CN1267113C - Extract of mulberry twig and its extracting process and novel usage - Google Patents

Extract of mulberry twig and its extracting process and novel usage Download PDF

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CN1267113C
CN1267113C CN02113004.3A CN02113004A CN1267113C CN 1267113 C CN1267113 C CN 1267113C CN 02113004 A CN02113004 A CN 02113004A CN 1267113 C CN1267113 C CN 1267113C
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extracting solution
ethanol
ramulus mori
extract
gout
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CN1382478A (en
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孔令东
杨澄
鲍建伟
张金龙
王业民
朱继孝
王颖
朱纲
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YANGYANTANG PHARMACEUTICAL CO Ltd ZHEJIANG KANG'ENBEI GROUP
Nanjing University
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YANGYANTANG PHARMACEUTICAL CO Ltd ZHEJIANG KANG'ENBEI GROUP
Nanjing University
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Abstract

The present invention relates to mulberry twig extract prepared into health food for preventing hyperuricemia and gout diseases and drugs for treating hyperuricemia and gout diseases. The mulberry twig extract is prepared from single drug mulberry twig by extraction by 0 to 95% of ethanol water solutions, or solvent of chloroform, acetone, ethyl acetate, carbinol, etc. or the combination of 0 to 95% of ethanol water solutions or solvent under the temperature condition of 10-100 DEG C for one time or more than once time by extracting solution combination and solvent recovery, or the mulberry twig extract is further purified by the following methods that the pH of the extracting solution is regulated into 7.5 to 9.5; the pH of supernatant after centrifugation is regulated into 4 to 6; the extracting solution is precipitated, washed by water to be neutral and dried; the extracting solution is degreased by petroleum ether or gasoline; after being degreased by the petroleum ether or the gasoline, the extracting solution is respectively or orderly extracted by the chloroform, or the ethyl acetate, or the acetone, or the carbinol or water; after being adsorbed by macroporous adsorption resin, the extracting solution is desorbed by 10 to 95% of ethanol water solutions; after being adsorbed by diatomaceous earth, alumina, etc. and eluted by the solvent of the chloroform, the ethyl acetate, the acetone, the carbinol, the ethanol, etc., the extracting solution is concentrated and dried; the extracting solution is separated by polyamide, or silica gel, or ion exchange resin, etc. in the way of column chromatography. The purification methods can be used in the mode of single use or combined use. The obtained mulberry twig extract comprises a large quantity of flavonoid components which comprises rutin, meletin, morin, etc.

Description

Ramulus Mori extract and extracting method thereof and purposes
Technical field:
The present invention relates to natural plant extracts and extraction process thereof and new purposes, particularly Ramulus Mori extract is prepared into the health food of prevention hyperuricemia and gout and the medicine of treatment hyperuricemia and gout.
Background technology:
Gout is the chronic lifelong participation disease of one group of purine metabolic disturbance, its pathogeny complexity, clinical characters is that hyperuricemia and the gouty arthritis that causes are thus shown effect repeatedly, tophus deposition, gouty chronic arthritis and joint deformity, often involves kidney and causes chronic interstitial nephritis and form uric acid renal calculus.
Hyperuricemia is meant the paranormal a kind of state of uric acid in the blood, and it is the prelude of gout, also is the important clinical performance of gout.Gouty arthritis be too much urate deposition at joint tissue, stimulate joint tissue also to cause that a series of inflammatory reaction causes, can be divided into acute and chronic two kinds.Acute gouty arthritis is red, swollen, hot with paroxysmal joint, pain and moving obstacle are outstanding behaviours, can disappear gradually after in a few days, and function of joint is also recovered normally not stay joint damage.Gouty arthritis,chronic is meant on the basis that acute arthritis is shown effect repeatedly, causes the destruction of articulation structure and soft tissue, and deformity and limitation of activity in various degree, the appearance tophus that has appear in the joint.After in a single day gouty arthritis,chronic forms, expendable often, the acute attack of outbreak repeatedly still can be arranged on chronic basis, make joint damage more obvious.
Along with the progress of human material progress and the significant change of dietary structure, gout in Europe, North America, Japan, Africa etc. area and country more and more general.As in the U.S., the gout sickness rate is near 1% among all groups, and sickness rate increases with advancing age, and over-65s male sickness rate surpasses 5%; In Europe, gout has been invaded 13% adult male and 5% adult female.The gout sickness rate is higher after women's menopause, and having report domestic is 69.2%, reaches 91% abroad.Gout has become one of world today's property commonly encountered diseases, difficult disease.
China's gout sickness rate increases trend clearly, and some big and medium-sized cities gout sickness rate are close with developed country.Scholarly forecast is arranged, and gout may become No. second metabolic disease that is only second to diabetes in China.
