypoglycemic.Do not study report for the method for extracting the total phenol of the wind-weed with to its purposes yet.
Therefore, the urgent exploitation in this area is based on the product and the technology of hypoglycemic efficient part in the wind-weed.
Summary of the invention
An object of the present invention is to provide a kind of method for preparing the total phenol extraction thing of the wind-weed, wherein by the aqueous solvent refluxing extraction, alcohol precipitation, column chromatography, wash-out filters and obtains efficient part.This total phenol extraction thing can be effectively hypoglycemic.
Another object of the present invention provides the total phenol extraction thing of the wind-weed and contains the composition of the total phenol extraction thing of the wind-weed.
In one aspect of the invention, provide a kind of method for preparing Rhizoma Anemarrhenae extract, it is characterized in that the method comprising the steps of:
(a) with the aqueous solvent refluxing extraction of 6-14 times of rhizoma ane marrhenae weight, obtain extracting solution, described aqueous solvent is the aqueous ethanolic solution or the water of 0-60% weight;
(b) filter the removal solid, concentrated filtrate obtains soup;
(c) regulate alcohol concn to 75% ± 15%, place 10-24 hour precipitation, layering is filtered and is obtained filtrate;
(d) reclaim ethanol from filtrate, and obtain concentrated medicament, thin up gets dilute liquid medicine to 2-50 times of medicinal material weight in soup;
(e) soup pH is transferred to 2-8;
(f) the gained soup is crossed the resin absorption post, described resin is selected from macroporous resin and anionite-exchange resin;
(g) wash post with water;
(h) with the basic solution wash-out of pH10-14, obtain elutriant;
(i) elutriant pH is transferred to 2-8, filter and obtain liquid Rhizoma Anemarrhenae extract.
In an embodiment aspect this, the consumption of the aqueous solvent in the step (a) preferably medicinal material weight 8-12 doubly, most preferably be 10 times; Aqueous solution is water preferably.
In another embodiment of the present invention, the refluxing extraction of this step (a) is divided and is carried out for 1-4 time, and each refluxing extraction 1-5 hour, united extraction liquid then.
In another embodiment of the present invention, this method also comprises step:
(j) water bath method solution then 50-100 ℃ of oven dry, obtains solid-state Rhizoma Anemarrhenae extract.
In another embodiment of the present invention, preferably pH being transferred to 5 ± 1 in this step (e), most preferably is to transfer to 5.
In another embodiment of the present invention, during the resin in this step (f) is selected from-the low-pole macroporous adsorbent resin, for example or strongly basic anion exchange resin, its consumption be medicinal material weight 0.5-5 doubly.
In another embodiment of the present invention, in this step (i), preferably pH being transferred to 5 ± 1, most preferably is 5, and step (h) neutral and alkali solution is NaOH solution, KOH, NaCO
3, K
2CO
3Solution or its mixing solutions are preferably used 0.5%NaOH solution.
Second aspect of the present invention provides a kind of Rhizoma Anemarrhenae extract, it is characterized in that, total phenol extraction substrate concentration is a 50%-95% weight in the said composition.
In the embodiment of the present invention aspect this, this extract is with the described method preparation of first aspect present invention.
Aspect the 3rd of the present invention, a kind of hypoglycemic pharmaceutical composition is provided, it contains 1-99% Rhizoma Anemarrhenae extract of the present invention and the pharmaceutically acceptable carrier of 1-99%.
Aspect the 4th of the present invention, provide Rhizoma Anemarrhenae extract of the present invention to be used to prepare the purposes of hypoglycemic medicine.
Embodiment
The present invention is through discovering that the aldehydes matter in the wind-weed is its main hypoglycemic effective constituent, has developed the method for the total phenol extraction thing of effective extraction wind-weed on this basis, thereby has finished the present invention.
The wind-weed is conventional Chinese medicinal materials, can be cut into medicine materical crude slice or grind to form meal to use.This medicinal material can be buied from each wind-weed place of production or Chinese medicine shop, for example Haozhou, Anhui medicinal material market, sky, Suzhou Ling Zhongyaoyinpianchang etc.Maybe can use other form, for example clean and cut into inch strips or be ground into the coarse particles form.
