CN102138941A - Precursor liposome of propolis flavone and preparation method thereof - Google Patents
Precursor liposome of propolis flavone and preparation method thereof Download PDFInfo
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- CN102138941A CN102138941A CN2011100800963A CN201110080096A CN102138941A CN 102138941 A CN102138941 A CN 102138941A CN 2011100800963 A CN2011100800963 A CN 2011100800963A CN 201110080096 A CN201110080096 A CN 201110080096A CN 102138941 A CN102138941 A CN 102138941A
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- propolis flavone
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Abstract
The invention relates to a precursor liposome of propolis flavone and a preparation method thereof. The method is characterized by comprising the following steps of: preparing a nanometer liposome suspension of the propolis flavone by a reverse-phase evaporation method, and freeze drying the nanometer liposome suspension of the propolis flavone to prepare the precursor liposome of the propolis flavone; and preparing a nanometer liposome of the propolis flavone with Tween 80 by taking the propolis flavone as cores and lecithin and cholesterol as membranes, and freeze drying the prepared nanometer liposome of the propolis flavone by the reverse-phase evaporation method to obtain the precursor liposome of the propolis flavone. Due to the adoption of the technical scheme, the stability of the propolis flavone in a strongly-acidic system of gastric acid can be enhanced to improve biological availability and achieve better health-care effect. The precursor liposome of the propolis flavone prepared by the freeze drying method can be convenient to store and convey and also can be applied in wide food systems.
Description
Technical field
The invention belongs to bee product technology field, be specifically related to a kind of propolis flavone pro-liposome and preparation method thereof.
Background technology
Propolis is the resin of Apis from plant plumule or trunk broken position, traumatic part collection, and sneaks into a kind of gluey solid content with fragranced that its mandibular gland secretions and Cera Flava etc. process.Propolis has antibiotic anticorrosion, antiviral, antioxidation, activating blood circulation to dissipate blood stasis, vessel softening, blood fat reducing, antitumor, enhancing body immunity, promote effect such as tissue regeneration;
The composition of propolis is quite complicated, and chromocor compound wherein is main active substance.Propolis has the good reputation of " chromocor compound treasure-house ", now from propolis the chromocor compound of isolation identification comprise kinds more than 70 such as chrysin, apigenin, acacetin, Quercetin, galangin, rutin, kaempferol, kaempferide, Fructus rhamni (Rhamnus davurica Pall.) element, Sciurus vulgaris element, wherein 5,7-dihydroxy-3,4-dimethyl flavone and 5-hydroxyl-4,7-dimethoxy flavanone is the endemic element of propolis.The propolis in the different places of production is because geographical environment is different with the season of weather and collection, and the content of its active component also has very big-difference.The functional activity of propolis and the content of effective ingredient chemical compound wherein, particularly flavones content is closely related.Therefore, Flavonoid substances content is the topmost quality index of propolis;
Propolis and propolis flavone thereof have very strong physiologically active, but it is insoluble in water, and light, heat, oxygen, metal ion and system pH are easy to have influence on the stability of flavone.The unstability of the dose-effect relationship of propolis physiologically active and flavones content and propolis, propolis flavone, the work that has determined researcheres mainly launches from aspects such as the protection of extraction and separation process, active component and the researchs of propolis novel form, in the hope of determining best extraction and guard method, make it avoid the influence of various unfavorable factors, thereby protect its physiological function composition to greatest extent, prolong shelf life;
Up to now, do not see the report for preparing relevant for the propolis flavone pro-liposome both at home and abroad.Liposomal formulation is the preparation based on amphiprotic substances such as phospholipid, is to be encapsulated in active substance to be encapsulated in the middle of the thin film of lipoids bilayer or the interior aqueous phase that is encapsulated in the lipoids bilayer obtains.Because active substance is encapsulated in the liposome, have advantages such as long-acting, targeting, Liposomal formulation shows great potentiality to be exploited aspect new product development.Many research institutions and drugmaker all are devoted to the Liposomal formulation Products Development, and one of them important use is the destruction that the active substance sealed of protection is not subjected to external environment.Different freeze drying protectants are joined in the Liposomal dispersion that makes, and lyophilization obtains the propolis flavone pro-liposome; can increase the physical and chemical stability of liposome, reduce the volume of product, be convenient to store and transportation; its application is expanded in instant food processing simultaneously and application.
