Pharmaceutical composition for treatment of cornea injury
FIELD: medicine, ophthalmology, pharmacy.
SUBSTANCE: invention proposes a pharmaceutical composition used in treatment of cornea injuries that comprises, mas.p.p.: sodium deoxyribonucleate, 14-15; sodium chloride, 0.8-0.9; lyophilized blood plasma, 74-75, and distilled water, the balance. Components are mixed in aseptic conditions and packaged. Invention provides anti-inflammatory, antiviral, antibacterial, reparative and regenerative effect, decreasing treatment period and declining post-operative complications.
EFFECT: improved, enhanced and valuable medicinal properties of pharmaceutical composition.
2 tbl, 5 ex
The technical field to which the invention relates.
The invention relates to medicine, in particular to ophthalmology, and can be used for the treatment of lesions of the cornea.
The level of technology
Known composition for the treatment of burns of the cornea by removing necrotic masses then close the resulting defect biological substrate, with the resulting defect diseased cornea is applied repeatedly until epithelialization freeze-dried blood plasma, dissolved in physiological solution (see autospid. The USSR №1344354, CL 61F 9/00, publ. 15.10.1987 year).
The disadvantage of this composition and method of treatment is a low effect, since the dried blood plasma itself is insufficient pronounced stimulant regenerative processes of the cornea, as well as long term treatment.
Renowned pharmaceutical composition for the treatment of inflammatory processes of viral etiology in the eyeball that contains the antiviral drug, while it additionally contains vegetable and/or animal phospholipids, sodium chloride one-deputizing, sodium phosphate disubstituted and water purified in the following ratio, wt.%:
|Antiviral medication||of 0.001-5|
|Vegetable and/or animal phospholipids||0,001-2|
|Sodium phosphate one-deputizing||of 0.001 to 0.2|
|Sodium phosphate disubstituted||of 0.001 to 0.2|
(see U.S. Pat. Of the Russian Federation No. 2245137, CL AC 31/00, 61F 9/00, F61P 27/02).
The disadvantage of this composition is a low efficiency and a prolonged effect in the treatment of lesions of the cornea.
The closest in technical essence and the achieved positive effect and adopted by the authors for the prototype is Derinat, including deoxyribonuclease, sodium chloride, distilled water (see the Handbook, VIDAL, drugs in Russia", B-402, 2005).
The disadvantage of this drug is a long term treatment, and he does not have a sufficiently strong antiviral, antimicrobial, reparative and regenerative action.
Disclosure of inventions
Object of the present invention to provide pharmaceutical compositions for the treatment of lesions of the cornea, which has anti-inflammatory, antiviral, antimicrobial, as well as reparative and regenerative action, the ability to normalize the condition of the tissue at distroficescimi in the cornea, the reduction of treatment time and reduce postoperative complications.
The technical result that can be achieved using the present invention is an anti-inflammatory, antiviral, antimicrobial, as well as reparative and regenerative action, the normalization condition of the tissues in degenerative changes, reducing treatment time and reduce postoperative complications.
The technical result is achieved by using a pharmaceutical composition for the treatment of lesions of the cornea, including deoxyribonuclease, sodium chloride and distilled water, it additionally contains freeze-dried blood plasma in the following ratio, wt.%:
|Freeze-dried blood plasma||74-75|
The essence of a pharmaceutical composition for the treatment of lesions of the cornea is as follows: the initial substance in wt.%, namely, deoxyribonuclease sodium 14-15, sodium chloride 0,8-0,9, freeze-dried blood plasma 74-75, the rest distilled water, mix in ASEP is practical conditions, and Packed.
Pharmacological action Derinat, namely, deoxyribonuclease, sodium chloride and distilled water is that it is an immunomodulator that affects cellular and humoral immunity, stimulates reparative processes and disorders (normalizes the number of granulocytes, lymphocytes, platelets), has anti-inflammatory, antiviral, antimicrobial, and reparative and regenerative action, normalizes the tissues in degenerative changes, has a poorly marked anticoagulant effect.
Freeze-dried blood plasma, biological dressing, improves the processes of regeneration, trophic corneal, restore osmotic equilibrium, at the same time having good adhesive properties, plasma provides biological isolation, performs a protective function: inhibits the growth of newly formed blood vessels, protects it from the tear fluid and pathogenic microflora, at the same time, lyophilized blood plasma by itself alone is not enough pronounced stimulant regenerative processes of the cornea.