Different with other diseases, the hazardness of gout does not often cause enough attentions.No matter be primary gout or part secondary gout, all be the property suffered from disease all the life, its course of disease is long, outbreak very easily repeatedly, the patient is often agonizing.Show that according to analysis of clinical 41% patient with gout has remarkable renal function injury, 25% patient with gout is died from renal failure.In addition, often occur together aged disease and metabolic diseases such as cardiovascular and cerebrovascular disease (as hypertension, coronary heart disease, atherosclerosis), hyperlipidemia, obesity, diabetes of patient with gout.Therefore, the hazardness that gout is extremely serious should cause our great attention.
The hyperuricemia animal model is generally to be used to estimate anti-hyperuricemia and the pharmaceutically-active experimental model of gout in the world.This model is with a kind of uricase inhibitor--and-oxonic acid potassium salt suppresses uricolysis as chemical inducer, increases serum uric acid level in the body.
Existing western medicine medicine has selectivity at patient with gout in the symptom that different times showed and uses curative effect.Be used for gout acute attack treatment as anti-inflammation analgesis medicament colchicine and indometacin, drainage uric acid medicine such as probenecid, benzbromarone are used for gap phase and chronic phase treatment, and it is synthetic that xanthine oxidase inhibitor medicine allopurinol (unique use clinically) suppresses uric acid.But because the said medicine toxic and side effects is big, the patient often can not adhere to taking for a long time, is difficult to disease controlling.Have the GI irritation effect as colchicine and indometacin, the former also produces side effect such as leucocytes reduction and alopecia; Probenecid, benzbromarone have gastrointestinal reaction, renal colic and excite side effect such as gout acute attack; Allopurinol causes allergy (incidence rate is up to 10-15%), super quick Stevena-Johnson syndrome (can cause 27.5% the maculopapule death occurs), bone marrow depression serious toxic and side effects such as (though more rare, consequence are serious).
The gout of modern medicine belongs to the traditional Chinese medical science " acute arthralgia ", " yellowish perspiration ", " gout ", " severe and migratory arthralgia " or " vitamin B1 deficiency " category, also have generally pass through with " arthromyodynia ".Chinese medicine is according to the principle of determination of treatment based on pathogenesis obtained through differentiation of symptoms and signs, differential diagnosis of diseases medication, and the treatment gout has obtained effect preferably, and few side effects, is easily accepted by the patient, thereby reaches long-term prescription, the purpose of final radical curing of disease.The anemofrigid-damp arthralgia person is based on dispelling cold by warming the meridian, eliminating dampness and dredging channels, and the available heart of a lotus seed subtracts with core Tonga or the treatment of Tetrandra and Poria Decoction plus-minus; Rheumatism pyretic arthralgia person is based on heat-clearing and toxic substances removing dampness removing, promoting blood circulation, removing blood stasis and relieving pain, and available white tiger guizhi decoction or SIMIAO WAN plus-minus are treated; The blockage of phlegm and blood stasis person regulates the flow of vital energy based on a surname's heap soil or fertilizer over and around the roots, and available pink drink adds the flavor treatment; The deficiency of YIN-blood person should hold concurrently with the dispelling wind damp eliminating based on nourshing blood and promoting blood circulation, and available SIWU TANG adds the flavor treatment.According to the principle of differential diagnosis of diseases medication, Rhizoma Smilacis Glabrae, silkworm excrement, Rhizoma Polygoni Cuspidati etc. have the increase urate excretion, reduce the effect of serum uric acid level, can be used for treating hyperuricemia; Caulis Piperis Kadsurae, Radix Clematidis, Semen Sinapis Albae, Radix Scrophulariae etc. can be used for treating stiff, the tophus of joint deformity; Herba Lysimachiae, Spora Lygodii etc. are used for the treatment of urinary stone.
