CN103735873A - Sterilizing liquid - Google Patents
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Abstract
The invention relates to sterilizing liquid for treating various skin diseases such as psoriasis. In the sterilizing liquid, the biological raw materials of plum-leaf crab, conch, oyster, clam shell, willow mushroom and centipede are adopted. The preparation can be used for killing various skin germs, particularly has a good treatment effect on intractable psoriasis, eczema, bromhidrosis, tinea unguium, scabies, pruritus and vegetative allergy, is free from toxic and side effects. The sterilizing liquid has the characteristics of high efficiency, rapidness and low relapse rate. A therapy method of cleaning an affected part by using a low-concentration liquid medicament and dressing by using a high-concentration liquid medicament is adopted. The sterilizing liquid is lower in cost, quick in response, and very easy and convenient for treating. The sterilizing liquid is an environment-friendly medicament since only water is taken as a leaching agent without any inorganic chemical additive.
Description
Technical field
The present invention relates to one and treats dermatosis bactericidal liquid.
Background technology
Psoriasis (psoriasis) is a kind of common skin stubborn disease easily repeatedly showing effect, and China belongs to district occurred frequently, and heals compared with refractory.Although this sick not threat to life, but make that patient's skin and flesh is scratched where it itches, squama comes off, even blood stain is full of stains or spots, has both affected the attractive in appearance of patient, brings unspeakable suffering to again patient's mind & body, many patients treat timely and effectively because can not get, with the passing of time the state of an illness is delayed, and health is weak, causes various complication, affect the functions such as the heart, liver, kidney, what have also entails offspring disease.
For psoriasis, the at present domestic methods that adopt the combination of Chinese and Western medicine are treated more, as decoct Chinese medicinal decoction and wash, and smear ointment liquid medicine, sleep therapy, therapy of blood-letting, injection etc. outward; Adopt in addition hormone therapy etc. abroad, though above-mentioned various Therapeutic Method has certain curative effect, but instant effect recurrence is also fast, and has certain side effect.
Multifunctional psora sterilizing liquid CN200310105020, the present invention relates to a kind of bactericidal liquid for the treatment of the multiple dermatosis such as psoriasis, i.e. Multifunctional psora sterilizing liquid.The many dermatosiss that resemble for a long time psoriasis and so on lack good medicine always, weak curative effect and have repeatability more.It is that the way that raw material is made biological preparation obtains good result that the present invention adopts multiple biology, and this is Multifunctional psora sterilizing liquid.The biological raw material adopting has Fructus mail micromali, Carnis Rapanae thomasianae, Concha Ostreae, Concha Meretricis Seu Cyclinae, Pholiota adiposa, Scolopendra.This preparation can be killed various skin pathogenic bacteria, especially to intractable psoriasis, also has eczema, bromhidrosis, tinea unguium, scabies, pruritus, vegetalitas allergy etc. to have good therapeutic effect and without any side effects.The present invention not containing any inorganic chemical additives and only water make leaching agent, be a kind of Green medicament.
Herba speranskiae tuberculatae is among the people with all herbal medicine, cures cold, traumatic injury, and skin infection, eczema, scabies are controlled in external.Fresh juice or the decocting liquid of root and leaf have strong toxicity to the larva of cabbage butterfly, housefly and Culex tritaeniorhynchus.Root is containing Herba speranskiae tuberculatae element (phrymarolin) and leptostachyol acetas (leptostachyol acetate), and the latter is insecticidal constituent.Among the peoplely with herb, decoct water and eliminate fly larvae and Pieris rapae.
Fructus Corni (formal name used at school: Cornus officinalis), Cornaceae machaka or dungarunga.Its mature fruit is Chinese medicine, another name Fructus Corni, medicine Fructus Jujubae, Fructus Corni, another name for Sichuan Province Ziziphi Spinosae, meat Fructus Jujubae, potato Fructus Jujubae, chicken foot, real Fructus Jujubae, Fructus Corni, medicine Fructus Jujubae, day wooden seed, Fructus Corni, real Fructus Jujubae; Fructus Corni sour in the mouth, adstringency, tepor.Return liver, kidney channel.Liver and kidney tonifying, essence astringing and desertion stemming.For vertigo and tinnitus, soreness of waist and knee joint, impotence and seminal emission, enuresis frequent micturition, bleeding not during menses, profuse sweating collapse, interior-heat is quenched one's thirst, and is astringent drug.