Wind-weed extracting method of the present invention adopts aqueous solvent as extracting reagent, and slightly to leach supernatant liquor different with the ethanol of routine, also different with other chloroform extraction method, and its effective constituent has the difference of essence on kind.
What use in the step (c) is conventional ethanol sedimentation step, obtains filtrate with conventional filtrations such as filter paper after the layering, and wherein determining alcohol is approximately 60-90%;
Recovery ethanol step in the step among the present invention (d) makes soup remove ethanol, effectively separates required component on the macroporous resin thereby can be adsorbed on.Recovery is undertaken by ordinary method, for example distills etc.
Column chromatography of the present invention uses the macroporous resin adsorption post of various routines, can adopt various pharmaceutical grade macroporous adsorbent resins.Macroporous resin commonly used be in-low-pole.Representational resin examples comprises: D101, AB-8, LSA-10, LSA-7, XDA-1.Preferred resin is available from the blue dark LSA-10 in Xi'an.Before use can first watering balance.Last sample, washing, elution speed is unrestricted, but preferred per kilogram resin elution speed most preferably was 60ml/min between 10-300ml/ minute.
Employed filtration unit can be various conventional filtration devices among the present invention (i).Filter paper vacuum suction filter for example, porous resin, ultra-filtration membrane, molecular sieve, packed column etc.
Total phenol extraction thing content is (as 50-95% more than 50% in the present composition, preferably be 60-90%), the efficient part yield is (based on the medicinal material gross weight) more than 0.5% (be that yield is the per-cent that extract and medicinal material drop into, yield 0.5% i.e. 1 kilogram of rhizoma ane marrhenae obtains the total phenol extraction thing of 5 grams).Detect through ultraviolet spectrophotometry or colorimetry, the content of efficient part is more than 50% in the extract.Simultaneously,, do not prepare expensive step such as TLC, electrophoresis, can be used for industrial production in enormous quantities because operation steps is simple.
The efficient part that the present invention obtains mainly is total phenol extraction thing, have hypoglycemic, reduce effects such as streptozotocin or suprarenin induced hyperglycemia.The main position toxicity that the present invention obtains is low, can high dosage uses and has no side effect.
Because the extract that the present invention obtained is the phenol extraction thing, therefore can simply use spectrophotometer, under the wavelength of 660nm, detect the concentration of its efficient part by ultraviolet spectrophotometry or colorimetry.
Rhizoma Anemarrhenae extract of the present invention can directly use, or on pharmacology acceptable carrier and additive, make pharmaceutical composition.For example add sweeting agent, seasonings, tinting material and sanitas, tablet, lozenge, water or oil suspension, dispersible powders or the particle of preparation oral administration, emulsion, hard or soft capsule or syrup or elixir.Tablet contains and the activeconstituents that is applicable to the nontoxic acceptable mixed with excipients of making tablet.These vehicle can be inert diluents for example, as lime carbonate, yellow soda ash, lactose, calcium phosphate or sodium phosphate; Granulating agent and disintegrating agent, for example alginic acid; Tackiness agent is as starch, gelatin or gum arabic; And lubricant, as Magnesium Stearate, stearic acid or talcum.Tablet can not have sugar-coat or uses the known technology dressing, postpones disintegration and absorption in gi tract, thereby continuous action is provided in a long time.For example, can use the time-delay material, as glyceryl monostearate or distearin.The preparation that orally uses also can be made into hard gelatin capsule, wherein activeconstituents and inert solid thinner mix as lime carbonate, calcium phosphate or kaolin, or make soft gelatin capsule, wherein activeconstituents and water or oil medium, for example peanut oil, liquid paraffin or mixed with olive oil.
The content of effective scope is 5-20mg/kg/ days in the pharmaceutical composition of the present invention, can determine route of administration and dosage according to clinical needs by experienced doctor.
