CN1223371C - Preparation of effective component from rhodiola crenulate - Google Patents
Preparation of effective component from rhodiola crenulate Download PDFInfo
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- CN1223371C CN1223371C CN 03128415 CN03128415A CN1223371C CN 1223371 C CN1223371 C CN 1223371C CN 03128415 CN03128415 CN 03128415 CN 03128415 A CN03128415 A CN 03128415A CN 1223371 C CN1223371 C CN 1223371C
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Abstract
The present invention relates to the extraction separation and a preparation of active components of leonurus integripetal rhodiola herb. The preparation of active components of leonurus integripetal rhodiola herb is various acceptable preparations in medicine and is prepared from the active components of total flavone and rhodioloside or the active components of 30% to 70% of amylase and 30% to 70% of medical auxiliary materials, the active components of total flavone and rhodioloside contain small molecular compounds and the active components of 30% to 70% of amylase and 30% to 70% of medical auxiliary materials contain macromolecular compounds. The preparation of active components of leonurus integripetal rhodiola herb has the advantages of exact component content, obvious curative effect and simple preparing methods, and can be selected and used according to different therapeutic purposes. The preparation of active components of leonurus integripetal rhodiola herb is suitable for industrialization production.
Description
Technical field
The present invention relates to the preparation and the preparation method of effective components of Chinese medicinal, be specifically related to Radix Rhodiolae effective ingredient preparation and preparation method.
Background technology
The Crassulaceae Rhodida plant is perennial herb or semishrub plant, be distributed in high and cold areas such as former Soviet Union the Far East Area and Northeast China, northwest, southwest, there is kind more than 90 in the whole world, China has kind more than 70, wherein Radix Rhodiolae (Rhodiola Crenulata) is very big at China's reserves, mainly is distributed in high and cold mountain areas such as Tibet, Sichuan, Yunnan.
There is the scholar that the composition of Radix Rhodiolae was made systematic study, find to have nearly more than 10 chemical compounds to exist at present, micromolecule glycoside such as rhodioloside (Salidroside) are wherein arranged, crenulatin (Crenulatin), the plain glycoside (rhodinin and rhodiosin) of flavonoid such as herbaceous stem is also arranged, Radix Rhodiolae glycoside (Crenuloside), Radix Rhodiolae element (flavonol combines with the phenylpropyl alcohol element), and other is as gallic acid (gallic acid), pyrogallol salt, butyl alcohol (tyrosol) etc., also have many macromolecular compounds such as polysaccharide, its structure mainly is made up of arabinose and glucose.
Because the existence of numerous compositions, make Radix Rhodiolae have physiologically active comparatively widely, it has resisting fatigue, radioprotective, anti-hypoxia, defying age, cold resistance and high temperature, blood sugar lowering and blood fat, regulates effects such as immunity and blood pressure, and this just clearly shows that it has a wide range of applications in sports medical science, aerospace medicine and geriatrics field.
Exploitation for Radix Rhodiolae, main at present capsule, electuary, oral liquid based on crude drug powder or crude extract, these qualities of the pharmaceutical preparations are stable inadequately, mouthfeel is not good, astringent taste is difficult to by hidden or elimination after the seasoning, and not with each active ingredient separation, quantitative, makes its effect indeterminate, only use as health product, usage degree is restricted.
Summary of the invention
The objective of the invention is to adopt modern separation means according to the relation of active ingredient and effect with micromolecular compound in the Radix Rhodiolae separated with macromolecular compound, purification, make the definite Radix Rhodiolae preparation of active ingredient, and can select to use according to different therapeutic purposes.
Radix Rhodiolae effective ingredient preparation disclosed by the invention be by
1. contain the medically acceptable all kinds of preparations that micromolecular compound extract 30～70% and 70～30% pharmaceutic adjuvants are formed.Wherein the micromolecular compound extract contains total flavones 25～40%, contains rhodioloside 4～8%.