Summary of the invention:
The bactericidal liquid medicament of the feature such as the object of the present invention is to provide a kind of dermatosis for fungal infection to have efficiently, quick, relapse rate is low, adopts that affected part low concentration medicinal liquid cleans, the therapy of working along both lines of high concentration medicinal liquid wiping.It not only saved money but also curative effect fast, treat very simple and convenient.
The parts by weight of bactericidal liquid of the present invention are composed as follows:
Rhizoma Gastrodiae powder 3-6 part, Fructus Corni extract 5-10 part, Herba speranskiae tuberculatae extract 6-12 part, fermentation of bacillus subtilis liquid extract 15-25 part, Radix Scutellariae extract 1-9 part, concentrated hydrochloric acid 8-13 part of containing hydrogen chloride 36%-38%, iodine tincture 1-3 part, water 9-25 part;
Described Herba speranskiae tuberculatae extract preparation method is as follows:
Take Herba speranskiae tuberculatae; Being crushed to particle diameter is below 2 millimeters, then in container, evenly mix and add the water of 3-6 times of weight, control temperature 70 C-90 ℃ and keep 2-4h, add the ethanol of 2-3 times of weight of mixed material and the mixture of propanol, control temperature to 60 ℃-78 ℃ of maintenance 3-4h, filter; Filtrate vacuum concentration postlyophilization obtains Herba speranskiae tuberculatae extract.
The mass ratio of described ethanol and propanol is 1:1-1.3; Concentration of alcohol is 95%; Propanol concentration is 100%;
The preparation method of described Fructus Corni extract is as follows:
Powder of Fructus Corni is broken to particle diameter to be 2 millimeters and to be placed in below container, add the water of 3-6 times of weight, control 40 ℃-60 ℃ of temperature and keep 2-4h, control temperature to 65 ℃-75 ℃ of maintenance 3-5h, add the ethanol of 2-3 times of weight of mixed material and the mixture of methanol, control temperature to 30 ℃-40 ℃ of maintenance 3-8h, filter; Filtrate vacuum concentration postlyophilization obtains Fructus Corni extract.
The mass ratio of described ethanol and methanol is 1:2, concentration of alcohol 85-95%.Methanol concentration 100%.
Described Radix Scutellariae extract preparation method is as follows:
Take the Radix Astragali; Being crushed to particle diameter is below 2 millimeters, then be placed in the water that container adds 3-6 times of weight, control 75 ℃～80 ℃ of temperature and keep 2～4h, then be cooled to 45-60 ℃, add mixing enzyme preparation to carry out enzymolysis, with newborn acid for adjusting pH value be 5.5-6.8, enzymolysis 2-4h, finally add the ethanol of mixed material 0.5-3 times weight and the mixture of propanol, control temperature to 60 ℃～78 ℃ and keep 3～4h, filter; Filtrate vacuum concentration postlyophilization obtains Radix Astragali extract.
Described mixing enzyme preparation addition is the 5-10% of mixed material gross weight.
The parts by weight of described mixing enzyme preparation consist of: endo-beta-glucanase 10-20 part, outer 1,4 beta-glucanase 10-20 part, beta-glucosidase 10-15 part, xylanase 15-20 part, beta amylase 10-15 part, neutral protease 10-15 part.
Described fermentation of bacillus subtilis liquid extract preparation method is as follows:
Described bacterial strain is specially bacillus subtilis (Bacillus subtilis) mutant HYX-008.This bacterial strain is preserved in China Committee for Culture Collection of Microorganisms's common micro-organisms center (be called for short CGMCC, address is: No. 3, No. 1, North Star West Road, Chaoyang District, BeiJing, China city institute) on July 15th, 2013, and preserving number is CGMCC7926.
Fungus characteristic is as follows:
1. somatic cells is Gram-positive raw in direct rod shape, spore or that near-end is raw.
2. colony characteristics is that white is circular, protuberance, and smooth surface, opaque, moistening, provoke adhesion.
3. can utilize various saccharides, as glucose, galactose, xylose, cellobiose, cottonseed sugar etc.; Produce amylase, protease, can gelatin hydrolysate, V.P, the C.I. 13020. positive.
4. 37 ℃ of the suitableeest cultivation temperature, optimum pH 7.0.
Described fermentation of bacillus subtilis liquid extract is the antibacterial substance containing in fermentation of bacillus subtilis liquid, and described antibacterial substance can effectively suppress psoriasic recurrence.The antibacterial substance solution of 1-2% concentration can effectively suppress psoriasic recurrence.