The implementation of the invention
Examples of specific performance of the receipt and testing of pharmaceutical compositions for the treatment of lesions of the cornea.
Example 1. Pharmaceutical composition (FC) th is ovat by mixing the components and dissolved in distilled water in the following ratio of components, wt.%:
|Freeze-dried blood plasma||65|
followed by the package.
The obtained composition was too liquid, as follows from the palpebral fissure, not acting as a biological dressing.
Example 2. Carried out analogously to example 1, but the composition is taken in the following ratio, wt.%:
|Freeze-dried blood plasma||74|
The resulting composition turned out to be consistency in the form of eye ointments, which are evenly located on the eyeball, performing the function of a biological dressing.
Example 3. Carried out analogously to example 1, but the composition is taken in the following ratio, wt.%:
|Leofiles the automated blood plasma||75|
The resulting composition is also turned consistency in the form of eye ointments, which are evenly located on the eyeball, performing the function of a biological dressing.
Example 4. Carried out analogously to example 1, but the composition is taken in the following ratio, wt.%:
|Freeze-dried blood plasma||80|
The resulting composition has a very thick consistency, which do not cover the affected area of the cornea.
Thus, the best was the composition according to examples 2 and 3.
Clinical studies using the proposed FC was performed in 70 patients (40 men and 30 women) aged from 18 to 73 years, among them, 21 patients with keratopathy, 22 patients with keratitis of various etiologies, 11 patients with AAD, 9 patients with burns, 7 patients with corneal opacities varying degrees. Assessment of corneal they had a way of ceratopogonidae, altimetry, refractometry, biomicroscopic activeassistant recovery of corneal transparency. All patients received treatment with 20% solution of Derinat, consisting of deoxyribonuclease, sodium chloride and distilled water, with dissolved lyophilized blood plasma. The control group consisted of 20 people receiving conventional treatment and holistic therapy (vitamins, antioxidants, detoxification and desensibiliziruyuschee therapy).
Example 5. The patient takes a horizontal position. Ever bred by wikopedia. On the affected cornea without anaesthetic, using eye glass sticks aplitsiruetsya pharmaceutical composition. Velodyne removed after 3-5 minutes. Impose monocular bandage on for 20-30 minutes, during which the patient remains in a horizontal position. This procedure is carried out 2 times a day. The best course of treatment was 5-6 days, when traditional methods of treatment, usually 10-12 days.
As a result of the treatment it is shown that the inflammatory process was alloreactive, after the start of treatment eyes became calm. In all cases, there is a strong link regenerated epithelium from underlying tissue of the cornea, which contributes to the conservation and restoration of transparency in affected areas, increased visual acuity, the average rose to 0.30+0.03 in (P<0.001) statistical and reliably, restored the sensitivity of the cornea statistically significantly (P<0,001).
Allergic reactions were not observed. Given the pronounced positive effect of therapy, this pharmaceutical composition may be recommended in treatment of lesions of the cornea doctors-ophthalmologists practical health care.
Assessment for each of the indicators in dystrophic diseases of the cornea are shown in tables 1 and 2.
|Day||Pachymetry before the treatment||Pachymetry Derinat + plasma||Pachymetry cent. the treatment|
|1st||The thickness of the cornea in the Central zone ranged from 1000 to 1200 microns||1100-1200||1000-1200|
|Diseases||Pachymetry is Einat + plasma||Pachymetry traditional treatment|
|Burns 1-2 degrees||600-650||650-700|
Visual acuity before treatment from 0,01-0,09 increased to 0.1 and 0.35 in the treatment of complex - Derinat + plasma, 0,05-0,1 traditional treatment.
The present invention in comparison with the prototype and other known technical solution has the following advantages:
- anti-inflammatory, antiviral, antimicrobial, as well as reparative and regenerative actions;
the normalization condition of the tissues in degenerative changes;
- reduction of treatment time and reducing postoperative complications;
the exception of allergic reactions.
