CN105012449A - Gardenia extract with antilipemic function - Google Patents
Gardenia extract with antilipemic function Download PDFInfo
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- CN105012449A CN105012449A CN201510509354.3A CN201510509354A CN105012449A CN 105012449 A CN105012449 A CN 105012449A CN 201510509354 A CN201510509354 A CN 201510509354A CN 105012449 A CN105012449 A CN 105012449A
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Abstract
The invention discloses gardenia extract with an antilipemic function. A preparing method for the extract comprises the steps that a, dry mature gardenia fruit is smashed; b, 70% ethyl alcohol is used for carrying out heat reflux extraction, and extracting solutions are combined and concentrated to be free from alcohol taste to obtain a concentrated solution; c, the concentrated solution in the step b is extracted through petroleum ether, ethyl acetate and water-saturated n-butyl alcohol in sequence to respectively obtain petroleum ether extract, ethyl acetate extract and n-butyl alcohol extract; d, the n-butyl alcohol extract is gathered through macroporous resin, 10% ethyl alcohol is used for washing in an eight-column-volume mode to remove polysaccharide, then 65% ethyl alcohol is used for eluting in a ten-column-volume mode, 65% eluant is collected, and volume concentrating and spray drying are carried out. The gardenia extract has the antilipemic function and can be used for preparing antilipemic medicine.
Description
Technical field
The present invention relates to the field of Chinese medicines, be specifically related to a kind of Fructus Gardeniae extract and containing the pharmaceutical preparation of this extract, this Fructus Gardeniae extract can be used for preparing the medicine of blood fat reducing.
Background technology
Fructus Gardeniae (Gardeniajasminoides Ellis.) is Rubiaceae (Rubiaceae) Gardenia Ellis evergreen shrubs, has another name called SHANZHIZI, Yellow Fructus Gardeniae, Yellow Fructus Gardeniae etc., main product in Zhejiang, Fujian, Jiangxi, Hunan, Guangdong.The dry mature fruit of Fructus Gardeniae is used as medicine and claims Fructus Gardeniae, and edition pharmacopeia is gone through by China and book on Chinese herbal medicine is all on the books, is conventional Chinese medicine.Its bitter in the mouth cold in nature, nontoxic, main GUIXIN, lung, tri-jiao channel, have the effects such as pathogenic fire purging relieving restlessness, clearing away heat and promoting diuresis, removing pathogenic heat from blood and toxic substance from the body.Clinically be widely used in that calentura is vexed, jaundice dark coloured urine, blood strangury and dry pain, heat in blood tell the cards such as nosebleed, conjunctival congestion and swelling pain, pathogenic fire,toxin and furuncles; Its root functionality heat clearing away, removing heat from blood, removing toxic substances, the treatment being mainly used in the diseases such as icterohepatitis, oedema due to nephritis, flu high heat, hemorrhage haematemesis, traumatic injury, sore swollen toxin among the people; Leaf can reducing swelling and alleviating pain, treatment traumatic injury; Flower can the blood fat reducing of clearing away lung-heat removing heat from blood, treatment cough due to lung-heat, epistaxis.
The research of Gardenia Ellis plant chemical ingredient starts from the twenties in 20th century, has distinctive compound and mainly comprises iridoids, organic acid and flavonoid.Containing a large amount of iridoids in Fructus Gardeniae, there are 4 kinds of main Types: iridoid alkanes, iridoid glycosides, iridoid two acetal esters and Secoiridoid Glycosides.Iridoid basic framework, through epoxidation, hydroxylating and the aromatic acid esterification that obtained by shikimic acid pathway, result in the multiformity of this compounds, also causes the multiformity of its pharmacologically active simultaneously.
Fructus Gardeniae is conventional Chinese medicine, and enjoy the attention of Chinese scholars, research work is more.Modern pharmacology experiment shows, the iridoid constituents in Fructus Gardeniae has obvious anti-inflammatory and antalgic, hepatic cholagogic, antithrombotic isoreactivity; Saffron glucoside constituents tool antiinflammatory, antithrombotic, Adjust-blood lipid in organic acid, resist myocardial ischemia isoreactivity.Along with the continuous progress of science and technology, there has also been many innovations and breakthrough to its research, especially to the research of iridoid constituents wherein, new compound and pharmacological action are constantly found, and pharmacological mechanism have also been obtained deep research.