The method of extracting antibacterial substance from fermentation liquid is organic solvent precipitation method:
First by fermentation liquid high speed centrifugation, temperature 0-4 ℃, rotating speed is 7000 revs/min, centrifugal ten minutes, retains supernatant; In supernatant, add equal-volume organic solvent (methanol: ethanol=1:3), fully dissolve and be placed in there-necked flask, 50 degrees Celsius of water-bath control temperature, mixing speed is 40rpm, the time is 4 hours, collects organic facies, adopts vacuum distillation method to remove organic solvent.Vapo(u)rizing temperature 45-65 ℃, distills 20 minutes, and organic solvent is removed.Collecting precipitation material and get final product.
This bacterium classification status is as follows: antibacterial territory (Bacteria); Firmicutes (Firmicutes); Bacillus cereus guiding principle (Bacillales); Bacillus cereus order (Bacillaceae); Bacillaceae (Bacillus); Bacillus (Bacillus); Bacillus subtilis strain (Bacillus subtilis).
The preparation method of described bactericidal liquid is as follows:
By proportionally mix homogeneously of above-mentioned several materials, fill, sterilization packaging.
Beneficial effect:
1, drug permeability is splendid, and sterilization face is dark and wide; Medicine is dissolved very competent, the toxin that fungus invasion produces, and moment is just disengaged; Reach efficiently, at a high speed.
Although 2, operation is strict, treatment is simple, under professional instructs, is easy to grasp.
3, medicine cheapness, needs only few dose during use, treat a position and only need 0.5-1g, and patient is economical and practical.
4, without any toxicity, and cure rate is very high.
5, product of the present invention adopts Chinese crude drug effectively to extract composition and fermentation of bacillus subtilis extract, carries out organic assembling, and combination product is evident in efficacy to treating the dermatosiss such as common psoriasis.
The specific embodiment
The following examples can make the present invention of those skilled in the art's comprehend, but do not limit the present invention in any way.
Bacterial strain is specially bacillus subtilis (Bacillus subtilis) mutant, and preserving number is CGMCC7926.
The wild mushroom LI002 that described bacterial strain is separated by acid ground obtains through ultraviolet mutagenesis and nitrosoguanidine mutagenesis screening repeatedly, and characteristic is that the enzyme activity of the high temperature resistant α-amylase of product is high, heat-resisting, acid resistance is strong.
The prepared high temperature resistant α-amylase enzyme activity of the present invention is 30000-35000u/ml; Applicable temperature scope is 105-115 ℃, and 110 ℃ of optimal reactive temperatures, at 110 ℃ of enzymes complete stability alive; Being suitable for pH value in reaction scope is 3.0-7.0, at pH value, is 3.0 o'clock enzyme complete stabilities alive, and optimal reaction pH value is 4.2.
Bacterial strain that the present invention obtains is when regularization condition ferments, and unexpected discovery contained efficient sterilizing material in fermentation broth extract, and Fungicidal substance has good prevention and the dermopathic effect for the treatment of psoriasis, inventor and then utilize the present invention to develop this kind of product.
Embodiment 1:
The parts by weight of bactericidal liquid are composed as follows:
5 parts of Rhizoma Gastrodiae powders, 8 parts of Fructus Corni extracts, 10 parts of Herba speranskiae tuberculatae extracts, 20 parts of fermentation of bacillus subtilis liquid extracts, 5 parts of Radix Scutellariae extracts, 10 parts of the concentrated hydrochloric acid of containing hydrogen chloride 36%-38%, 2 parts, iodine tincture, 20 parts, water;
Described Herba speranskiae tuberculatae extract preparation method is as follows:
Take Herba speranskiae tuberculatae; Being crushed to particle diameter is below 2 millimeters, then in container, evenly mixes and add the water of 5 times of weight, controls 80 ℃ of temperature and keeps 3h, adds the mixture of 3 times of weight ethanol of mixed material and propanol, controls temperature to 75 ℃ and keeps 3h, filters; Filtrate vacuum concentration postlyophilization obtains.
The mass ratio of described ethanol and propanol is 1:1; Concentration of alcohol is 95%; Propanol concentration is 100%;
The preparation method of described Fructus Corni extract is as follows:
To be crushed to particle diameter is that 2 millimeters of following Fructus Corni are placed in container, adds the water of 5 times of weight, controls temperature 50 C and keeps 3h, control temperature to 70 ℃ and keep 4h, add the mixture of 3 times of weight ethanol of mixed material and methanol, control temperature to 35 ℃ and keep 5h, filter; Filtrate vacuum concentration postlyophilization obtains Fructus Corni extract.