2. contain the medically acceptable all kinds of preparations that macromolecular compound extract 30～70% and 70～30% pharmaceutic adjuvants are formed.Wherein contain polysaccharide 50～90% in the macromolecular compound extract.
Pharmaceutic adjuvant of the present invention is that pharmaceutic adjuvant such as the used adjuvant of solid preparation commonly used in the pharmacy industry is starch, dextrin, microcrystalline Cellulose, calcium hydrogen phosphate, calcium sulfate, carboxymethyl cellulose, sucrose, hyprolose, magnesium stearate, white carbon black, Pulvis Talci, pyrrolidone, lactose, sucrose, HPMC etc.The used adjuvant of liquid preparation is water, sucrose, A Si BATANG, citric acid, sodium chloride, vitamin C, propylene glycol, glycerol etc.
Another object of the present invention provides the preparation method of above-mentioned Radix Rhodiolae effective ingredient preparation.
Radix Rhodiolae effective ingredient preparation of the present invention makes by following method:
1. the preparation of total extractum
The Radix Rhodiolae root is through section, and water-pure intermixture water: ethanol=99: 1～1: 99 reflux, extract, 1-3 time, total solvent amount are 10 to 30 times of crude drug, always 2 to 6 hours extraction times.The medicinal liquid that obtains can obtain total extract extractum through concentrated, vacuum drying.
The extractum extraction ratio is 20～40%, wherein contains total flavones 10～20%, rhodioloside 1～3%, polysaccharide 10～50%.
2. the separation of active ingredient, purification
(1) separation of micromolecular compound.
Get 1 part of above-mentioned total extractum, add 5～10 parts of dissolvings of water, medicinal liquid is flowed through and is filled with the chromatographic column of the separating medium of not being with ion-exchange group, and 10～40 parts of eluting of water are clean, and eluent is pending; Use 30～90% ethanol elution chromatographic columns then, concentrate ethanol elution, get the micromolecular compound extract, wherein contain total flavones 25～40%, rhodioloside 4～8%.
(2) separation of macromolecular compound
Get 20 parts of the above-mentioned micromolecular compound water elution liquid in separating, be concentrated into 1 part, stirring adds 1～3 part of 95～100% ethanol down, and static spending the night filtered, filter cake is with 1 part of water dissolution, the active carbon that adds 2% amount refluxed filtered while hot, filtrate cold drying or spray drying 15 minutes, get light brown powder and be macromolecular compound, polyoses content 50～90%.
3. the preparation of effective ingredient preparation
(1) get above-mentioned micromolecular compound extract 30～70%, add pharmaceutic adjuvant 70～30%, formulation method makes tablet, capsule, granule, oral liquid or injection etc. routinely.
(2) get above-mentioned macromolecular compound extract 30～70%, add pharmaceutic adjuvant 70～30%, formulation method makes tablet, capsule, granule, oral liquid or injection etc. routinely.
Of the present inventionly be not meant macroporous resin, silica gel, polyamide, aluminium oxide, cellulose, kieselguhr, activated carbon etc. with the separating medium of ion-exchange group.
Micromolecular compound of the present invention is meant that molecular weight in the Radix Rhodiolae crude extract less than 2000 all kinds of glycosides and glycoside unit, is mainly butyl alcohol, rhodioloside, flavonoid glycoside, gallic acid, protocatechuic acid, 3-O-times of hydrochlorate of (one) epi-nutgall catechuic acid and derivant thereof etc.
Find according to modern study, prolyl endopeptidase (PEP) is active in Alzheimer's disease (senile dementia early) patient's cerebral tissue increases, and this enzyme participates in the distinctive amyloid generation of Alzheimer's disease, therefore expects that the PEP inhibitor can become this sick curative.The gallic acid that contains in the Radix Rhodiolae, accompany hydrochlorate, protocatechuic acid etc., PEP is had remarkable inhibitory action, wherein times acids activity can be equal to mutually with the strongest active natural non-peptide inhibitor D-82041 DEISENHOFEN of having reported that is.The raising learning memory effect of PEP inhibition active component and these product has substantial connection in the Radix Rhodiolae.
