CN102389548A - Application of Smilax glabra flavonoid extract in preparing drugs for treating hyperuricemia - Google Patents
Application of Smilax glabra flavonoid extract in preparing drugs for treating hyperuricemia Download PDFInfo
- Publication number
- CN102389548A CN102389548A CN2011103138081A CN201110313808A CN102389548A CN 102389548 A CN102389548 A CN 102389548A CN 2011103138081 A CN2011103138081 A CN 2011103138081A CN 201110313808 A CN201110313808 A CN 201110313808A CN 102389548 A CN102389548 A CN 102389548A
- Authority
- CN
- China
- Prior art keywords
- rhizoma smilacis
- smilacis glabrae
- extract
- smilax glabra
- xod
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
Landscapes
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
Abstract
The invention discloses the application of a Smilax glabra flavonoid extract in preparing drugs for treating and preventing hyperuricemia. The Smilax glabra flavonoid extract is prepared by the following steps: extracting Smilax glabra by percolation with 50% to 90% ethanol (6 to 12 times the weight of Smilax glabra), recovering the percolate until the crude drug concentration is 1 g/mL, extracting with chloroform (2 to 5 times the volume of the percolate), extracting the water layer with ethyl acetate (2 to 5 times the volume of the water layer) for 3 to 5 times, concentrating the extraction solution, and drying at 60 DEG C under reduced pressure. The purity of flavones in the Smilax glabra flavonoid extract reaches 92%, accounting for 62.5% of Smilax glabra total flavones. The flavonoid extract is better in inhibitory effect on xanthine oxidase (XOD) activity than Smilax glabra crude drug, and has an XOD activity inhibition rate of 50% to 60% within the range of 125 Mug to 500 Mug. Therefore, the Smilax glabra flavonoid extract can be applied to pharmaceuticals for preventing and treating hyperuricemia.
Description
Technical field
The present invention relates to the new purposes of Chinese medicine extract in medical science, specifically relate to the application of Rhizoma Smilacis Glabrae flavone extract in preparation treatment antihyperuricemic disease drug.
Background technology
Hyperuricemia is a key factor of gout development, and blood uric acid produces too much, the comprehensive function of kidney uric acid excretion disorder or two factors causes the generation of hyperuricemia, and the sickness rate height has become the second largest metabolic disease that is only second to diabetes at present.(Xanthine Oxidase, XOD) ability catalysis xanthine and hypoxanthine oxidation generate uric acid to xanthine oxidase, and produce peroxide radical.Therefore the XOD inhibitor reduces uricopoiesis through the activity that suppresses XOD, as the uric acid resisting medicine.XOD inhibitor allopurinol commonly used clinically at present and promotion urate excretion medicine narcaricin; Since the side effect limitations its application clinically, therefore from Chinese herbal medicine and crude drug, seek the focus that novel anti gout and antihyperuricemic disease drug become study of pharmacy.
Rhizoma Smilacis Glabrae, another name Tu Rhizoma Dioscoreae Septemlobae is the liliaceous plant smilacis glabra
Smilax glabraRoxb. dry rhizome has the effect of dehumidifying, detoxifcation, easing joint movement.Clinical Rhizoma Smilacis Glabrae treatment hyperuricemia commonly used and ventilation [Chinese Pharmacopoeia. [S] .2005:14; ] [Ni Qing, Ren Jianwen, Shi Zhensheng. treatment gouty nephropathy 47 routine clinical summaries [J]. Beijing traditional Chinese medical science, 1997, (1): 3.].Research shows that Rhizoma Smilacis Glabrae has the uric acid resisting effect, to external XOD activity have the obvious suppression effect [Luo Liubao, Xie Zhipeng. xanthine oxidase inhibitory activity research [J] in the determined by ultraviolet spectrophotometry Chinese medicine. chemistry world, 2009,5:273].Rhizoma Smilacis Glabrae mainly contains compositions such as flavones ingredient and organic acid; Wherein flavones ingredient has tangible antiinflammatory, analgesic activity [Qin Rulan; Yellow Tian Yu, high fine jade. total flavone extracting method and pharmacological action [J] in the Rhizoma Smilacis Glabrae. Tonghua Teachers College's journal, 2010; 31 (2): 39.], but the Rhizoma Smilacis Glabrae uric acid resisting, to suppress the active material base of XOD unclear.