Summary of the invention
Purpose of the present invention is in order to strengthen the stability of propolis flavone in gastric juice strong acid system, reach and improve bioavailability and better health-care effect, and store more easily, transport and using in the food system widely, a kind of propolis flavone pro-liposome and preparation method thereof is provided.
For achieving the above object, the technical solution adopted in the present invention is: this kind propolis flavone pro-liposome is characterized in that: be core with the propolis flavone, the wall material is made up of lecithin, cholesterol and Tween 80, and the propolis flavone pro-liposome of making is a powdery;
Described a kind of propolis flavone pro-liposome is characterized in that: lecithin and cholesterol weight ratio are more than or equal to 4; Tween 80 and lecithin weight ratio are more than or equal to 1.2;
Described a kind of propolis flavone pro-liposome is characterized in that: core propolis flavone content be lecithin content 2.5 weight % or more than;
The preparation method of described a kind of propolis flavone pro-liposome is characterized in that may further comprise the steps:
(1) preparation propolis flavone aqueous solution: in the propolis flavone deionized water, the concentration of control propolis flavone gets water smaller or equal to 12mg/mL;
(2) preparation lipid anhydrous ether solution: lecithin, cholesterol are dissolved in absolute ether by mass ratio 10:1-10:2, and the concentration of control lecithin in its diethyl ether solution gets organic facies smaller or equal to 25 mg/mL;
(3) emulsion forms: propolis flavone aqueous solution and lipid anhydrous ether solution are mixed, ultrasonicly in ice-water bath again become single-phase dispersion to mixture, promptly formed Water in Oil emulsion;
(4) liposome generates: the Water in Oil emulsion that forms is transferred in the rotary evaporation flask, rotary evaporation is removed absolute ether in 30 ℃ of-40 ℃ of water-baths, there is gel to generate when removing absolute ether, be further rotated evaporation, gel subsides, and behind the formation suspension, adds the aqueous solution of tween 80, making the Tween 80 that contains in the solution and the mass ratio of lecithin is 1:4-1.25:1, and this system is rotated hydration 15-30 minute under 30 ℃ of-40 ℃ of water-baths; Promptly get the propolis flavone nano liposome dispersion liquid;
(5) adding of freeze drying protectant: with the freeze drying protectant of one of glucose, mannose, lactose, sucrose, maltose or trehalose, with freeze drying protectant: the phospholipid mass ratio adds in the propolis flavone nano liposome dispersion liquid more than or equal to the amount of 6:1;
(6) generation of pro-liposome: the propolis flavone nano liposome dispersion liquid that will add freeze drying protectant is in freezer dryer, in below-39 ℃, more than following lyophilization 24 h of vacuum 23.8Pa; Lyophilisation product is powdery propolis flavone pro-liposome;
(7) rehydration of propolis flavone pro-liposome: get 0.2g propolis flavone prerequisite liposome, in the PBS of PH 6.8-PH7.0 buffer, disperse, form dispersion liquid; After the rehydration, seal the retention rate 81%-96% of core; Mean diameter is 20nm-400nm.
The present invention's beneficial effect compared with prior art:
Because with the propolis flavone is core, lecithin, cholesterol are the film material, are aided with Tween 80, adopt reverse phase evaporation to prepare the propolis flavone nanometer liposome, and propolis flavone prerequisite liposome is prepared in lyophilization then.Can strengthen the stability of propolis flavone in the highly acid system of gastric acid, reach and improve bioavailability and better health-care effect.Make the propolis flavone pro-liposome with freeze-drying, make it can store more easily, transport and use in the food system widely.
The specific embodiment
Embodiment 1
Claim lecithin 200 mg, cholesterol 20 mg, be dissolved in the 30 mL absolute ethers (organic facies); Claim 20 mg propolis flavones to be dissolved in the 10 mL deionized waters (water).Gained organic facies and water are mixed, form the Water in Oil emulsion of stable uniform through supersound process, be transferred in the 250 mL rotary evaporation bottles, rotary evaporation is removed ether in 35 ℃ of water-baths, had gel in 5-10 minute and generate, along with rotary evaporation continues, gel subsides, keep decompression 30 minutes, to remove remaining ether.Add the aqueous solution 10mL that contains 250 mg Tween 80s in evaporative flask, the rotation hydration is 30 minutes in 35 ℃ of water-baths.After the hydration, promptly get the propolis flavone nano liposome dispersion liquid.Pipette propolis flavone nano liposome dispersion liquid 20mL in surface plate, containing the phospholipid quality in the dispersion liquid is 200mg, adds the 4g glucose and does freeze drying protectant.At first, then sample is put into, be evacuated to 23.8Pa in-39 ℃ of pre-cooling freezer dryers 20 minutes, freezing 24h, the gained sample preservation is in exsiccator.The retention rate of sealing core is 81.2%, and mean diameter is between 30-400 nm.