Summary of the invention
The object of the present invention is to provide a kind of Fructus Gardeniae extract, the preparation method of this extract and liquid phase analysis method, the pharmaceutical preparation containing this extract and utilize this extract to prepare the purposes of the medicine of blood fat reducing.
Above-mentioned purpose of the present invention is achieved by technical scheme below:
A kind of Fructus Gardeniae extract, this extract is prepared by following methods:
A the Fructus Gardeniae mature fruit of drying is pulverized by (); B () extracts with 70% alcohol heat reflux, merge extractive liquid, is concentrated into and obtains concentrated solution without alcohol taste; C () uses petroleum ether, ethyl acetate and water saturated n-butanol extraction successively to step (b) concentrated solution, obtain petroleum ether extract, acetic acid ethyl ester extract and n-butyl alcohol extract respectively; D () n-butyl alcohol extract macroporous resin enrichment, first with 10% alcohol flushing, 8 column volume removing polysaccharide, then uses 65% ethanol elution, 10 column volumes, collects 65% eluent, concentrating under reduced pressure, spraying dry.
Further, macroporous resin described in step (d) is AB-8 type macroporous resin.
In order to be controlled Fructus Gardeniae extract differences between batches prepared by Different sources, batch medical material by the method setting up HPLC finger printing, the liquid phase analysis method of described extract is:
Chromatographic column: Agilent ZORBAX SB C18 post (4.6mm × 250mm, 5 μm);
Mobile phase: A is acetonitrile, and B is 0.1% phosphate aqueous solution;
Gradient elution program: 0.01 ~ 15min, A25% ~ 35%; 15 ~ 25min, A 35% ~ 50%; 25 ~ 32min, A 50%; 32 ~ 35min, A 50% ~ 25%; 35 ~ 40min, A25%;
Flow rate of mobile phase: 1.0mLmin
-1; Determined wavelength: 252nm; Column temperature: 30 DEG C; Sample size: 10 μ L.
Pharmaceutical preparation, the described Fructus Gardeniae extract containing treatment effective dose and pharmaceutically acceptable carrier.
The application of described Fructus Gardeniae extract in the medicine of preparation blood fat reducing.
The application of described pharmaceutical preparation in the medicine of preparation blood fat reducing.
When extract of the present invention is used as medicine, directly can uses, or use in the form of a pharmaceutical preparation.
Described pharmaceutical preparation contains the Fructus Gardeniae extract of the present invention for the treatment of effective dose, and all the other are acceptable on materia medica, nontoxic to humans and animals and pharmaceutically suitable carrier of inertia and/or excipient.
Described pharmaceutically suitable carrier or excipient are that one or more are selected from solid, semisolid and liquid diluent, filler and pharmaceutical preparation adjuvant.Pharmaceutical preparation of the present invention is used with the form of per weight dose.Extract of the present invention is applied to by form that is oral or injection the patient needing treatment.For time oral, tablet, slow releasing tablet, controlled release tablet, capsule, drop pill, micropill, suspensoid, Emulsion, powder or granule, oral liquid etc. can be made into; During for injecting, can be made into aqueous or oily solution, aseptic powder injection, liposome or the Emulsion etc. of sterilizing.
Advantage of the present invention: the present invention by extracting Fructus Gardeniae, remove impurity, enrichment process, the extract with effect for reducing blood fat can be obtained; Liquid phase analysis method provided by the invention may be used for the finger printing setting up this extract, for controlling Different sources, the differences between batches of extract prepared by batch medical material.
Accompanying drawing explanation
Fig. 1 is Fructus Gardeniae extract HPLC chromatograms.
Detailed description of the invention
Further illustrate essentiality content of the present invention below in conjunction with embodiment, but do not limit scope with this.Although be explained in detail the present invention with reference to preferred embodiment, those of ordinary skill in the art should be appreciated that and can modify to technical scheme of the present invention or equivalent replacement, and does not depart from essence and the scope of technical solution of the present invention.
Embodiment 1: prepared by Fructus Gardeniae extract
Main agents and material: Fructus Gardeniae is purchased from Hui nationality's Chinese Medicinal Materials Markets; Food-grade ethanol is purchased from Shanghai Ling Feng chemical reagent company limited; Pharmaceutical grade AB-8 macroporous resin is purchased from sky tunami letter resin company limited; Acetonitrile is HPLC level, is purchased from TEDIA; 85% phosphoric acid is HPLC level, is purchased from TEDIA; Chromatographic grade pure water is heartily pure water.