Summary of the invention
The present invention is an index with external XOD activity; System's separation screening Rhizoma Smilacis Glabrae suppresses the XOD active effective part; Reuse colorimetrically analysing effective site flavones content, it is flavones ingredient that clear and definite Rhizoma Smilacis Glabrae suppresses the active material base of XOD, can be used for preparation treatment antihyperuricemic disease drug.The object of the present invention is to provide the new purposes of Rhizoma Smilacis Glabrae flavone extract in medical science, relate to the application of Rhizoma Smilacis Glabrae flavone extract in preparation treatment antihyperuricemic disease drug specifically.
The objective of the invention is to realize through following technical scheme:
(1) Rhizoma Smilacis Glabrae suppresses the screening of XOD active site
1, method for preparing
(1) Rhizoma Smilacis Glabrae extract
The rhizoma smilacis glabrae medicinal material powder, with the 50%-90% ethanol percolate extraction of 6-12 times of medical material amount, treat that percolation finishes after, reclaim percolate to 1g crude drug/mL, be Rhizoma Smilacis Glabrae extract (T);
(2) four kinds of solvent extractive parts
Fetching earth, (T) is a certain amount of for Poria extract; Respectively with chloroform, ethyl acetate, the n-butanol extraction of 2-5 times of volume 3-5 time; Three extracts and water layer are recycled to dried; With 70% dissolve with ethanol and be settled to 10 mL, Rhizoma Smilacis Glabrae chloroform extract (TC), ethyl acetate extract (TE), n-butanol portion (TB) and water position (TH) extract.
2, suppress the XOD activity rating
Draw 1.2 mL pH7.9 phosphate buffers in 5 mL volumetric flasks, add 1.5 mL, 2.0 mmolL more respectively
-1Xanthine solution and 0.2 mL1.8 mgmL
-1Chlorination nitro blue tetrazolium (NBT) shakes up, for blank solution is used in the XOD determination of activity.Fetch earth Poria (T) and each solvent extractive part (C, E, B, H) in right amount, volatilize solvent, the DMSO dissolving, a certain amount of blank solution that places of accurate absorption is a sample determination liquid.
In this blank solution and sample determination liquid, add 1.2 mL XOD solution (0.08UmL
-1), shake up, pick up counting with the adding of XOD solution, behind 25 ℃ of reaction 30 min, place the UV-2401 ultraviolet spectrophotometer, with the blank solution contrast, detecting wavelength is 560 nm, measures XOD solution absorbency A
0, measure Rhizoma Smilacis Glabrae and respectively extract position absorbance A s.
Suppression ratio %=(A
0-As)/A
0* 100%
The result sees table 1.Rhizoma Smilacis Glabrae extract has stronger inhibitory action to the XOD activity.In each extractant position, it is active that only ethyl acetate extract has the XOD of inhibition, and be better than Rhizoma Smilacis Glabrae, shows that ethyl acetate extract is that Rhizoma Smilacis Glabrae suppresses the active effective site of XOD.
Table 1 Rhizoma Smilacis Glabrae reaches and respectively extracts the active result (n=3) of position inhibition XOD
| The sample name | Crude drug amount (mg) | Absorbance | Suppression ratio (%) |
| XOD | —— | 0.116 | —— |
| The Rhizoma Smilacis Glabrae sample solution | 16.75 | 0.091 | 21.55 |
| Chloroform extract | 16.75 | 0.191 | -64.66 |
| Ethyl acetate extract | 16.75 | 0.065 | 43.96 |
| N-butanol portion | 16.75 | 0.118 | -1.72 |
| The water position | 16.75 | 0.117 | -0.86 |
3, ethyl acetate extract suppresses XOD live vol effect relationship
Get high, medium and low three parts of ethyl acetate extract respectively, the DMSO dissolving, operational approach is the same, measures absorbance, and the result sees table 2.Ethyl acetate extract has dose-effect relationship to the active inhibitory action of XOD in 125 ~ 500ug scope.
Table 2 ethyl acetate extract is to the active inhibitory action (n=3 of XOD
| Sample | Sampling amount (ug) | Absorbance | Suppression ratio (%) |
| XOD | —— | 1.424 | —— |
| 1 | 125 | 0.702 | 50.70 |
| 2 | 250 | 0.678 | 52.39 |
| 3 | 500 | 0.561 | 60.60 |
Through system's separated from solvent, XOD activity rating, ethyl acetate extract is that Rhizoma Smilacis Glabrae suppresses the active effective site of XOD in the Rhizoma Smilacis Glabrae.