Embodiment 2
Claim lecithin 200 mg, cholesterol 20 mg, be dissolved in the 30 mL absolute ethers (organic facies); Claim 20 mg propolis flavones to be dissolved in the 10 mL deionized waters (water).Gained organic facies and water are mixed, form the Water in Oil emulsion of stable uniform through supersound process, be transferred in the 250 mL rotary evaporation bottles, rotary evaporation is removed ether in 35 ℃ of water-baths, had gel in 5-10 minute and generate, along with rotary evaporation continues, gel subsides, keep decompression 30 minutes, to remove remaining ether.Add the aqueous solution 10mL that contains 250 mg Tween 80s in evaporative flask, the rotation hydration is 30 minutes in 35 ℃ of water-baths.After the hydration, promptly get the propolis flavone nano liposome dispersion liquid.Pipette propolis flavone nano liposome dispersion liquid 20mL in surface plate, containing the phospholipid quality in the dispersion liquid is 200mg, adds the 4g mannose and does freeze drying protectant.At first, then sample is put into, be evacuated to 23.8Pa in-39 ℃ of pre-cooling freezer dryers 20 minutes, freezing 24h, the gained sample preservation is in exsiccator.The retention rate of sealing core is 82.6%, and mean diameter is between 20-380 nm.
Embodiment 3
Claim lecithin 300 mg, cholesterol 60 mg, be dissolved in (organic facies) in the 30mL absolute ether; Claim 30 mg propolis flavones to be dissolved in (water) in the 10mL deionized water.Gained organic facies and water are mixed, form the Water in Oil emulsion of stable uniform through supersound process, be transferred in the 250mL rotary evaporation bottle, rotary evaporation is removed ether in 40 ℃ of water-baths, had gel in 5-10 minute and generate, along with rotary evaporation continues, gel subsides, keep decompression 30 minutes, to remove remaining ether.Add deionized water solution 20 mL that contain 300 mg Tween 80s in evaporative flask, the rotation hydration is 30 minutes in 40 ℃ of water-baths.Promptly get the propolis flavone nano liposome dispersion liquid after the hydration.The gained dispersion liquid is deposited in 4 ℃ of refrigerators.Pipette propolis flavone nano liposome dispersion liquid 20mL in surface plate, containing the phospholipid quality in the dispersion liquid is 200mg, adds the 4g lactose and does freeze drying protectant.At first, then sample is put into, be evacuated to 23.8Pa in-39 ℃ of pre-cooling freezer dryers 20 minutes, freezing 24h, the gained sample preservation is in exsiccator.The retention rate of sealing core is 86.6%, and mean diameter is between 30-300 nm.
Embodiment 4
Claim lecithin 250 mg, cholesterol 30 mg, be dissolved in (organic facies) in the 24mL absolute ether; Claim 20 mg propolis flavones to be dissolved in the 8 mL deionized waters (water).Gained organic facies and water are mixed, form the Water in Oil emulsion of stable uniform through supersound process, be transferred in the 250mL rotary evaporation bottle, rotary evaporation is removed ether in 40 ℃ of water-baths, had gel in 5-10 minute and generate, along with rotary evaporation continues, gel subsides, keep decompression 30 minutes, to remove remaining ether.Add the aqueous solution 12mL that contains 200 mg Tween 80s in evaporative flask, the rotation hydration is 30 minutes in 35 ℃ of water-baths.Promptly get the propolis flavone nano liposome dispersion liquid.Pipette propolis flavone nano liposome dispersion liquid 20mL in surface plate, containing the phospholipid quality in the dispersion liquid is 200mg, adds 4g sucrose and does freeze drying protectant.At first, then sample is put into, be evacuated to 23.8Pa in-39 ℃ of pre-cooling freezer dryers 20 minutes, freezing 24h, the gained sample preservation is in exsiccator.The retention rate of sealing core is 88.5%, and mean diameter is between 30-400 nm.