Preparation method: the Fructus Gardeniae mature fruit of 10kg drying is pulverized, extract (25L × 3 time) with 70% alcohol heat reflux, merge extractive liquid, is concentrated into and obtains concentrated solution (6L) without alcohol taste, petroleum ether (6L × 3 time) is used successively to concentrated solution, ethyl acetate (6L × 3 time) and water saturated n-butyl alcohol (6L × 3 time) extraction, obtain petroleum ether extract respectively, acetic acid ethyl ester extract and n-butyl alcohol extract 305g, with AB-8 macroporous resin (2kg after n-butyl alcohol extract dissolves with 5L distilled water, column volume 1.5L) enrichment, 10% alcohol flushing, 8 column volumes (12L) are first used to remove polysaccharide, use 65% ethanol elution, 10 column volumes (15L) again, collect 65% eluent, concentrating under reduced pressure, spraying dry obtains Fructus Gardeniae extract extract powder 185g.
Embodiment 2: liquid-phase chromatographic analysis
Need testing solution is prepared: in the brown volumetric flask of extract extract powder 5mg to 50mL that Example 1 method is obtained, add 30mL 20% acetonitrile solution ultrasonic dissolution, after being cooled to room temperature, continue to add 20% acetonitrile solution standardize solution.
Analytical method:
Chromatograph of liquid: Agilent 1260, binary pump;
Chromatographic column: Agilent ZORBAX SB C18 post (4.6mm × 250mm, 5 μm);
Mobile phase: A is acetonitrile, and B is 0.1% phosphate aqueous solution;
Gradient elution program: 0.01 ~ 15min, A25% ~ 35%; 15 ~ 25min, A 35% ~ 50%; 25 ~ 32min, A 50%; 32 ~ 35min, A 50% ~ 25%; 35 ~ 40min, A25%;
Flow rate of mobile phase: 1.0mLmin
-1; Determined wavelength: 252nm; Column temperature: 30 DEG C; Sample size: 10 μ L.
Analyze the extract prepared with 10 batches of Different sources, batch Fructus Gardeniae, set up finger printing and mate, 1 ~ No. 11 peak all occurs in 10 batch sample chromatograms as a result.Therefore demarcate 11 peaks for total fingerprint peaks, set up the HPLC finger printing of this extract accordingly, the results are shown in Figure 1.
Embodiment 3: Fructus Gardeniae extract pharmacological testing
One, material
1, laboratory animal: Wistar rat, ICR mice, provides by Beijing Vital River Experimental Animals Technology Co., Ltd., the quality certification number: SCXK (capital) 2002-0003.
2, reagent: HDL-C (HDL-C) testing cassete (lot number 050919), low-density lipoprotein cholesterol (LDL-C) testing cassete (lot number 050707), T-CHOL (TC) testing cassete (lot number 050523), triglyceride (TG) testing cassetes (lot number 050914) etc., provide above by the safe clinical reagent company limited of Beijing Northization.
3, instrument: ZS-3 type semiautomatic biochemistry analyzer, Beijing Zhong Sheng biotechnology company.
4, medicine: Fructus Gardeniae extract is by the preparation of embodiment 1 method.
5, statistical procedures: experimental data is through one factor analysis of variance and carry out many group t inspection.
Two, method
1, the auxiliary antilipemic experiment of Fructus Gardeniae extract
(1) experiment grouping
Wistar male rat 50, body weight (200 ± 20) g, be divided into 2 groups, blank group 10 only gives normal diet and feeds, and all the other 40 is high fat group, give high lipid food (3.5% cholesterol, 11.0% Adeps Sus domestica, 0.5% cholate, 85.0% normal diet) feed 7d, 8d (fasting 16h) high fat treated animal orbital venous plexus blood sampling 1mL, measures serum TC and TG.According to blood lipid level, be divided into hyperlipidemia model group and the large, medium and small dosage group of Fructus Gardeniae extract (often organizing 10), divide into groups to start administration the same day, blank group and hyperlipidemia model group give the distilled water with administration group same volume.High fat group continues to give high lipid food and feeds, successive administration 1 month.