Patent Office of the People's Republic of China successively discloses the Chinese medicine compound goods of a plurality of treatment gouts, as disclose " gout miraculous cure wine and preparation technology thereof " [publication number CN 1144676A on March 12nd, 1997, application number 961028513], " medicated wine of treatment gout " [publication number CN 1178689 was disclosed on April 15th, 1998, application number 97107775.4], " compound sanbi medicinal liquor for treating gout " [publication number CN 1229658 was disclosed on the 29th in JIUYUE, 1999, application number 98111903.4], " a kind of Chinese medicine for the treatment of sciatica and goat " [publication number CN1080868A was disclosed on January 19th, 1994, application number 93111076.9], " a kind of Chinese medicine for the treatment of sciatica and goat " [publication number CN 1116528 was disclosed on February 14th, 1996, application number 94109349.2], " Chinese medicine of treatment goat " [publication number CN 1189374A was disclosed on August 5th, 1998, application number 98110667.6], " a kind of Chinese medicine for the treatment of goat " [publication number CN 1249192A was disclosed on April 5th, 2000, application number 98121618.8], " a kind of Chinese medicine for the treatment of goat " [publication number CN 1250664A, application number 991187725] disclosed on April 19th, 2000.Above-mentioned various gout Chinese medicine compound, contained taste of Chinese medicine number is more, and is crude preparation by using, and wherein main medicine and effective site and composition are not outstanding or not clear, the quality standard that does not have science.The partial monopoly product is a wine product, and modern study shows, various drinks and processed with wine food all can cause the gout outbreak, its reason has: the ethanol in the wine can directly be accelerated purine aggregate velocity in the body, purine output is increased, and can stimulate the interior lactic acid of human body synthetic, and lactic acid can suppress the function of renal excretion uric acid, causes urinary system calculus easily; Some drinks, especially medicated beer can produce a large amount of purine during the fermentation.Therefore on the modern science angle, the above-mentioned patent Chinese medicine discomfort that relates to medicated wine treatment gout is preferably patient with gout or anemia of pregnant woman patient, or takes with patients such as heart disease, hypertension.
Ramulus Mori is the twig of moraceae plants Morus alba L., is the medicine-food two-purpose conventional Chinese medicine.Ramulus Mori energy wind-damp dispelling, sharp joint, row aqueous vapor are controlled anemofrigid-damp arthralgia, spasm of the limbs, and beriberi edema, the human body pruritus due to wind pathogen can be used for treating brachialgia, aqueous vapor beriberi, hypertension, lichen planus.Ramulus Mori contains tannin, saccharide and polyhydroxylated alkaloid etc., and contains a large amount of flavone compounds, as rutin (Rutin), Quercetin (Quercetin) and morin (Morin) etc.The Ramulus Mori compound preparation is mainly used in diseases such as treatment apoplexy sequela, type ii diabetes clinically, but does not see the relevant report that treatment hyperuricemia and gout are arranged.Patent Office of the People's Republic of China once disclosed " Chinese medicine of treatment goat " [publication number CN 1192916 on the 16th JIUYUE in 1998, application number 97107394.5], this Chinese medicine compound is to contain by the Ramulus Mori stir-fry to process product and other 22 kinds of Chinese herbal medicine, modern study confirms that Ramulus Mori is after parch, the main component general flavone content obviously reduces, in addition, this compound effective component and effective ingredient are all indeterminate.
Summary of the invention
Purpose of the present invention is exactly in order to address the above problem, propose a kind of Ramulus Mori extract and extraction process thereof and new medicine use, this new purposes is application and the application in treatment hyperuricemia and gout medication preparation of Ramulus Mori extract in the health food preparation that prevents and treats hyperuricemia and gout.
One of technical solution of the present invention is: Ramulus Mori extract, system extracts more than 1 time or 1 time with organic solvents such as 0~95% ethanol water or chloroform, acetone, ethyl acetate, methanol or their extraction solvent that is combined as by single medical material Ramulus Mori under 10-100 ℃ of temperature conditions, merge extractive liquid, reclaims solvent promptly.
Two of technical solution of the present invention is: the extracting method of Ramulus Mori extract, single medical material Ramulus Mori extracts more than 1 time or 1 time with 0~95% ethanol water, chloroform, acetone, ethyl acetate, methanol equal solvent or their extraction solvent that is combined as under 10-100 ℃ of temperature, merge extractive liquid, reclaims solvent promptly.
Extract obtained available following method is further purified: 1. the gained extracting solution is transferred pH to 7.5-9.5 with alkaline matters such as sodium hydroxide, leaves standstill, and centrifugal back supernatant is transferred pH to 4-6 with hydrochloric acid, leave standstill, get precipitation, be washed to neutrality, be drying to obtain, general flavone content is 10-60%; 2. the gained extracting solution with defats such as petroleum ether or gasoline after, concentrate, be drying to obtain, general flavone content is 1-5%; 3. the gained extracting solution with defats such as petroleum ether or gasoline after, extract respectively or in turn with chloroform, ethyl acetate, acetone, methanol or water, each extracting solution concentrates respectively, is drying to obtain, and general flavone content is followed successively by 5-20%, 10-30%, 5-30%, 1-10%, 1-10%; 4. gained extracting solution macroporous resin adsorption is adsorbed with 10~95% ethanolysis, and desorbed solution concentrates, and is drying to obtain, and general flavone content is 20-80%; 5. the gained extracting solution further after adsorbents adsorb such as kieselguhr, aluminium oxide, behind organic solvent eluting such as chloroform, ethyl acetate, acetone, methanol, ethanol, reclaims solvent, concentrate drying, that is and, general flavone content is 10-80%; 6. being filler with polyamide, silica gel, ion exchange resin, using column chromatography, is eluant with organic solvent such as chloroform, ethyl acetate, acetone, methanol, ethanol or water, and eluent concentrates, and is drying to obtain, and general flavone content is 10-80%; 7. the combination of several method can singly be used or adopt to above purification process, and general flavone content is 0.5-80% in the Ramulus Mori extract, wherein contains rutin, Quercetin and morin etc.