Pharmaceutical composition of the present invention also can contain other blood sugar reducing components, for example conventional antidiabetic drug that can get at present etc. or other Chinese medicine preparation.
Pharmaceutical composition of the present invention can be used with various approach, and for example oral, injection, Transdermal absorption are preferably oral.
The invention effect
Use preparation method of the present invention, can obtain having the Rhizoma Anemarrhenae extract of total phenol extraction thing of high density.Rhizoma Anemarrhenae extract of the present invention has significant hypoglycemic activity, and is suitable with the diabetes pill effect.
Embodiment
The extraction and the detection of the total phenol of the embodiment 1-4 wind-weed
1. material and instrument
Material: the wind-weed that originates from Chinese Jilin.
Resin: LSA-10 is blue dark available from Xi'an
Instrument: multi-function extractor (500L), Wujin, Changzhou Meng He pharmaceutical machine factory
Embodiment 1
Get the Powdered rhizoma ane marrhenae of 50kg,, extract altogether twice with 10 times of rhizoma ane marrhenae amount 30% alcohol reflux, each 1 hour, merge ethanol extract twice, filter, reclaim ethanol, concentrate, alcohol is sink to and contains alcohol 70%, reclaim ethanol, thin up to 10 times medicinal material weight obtains the 500kg dilute liquid medicine, transfers pH to 5, the gained soup is crossed resin absorption post (LSA-10), resin demand is 2 times of medicinal material amount, and soup has been crossed and washed post bed to effluent liquid with water behind the post bed is colourless, and it is colourless that 0.4%NaOH is eluted to effluent liquid, collect the alkali elutriant, transfer pH to 4, filter water bath method, 60 ℃ of vacuum dryings get total phenol extraction thing.
Embodiment 2
Get 50kg particulate state rhizoma ane marrhenae,, extract altogether twice with 9 times of medicinal material amount 10% alcohol reflux, each 1.5 hours, merge ethanol extract twice, filter, reclaim ethanol, concentrate, alcohol is sink to and contains alcohol 80%, reclaim ethanol, thin up to 8 times medicinal material weight obtains the 400kg dilute liquid medicine, transfer PH4, the gained soup is crossed resin absorption post (LSA-10), and resin demand is 3 times of medicinal material amount, soup has been crossed and washed post bed to effluent liquid with water behind the post bed is colourless, it is colourless that 0.6%NaOH is eluted to effluent liquid, collects the alkali elutriant, transfers PH5, filter, water bath method, 70 ℃ of vacuum dryings get total phenol extraction thing.
Embodiment 3
Get 50kg sheet rhizoma ane marrhenae,, extract altogether twice with 12 times of medicinal material amount 5% alcohol reflux, each 2 hours, merge ethanol extract twice, filter, reclaim ethanol, concentrate, alcohol is sink to and contains alcohol 75%, reclaim ethanol, thin up to 9 times medicinal material weight obtains the 450kg dilute liquid medicine, transfers PH6, the gained soup is crossed resin absorption post (LSA-10), resin demand is 1 times of medicinal material amount, and soup has been crossed and washed post bed to effluent liquid with water behind the post bed is colourless, and it is colourless that 0.5%NaOH is eluted to effluent liquid, collect the alkali elutriant, transfer PH6, filter water bath method, 80 ℃ of vacuum dryings get total phenol extraction thing.
The detection of total phenol content in embodiment 4 Rhizoma Anemarrhenae extracts
1. instrument: ultraviolet-visible pectrophotometer (UV-1240, day island proper Tianjin Suzhou factory of company)
2. experimental technique:
Conventional phenols colorimetry detects the total phenol content in the Rhizoma Anemarrhenae extract solution under 660nm.
3. the result is as follows:
Total phenol content is measured
Embodiment 1 55%
Embodiment 2 53%
Embodiment 3 58%
Three batches 50 kilograms medicinal material is through extracting, the final extraction yield of the main total phenol of the efficient part wind-weed is (the total phenol that contains in medicinal material is 100%) more than 60%, the efficient part yield is (relatively medicinal material input amount) more than 0.3%, after testing in the extract content of efficient part at (phenol shared weight percent in extract always) 50% or more.