(2) blood is got
Fasting 16h after last administration, abdominal aortic blood, 3000rmin
-1centrifugal 15min, divides and gets serum.
(3) index determining measures TC, TG, HDL-C, LDL-C in serum.
2, Mouse Acute Toxicity experiment
ICR mice, body weight (20 ~ 22) g, fasting 16h, be divided into 2 groups at random by body weight, blank group and Fructus Gardeniae extract 10.67gkg
-1group, often organizes 20 animals, male and female half and half, after single oral gavage administration, and Continuous Observation 2 weeks, record animal poisoning symptom and death condition.
3, rat 30d feeds experiment
After Animal adaptability feeds 3d, observe 1 week before administration, the situations such as animal activity, feed, feces are all without exception.Animal is divided into 4 groups at random by body weight, be respectively Fructus Gardeniae extract small dose group: 0.15g extract/kg, middle dosage group: 0.30g extract/kg, heavy dose of group: 0.60g extract/kg, if Normal group, often organizes 20, male and female half and half, gavage 1mL/100g body weight, awards ordinary water by equivalent capability, successive administration 4 weeks.Observe every day and record the activity of animal, hair, feces, feed; Rat is weighed weekly 1 time, record food-intake, and according to body weight adjustment dosage.Administration is after 4 weeks, and orbital venous plexus blood sampling detects blood biochemical analysis index, hematological indices.Carry out system after Animal Anesthesia to become celestial and histopathological examination, and weigh the heart, liver, spleen, lung, kidney, thymus, testis, ovary, uterus, stomach weight, calculate organ coefficient (Organ Wet Weight g100g
-1body weight).
Three, result
1, on the impact of hyperlipidemia rats blood fat
The each dosage group of Fructus Gardeniae extract significantly can reduce TC, TG, and heavy dose of group reduces LDL-C; But HDL-C is had no significant effect, the results are shown in Table 1.
The table 1 Fructus Gardeniae extract administration impact on hyperlipidemia rats blood fat in 4 weeks (x ± s, n=10)
Note: compare 1 with model group) P<0.05,2) P<0.01.
2, acute toxicity testing
There is obvious poisoning symptom in Fructus Gardeniae extract 10.67g extract/kg.Fructus Gardeniae extract is 10.67g extract/kg to mice maximum tolerated dose.
3,30d feeds experiment
(1) on the impact of rat body weight and food-intake
Fructus Gardeniae extract is little, in, heavy dose of group in administration 3 weeks, 4 weeks time buck feed consumption lower than matched group.Fructus Gardeniae extract is little, in, heavy dose of group administration 3 weeks, 4 weeks time male rat body weight be starkly lower than concurrent control group (P<0.05 or P<0.01).Jenny feed consumption and body weight are had no significant effect, the results are shown in Table 2.
The table 2 Fructus Gardeniae extract successive administration impact on rat body weight in 4 weeks (x ± s, n=10)
Note: compare 1 with matched group) P<0.05,2) P<0.01 (lower same).
(2) blood parameters inspection
Difference (P>0.05) that Fructus Gardeniae extract 3 dosage successive administrations 4 weeks blood parameters compare with matched group that there are no significant.The results are shown in Table 3.
The table 3 Fructus Gardeniae extract successive administration impact on rat blood biochemical indicator in 4 weeks (x ± s, n=10)
(3) on the impact of Rats Organs and Tissues weight
Fructus Gardeniae extract successive administration 4 weeks, each organ coefficient value is all within normal range.The results are shown in Table 4 ~ 5.
The table 4 Fructus Gardeniae extract successive administration impact on male rat organ coefficient in 4 weeks (x ± s, n=10)
Note: * organ coefficient unit: g tissue wet/100g body weight (lower same).
The table 5 Fructus Gardeniae extract successive administration impact on female rats organ coefficient in 4 weeks (x ± s, n=10)
(4) on hematological impact
Fructus Gardeniae extract successive administration 4 weeks rat blood indices, all within normal range, react without overt toxicity.The results are shown in Table 6.
The table 6 Fructus Gardeniae extract successive administration impact on rat blood in 4 weeks (x ± s, n=20)
(5) pathologic examination
Gross anatomy internal organs are showed no exception.Pathologic examination, each treated animal heart of Fructus Gardeniae extract, liver, spleen, lung, kidney, adrenal gland, thymus, stomach, testis, epididymis, prostate, ovary, uterus, brain, myelopathy Neo-Confucianism check that structure is normal, have no the change of obvious pathological change.