Three of technical solution of the present invention is: Ramulus Mori extract and be the medicine made of active component (comprising oral, ejection preparation and external preparation) and the application of health food in diseases such as prevention and treatment hyperuricemia and gout with the Ramulus Mori extract.
Advantage of the present invention is, remedies the deficiencies in the prior art, and a kind of medicine-food two-purpose is provided, and has the new medicine preparation or the health food of prevention or treatment hyperuricemia and gout effect.
The Ramulus Mori extract of gained of the present invention does not have influence to normal serum uric acid level, has good safety, and the patient who is suitable for all ages and classes level and all types of hyperuricemia and patient with gout and the diseases such as cardiovascular and diabetes that occur together uses.
The present invention scientifically estimates on hyperuricemia model and the effectiveness of definite Ramulus Mori extract treatment hyperuricemia and gout, and be index with the content of total flavones and rutin, Quercetin, morin, set up the quality standard of science, drug effect is reliable and stable, and the technological process controllability is strong.
The treatment hyperuricemia that is obtained by the present invention and the medicament sources of goat are single, the main pharmacodynamics definite ingredients.
The present invention adopts the different solvents system to extract or obtains to have treatment hyperuricemia and gout drug effect extract through the macroporous adsorbent resin enrichment, and technology is reasonable, and strong operability is convenient to commercial production.
Along with the change of social development, dietary structure and the prolongation of human longevity, the gout sickness rate raises year by year, has become one of world today's property commonly encountered diseases, difficult disease.Its hazardness is on the rise, in the international coverage demand of preventing and treating the gout medicine is increased sharply, the invention solves the problem that lacks efficient, of fine quality, safe, controlled gout Chinese medicine on the at present domestic and international drug market, the product that obtains has good effect, safe and reliable, quality controllable, be easy to industrialization, and the incomparable advantage of multiple Western medicine such as be easy to take, cheap, can make the lot of domestic and foreign patient break away from slight illness, improve the quality of living, have vast market prospect.
Experimentation of the present invention fully proves, the xanthine oxidase inhibitor medicine allopurinol of unique use clinically, though can very significant inhibition uric acid synthetic, reduce hyperuricemia animal serum uric acid level, simultaneously the Normal animal serum level is also had remarkable reduction effect.Ramulus Mori extract has the effect that reduces hyperuricemia mice and rat blood serum uric acid level, but intact animal's uric acid level is not made significant difference.And Ramulus Mori extract can significantly suppress the gouty arthroncus that rat uric acid sodium causes, K in the reduction inflammation tissue +Content and peripheral neurotransmitter, protection inflammation tissue alleviates collagen fiber fracture situation.Therefore, compare with allopurinol, the new product of Chinese medicine that is prepared by Ramulus Mori extract has the advantage safe, that effectiveness is good.
Each extract of Ramulus Mori that obtains with the present invention can be used for the treatment of diseases such as hyperuricemia and gout as the main medicine of effective ingredient.Each extract of Ramulus Mori is equipped with proper pharmaceutical excipients, can be made into preparations such as tablet, granule, capsule or injection.These dosage forms are according to the known method preparation of those skilled in the art.The adjuvant that is used to make moulding processs such as tablet, granule, capsule is the auxiliary agent of using always, for example starch, gelatin, arabic gum, Polyethylene Glycol, or the like, surfactant, lubricant, disintegrating agent, antiseptic, correctives, pigment, or the like.The solvent that is used to make injection is the water for injection used always, oil for injection, ethanol, glycerol, propylene glycol, Polyethylene Glycol, benzyl benzoate, dimethyl acetylamide etc., also contain other additives in the preparation, for example solubilizing agent, wetting agent, emulsifying agent, buffer agent and suspensoid etc.
The specific embodiment
Following examples are only as the usefulness of further setting forth invention, can not be used for limiting the present invention.
Embodiment 1: the influence of the gouty arthritis that Ramulus Mori extract WE causes hyperuricemia and normal mouse and rat blood serum uric acid level and rat uric acid sodium
Get exsiccant Ramulus Mori decoction pieces, decoct three times each 1 hour respectively with 10 times, 8 times, 6 times water gagings.Merge decoction liquor, concentrating under reduced pressure, drying promptly get WE, and yield is 11.52%, and general flavone content is 1.1%.