The experimental study of embodiment 5 wind-weed hypoglycemic efficient parts
This experiment with the hypoglycemic main efficient part of the streptozotocin inductive diabetic mice model discrimination wind-weed, and is further inquired into its blood sugar reducing function and mechanism thereof by extracting three efficient parts of the wind-weed.
Experiment material
1. medicine and reagent
Anemaran, total phenol, saponin extract:
(1) to adopt conventional polyoses producing method be water extraction and alcohol precipitation method to anemaran.
(2) extract of total phenol extraction thing: embodiment 3.
(3) zhimusaponin prepares according to literature method, promptly ethanol and macroporous resin backflow washing method (Chinese experimental pharmacology of traditional Chinese medical formulae magazine, February calendar year 2001, Vol.7, No.1, pp.9-11).
Citrate buffer solution (pH 7.4): citric acid 2.1g, Trisodium Citrate 2.94g is analytical pure, respectively with physiological saline quantitatively to 100ml, making its concentration separately is 0.1mM.Get above-mentioned citric acid solution 65.1ml, sodium citrate solution 54.9ml, abundant mixing, 4 ℃ of refrigerators are preserved standby
Streptozotocin (streptozotocin, STZ): Sigma company, face with preceding that to be made into concentration with citrate buffer solution be 1% STZ solution
Blood sugar detection test kit: Shanghai Biological Products Inst., Ministry of Public Health, lot number: 20039218
Suprarenin: Tian Feng pharmaceutical factory, Shanghai product, lot number: 020616
Diabetes pill: Guangzhou No.1 Chinese Pharmacy Factory product, lot number: 20030316.
2. animal
Kunming mouse, male and female half and half, body weight 20 ± 2g, medical college of University Of Suzhou Experimental Animal Center provides, laboratory animal occupancy permit number: SYXK (Soviet Union) 2002-0037, laboratory animal production licence number: SCXK (Soviet Union) 2002-008.
3. instrument
Spectrophotometer, 752 types: Shanghai the 3rd analytical instrument factory
Electric heating constant temperature tank: Shanghai Medical Instruments seven factories
4. data processing
Adopt SPSS10.0 version statistical analysis software bag to handle experimental data.Employing ± s represents, organizes the test of significance of dosage data with one-way analysis of variance (ANOVA) with compare (LSD) method in twos more.
Experimental technique and result
1, anemaran, the total phenol of the wind-weed, zhimusaponin are to the research of STZ inductive diabetic mice blood sugar reducing function
Get 70 of healthy mices, abdominal injection 100mg/kg STZ, eyeground vein is got blood with determination of glucose oxidase blood sugar behind the 72h, gets blood glucose value 〉=13.88mmol/L person and classifies diabetic mice as.Be divided into 6 groups at random, that is: diabetic model group (DM); Anemaran group (300mg/kg), total phenol (the 100mg/kg of the wind-weed, herein and consumption hereinafter be meant extract the milligram number), zhimusaponin group (100mg/kg), three position dosages are equivalent to equal crude drug amount (5g/kg), decocting 5g/kg, positive control diabetes pill group (1500mg/kg); Other gets 8 mouse and makes the normal control group.Irritate stomach every day 1 time, successive administration 10 days, the normal control group is irritated stomach and is given equivalent physiological saline.Animal fasting 10h after the last administration, the determination of glucose oxidase mouse blood sugar is used in the eyeground vein blood sampling.