Flavone compound has the rat of raising erythrocyte in the Radix Rhodiolae, the activity of liver SOD, and the active trend of the myocardium superoxide dismutase (SOD) of increasing is arranged, reduce lipid peroxide (LPO) content in rat blood plasma, cardiac muscle, the cerebral tissue.In recent years research thinks that LPO is by the coup injury blood vessel endothelium and make prostaglandin metabolism loss of equilibrium etc. participate in arteriosclerotic generation, evolution, SOD activity improving, and reducing the LPO level has certain effect to treatment coronary heart disease.
Butyl alcohol, rhodioloside etc. have oxygen lack resistant function in the Radix Rhodiolae, can obviously prolong the mice hypobaric hypoxia following time-to-live of environmental condition.Have antifatigue effect, its can significant prolongation mice swimming with a load attached to the body time and makes and grab rod and drop the time significant prolongation, can also increase oxygen content in the blood in addition, eliminates in the blood, in the muscle lactic acid and piles up, and reaches the purpose of allaying tiredness.
Macromolecular compound of the present invention is meant polysaccharide and the tannin of molecular weight between 3000 to 30000 in the Radix Rhodiolae total extract, and polysaccharide mainly is made up of arabinose and glucose.
Radix Rhodiolae polysaccharide all shows promotion and restitution preferably to lymphocyte transfer reaction and NKT (NK) activity of normal and immunocompromised mice, point out it to be-kind of new biological response modifier, significant in the treatment of immunocompromised disease (as tumor).
Radix Rhodiolae polysaccharide significantly reduces the various experimental hyperglycemia due to mice euglycemia and epinephrine, the glucose etc., and effect has dose dependent, and it also has the reduction effect to blood fat simultaneously.
Radix Rhodiolae polysaccharide has radiation resistance, can significantly improve the survival rate that is subjected to according to mice, and the hemopoietic system of organism is also had obvious protective effect, and its significant degree can be compared with the positive drug Benzalkonium, and this seldom sees in Chinese medicine and natural drug.
Carry out the test of safety toxicological evaluation with medicine containing active components of Rhodiola crennulata root of the present invention by the GB15193-94 pertinent regulations, the result shows:
1. acute toxicity test: sample is to the acute oral LD of male and female white mice
50All,, belong to nontoxic level material with acute toxicity median lethal dose(LD 50) toxicity grading greater than 21500mg/kg.
2. micronucleus test result: feminine gender.
3. sperm malformation test result: feminine gender.
4.Ames result of the test: feminine gender.
5.30 it feeding trial: the laboratory animal growing state is good, hematological examination, and biochemical analysis, main dirty body is histological examination result when compare with matched group, all no significant difference.
Carry out function test with medicine containing active components of Rhodiola crennulata root of the present invention, the result shows to have antifatigue effect and radiation resistance.
1. antifatigue effect:
1.1. sample: sample is modulated into each concentration for examination with distilled water.
1.2. dosage design: this product human body recommended dose is that basic, normal, high three dosage groups are established in this experiment of 2.2g/60kg. every day, is respectively 0.19,0.37,1.1g/kg, and other establishes blank group (giving distilled water).
1.3. laboratory animal: Kunming kind white mice, male, 20-22 gram, by the cleaning level animal that Shanghai Medical Univ's Animal House provides, the quality certification number: 02--22-1.
1.4. give the sample approach: irritate stomach.
1.5. experimental technique and result:
1.5.1 swimming with a load attached to the body test:
Get 40 of Kunming kind white mice, be divided into four groups at random, 10 every group.Design was according to dosage fed 28 days continuously, after last is irritated stomach 30 minutes, mice is put into water swims, depth of water 30cm, 25 ± 0.5 ℃ of water temperatures, the load sheet lead of 5% body weight of Mus root of the tail portion, the record mice from the swimming beginning to the dead time.