Above-mentioned result of study shows, Fructus Gardeniae, as health product, has hypolipemic function, improves blood lipid level to hyperlipemia, and the sickness rate reducing cardiovascular and cerebrovascular disease has positive role, and without obvious toxic and side effects.
Embodiment 4: the preparation of tablet
By embodiment 1 method first obtained extract, add excipient, pelletizing press sheet in itself and excipient weight than the ratio for 1:10.
Embodiment 5: the preparation of oral liquid
By embodiment 1 method first obtained extract, oral liquid method for making makes oral liquid routinely.
Embodiment 6: the preparation of capsule or granule
By embodiment 1 method first obtained extract, add excipient in itself and excipient weight than the ratio for 1:9, make capsule or granule.
Embodiment 7: the preparation of injection
Obtain extract by embodiment 1 method, inject with water, fine straining, injection is made in embedding sterilizing.
Embodiment 8: the preparation of aseptic powder injection
By embodiment 1 method first obtained extract, be dissolved in sterile water for injection, stirring makes molten, filters with aseptic suction funnel, more aseptic fine straining, and be sub-packed in ampoule, after frozen drying, aseptic sealing by fusing obtains injectable powder.
Claims (6)
1. a Fructus Gardeniae extract, is characterized in that described extract is prepared by following methods:
A the Fructus Gardeniae mature fruit of drying is pulverized by (); B () extracts with 70% alcohol heat reflux, merge extractive liquid, is concentrated into and obtains concentrated solution without alcohol taste; C () uses petroleum ether, ethyl acetate and water saturated n-butanol extraction successively to step (b) concentrated solution, obtain petroleum ether extract, acetic acid ethyl ester extract and n-butyl alcohol extract respectively; D () n-butyl alcohol extract macroporous resin enrichment, first with 10% alcohol flushing, 8 column volume removing polysaccharide, then uses 65% ethanol elution, 10 column volumes, collects 65% eluent, concentrating under reduced pressure, spraying dry.
2. extract according to claim 1, is characterized in that: macroporous resin described in step (d) is AB-8 type macroporous resin.
3. Fructus Gardeniae extract according to claim 1, is characterized in that: the liquid phase analysis method of described extract is:
Chromatographic column: Agilent ZORBAX SB C18 post (4.6mm × 250mm, 5 μm);
Mobile phase: A is acetonitrile, and B is 0.1% phosphate aqueous solution;
Gradient elution program: 0.01 ~ 15min, A 25% ~ 35%; 15 ~ 25min, A 35% ~ 50%; 25 ~ 32min, A 50%; 32 ~ 35min, A 50% ~ 25%; 35 ~ 40min, A 25%;
Flow rate of mobile phase: 1.0mLmin
-1;
Determined wavelength: 252nm;
Column temperature: 30 DEG C;
Sample size: 10 μ L.
4. pharmaceutical preparation, is characterized in that: the Fructus Gardeniae extract according to claim 1 containing treatment effective dose and pharmaceutically acceptable carrier.
5. the application of Fructus Gardeniae extract according to claim 1 in the medicine of preparation blood fat reducing.
6. the application of pharmaceutical preparation according to claim 4 in the medicine of preparation blood fat reducing.
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN113041294A (en) * | 2019-12-26 | 2021-06-29 | 大江生医股份有限公司 | Application of plant fermentation liquor in preparation of lipid-lowering composition |
Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101057674A (en) * | 2007-06-13 | 2007-10-24 | 深圳市生物谷科技有限公司 | Composition for preventing and curing diabetes |
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2015
- 2015-08-19 CN CN201510509354.3A patent/CN105012449A/en active Pending
Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101057674A (en) * | 2007-06-13 | 2007-10-24 | 深圳市生物谷科技有限公司 | Composition for preventing and curing diabetes |
Non-Patent Citations (1)
| Title |
|---|
| 杨庆等: "红花提取物对高脂血症模型大鼠降血脂作用和安全性实验研究", 《中国实验方剂学杂志》 * |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN113041294A (en) * | 2019-12-26 | 2021-06-29 | 大江生医股份有限公司 | Application of plant fermentation liquor in preparation of lipid-lowering composition |
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Application publication date: 20151104 |