Pharmaceutical composition for the treatment of lesions of the cornea, including deoxyribonuclease, sodium chloride and distilled water, characterized in that it further comprises a freeze-dried plasma in the following ratio, wt.%:
|Freeze-dried blood plasma||74-75|
SUBSTANCE: method involves covering internal surface of soft contact lens with 0.2 ml of visco-elastic and silicon-dried amnion having its diameter equal to lens diameter is superposed over it. The lens is placed onto injured cornea for 6-7 days. When needed, the manipulation is repeated depending on disease severity.
EFFECT: enhanced effectiveness of treatment; excluded unpleasant feelings and surgical intervention application.
SUBSTANCE: method involves instilling derinate solution into eye 30 min before treating eyelid edges with acaricide preparation during the whole 3 weeks long treatment course. To prevent disease recurrence, the derinate instillation course is repeated in spring and autumn during 10-14 days after having finished treatment course.
EFFECT: reduced toxico-allergic complications frequency.
SUBSTANCE: method involves treating trabecule zone with laser radiation of 532 nm wavelength. Laser radiation treatment is carried out with unit pulses of 3 ns duration in the amount of 40-50 per trabecular network of temporal or nasal segment spread over 180°. Pulse energy is equal to 0.8-1.5 mJ. Focal spot diameter is 300-400 mcm. Then, medical mixture is introduced into lymphatic orbit region. The mixture contains lidocaine 50-100 mg; histochrome 10-20 mg; Euphillin 12-24 mg; Glutoxim 5-10 mg; Lidase 16-32 U. The mixture is introduced daily by doing pterygopalatine blocks on injured eye side in 3-5 blocks long course.
EFFECT: enhanced effectiveness of treatment; reduced risk of adverse side effects.
SUBSTANCE: method involves delivering drugs directly to optic nerve via infusion system placed in retrobulbar space. 4 mg of Dexamethason in 1 ml of solution is introduced beginning from 8 o'clock in the morning every 30 min. The first three days, it takes place 5 times at daily dose of 20 mg. Daily dose of 16 mg is given four times at the fourth and fifth day. Daily dose of 12 mg is administered three times at the sixth and seventh day. Daily dose of 8 mg is given twice at the eighth and ninth day. Daily dose of 4 mg is given once at the tenth day. Trental, Emoxipin, Solcoseryl and Taufon are delivered to the optic nerve afternoons at a dose of 0.5 ml each, beginning from 12 o'clock with 2 h long pause via infusion system.
EFFECT: accelerated treatment course; reduced risk of traumatic complications.
SUBSTANCE: method involves administering vitaminotherapy. Peptide bioregulator of Retinalamine is additionally introduced at a dose of 5 mg every other day in inactive tuberculous inflammation phase. The total treatment course is 10 procedures long.
EFFECT: activated mediatory interaction between retinal cells; intensified oxygen supply; retained or increased vision acuity.
SUBSTANCE: method involves intravenously introducing chlorine row photosensitizer at a dose of 0.8-1.0 mg/kg, 70% of the dose being drop-by-drop infused during 10 min. The remaining 30% are introduced as bolus dose 1.5 h later. Photosensitizer introduction being finished, the neoplasm is transpupillarily exposed to laser radiation at wavelength corresponding to photosensitizer light absorption maximum with some overlap of neighboring fields. Tumor periphery is irradiated with intact tissue being caught not less than 1 mm far from ophthalmoscopically visible tumor boundary at power density of 25-40 J/cm2. Then, the whole tumor surface is concentrically irradiated from periphery to the center by applying power density of 110-120 J/cm2. 2 weeks later, photosensitizer intravenous injection is repeated and transpupillary laser radiation treatment of tumor surface. 2-4 intravenous photosensitizer injection sessions and tumor surface irradiation are given to achieve its complete resorption.
EFFECT: minimized adjacent intact tissue exposure; excluded recurrence and tumor dissemination in remote postoperative period.
FIELD: organic chemistry, medicine, ophthalmology.
SUBSTANCE: invention relates to an ophthalmic solution containing compound of the following chemical formula (I): wherein R1 represents group of the formula (Ia): or its pharmaceutically acceptable salt or derivative, cationic antiseptic agent and basic amino-compound wherein cationic antiseptic agent represents quaternary ammonium salt or chlorhexidine, and basic amino-compound represents at least one compound chosen from trometamol, lidocaine and nicotinamide. Quaternary ammonium salt represents benzalconium chloride, benzetonium chloride, cetylpyridinium or cetrimide. Ophthalmic solution can contain additionally boric acids, ethylenediaminetetraacetic acids. Owing the presence of additives the invention provides enhancing antiseptic effect of ophthalmic solutions.