WE sees Table 1 and table 2 respectively to the result that influences of hyperuricemia and normal mouse and rat uric acid level.Under this experiment condition, the high, medium and low dosage group of WE all reduces hyperuricemia mice and rat blood serum uric acid level very significantly, and wherein WE height, middle dosage group can return to hyperuricemia mice serum uric acid level normally; Among the WE, low dosage and high dose group all can return to hyperuricemia rat blood serum uric acid level normally, WE high dose group action intensity is close with the positive drug allopurinol.
The high, medium and low dosage group of WE does not have the significance influence to normal mouse and rat blood serum uric acid level, and the allopurinol group can reduce normal mouse and rat blood serum uric acid level significantly.
Table 1 Ramulus Mori extract WE is to the influence of hyperuricemia mice and rat uric acid level (x ± s)
Group Dosage (mg/kg) Serum uric acid level (mg/100ml)
Mice Rat
The blank group of normal saline group model WE group purine alcohol group - - 26 52 104 10 8.87±0.17 12.39±0.28 9.64±0.19 * 8.72±0.12 ** 8.62±0.10 *** 7.62±0.17 *** 10.26±0.27 12.58±0.28 10.26±0.18 ** 10.53±0.12 *** 9.63±0.28 *** 9.36±0.17 ***
Compare with the blank group of model: P<0.05, *P<0.01, * *P<0.005
Table 2 Ramulus Mori extract WE is to the influence of Normal animal serum uric acid level (x ± s)
Group Dosage mg/kg Serum uric acid level (mg/100ml)
Mice Rat
Normal saline group WE allopurinol - 26 52 104 10 3.52±0.12 3.38±0.15 3.16±0.16 2.91±0.19 1.99±0.16 *** 3.40±0.26 3.54±0.34 3.37±0.12 3.20±0.29 2.21±0.16 ***
Compare with the normal saline group: *P<0.05, *P<0.01, * *P<0.001
Ramulus Mori extract WE causes that to rat uric acid sodium the result that influences of gouty arthritis model shows that WE suppresses the rat gouty arthritis swelling that uric acid sodium causes significantly,, effect and colchicine be (following see Table 3) quite.Tried periarticular soft tissues Histological research and show, WE and colchicine group can be protected the inflammation tissue, alleviate collagen fiber fracture situation.WE can reduce K in the inflammation tissue significantly +Content (P<0.001) and 5-HT content, action intensity and colchicine is (following see Table 3) quite.And the content that is tried NE in the periarticular soft tissues, DA is had certain reduction effect, but there is not significance influence (data slightly).
Table 3 Ramulus Mori extract WE causes the influence of gouty arthritis swelling degree to rat uric acid sodium
( x±s)
Dosage (mg/kg) Ankle joint Zhou Jing (cm) K +Content (mol/mg) 5-HT content (g/g)
The blank group of normal saline group model WE group colchicine group - - 52 12.5 2.22±0.08 3.28±0.12 2.76±0.09 2.56±0.14 2.10±0.02 7.42±0.06 3.02±0.04 * 3.72±0.03 * 382±2.14 723±9.45 395±4.54 * 401±5.12 *
Compare with the blank group of model: *P<0.001
Embodiment 2: Ramulus Mori extract AE-PH is to the influence of hyperuricemia and normal mouse serum uric acid level
Get exsiccant Ramulus Mori decoction pieces, extract each 2 hours three times with the 80% ethanol water hot reflux of 8 times, 8 times, 6 times amounts respectively.Extracting solution merges, and decompression recycling ethanol is transferred pH8.5 with 10%NaOH, and is centrifugal behind the sufficient standing, and it is 4-6 that supernatant transfers to pH with hydrochloric acid, leaves standstill, and gets precipitation, is washed with distilled water to neutrality, promptly gets AE-PH after the drying, and yield is 0.5%, and general flavone content is 23%.
AE-PH can significantly reduce hyperuricemia mice serum uric acid level, has good dose-effect relationship (table 4), and to normal mice serum uric acid level do not make significant difference (data slightly).
Table 4 Ramulus Mori extract AE-PH is to the influence of hyperuricemia mice serum uric acid level (x ± s)
Group Dosage (mg/kg) Serum uric acid level (mg/100ml)
The blank group of normal saline group model AE-PH allopurinol group - - 68 34 17 10 8.30±1.25 12.01±1.19 7.97±1.05 *** 8.17±2.64 *** 10.44±2.01 * 7.01±0.13 ***
Compare with the blank group of model: P<0.05, *P<0.01, * *P<0.001
Embodiment 3: Ramulus Mori extract AE1, AE1-PE and AE1-1~AE1-4 are to the influence of hyperuricemia and normal mouse serum uric acid level
Get exsiccant Ramulus Mori decoction pieces, extract each 2 hours three times with the 80% ethanol water hot reflux of 8 times, 8 times, 6 times amounts respectively.Extracting solution merges, and concentrating under reduced pressure promptly gets AE1 after the drying, and yield is 4.18%, and general flavone content is 2.0%.