Table 1 administration group is to the influence of streptozotocin inductive blood glucose in diabetic mice
Group dosage (mg/kg) blood sugar (mmol/L)
The normal control group--8 6.16 ± 0.53
Diabetic groups--8 15.48 ± 1.35
*
Anemaran 300 8 13.61 ± 1.77
#
The total phenol 100 9 11.96 ± 1.18 of the wind-weed
##
Zhimusaponin 100 10 14.16 ± 2.09
Wind-weed decocting 5,000 10 11.86 ± 1.11
##
Diabetes pill 1,500 9 11.67 ± 1.08
##
*P＜0.01 (with respect to contrast);
#P＜0.05,
##P＜0.01 (with respect to contrast)
The result shows that the wind-weed decocting liquid of equal crude drug amount, total phenol all have high significantly hypoglycemic activity (P＜0.01), and anemaran also has certain hypoglycemic activity (P＜0.05).Wherein the total phenol blood sugar reducing function of the wind-weed is than polysaccharide tool significant difference, and its blood sugar reducing function and wind-weed decocting liquid and diabetes pill are suitable, and consumption is few, and the total phenol of the prompting wind-weed is the hypoglycemic main efficient part of the wind-weed.
2, the total phenol of the wind-weed is to the influence of normal mouse fasting plasma glucose and postprandial blood sugar
50 of mouse are divided into 5 groups at random, the normal control group, and diabetes pill control group (1500mg/kg), the total phenol of the wind-weed (A) small dose group (50mg/kg), middle dosage group (100mg/kg) and heavy dose of group (200mg/kg) are irritated stomach every day 1 time, successive administration 10 days.Animal fasting 10h after the last administration, the determination of glucose oxidase mouse blood sugar is used in the eyeground vein blood sampling.The result shows: three dosage of the total phenol of the wind-weed all can obviously reduce the fasting plasma glucose (P＜0.05) of normal mouse.Other gets 72 mouse, and grouping, administration, get blood, detect all identically with the front, be the preceding non-fasting of last administration.The result shows: three total phenol of the dosage wind-weed all have remarkable reduction effect (P＜0.05) (table 3) to the normal mouse postprandial blood sugar.
Table 2 administration group is to the influence of normal empty stomach mouse blood sugar
Group dosage (mg/kg) blood sugar (mmol/L)
Contrast 06 5.86 ± 0.49
Diabetes pill 1,500 8 4.52 ± 0.75
*
A(L) 200 10 4.06±0.41
**
A(M) 100 9 4.15±0.63
**
A(S) 50 9 4.82±0.78
*
*P＜0.05,
*P＜0.01 (with respect to contrast)
Table 3 administration group is to the influence of mouse postprandial blood sugar
Group dosage (mg/kg) blood sugar (mmol/L)
Contrast 08 6.34 ± 0.71
Diabetes pill 1,500 10 5.19 ± 0.36
*
A(L) 250 10 4.97±0.51
**
A(M) 125 9 4.95±0.66
**
A(S) 62.5 9 5.34±0.32
*
*P＜0.05,
*P＜0.01 (with respect to contrast)
The total phenol of 3 wind-weeds is to the influence of STZ inductive blood glucose in diabetic mice
58 of mouse, get 8 at random and be the normal control group, all the other tail vein injection streptozotocin 100mg/kg cause the diabetic mice model, fasting 12h (spending the night) before the injection, injection back 72h eyeground vein measuring blood sugar of blood extracting, remove and do not cause diabetes model (fasting 12h, blood sugar is lower than 13.88mmol/L) person, be divided into 8 groups at random, model group and the blank water of irritating the stomach corresponding dosage of organizing, the administration group is irritated the total phenol of the low dose of wind-weed of stomach [A (S) respectively, 50mg/kg], middle dosage [A (M), 100mg/kg] and heavy dose of [A (L), 200mg/kg], the positive drug group is irritated stomach diabetes pill (1500mg/kg).Every day, gastric infusion was 1 time, and continuous 14 days, fasting 10h after the last administration, eyeground vein got blood and survey blood sugar, and method is the same.The result shows: compare with diabetic model group, blood sugar all significantly reduces (middle dosage P＜0.05 after the total phenol heavy dose of the wind-weed, the middle dosage treatment; Heavy dose of P＜0.01) (Tab4).