EFFECT: improved and valuable medicinal properties of ophthalmic solutions.
11 cl, 4 tbl, 2 dwg, 7 ex
SUBSTANCE: method involves intravitreally smoothing out central retina zone by means of perfluororganic substance. Circular retinotomy is carried out after perfluororganic substance smoothing-out. The, conjugate of chlorine e6 bound in covalent way with albumin in 1:2.5 proportion. Later on, the rest of perfluororganic substance is introduced. Retina is circularly intravitreally irradiated in accumulated conjugate projection zone with continuous laser beam of 400 mW power field-by-field during 2 min. Wavelength corresponds to chlorine e6 light absorption maximum. Irradiation being finished, the perfluororganic substance is substituted with silicon oil.
EFFECT: complete anatomical adjacency of retina and sublayer tissues; avoided cases of perfluororganic substance or silicon being found under retina without necessity of mechanical retina attachment to eye tunics; reliable retina detachment recurrence prophylaxis in remote postoperative period.
FIELD: medicine, ophthalmology, pharmacy.
SUBSTANCE: invention proposes ophthalmic ointment containing ciprofloxacin hydrochloride monohydrate equivalent to the concentration 0.1-0.9% as a free base, preserving agent, for example nipagin or nipasol, and a sterile base consisting of vaseline melt of sort "for ophthalmic ointments" and anhydrous lanolin. Ophthalmic ointment is prepared by melting vaseline and anhydrous lanolin and step-by-step mixing milled ciprofloxacin hydrochloride and preserving agent with sterile base. Invention provides producing a sterile and stable ciprofloxacin-containing ophthalmic ointment used for local treatment of eye inflammatory diseases and of bacterial etiology.
EFFECT: improved preparing method, valuable medicinal properties of composition and ointment.
2 cl, 3 ex
FIELD: medicine, ophthalmology, pharmacy, preparative biochemistry.
SUBSTANCE: method involves isolation of undamaged lens from fresh enucleated eyes of vertebrate animals. Lenses are kept in an aqueous-saline solution, tissue extract is centrifuged and supernatant is collected. Supernatant is separated by its dissolving in 100% ammonium sulfate solution, dialyzed up to the complete removal of ammonium sulfate ions and separated by isoelectrofocusing method in sucrose gradient density at pH 3.5-10.0. Fractions of acid proteins are collected, dialyzed up to the complete removing sucrose and ampholines and prepared protein aqueous solution is dried and purified by electrophoresis in 7.5-15% polyacrylamide gel. Low-molecular fraction with a value Rf = 0.9-0.95 is eluted from gel with deionized water at temperature 4-7°C for 2-5 days, and prepared fraction is dialyzed in dialysis sacs with penetration limit value of pores 2-8 kDa at temperature 4-7°C for 7-10 days. Prepared regulatory peptide is dried and dissolved in physiological aqueous-saline solution. Invention provides the enhanced percent yield of regulatory peptides possessing biological activity in superlow doses and isolation of highly purified polypeptides of low molecular mass.
EFFECT: improved preparing method.
FIELD: medicine, pharmacy.
SUBSTANCE: invention describes a pharmaceutical composition for its using in treatment of symptoms of catarrhal diseases and influenza, or as an analgesic drug. Composition represents a single dose as an aqueous medicinal formula of the total volume less 100 ml and containing paracetamol and a masking component that attenuates or masks its bitter taste and consisting of at least two components chosen from the following groups (i)-(vi): (i) magnesium chloride, gluconate, hydroxide, carbonate or hydrocarbonate, and/or a mixture of magnesium chloride and magnesium sulfate in the molar ratio from 0.0001:1 to 0.01:1 measured as the ratio magnesium atoms : paracetamol; (ii) gingerin and/or ginger oil in the weight ratio from 1:20000 to 1:10000 with respect to paracetamol; (iii) sweetening agent of the sustained onset effect and prolonged effect chosen from taumatin, neohesperidin DC and/or glycyrrhizin taken in the weight ratio from 1:20000 to 1:100 with respect to paracetamol; (iv) soluble starch taken in the weight ratio from 1:10 to 1:1 with respect to paracetamol; (v) sucrose ester having value of hydrophilic-lipophilic balance (HLB) above 10 and taken in the weight ratio from 1:10 to 2:1 with respect to paracetamol, and (vi) glycine taken in the molar ratio from 1:4 to 2:1 with respect to paracetamol. Proposed composition provides effective taste masking or bitterness of paracetamol being especially in liquid preparations containing less 100 ml of water, and in using the combination of different masking agents also.