After AE1 and kieselguhr mix and stir, adopt the Soxhlet circumfluence method, behind defat with petroleum ether, be drying to obtain AE1-PE, yield is 3.2%, and general flavone content is 3.5%.
After AE1 and kieselguhr mix and stir, adopt the Soxhlet circumfluence method, behind defat with petroleum ether, use chloroform, ethyl acetate, methanol and water extraction to colourless successively, each extracting solution is concentrated, dry respectively, promptly get AE1-1, AE1-2, AE1-3 and AE1-4, yield is respectively 3.16%, 9.85%, 64.17%, 0.7%, and general flavone content is respectively 10.1%, 12.7%, 6.0% and 5.6%.
AE1, AE-1-PE and AE1-1~AE1-4 all can significantly reduce hyperuricemia mice serum uric acid level (table 5), and to normal mice serum uric acid level all do not make significant difference (data slightly).
Table 5 Ramulus Mori extract AE1, AE-1-PE and AE1-1~AE1-4 are to hyperuricemia mice blood
The influence of clear uric acid level (x ± s)
Group Dosage (mg/kg) Serum uric acid level (mg/100ml)
The blank group of physiological saline group model AE1 AE-1-PE AE1-1 AE1-2 AE1-3 AE1-4 allopurinol group - - 550 360 17 54 350 4.5 10 8.10±1.45 11.70±1.09 8.02±1.15 *** 7.81±1.10 *** 7.87±2.94 *** 10.24±4.04 * 8.09±2.25 *** 8.70±1.92 *** 7.21±0.16 ***
Compare with the blank group of model: P<0.05, *P<0.01, * *P<0.001
Embodiment 4: Ramulus Mori extract AE2~AE6 is to the influence of hyperuricemia and normal mouse uric acid level
Get exsiccant Ramulus Mori decoction pieces, with 4 times of volume 20% ethanol waters, hot reflux three times, each 2 hours, extracting solution merged, and concentrating under reduced pressure is drying to obtain AE2, and yield is 8.8%, and general flavone content is 2.3%; Get exsiccant Ramulus Mori decoction pieces, with 4 times of volume 40% ethanol waters, hot reflux three times, each 2 hours, extracting solution merged, and concentrating under reduced pressure is drying to obtain AE3, and yield is 5.8%, and general flavone content is 3.3%; The Ramulus Mori decoction pieces, with 4 times of volume 60% ethanol waters, hot reflux 2 times, each 1 hour, extracting solution merged, and concentrating under reduced pressure is drying to obtain AE4, and yield is 4.8%, and general flavone content is 3.2%; The Ramulus Mori decoction pieces, with 4 times of volume 70% ethanol waters, hot reflux 1 time, each 1.5 hours, extracting solution merged, and concentrating under reduced pressure is drying to obtain AE5, and yield is 2.7%, and general flavone content is 2.5%; The Ramulus Mori decoction pieces, with 4 times of volume 95% ethanol waters, hot reflux 1 time, each 1.5 hours, extracting solution merged, and concentrating under reduced pressure is drying to obtain AE6, and yield is 2.1%, and general flavone content is 2.4%.
Ramulus Mori extract AE2~AE6 all reduces hyperuricemia mice serum uric acid level very significantly, and returns to normal (table 6), and to normal mice serum uric acid level do not make significant difference (data slightly).
Table 6 Ramulus Mori extract AE2~AE6 is to the influence of hyperuricemia mice uric acid level (x ± s)
Group Dosage (mg/kg) Serum uric acid level (mg/100ml)
The blank group of physiological saline group model AE2 AE3 AE4 AE5 AE6 allopurinol group - - 712 605 500 283 240 5 10 7.78±1.39 9.77±7.55 8.15±0.19 ** 7.05±0.19 ** 7.18±0.45 ** 7.15±0.66 ** 7.25±0.66 ** 5.90±1.20 *** 5.31±2.34 ***
Compare with the blank group of model: P<0.05, *P<0.01, * *P<0.005
Embodiment 5: the influence of 10 pairs of hyperuricemias of Ramulus Mori extract AE7~AE and normal mouse uric acid level
Get exsiccant Ramulus Mori decoction pieces, with 7 times of volume 60% ethanol waters, hot reflux three times, each 2 hours.Extracting solution merges, concentrating under reduced pressure, remove ethanol after by absorption with macroporous adsorbent resin, resolve with 20%, 40%, 60%, 80% ethanol successively, desorbed solution concentrates respectively, promptly get AE7, AE8, AE9, AE10 after the drying, yield is respectively 1.3%, 0.91%, 0.57%, 0.34%, and general flavone content is respectively 27.6%, 54.2%, 60.0%, 67.9%.