(2) Rhizoma Smilacis Glabrae ethyl acetate extract flavones content is analyzed
1, methodological study
(1) control substance of Rutin solution
It is an amount of to get control substance of Rutin, is dried to constant weight, and accurate the title decides, and adds 70% alcoholic solution and is mixed with 0.2320 mgmL
-1Control substance of Rutin solution.
(2) mensuration of maximum absorption wavelength
Get control substance of Rutin solution and above-mentioned Rhizoma Smilacis Glabrae sample (T) solution respectively in right amount in 25 mL volumetric flasks, add 5% sodium nitrite solution 1mL, shake up; Place 6 min, add 10% aluminum nitrate solution, 1 mL, shake up; Place 6 min, add 10% sodium hydroxide solution, 10 mL again, adding distil water is settled to scale; Shake up; Behind 15 min in the UV-2401 ultraviolet spectrophotometer sweep measuring, reference substance and T sample solution all have maximum absorption band at 500 nm places, confirm that thus Rhizoma Smilacis Glabrae total flavones optimum detection wavelength is 500 nm.
(3) set up standard curve
Precision is measured control substance of Rutin solution 0 mL, 2.0 mL, 2.5 mL, 3.0 mL, 3.5 mL, 4.0 mL, 4.5 mL and is placed 25 mL volumetric flasks respectively; Press the assay method of maximum absorption wavelength; Measure trap at 500 nm places; Calculate according to reference substance amount (X) and absorbance (Y), get regression equation: Y=0.5308X-0.0054, r
2=0.9998 (n=6), the range of linearity is 0.4640 ~ 1.0440 mg.
(4) precision is investigated
Precision is measured 5 parts of Rhizoma Smilacis Glabrae sample (T) solution, presses the maximum absorption wavelength assay method and measures absorbance, and RSD is 2.36%, shows that this method precision is good.
(5) repeatability is investigated
Precision takes by weighing 5 parts of rhizoma smilacis glabrae medicinal materials, according to the preparation of T sample preparation methods, presses the assay method of maximum absorption wavelength and measures absorbance, and the RSD value is 2.48%, shows that this method repeatability is good.
(6) study on the stability
The a certain amount of T sample solution of accurate absorption according to the assay method of maximum absorption wavelength, is measured absorbance respectively at 0 min, 5 min, 10 min, 15 min, 30 min, 60 min, and RSD is 0.76%, shows that this method is stable in 60 min.
(7) average recovery is investigated
Precision takes by weighing rhizoma smilacis glabrae medicinal material 1.0 g of 6 parts of known content; A certain amount of control substance of Rutin solution of accurate respectively adding prepares need testing solution by the T sample preparation methods, presses the assay method of maximum absorption wavelength and measures content; The result sees table 3, shows that this method is accurate.
Table 3 average recovery result of calculation
2, ethyl acetate extract assay
Fetch earth respectively Poria sample (T) and ethyl acetate extract (TE) sample are measured flavones content by preceding method.The result shows that the flavone total amount is 62.50% of a medical material in the Rhizoma Smilacis Glabrae ethyl acetate extract, and purity is 92.13%.Show that ethyl acetate extract is mainly flavones ingredient.
Table 4 ethyl acetate extract flavones content is measured the result
| Sample | Content (mg) | Relative quantity (%) | Purity (%) |
| T | 150.93 | 100 | —— |
| TE | 94.34 | 62.5 | 92.13 |
Show that through the flavones content analysis it is flavones ingredient that Rhizoma Smilacis Glabrae suppresses the XOD active site.
Beneficial effect
Show that through system each solvent position of separation, external XOD activity rating, flavone macroanalysis it is flavones ingredient that Rhizoma Smilacis Glabrae suppresses the active material base of XOD, can be used for preparation treatment and prevention antihyperuricemic disease drug.
The specific embodiment
The extraction of embodiment 1 Rhizoma Smilacis Glabrae flavones ingredient
(1) takes by weighing rhizoma smilacis glabrae medicinal material powder 1kg,, after percolation finishes, reclaim percolate and get Rhizoma Smilacis Glabrae extract to 1g crude drug/mL with 6L 85% ethanol percolate extraction;
(2) fetch earth Poria extract with 2L chloroform extraction 5 times, chloroform layer discards, and water layer divides 4 extractions with the 2L ethyl acetate extraction, and ethyl acetate layer merges and reclaims solvent, and 60 ℃ of drying under reduced pressure get the Rhizoma Smilacis Glabrae flavone extract.