Table 4 administration group is to the influence of streptozotocin inductive blood glucose in diabetic mice
Group dosage (mg/kg) blood sugar (mmol/L)
Contrast 08 6.03 ± 0.51
Model group 08 14.91 ± 2.82
*
Diabetes pill 1,500 10 10.93 ± 2.16
▲ ▲
A(L) 200 10 11.34±2.33
▲▲
A(M) 100 8 12.69±1.06
▲
A(S) 50 9 12.90±1.76
*P＜0.01 (with respect to contrast)
▲P＜0.05,
▲ ▲P＜0.01 is with respect to model group
The total phenol of 4 wind-weeds is to the influence of suprarenin induced hyperglycemia mice blood sugar
60 of normal mouses are divided into 6 groups at random, and grouping and medication are the same.2h after the last administration, except that the blank group, all the other abdominal injection suprarenin 20 μ g/kg, blank group intraperitoneal injection of saline is got blood and is surveyed blood sugar behind the 30min.The result shows: compare with the suprarenin model group, three dosage of the total phenol of the wind-weed all can obviously reduce suprarenin induced hyperglycemia (little, middle dosage P＜0.05; Heavy dose of P＜0.01) (table 5).
Table 5 administration group is to the influence of suprarenin inductive blood glucose in diabetic mice
Group dosage (mg/kg) blood sugar (mmol/L)
Contrast 08 5.98 ± 0.56
Model group 08 12.76 ± 1.84
*
Diabetes pill 1,500 9 10.18 ± 1.23
▲
A(L) 200 10 9.24±2.09
▲▲
A(M) 100 8 10.71±1.93
▲
A(S) 50 9 10.96±1.47
▲
*P＜0.01 (with respect to contrast)
▲P＜0.05,
▲ ▲P＜0.01 pair model group
The total phenol of 5 wind-weeds is to the influence of glucose induced hyperglycemia mice blood sugar
45 of normal mouses, be divided into 6 groups at random, grouping, administration and dosage are the same, administration 15 days, fasting 10h, after each organizes the mouse administration, get hematometry fasting plasma glucose (0h), all mouse are all irritated stomach glucose solution (2g/kg) behind the 2h, eyeground vein is got hematometry and is given the blood glucose value behind 0.5h, the 1h and 2h behind the glucose, the result shows: blood glucose value all obviously reduces than control group after gavaging the mouse oral glucose of three total phenol of the dosage wind-weed, shows that the total phenol of the wind-weed can improve mouse sugar tolerance (table 6).
Table 6 administration group is to the influence of the blood glucose in diabetic mice of glucose induction
Blood sugar (mmol/L)
Group N
0h 0.5h
Normal control group 6 5.78 ± 0.53 6.06 ± 0.81
Glucose group 7 5.91 ± 0.27 9.35 ± 1.36
*
A(L) 7 5.51±0.71 6.99±2.32
▲
A(M) 8 5.68±0.31 6.54±1.79
▲▲
A(S) 8 5.80±0.64 6.19±1.92
▲▲
Diabetes pill 8 6.13 ± 0.49 6.24 ± 1.89
▲ ▲
*P＜0.05,
*P＜0.01 (with respect to glucose group)
The total phenol of 6 wind-weeds is to the acute toxicity test of mouse
The total phenol of the wind-weed is used for the treatment of diabetes.For the dosage range of experimental safe medication is provided, the total phenol of the wind-weed has been carried out acute toxicity test.
In pilot study, the heavy dose of SY-2 extract of mouse gavaging bulk drug does not also cause death, and can not measure medium lethal dose, so adopt the maximum dosage-feeding test.
Be subjected to the total phenol of the reagent thing wind-weed, Si Yuan medical sci-tech development company provides by Suzhou.
Preparation: adding distil water, be mixed with the suspension that every ml contains the 20mg bulk drug, use for irritating stomach.
Laboratory animal
40 of ICR mouse, 18～21g, male and female half and half provide conformity certification by preclinical medicine institute of University Of Suzhou Experimental Animal Center: SCXK (Soviet Union) 2001-0001 number.