Table 1 sample is to the influence of mice swimming with a load attached to the body time (X ± SD)
| Group | Number of animals (only) | The swimming with a load attached to the body time (second) |
| Dosage high dose in the blank low dosage | 10 10 10 10 | 370±1172 408±255 448±225 666±183 * |
*(through variance analysis) compared with the blank group in P＜0.05
As seen from the above table, the sample high dose group is compared with the blank group, and significant difference is arranged.
1.5.2 liver glycogen is measured:
Get 40 of Kunming kind white mice, be divided into four groups at random, 10 every group.Design was according to dosage fed 28 days continuously, and after last is irritated stomach 30 minutes, mice is put into 30 ℃ of water swimming 90 minutes, take out, it is dirty to get Hepar Mus at once, surveys liver glycogen.
Table 2 sample to mouse movement after the influence (X ± SD) of liver glycogen
| Group | Number of animals (only) | Liver glycogen (mg/100g) |
| Dosage high dose in the blank low dosage | 10 10 10 10 | 433±53 479±88 618±147 746±124 * |
*(through variance analysis) compared with the blank group in P＜0.05
As seen from the above table, the sample high dose group is compared with the blank group, and significant difference is arranged.
1.5.3 blood lactic acid is measured: get 40 of Kunming kind white mice, be divided into four groups at random, 10 every group.Design is according to dosage fed continuously and is surveyed blood lactic acid after 28 days and after last is given sample 30 minutes, and 2% (body weight) of bearing a heavy burden is at temperature 25-30
*(swimming is 60 minutes in 2 water, takes out, and surveys blood lactic acid, and hot-air seasoning after quiet 30 minutes, is surveyed blood lactic acid again.
Table 3 sample is to the influence of mouse movement bleeding from anus lactic acid (X ± SD)
| Group | Number of animals (only) | Blood lactic acid (mmol/l) | ||
| Before the experiment | Tested back 0 minute | Tested back 30 minutes | ||
| Dosage high dose in the blank low dosage | 10 10 10 10 | 4.5±0.2 4.4±0.3 4.4±0.1 4.3±0.3 | 10.5±3.7 10.7±2.0 10.5±0.8 11.0±1.0 | 7.6±1.4 7.9±1.2 7.0±1.0 5.2±0.7 * |
*(through variance analysis) compared with the blank group in P＜0.05
As seen from the above table, the sample high dose group is tested back 30 minutes animal blood lactic acid contents and is compared with the blank group, and significant difference is arranged.
1.5.4 determination of urea nitrogen:
Get 40 of Kunming kind white mice, be divided into four groups at random, 10 every group.Design was according to dosage fed 28 days continuously, and after last is irritated stomach 30 minutes, mice is put into 30 ℃ of water swimming 90 minutes, take out, hot-air seasoning makes peace and quiet, gets Mus blood, and is centrifugal, gets serum and surveys blood urea nitrogen.
Table 4 sample to mouse movement after the influence (X ± SD) of serum urea nitrogen
| Group | Number of animals (only) | Blood urea nitrogen (mmol/l) |
| Dosage high dose in the blank low dosage | 10 10 10 10 | 7.8±0.6 7.7±1.1 7.1±1.7 5.8±1.8 * |
*(through variance analysis) compared with the blank group in P＜0.05
As seen from the above table, the sample high dose group is compared with the blank group, and significant difference is arranged.
2. radiation resistance:
2.1. sample: sample is modulated into each concentration with distilled water, for examination.
2.2. animal origin: Kunming kind white mice, body weight 18-22 gram, male, by the cleaning level animal that laboratory animal portion of Shanghai Medical Univ provides, the quality certification number: 02-22-1.
2.3. dosage design: this sample human body recommended dose is 2.2g/60kg every day, and basic, normal, high three dosage groups are set up in this experiment, is respectively 0.19,0.37,1.1g/kg, and normal control group and radiation matched group are all drunk distilled water.