The extraction of embodiment 2 Rhizoma Smilacis Glabrae flavones ingredients
(1) takes by weighing rhizoma smilacis glabrae medicinal material powder 1kg,, after percolation finishes, reclaim percolate and get Rhizoma Smilacis Glabrae extract to 1g crude drug/mL with 9L 70% ethanol percolate extraction.
(2) Rhizoma Smilacis Glabrae extract is with 3.5L chloroform extraction 4 times, and chloroform layer discards, and water layer divides 4 extractions with the 3L ethyl acetate, and ethyl acetate layer merges and reclaims solvent, and 60 ℃ of drying under reduced pressure get the Rhizoma Smilacis Glabrae flavone extract.
The extraction of embodiment 3 Rhizoma Smilacis Glabrae flavones ingredients
(1) takes by weighing rhizoma smilacis glabrae medicinal material powder 1kg,, after percolation finishes, reclaim percolate and get Rhizoma Smilacis Glabrae extract to 1g crude drug/mL with 12L 50% ethanol percolate extraction.
(2) Rhizoma Smilacis Glabrae extract is with 5L chloroform extraction 3 times, and chloroform layer discards, and water layer divides 2 extractions with the 5L ethyl acetate, and ethyl acetate layer merges and reclaims solvent, and 60 ℃ of drying under reduced pressure get the Rhizoma Smilacis Glabrae flavone extract.
Embodiment 4 Rhizoma Smilacis Glabrae flavone extracts are to the active inhibitory action of XOD
(1) draws 1.2 mL pH7.9 phosphate buffers in 5 mL volumetric flasks, add 1.5 mL, 2.0 mmolL more respectively
-1Xanthine solution and 0.2 mL1.8 mgmL
-1Chlorination nitro blue tetrazolium (NBT) shakes up, for blank solution is used in the XOD determination of activity;
(2) the Poria flavone extract 150 μ g that fetch earth, the DMSO dissolving is mixed with 50 μ g μ L
-1Solution, adding in the blank solution is sample determination liquid;
(3) in XOD blank solution and sample solution, add 1.2 mL XOD solution (0.08UmL
-1), shake up, for XOD solution is used in the XOD determination of activity;
(4) adding with XOD solution picks up counting, and behind 25 ℃ of reaction 30 min, places the UV-2401 ultraviolet spectrophotometer, is contrast with the blank solution, and detecting wavelength is 560 nm, measures XOD solution and Rhizoma Smilacis Glabrae flavone extract absorbance.
Claims (2)
1. the application of Rhizoma Smilacis Glabrae flavone extract in preparation prevention and treatment antihyperuricemic disease drug; Wherein the method for distilling of Rhizoma Smilacis Glabrae flavone extract is following:
(1) takes by weighing rhizoma smilacis glabrae medicinal material powder 1kg,, after percolation finishes, reclaim percolate and get Rhizoma Smilacis Glabrae extract to 1g crude drug/mL with 6-12L 50%-90% ethanol percolate extraction;
(2) fetch earth Poria extract with 2-5L chloroform extraction 3-5 time, chloroform layer discards, and water layer divides 2-5 extraction with the 2-5L ethyl acetate extraction, and ethyl acetate layer merges and reclaims solvent, and 60 ℃ of drying under reduced pressure get the Rhizoma Smilacis Glabrae flavone extract.