Embodiment 5
Claim lecithin 250 mg, cholesterol 30 mg, be dissolved in (organic facies) in the 24mL absolute ether; Claim 20 mg propolis flavones to be dissolved in the 8 mL deionized waters (water).Gained organic facies and water are mixed, form the Water in Oil emulsion of stable uniform through supersound process, be transferred in the 250mL rotary evaporation bottle, rotary evaporation is removed ether in 40 ℃ of water-baths, had gel in 5-10 minute and generate, along with rotary evaporation continues, gel subsides, keep decompression 30 minutes, to remove remaining ether.Add the aqueous solution 12mL that contains 200 mg Tween 80s in evaporative flask, the rotation hydration is 30 minutes in 35 ℃ of water-baths.After the hydration, promptly get the propolis flavone nano liposome dispersion liquid.Pipette propolis flavone nano liposome dispersion liquid 20mL in surface plate, containing the phospholipid quality in the dispersion liquid is 200mg, adds 4g maltose and does freeze drying protectant.At first, then sample is put into, be evacuated to 23.8Pa in-39 ℃ of pre-cooling freezer dryers 20 minutes, freezing 24h, the gained sample preservation is in exsiccator.The retention rate of sealing core is 83.8%, and mean diameter is between 20-350 nm.
Embodiment 6
Claim lecithin 250 mg, cholesterol 30 mg, be dissolved in (organic facies) in the 24mL absolute ether; Claim 20 mg propolis flavones to be dissolved in the 8 mL deionized waters (water).Gained organic facies and water are mixed, form the Water in Oil emulsion of stable uniform through supersound process, be transferred in the 250mL rotary evaporation bottle, rotary evaporation is removed ether in 40 ℃ of water-baths, had gel in 5-10 minute and generate, along with rotary evaporation continues, gel subsides, keep decompression 30 minutes, to remove remaining ether.Add the aqueous solution 12mL that contains 200 mg Tween 80s in evaporative flask, the rotation hydration is 30 minutes in 35 ℃ of water-baths.Pipette propolis flavone nano liposome dispersion liquid 20mL in surface plate, containing the phospholipid quality in the dispersion liquid is 200mg, adds the 4g trehalose and does freeze drying protectant.At first, then sample is put into, be evacuated to 23.8Pa in-39 ℃ of pre-cooling freezer dryers 20 minutes, freezing 24h, the gained sample preservation is in exsiccator.The retention rate of sealing core is 96.2%, and mean diameter is between 30-400 nm.
Claims (4)
1. propolis flavone pro-liposome, it is characterized in that: be core with the propolis flavone, the wall material is made up of lecithin, cholesterol and Tween 80, and the propolis flavone pro-liposome of making is a powdery.
2. a kind of propolis flavone pro-liposome according to claim 1 is characterized in that: lecithin and cholesterol weight ratio are more than or equal to 4; Tween 80 and lecithin weight ratio are more than or equal to 1.2.
3. a kind of propolis flavone pro-liposome according to claim 1 is characterized in that: core propolis flavone content be lecithin content 2.5 weight % or more than.