2.4. give the sample approach: irritate stomach
2.5. experimental technique and result:
After animal is given around the sample continuously, tried thing group and radiation matched group all once with the 60Co irradiation, dosage is 8GY, irradiation time is 8 minutes, irradiation distance is 2 meters, shine back second day half animal and get blood and do numeration of leukocyte, second half animal is observed mean survival time or 30 days survival rates, and the normal control group is got blood and done numeration of leukocyte and observe 30 days survival rates.
Table 5 mice mean survival time (X ± SD)
| Group | Number of animals (only) | Mean survival time (my god) | Survival rate (%) |
| Dosage high dose in the normal control radiation contrast low dosage | 10 10 10 10 10 | 30.0±0 * 10.8±0.6 10.6±0.8 13.9±5.3 14.8±5.9 * | 100 0 0 0 10 |
*(through variance analysis) compared with the radiation matched group in P＜0.05
Learn check by statistics, the sample high dose group is compared with the radiation matched group, and significant difference is arranged.
Table 6 murine interleukin counting (X ± SD)
| Group | Number of animals (only) | Numeration of leukocyte (10 9/L) |
| Dosage high dose in the normal control radiation contrast low dosage | 10 10 10 10 10 | 9.72±0.52 * 2.25±1.79 2.33±1.82 2.03±0.95 3.61±1.14 * |
*(through variance analysis) compared with the radiation matched group in P＜0.05
Learn check by statistics, the numeration of leukocyte of sample high dose group is compared with the radiation matched group, and significant difference is arranged.
Result of the test shows: sample can prolong the animal swimming with a load attached to the body time, reduces animal movement bleeding from anus lactic acid and urea nitrogen levels, promotes the plain deposit of glycogen; Can prolong mean survival time and the survival rate of animal after the disposable irradiation of higher dosage, the leukocyte increasing sum.Illustrate that this sample has resisting fatigue and radiation resistance.
Radix Rhodiolae effective ingredient preparation disclosed by the invention, effective component content is definite, and is evident in efficacy, can select to use according to different therapeutic purposes.The preparation method of Radix Rhodiolae effective ingredient preparation disclosed by the invention is easy, is applicable to suitability for industrialized production.
The specific embodiment:
The preparation of the total extractum of embodiment 1 Radix Rhodiolae active ingredient
Get Radix Rhodiolae 10Kg section, with 70% ethanol 80Kg reflux, extract, 3 hours, inclining medicinal liquid, adds 70% ethanol 70Kg in the medicinal residues again, refluxed 2.5 hours, and merged medicinal liquid, concentrate, vacuum drying obtains total extract extractum 3Kg, wherein contain total flavones 17%, rhodioloside 2.5%, polysaccharide 48%.
The preparation of embodiment 2 Radix Rhodiolae active ingredient micromolecular compound extracts
Get 1 part of embodiment 1 total extract extractum, add 10 parts of dissolvings of water, medicinal liquid is flowed through and is filled with the macroporous resin chromatographic column, and 30 parts of eluting of water are clean, and eluent is pending; Use 90% ethanol elution chromatographic column then, concentrate ethanol elution, promptly get the micromolecular compound extract, wherein contain total flavones 37%, rhodioloside 7%;
The preparation of embodiment 3 Radix Rhodiolae active ingredient macromolecular compound extracts
Get 20 parts of pending eluents among the embodiment 2, be concentrated into 1 part, stirring adds 3 parts of 98% ethanol down, and static spending the night filtered, filter cake is with 1 part of water dissolution, the active carbon that adds 2% amount refluxed filtered while hot, filtrate cold drying or spray drying 15 minutes, get light brown powder and be the macromolecular compound extract, polyoses content 75%;
Embodiment 4
Get the micromolecular compound extract 40% of embodiment 2 by the preparation total amount, add pharmaceutic adjuvant microcrystalline Cellulose 20%, calcium sulfate 10%, dextrin 10%, starch 14.7%, hyprolose 4%, magnesium stearate 0.8%, white carbon black 0.5% is fully granulated behind the mixing, presses required dosage and divides packing promptly to get granule.