2. application according to claim 1, the purity that it is characterized in that flavone in the described Rhizoma Smilacis Glabrae flavone extract is 90%.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2011103138081A CN102389548A (en) | 2011-10-17 | 2011-10-17 | Application of Smilax glabra flavonoid extract in preparing drugs for treating hyperuricemia |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2011103138081A CN102389548A (en) | 2011-10-17 | 2011-10-17 | Application of Smilax glabra flavonoid extract in preparing drugs for treating hyperuricemia |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN102389548A true CN102389548A (en) | 2012-03-28 |
Family
ID=45857013
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN2011103138081A Pending CN102389548A (en) | 2011-10-17 | 2011-10-17 | Application of Smilax glabra flavonoid extract in preparing drugs for treating hyperuricemia |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN102389548A (en) |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105056027A (en) * | 2015-08-05 | 2015-11-18 | 贵阳中医学院 | Red-section Smilax ocreafa A. DC effective part and use thereof |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101204500A (en) * | 2007-12-14 | 2008-06-25 | 南京中医药大学 | Preparation method and composition and Chinese traditional medicine preparation for preventing and curing hyperuricaemia |
| CN101879264A (en) * | 2009-05-05 | 2010-11-10 | 戴永青 | Chinese medicinal composition for reducing blood uric acid |
-
2011
- 2011-10-17 CN CN2011103138081A patent/CN102389548A/en active Pending
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101204500A (en) * | 2007-12-14 | 2008-06-25 | 南京中医药大学 | Preparation method and composition and Chinese traditional medicine preparation for preventing and curing hyperuricaemia |
| CN101879264A (en) * | 2009-05-05 | 2010-11-10 | 戴永青 | Chinese medicinal composition for reducing blood uric acid |
Non-Patent Citations (5)
| Title |
|---|
| 张永太等: "土茯苓中有效成分提取工艺考察", 《中成药》 * |
| 张白嘉等: "土茯苓及落新妇苷抗炎、镇痛、利尿作用研究", 《中药药理与临床》 * |
| 林耿丰等: "正交设计法优选土茯苓中落新妇苷及总黄酮的提取工艺", 《亚太传统医药》 * |
| 陈广耀，沈连生，江佩芬: "土茯苓化学成分的研究", 《北京中医药大学学报》 * |
| 陈雪等: "土茯苓对小鼠高尿酸血症的实验研究", 《吉林医药学院学报》 * |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105056027A (en) * | 2015-08-05 | 2015-11-18 | 贵阳中医学院 | Red-section Smilax ocreafa A. DC effective part and use thereof |
| CN105056027B (en) * | 2015-08-05 | 2019-12-17 | 贵州中医药大学 | Effective part of glabrous greenbrier rhizome with red section and application thereof |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN1895408B (en) | Radix saposhnikoviae Tong Sheng capsules, preparation and quality control method thereof | |
| CN102183517A (en) | Method for determining general flavone content | |
| CN101274025A (en) | Chinese medicinal composition with functions of reducing fever, purging the intense heat and detoxicating and preparation method thereof and quality control method | |
| CN101480422A (en) | Tibetan oriental wormwood extract as well as preparation method and use thereof | |
| CN101496870B (en) | Chinese medicinal composition for resolving phlegm and suppressing cough as well as preparation method and quality control method thereof | |
| CN102228517A (en) | Plantain seed extract and application thereof | |
| CN1969923A (en) | Antivirus extract of isatis root, its extraction method, use and content assaying method | |
| CN101040906B (en) | Injection for treating cardiovascular or cerebrovascular disease and the preparing method and the quality control method | |
| CN104546995B (en) | A kind of medicinal usage of emblic extract | |
| CN105125941A (en) | Preparation method of assistant hypoglycemic corn-stigma extractive capsules | |
| CN103040945B (en) | Preparation and detection method of gangsong general flavones | |
| CN101766664B (en) | Detection method of total saponin of Radix Ilicis Asprellae | |
| CN102389548A (en) | Application of Smilax glabra flavonoid extract in preparing drugs for treating hyperuricemia | |
| CN101474314A (en) | Application of catechin in pharmacy | |
| CN103739652A (en) | New 23, 29-drop oleanolic acid compound, preparation method thereof and application in preparation of glucosidase inhibitor medicines | |
| CN100478015C (en) | Composition of Chinese traditional medicine for treating allergic rhinitis, and preparation method | |
| ItharatPhD | Determination of cytotoxic compounds of Thai traditional medicine called Benjakul using HPLC | |
| CN103356812B (en) | A kind of Radix Wikstroemae granule | |
| CN107913277A (en) | The purposes of the anti-uric acid nephropathy of tanshinone | |
| CN113820411A (en) | Method for measuring contents of schizandrol A and schizandrol B in preparation for strengthening body resistance and removing blood stasis | |
| CN103372038A (en) | Novel application of centella extract | |
| CN103083388A (en) | Preparation method of fructus gleditsiae total saponins | |
| CN105106254A (en) | Anti-influenza loosestrife extract | |
| Kitagawa et al. | Quantitative determination of principal alkaloid and flavonoid constituents in wintersweet, the flower buds of Chimonanthus praecox | |
| Chen et al. | Determination of two major xanthone glycosides in rhizome of Anemarrhena asphodeloides using high performance capillary electrophoresis |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C12 | Rejection of a patent application after its publication | ||
| RJ01 | Rejection of invention patent application after publication |
Application publication date: 20120328 |