4. the preparation method of a kind of propolis flavone pro-liposome according to claim 1 is characterized in that may further comprise the steps:
(1) preparation propolis flavone aqueous solution: in the propolis flavone deionized water, the concentration of control propolis flavone gets water smaller or equal to 12mg/mL;
(2) preparation lipid anhydrous ether solution: lecithin, cholesterol are dissolved in absolute ether by mass ratio 10:1-10:2, and the concentration of control lecithin in its diethyl ether solution gets organic facies smaller or equal to 25 mg/mL;
(3) emulsion forms: propolis flavone aqueous solution and lipid anhydrous ether solution are mixed, ultrasonicly in ice-water bath again become single-phase dispersion to mixture, promptly formed Water in Oil emulsion;
(4) liposome generates: the Water in Oil emulsion that forms is transferred in the rotary evaporation flask, rotary evaporation is removed absolute ether in 30 ℃ of-40 ℃ of water-baths, there is gel to generate when removing absolute ether, be further rotated evaporation, gel subsides, and behind the formation suspension, adds the aqueous solution of tween 80, making the Tween 80 that contains in the solution and the mass ratio of lecithin is 1:4-1.25:1, and this system is rotated hydration 15-30 minute under 30 ℃ of-40 ℃ of water-baths; Promptly get the propolis flavone nano liposome dispersion liquid; (5) adding of freeze drying protectant: with the freeze drying protectant of one of glucose, mannose, lactose, sucrose, maltose or trehalose, with freeze drying protectant: the phospholipid mass ratio adds in the propolis flavone nano liposome dispersion liquid more than or equal to the amount of 6:1; (6) generation of pro-liposome: the propolis flavone nano liposome dispersion liquid that will add freeze drying protectant is in freezer dryer, in below-39 ℃, more than following lyophilization 24 h of vacuum 23.8Pa; Lyophilisation product is powdery propolis flavone pro-liposome; (7) rehydration of propolis flavone pro-liposome: get 0.2g propolis flavone prerequisite liposome, in the PBS of PH 6.8-PH7.0 buffer, disperse, form dispersion liquid; After the rehydration, seal the retention rate 81%-96% of core; Mean diameter is 20nm-400nm.
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| Application Number | Priority Date | Filing Date | Title |
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| CN2011100800963A CN102138941A (en) | 2011-03-31 | 2011-03-31 | Precursor liposome of propolis flavone and preparation method thereof |
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| CN2011100800963A CN102138941A (en) | 2011-03-31 | 2011-03-31 | Precursor liposome of propolis flavone and preparation method thereof |
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103191157A (en) * | 2012-06-20 | 2013-07-10 | 南京农业大学 | Propolis flavone liposome for improving livestock and poultry immunity and preparation method thereof |
| WO2017089842A1 (en) * | 2015-11-23 | 2017-06-01 | Apivita S.A. | Method for preparing a stable controlled-release propolis colloidal dispersion system for various uses |
| CN110478379A (en) * | 2019-06-21 | 2019-11-22 | 福建医科大学 | A kind of total biflavone proliposome of selaginella doederlleini and preparation method thereof |
| CN113812547A (en) * | 2021-09-22 | 2021-12-21 | 云南省农业科学院蚕桑蜜蜂研究所 | Feed for promoting growth of bee colonies and preparation method thereof |
-
2011
- 2011-03-31 CN CN2011100800963A patent/CN102138941A/en active Pending
Non-Patent Citations (3)
| Title |
|---|
| 《中国优秀硕士学位论文全文数据库》 20091115 张冰慧 蜂胶脂质体的制备及其对高血糖小鼠降糖作用的研究 第10-26页 4 , 第11期 * |
| 张冰慧: "蜂胶脂质体的制备及其对高血糖小鼠降糖作用的研究", 《中国优秀硕士学位论文全文数据库》 * |
| 汤志勇等: "蜂胶脂质体的制备", 《食品工业科技》 * |
Cited By (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103191157A (en) * | 2012-06-20 | 2013-07-10 | 南京农业大学 | Propolis flavone liposome for improving livestock and poultry immunity and preparation method thereof |
| CN103191157B (en) * | 2012-06-20 | 2014-12-10 | 南京农业大学 | Propolis flavone liposome for improving livestock and poultry immunity and preparation method thereof |
| WO2017089842A1 (en) * | 2015-11-23 | 2017-06-01 | Apivita S.A. | Method for preparing a stable controlled-release propolis colloidal dispersion system for various uses |
| CN110478379A (en) * | 2019-06-21 | 2019-11-22 | 福建医科大学 | A kind of total biflavone proliposome of selaginella doederlleini and preparation method thereof |
| CN110478379B (en) * | 2019-06-21 | 2022-02-18 | 福建医科大学 | Selaginella chinensis total biflavone precursor liposome and preparation method thereof |
| CN113812547A (en) * | 2021-09-22 | 2021-12-21 | 云南省农业科学院蚕桑蜜蜂研究所 | Feed for promoting growth of bee colonies and preparation method thereof |
| CN113812547B (en) * | 2021-09-22 | 2024-02-23 | 云南省农业科学院蚕桑蜜蜂研究所 | Feed for promoting growth of bee colony and preparation method thereof |
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Application publication date: 20110803 |