Embodiment 5
With the granule that embodiment 4 makes, press the required dosage fill and in the hungry area shell, promptly get capsule.
Embodiment 6
With the granule direct compression that embodiment 4 makes, film coating promptly gets tablet.
Embodiment 7
Get the macromolecular compound extract 55% of embodiment 3 by preparing total amount, add pharmaceutic adjuvant microcrystalline Cellulose 20%, starch 10%, lactose 7.7%, hyprolose 6%, magnesium stearate 0.8%, white carbon black 0.5% is fully granulated behind the mixing, presses required dosage and divides packing promptly to get granule.
Embodiment 8
With the granule that embodiment 7 makes, press the required dosage fill and in the hungry area shell, promptly get capsule.
Embodiment 9
With the granule direct compression that embodiment 7 makes, film coating promptly gets tablet.
Claims (3)
1, a kind of preparation of Radix Rhodiolae active ingredient is characterized in that this effective ingredient preparation is by containing the medically acceptable all kinds of preparations that Radix Rhodiolae macromolecular compound extract 30～70% and 70～30% pharmaceutic adjuvants are formed;
Wherein Radix Rhodiolae macromolecular compound extract be by the Radix Rhodiolae root through water-pure intermixture reflux, extract,, concentrate, flowing through behind the water dissolution is filled with the chromatographic column of macroporous resin, the water eluting; Get 20 parts of eluents, be concentrated into 1 part, stirring adds 1～3 part of 95～100% ethanol down, and static spending the night filtered, and filter cake refluxed 15 minutes with 1 part of water dissolution, the active carbon that adds 2% amount, filtered while hot, and filtrate cold drying or spray drying, promptly.
2, a kind of preparation of Radix Rhodiolae active ingredient as claimed in claim 1 is characterized in that containing polysaccharide 50～90% in the wherein said Radix Rhodiolae macromolecular compound extract, and molecular weight is between 3000 to 30000.
3, a kind of preparation method of Radix Rhodiolae effective ingredient preparation as claimed in claim 1 is characterized in that said preparation is made by following step:
(1) preparation of total extractum
The Radix Rhodiolae root is through section, and water-pure intermixture water: ethanol=99: 1～1: 99 reflux, extract, 1-3 time, total solvent amount are 10 to 30 times of crude drug, and total 2 to 6 hours extraction times, medicinal liquid obtains total extract extractum through concentrated, vacuum drying; Wherein contain total flavones 10～20%, rhodioloside 1～3%, polysaccharide 10～50%;
(2) separation of active ingredient, purification
Get 1 part of above-mentioned total extractum, add 5～10 parts of dissolvings of water, medicinal liquid is flowed through and is loaded the chromatographic column of macroporous resin, 10～40 parts of eluting of water;
Get 20 parts of eluents, be concentrated into 1 part, stirring adds 1～3 part of 95～100% ethanol down, and static spending the night filtered, filter cake is with 1 part of water dissolution, the active carbon that adds 2% amount refluxed filtered while hot, filtrate cold drying or spray drying 15 minutes, get light brown powder and be macromolecular compound, polyoses content 50～90%;
(3) preparation of effective ingredient preparation
Get above-mentioned macromolecular compound extract 30～70%, add pharmaceutic adjuvant 70～30%, formulation method makes tablet, capsule, granule, oral liquid or injection routinely.
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN 03128415 CN1223371C (en) | 2003-04-23 | 2003-04-23 | Preparation of effective component from rhodiola crenulate |
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| Application Number | Priority Date | Filing Date | Title |
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| CN 03128415 CN1223371C (en) | 2003-04-23 | 2003-04-23 | Preparation of effective component from rhodiola crenulate |
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| CNB001154230A Division CN1150917C (en) | 2000-04-21 | 2000-04-21 | Medicine containing active components of Rhodiola crennulata root and preparing process thereof |
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| CN1223371C true CN1223371C (en) | 2005-10-19 |
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