Test method and result
Get 20 of healthy mices, male and female half and half, the 0.4ml/10g gastric infusion was irritated stomach 3 times in one day, observed continuously 7.
Mouse administration same day, slow in action, feed reduces.Recover appetite next day, movable, stool is all normal.Observe and do not see death on the 7th, empty stomach 12h weighed on 1, became celestial and did not see that main organs has unusually.Body weight change sees Table 7.
Table 7 A-2 acute toxicity test in mice body weight change (X ± SD)
Rate of increase
The sex mouse is counted preceding body weight (g) the 8th daily weight (g) of administration
(％)
♂ 10 20.0±0.82 27.6±1.55
*** 38.23
♀ 10 19.6±0.84 24.9±1.61
*** 27.01
With ratio before the administration,
* *P＜0.01
By table 7 as seen, the total phenol of the mouse gavaging wind-weed, body weight normal growth.
In acute toxicity test in mice, adopt the maximum dosage-feeding method, the total phenol dosage of wind-weed 2400mg/kg is about 240 times of clinical people's consumption 10mg/kg.Observed 7 after the administration, dead and obvious toxic reaction, body weight normal growth all do not occur.
Conclusion
1. total phenol of the wind-weed and polysaccharide all have hypoglycemic activity, and wherein the blood sugar reducing function of the total phenol of the wind-weed is particularly remarkable and suitable with wind-weed decocting liquid and diabetes pill, and the total phenol of the prompting wind-weed is the main efficient part of wind-weed hypoglycemic.
2. the total phenol of the wind-weed can obviously reduce STZ inductive blood glucose in diabetic mice, shows that the total phenol of the wind-weed can improve the symptom of diabetes by weakening streptozotocin to the damage of islet cells or improve the function of the β cell of damaged;
3. the total phenol of the wind-weed can obviously reduce the suprarenin induced hyperglycemia, adrenergic promotes the decomposition of liver starch and muscle glycogen and causes blood sugar increasing, but and the adrenergic blood glucose increasing effect of the total phenol antagonism of the wind-weed, this prompting wind-weed total phenol hypoglycemic activity also may with suppress the decomposition of human body and utilize relevant glycogen.
4,5 by this experimental result as can be known: the total phenol of the wind-weed all has blood sugar reducing function preferably to the animal hyperglycemia that multiple reason causes, and curative effect is close with diabetes pill.
6. the acute toxicity result shows, the total phenol toxicity of the wind-weed is little, drug safety.
As mentioned above, method of the present invention can be extracted the total phenol extraction thing of the wind-weed efficiently, and technology is simple, the active constituent content height.In addition, resulting product all has remarkable effect in the hypoglycemic for the diabetes of different mechanisms.
Product of as above having described Rhizoma Anemarrhenae extract preparation method disclosed in this invention and having made and uses thereof.But need to understand and wherein comprised variation understood by one of ordinary skill in the art and change.These variations and change are also contained in the scope of the present invention that claim limits.
Claims (10)
1. method for preparing Rhizoma Anemarrhenae extract is characterized in that the method comprising the steps of:
(a) with the aqueous solvent refluxing extraction of 6-14 times of rhizoma ane marrhenae weight, obtain extracting solution, described aqueous solvent is the aqueous ethanolic solution or the water of 0-60% weight;
(b) filter the removal solid, concentrated filtrate obtains soup;
(c) regulate alcohol concn to 75% ± 15%, place 10-24 hour precipitation, layering is filtered and is obtained filtrate;
(d) reclaim ethanol from filtrate, and obtain concentrated medicament, thin up gets dilute liquid medicine to 2-50 times of medicinal material weight in soup;
(e) soup pH is transferred to 2-8;
(f) the gained soup is crossed the resin absorption post, described resin is selected from macroporous resin or anionite-exchange resin;
(g) wash post with water;
(h) with the basic solution wash-out of pH10-14, obtain elutriant;
(i) elutriant pH is transferred to 2-8, filter and obtain liquid Rhizoma Anemarrhenae extract.