AE7~AE10 can significantly reduce hyperuricemia mice serum uric acid level (table 7), but to there are no significant the influence (data slightly) of normal mice serum uric acid level.
Table 7 Ramulus Mori extract AE7~AE10 is to the influence of hyperuricemia mice uric acid level
( x±s)
Group Dosage (mg/kg) Serum uric acid level (mg/100ml)
The blank group of normal saline group model AE7 AE8 AE9 AE10 allopurinol group - - 94 67 39 24 5 10 4.85±0.04 5.70±0.06 5.37±0.03 * 4.34±0.02 *** 4.92±0.06 *** 5.13±0.05 ** 4.97±0.05 ** 4.01±0.06 ***
Compare with the blank group of model: *P<0.05, *P<0.01, * *P<0.001
Embodiment 6: Ramulus Mori extract AE11~AE13 is to the influence of hyperuricemia and normal mouse uric acid level
Get exsiccant Ramulus Mori decoction pieces, with 7 times of volume 60% ethanol waters, hot reflux three times, each 2 hours.Extracting solution merges, concentrating under reduced pressure, remove ethanol after behind the diatomite adsorption, with organic solvents such as chloroform, ethyl acetate, methanol eluting successively, reclaim solvent respectively, concentrate drying promptly gets AE11~AE13, yield is respectively 1.3%, 5.69%, 2.57%, and general flavone content is respectively 10.6%, 54.2%, 36.0%.
AE11~AE13 can significantly reduce hyperuricemia mice serum uric acid level (table 8), but to there are no significant the influence (data slightly) of normal mice serum uric acid level.
Table 8 Ramulus Mori extract AE11~AE13 is to the influence of hyperuricemia mice uric acid level
( x±s)
Group Dosage (mg/kg) Serum uric acid level (mg/100ml)
The blank group of normal saline group model AE11 AE12 AE13 allopurinol group - - 44 192 87 5 10 6.85±0.03 8.70±0.02 7.37±0.05 * 6.98±0.03 *** 7.45±0.05 *** 6.98±0.04 ** 5.59±0.07 ***
Compare with the blank group of model: *P<0.05, *P<0.01, * *P<0.001
Embodiment 7: Ramulus Mori extract AE14~AE17 is to the influence of hyperuricemia and normal mouse uric acid level
Get exsiccant Ramulus Mori decoction pieces, with 7 times of volume 60% ethanol waters, hot reflux three times, each 2 hours.Extracting solution merges, concentrating under reduced pressure, remove ethanol after polyamide column chromatography, water, 30% ethanol, 60% ethanol and 80% ethanol is eluting successively, reclaim solvent respectively, concentrate drying promptly gets AE14~AE17, yield is respectively 2.52%, 8.48%, 7.67%, 8.88%, and general flavone content is respectively 1.1%, 48.5%, 63.6%, 21.3%.
AE14~AE17 can significantly reduce hyperuricemia mice serum uric acid level (table 9), but to there are no significant the influence (data slightly) of normal mice serum uric acid level.
Table 9 Ramulus Mori extract AE14~AE17 is to the influence of hyperuricemia mice uric acid level (x ± s)
Group Dosage (mg/kg) Serum uric acid level (mg/100ml)
The blank group of normal saline group model AE14 AE15 AE16 AE17 allopurinol group - - 56 188 170 197 5 10 5.85±0.04 7.87±0.02 7.47±0.02 6.68±0.03 * 5.75±0.06 *** 6.80±0.05 * 5.98±0.03 ** 5.07±0.05 ***
Compare with the blank group of model: *P<0.05, *P<0.01, * *P<0.001
Embodiment 8: the preparation of shaped preparation
Get above-mentioned Ramulus Mori extract 1000g, add 500-5000g starch or dextrin, mix homogeneously is granulated, and tabletting makes tablet; Get above-mentioned Ramulus Mori extract 1000g, add 500-5000g starch, mix homogeneously, encapsulated, promptly get capsule; Get above-mentioned Ramulus Mori extract 1000g, add macromolecule adjuvants such as 1000-10000g vegetable oil or Polyethylene Glycol, mix homogeneously is made soft capsule.