EFFECT: improved and valuable properties of composition.
10 cl, 3 tbl, 3 ex
FIELD: medicine, pharmacy.
SUBSTANCE: invention relates to a pharmaceutical composition comprising enalapril maleate, lactose, potato starch, polyvinylpyrrolidone and magnesium stearate wherein a medium-molecular polyvinylpyrrolidone aqueous solution is used as polyvinylpyrrolidone, and the pharmaceutical composition comprises indapamide and aerosil additionally. The pharmaceutical composition is made as an enveloped-covered core in the following ratio of the core components, wt.-%: enalapril maleate is taken in the therapeutically effective dose; indapamide, 1.1-2.6; aerosil, 0.2-0.4; potato starch, 20.0-40.0; medium-molecular polyvinylpyrrolidone aqueous solution with the concentration 12%, 1.7-2.3; magnesium stearate, 0.8-1.0, and lactose, the balance. The claimed composition provides enhancing the hypotensive effect based on design of the composition comprising enalapril maleate and indapamide. The selected qualitative and quantitative composition excludes sticking tablet mass on equipment in tablets making process, loss of active substance and change of the ratio mass of components in ready tablets as compared to the measured one. Invention provides high strength and stability of tablets in storage.
EFFECT: improved and valuable properties of pharmaceutical composition, improved preparing method.
SUBSTANCE: invention relates to system for controlling oral or anal administering of biological component and method for prolonged bacteria administering into living body. Claimed system contains hydrophilic agent and bacterium; or viscosity modifier, hydrophilic agent and bacterium; or viscosity modifier, electrolytic agent, hydrophilic agent and bacterium. Hydrophilic agent provides viscosity from 4000 mPa to 15000 mPa and is selected from at least one group comprising: a) gum, such as tragacant gum, locust gum, acacia gum, guar gum, xantan gum, etc.; b) polysaccharide such as pectin and maltodextrin; c) cellulose derivative such as methylcellulose, carboxymethyl cellulose, methylcellulose hydroxyethyl, methylcellulose hydroxypropyl, etc.; d) polypeptide such as gelatin, collagen, casein of heterogeneous protein mixture. Claimed systems are obtained by component drying, blending and pressing. Bacterium/hydrophilic agent ratio is from 1:0.33 to 1:1.33.
EFFECT: system for controlling administering of biological component in desired regions of gastrointestinal tract.
32 cl, 13 ex, 13 tbl, 13 dwg
FIELD: medicine, ophthalmology.
SUBSTANCE: invention proposes a solution used in treatment of cornea injuries that comprises sulfated glycosaminoglycans, cellulose water-soluble derivatives and a saline solution wherein chondroitin sulfate is used as sulfated glycosaminoglycans, hydroxyethylcellulose is used as cellulose water-soluble derivatives, and balanced saline solution is used as a saline solution wherein these components are taken in the definite ratio, g/100.0 ml. Invention provides improving rheological and viscous properties of the solution that allows obtaining anti-inflammatory and anti-edematous effect of the preparation and to accelerate the cornea tissue reparation processes.
EFFECT: improved and valuable medicinal properties of solution.
1 tbl, 3 ex
FIELD: chemical-pharmaceutical industry.
SUBSTANCE: the present innovation refers to the area of medicinal preparations, to new thyxotropic butyric carrier, in particular, that contains about 0.2-5 weight% colloid silicon dioxide and about 0.2-5 weight% hydrophilic polymer in edible oil. Interaction between hydrophilic polymer and colloid silicon dioxide in case of their application in the above-mentioned range of concentrations enables to keep low concentration of the last component, but, moreover, provides the chance to obtain solution at sufficient thyxotropy and low viscosity in case of shift. Moreover, the present innovation deals with capsules filled with a thyxotropic carrier and a pharmaceutically active substance.
EFFECT: higher efficiency.
17 cl, 11 ex, 1 tbl
SUBSTANCE: liquid preparation forms of improved light resistance contain aqueous Sitafloxacin and sodium chloride solution.