2. the method for claim 1 is characterized in that, the consumption of the aqueous solvent in the described step (a) is 8-12 a times of medicinal material weight.
3. the method for claim 1 is characterized in that, in the described step (a), refluxing extraction is divided and carried out for 1-4 time, each refluxing extraction 1-5 hour, and united extraction liquid then.
4. the method for claim 1 is characterized in that, described method also comprises step:
(j) water bath method solution then 50-100 ℃ of oven dry, obtains solid-state Rhizoma Anemarrhenae extract.
5. the method for claim 1, it is characterized in that, in the step (e) pH transferred to 5 ± 1 (with power 2), and the resin in the described step (f) be selected from-low-pole macroporous adsorbent resin or strongly basic anion exchange resin, consumption be medicinal material weight 0.5-5 doubly.
6. the method for claim 1 is characterized in that, in described step (i), pH is transferred to 5 ± 1, and step (h) neutral and alkali solution is NaOH solution, KOH, NaCO
3, K
2CO
3Solution or its mixing solutions.
7. a Rhizoma Anemarrhenae extract is characterized in that, total phenol extraction substrate concentration is a 50%-95% weight in the said composition.
8. Rhizoma Anemarrhenae extract as claimed in claim 7 is characterized in that, this extract prepares with the described method of claim 1.
9. a hypoglycemic pharmaceutical composition is characterized in that, it contains described Rhizoma Anemarrhenae extract of 1-99% claim 7 and the pharmaceutically acceptable carrier of 1-99%.
10. the purposes of the described Rhizoma Anemarrhenae extract of claim 7 is characterized in that, said composition is used to prepare hypoglycemic medicine.
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| CNB031508790A CN100366588C (en) | 2003-09-10 | 2003-09-10 | Rhizoma Anemarrhenae total phenol extract, its preparation method and application |
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|---|---|---|---|
| CNB031508790A CN100366588C (en) | 2003-09-10 | 2003-09-10 | Rhizoma Anemarrhenae total phenol extract, its preparation method and application |
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| Publication Number | Publication Date |
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| CN1594246A true CN1594246A (en) | 2005-03-16 |
| CN100366588C CN100366588C (en) | 2008-02-06 |
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| Application Number | Title | Priority Date | Filing Date |
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101229316B (en) * | 2008-01-31 | 2010-08-25 | 广东药学院 | Rhizoma anemarrhenae extrac |
| CN105582104A (en) * | 2015-12-30 | 2016-05-18 | 中国中医科学院中医临床基础医学研究所 | Traditional Chinese medicine composition for treating rheumatoid arthritis |
| CN105768099A (en) * | 2016-03-04 | 2016-07-20 | 吴正锋 | Blood sugar-reducing healthcare food with addition of rhizoma anemarrhenae extract |
| CN107753500A (en) * | 2017-11-03 | 2018-03-06 | 黑龙江中医药大学 | The Preparation method and use of oligosaccharide compositions in the wind-weed |
Family Cites Families (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1203083C (en) * | 2003-02-25 | 2005-05-25 | 广州莱泰制药有限公司 | Rhizoma anemarrhenae extract and preparation process and use thereof |
-
2003
- 2003-09-10 CN CNB031508790A patent/CN100366588C/en not_active Expired - Lifetime
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101229316B (en) * | 2008-01-31 | 2010-08-25 | 广东药学院 | Rhizoma anemarrhenae extrac |
| CN105582104A (en) * | 2015-12-30 | 2016-05-18 | 中国中医科学院中医临床基础医学研究所 | Traditional Chinese medicine composition for treating rheumatoid arthritis |
| CN105768099A (en) * | 2016-03-04 | 2016-07-20 | 吴正锋 | Blood sugar-reducing healthcare food with addition of rhizoma anemarrhenae extract |
| CN107753500A (en) * | 2017-11-03 | 2018-03-06 | 黑龙江中医药大学 | The Preparation method and use of oligosaccharide compositions in the wind-weed |
Also Published As
| Publication number | Publication date |
|---|---|
| CN100366588C (en) | 2008-02-06 |
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