Get above-mentioned Ramulus Mori extract 1000g, add 1000-10000g Icing Sugar or lactose or soluble starch etc., mix homogeneously is granulated, and gets granule.Get above-mentioned Ramulus Mori extract 1000g, add water and an amount of solubilizing agent dissolving, packing, oral liquid is made in sterilization
Get above-mentioned Ramulus Mori extract 1000g, add injection additive and water for injection to 1000ml, dissolving is filtered, and filtrate is transferred pH 5.0-9.0, and sterilization promptly gets injection.
Get above-mentioned Ramulus Mori extract 1000g, use dissolve with ethanol, be prepared into liniment or spray;
Get above-mentioned Ramulus Mori extract 1000g, use dissolve with ethanol, make aerosol behind the adding propellant.
Get above-mentioned Ramulus Mori extract 1000g, with auxiliary materials and mixing such as an amount of water-soluble base or rubber, cataplasma or rubber agent are made in coating.

Claims (3)

1. Chinese medicine Ramulus Mori extract, it is characterized in that this extract is to extract purification with following method to obtain: get exsiccant Ramulus Mori decoction pieces, with 7 times of volume 60% ethanol waters, hot reflux three times, each 2 hours, extracting solution is merged, concentrating under reduced pressure is removed extracting solution behind the ethanol through polyamide column chromatography, and water, 30% ethanol, 60% ethanol and 80% ethanol is eluting successively, reclaim solvent respectively, concentrate drying.
2. the method for extraction and purification of a Chinese medicine Ramulus Mori extract, it is characterized in that this method comprises the steps: to get exsiccant Ramulus Mori decoction pieces, with 7 times of volume 60% ethanol waters, hot reflux three times, each 2 hours, extracting solution is merged, concentrating under reduced pressure is removed extracting solution behind the ethanol through polyamide column chromatography, and water, 30% ethanol, 60% ethanol and 80% ethanol is eluting successively, reclaim solvent respectively, concentrate drying.
3. the Ramulus Mori extract of claim 1 is in preparation prevention or the health product of treatment hyperuricemia and the purposes in the medicine.
CN02113004.3A 2002-05-14 2002-05-14 Extract of mulberry twig and its extracting process and novel usage Expired - Fee Related CN1267113C (en)

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Publication number Priority date Publication date Assignee Title
CN1313458C (en) * 2005-04-30 2007-05-02 苏州大学 Extraction process of total mulberry twig flavone
CN100384831C (en) * 2006-05-10 2008-04-30 江南大学 Extraction separation purification and identification of flavonoid monomers in oriental blueberry melanin
CN101869605B (en) * 2010-07-02 2012-05-02 安徽济人药业有限公司 Method for extracting and separating mulberry leaf flavone and alkaloid composite
CN102154876B (en) * 2011-02-21 2012-05-09 浙江理工大学 Method for extracting and separating components in ramulus mori skin
CN102429897B (en) * 2011-09-16 2013-03-13 四川大学 Pharmaceutical composition for improving oral bioavailability of morin
CA2915822C (en) * 2013-06-17 2016-11-29 Unigen, Inc. Compositions and methods for joint health
CN103583952B (en) * 2013-11-15 2015-04-29 郑州轻工业学院 Ramulus mori polysaccharide oral liquid and preparation method thereof
CN104644753A (en) * 2015-02-04 2015-05-27 新疆医科大学 Medical application and preparation method of mulberry twig alpha-glycosidase inhibitory activity extract and composition of mulberry twig alpha-glycosidase inhibitory activity extract
KR101876805B1 (en) * 2016-10-18 2018-07-10 고려대학교 산학협력단 The composition containing the ethanol extracts of Ramulus mori for preventing or treating obesity
CN108395461B (en) * 2017-02-07 2022-01-07 浙江康恩贝制药股份有限公司 7, 4' -O-beta-D-glucose morin glycoside, preparation thereof and application thereof in preparing anti-inflammatory drugs
CN111358882A (en) * 2020-05-07 2020-07-03 袁晓明 Traditional Chinese medicine composition for treating gouty arthritis, preparation method, preparation and application
CN113262886B (en) * 2021-05-24 2022-09-16 衢州职业技术学院 Mulberry leaf essential oil extraction and purification method and centrifuge for extraction
CN114190542A (en) * 2021-12-06 2022-03-18 广东省农业科学院蚕业与农产品加工研究所 Uric acid-reducing health-preserving soup package material and preparation method thereof
CN114176212A (en) * 2021-12-06 2022-03-15 广东省农业科学院蚕业与农产品加工研究所 Chafing dish ingredient capable of reducing uric acid and improving immunity and preparation method thereof
CN114989124A (en) * 2022-06-15 2022-09-02 华中科技大学同济医学院附属协和医院 Method for extracting quercetin from herba Hedyotidis Diffusae, and preparation method and liposome product of liposome thereof

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