EFFECT: wide range of functional applications.
22 cl, 2 tbl
FIELD: medicine, pharmacology, pharmacy.
SUBSTANCE: invention relates to a method for enhancing availability of epotilone analog. Method involves oral administration of epotilone analog or its pharmaceutically acceptable salt, solvate, clathrate, hydrate or a prodrug in mammal in combination with pharmaceutically acceptable buffer neutralizing acid in the amount providing at least 20 mequiv. of capacity for acid neutralization that results to improving bioavailability of epotilone.
EFFECT: improved properties of formulations of drug.
20 cl, 5 tbl, 2 dwg, 9 ex
SUBSTANCE: invention relates to ophthalmologic agent composition containing therapeutically active component, namely 5-bromo-6-(2-imidazolin-2-yl-amino)-quinoxaline, solubility-increasing components except for cyclodextrins, and chloroxy derivatives, preferably effective as conserving agents. Chloroxy derivatives are preferable for conserving of therapeutically active components. Solubility-increasing components includes carboxymethyl cellulose.
EFFECT: improves composition for ophthalmology.
6 cl, 7 tbl, 1 dwg, 3 ex
FIELD: pharmaceutical industry.
SUBSTANCE: invention relates to aqueous autoclaving composition for parantheral administration useful in treatment of pathological heart conditions. Claimed composition contains methyl-3-[4-(2-hydroxy-3-isopropylamino)propoxy]-phenylpropionate hydrochloride (esmolol hydrochloride), buffer agent and osmotic controlling agent. Composition has pH between 4.5 and 5.5. Also disclosed is method for production of abovementioned esmolol hydrochloride composition including step of composition autoclaving in sealed container for time sufficient to sterilize the composition. After autoclaving composition is stable for at least 24 months.
EFFECT: composition of prolonged storage time.
11 cl, 3 ex
SUBSTANCE: the present innovation deals with stable peroral solid composition of ramosetron or its pharmaceutically acceptable salt. The composition includes one or several components chosen out of the groups including aliphatic carboxylic acid or its ester, hydroxycarboxylic acid or its ester, acidic amino acid, enolic acid, aromatic carboxylic compound or its ester and high-molecular substance that contains carboxylic group. Such an innovation refers to the method for stabilizing ramosetron composition. The suggested innovation provides stability of ramosetron composition under conditions of increased temperature and humidity, especially at low content of active component.
EFFECT: higher efficiency of application.
14 cl, 10 ex, 6 tbl
FIELD: medicine, pharmaceutical industry.
SUBSTANCE: invention proposes a method for making tablets of alpha-fetoprotein preparation. Method involves mixing alpha-fetoprotein with a stabilizing agent, addition of a filling agent consisting of microcrystalline cellulose and potato starch or lactose, coating the prepared mixture by envelope consisting of acetylphthalylcellulose in the following ratio of components, weight%: alpha-fetoprotein, 0.01-0.03; stabilizing agent, 8.57-14,28; microcrystalline cellulose, 40-45.7; potato starch or lactose, 40-45.7, and acetylphthalylcellulose, 1.14-2.85. Rheopolyglucin is used as a stabilizing agent, and alpha-fetoprotein is obtained from abortive, cord or placental blood by separation of inert proteins from blood by centrifugation, fractionation of supernatant with ammonium sulfate or sodium sulfate, or ethyl alcohol for 1 h, repeated centrifugation of solution, carrying out the first chromatography treatment of supernatant on an affinity sorbent using Sepharose CL-4B, elution of alpha-fetoprotein with 0.1 M glycine-HCl buffer solution at pH 2.5 up to obtaining zero values of optical density at wavelength 280 nm, and the following chromatography treatment on an affinity sorbent using Sepharose CL-4B containing immobilized antibodies against human IgG and dialysis of alpha-fetoprotein solution against 0.15 mole/l of sodium chloride solution. The end product is diluted with rheopolyglucin up to the concentration alpha-fetoprotein 0.9 mg/ml and that of dextran 10 mg/ml. Invention provides increasing yield of alpha-fetoprotein and maximal purity of the end product, increasing stability of alpha-fetoprotein in storage, expanding region of its using and design of preparation of prolonged effect.
EFFECT: improved preparing method.