The mass ratio of described ethanol and methanol is 1:2, concentration of alcohol 85%, methanol concentration 100%.
Radix Scutellariae extract preparation method is as follows:
Take the Radix Astragali; Being crushed to particle diameter is below 2 millimeters, then in container, add the water of 5 times of weight, control 75 ℃ of temperature and keep 3h, then be cooled to 55 ℃, add mixing enzyme preparation to carry out enzymolysis, with newborn acid for adjusting pH value be 6, enzymolysis 3h, finally add the mixture of 2 times of weight ethanol of mixed material and propanol, control temperature to 70 ℃ and keep 3h, filter; Filtrate vacuum concentration postlyophilization obtains.
Described mixed enzyme addition is 6% of mixed material gross weight.
The parts by weight of described mixed enzyme consist of: 13 parts of endo-beta-glucanases, 15 parts of outer 1,4 beta-glucanases, 12 parts of beta-glucosidases, 16 parts of xylanase, 12 parts of beta amylases, 12 parts of neutral protease.
Fermentation of bacillus subtilis liquid extract preparation method is as follows:
Bacterial strain is specially bacillus subtilis (Bacillus subtilis).This bacterial strain is preserved in China Committee for Culture Collection of Microorganisms's common micro-organisms center (be called for short CGMCC, address is: No. 3, No. 1, North Star West Road, Chaoyang District, BeiJing, China city institute) on July 15th, 2013, and preserving number is CGMCC7926.
(1) actication of culture
The slant strains of intact bacillus subtilis is inoculated in to slant medium, cultivates 30h for 40 ℃ and carry out actication of culture, so activate 2-3 time;
Described slant medium consists of: Carnis Bovis seu Bubali cream 6g, sodium chloride 10g, peptone 15g, glucose 3g, agar 18g, distilled water l000mL, 6,121 ℃ of sterilizing 20min of pH value;
(2) liquid seeds amplification culture
1. first order seed is cultivated: the slant strains 1-2 articulating after step (1) activation is entered in 500 ml shake flasks to 100 milliliters of culture medium loading amounts, 180 revs/min of rotary shaking tables, 40 ℃ of cultivation temperature, incubation time 12h;
2. secondary seed is cultivated: by first order seed, according in 500 milliliters of secondary seed shaking flasks of inoculum concentration access of 10%, condition of culture is identical with first order seed;
3. three grades of seed culture: secondary seed is accessed in 5000 milliliters of three grades of seed shaking flasks to 1000 milliliters of culture medium loading amounts, 100 revs/min of rotary shaking tables, 40 ℃ of cultivation temperature, incubation time 12h with 10% inoculum concentration;
4. first class seed pot is cultivated: the first class seed pot by three grades of seeds take 10% inoculum concentration access total measurement (volume) as 150L, fermentation medium loading amount 100L, 40 ℃ of cultivation temperature, mixing speed 300rpm, ventilation (V/V) 1:1, tank pressure 0.05Mpa, incubation time 15h;
Described one-level, secondary, three grades of seed culture medium weight consist of:
Yeast powder 0.3%, glucose 1.5%, peptone 0.3%, Carnis Bovis seu Bubali cream 0.5%, dipotassium hydrogen phosphate 0.8%, calcium sulfate 1g, magnesium chloride 2g, sodium citrate 1g, insufficient section pure water is supplied, 5,121 ℃ of sterilizing 30min of pH value.
Described seed tank culture basic weight amount consists of:
Maltodextrin 10%, yeast powder 0.4%, peptone 0.3%, Semen Maydis pulp 0.4%, dipotassium hydrogen phosphate 1.0%, magnesium sulfate 0.08%, sodium citrate 0.3%, insufficient section pure water is supplied, 6,121 ℃ of sterilizing 30min of pH value.
Described seed tank fermentation liquid cell concentration is 7.0x108/ml;
(3) ferment tank
First class seed pot fermentation liquid in step (2) is accessed to fermentation tank, 36 ℃ of cultivation temperature, mixing speed 300r/m, ventilation (V/V) 1:2, incubation time 36h with 6% inoculum concentration.
Dissolved oxygen control: by adjusting speed of agitator and ventilation, control dissolved oxygen 20%;
PH controls: by mending ammonia or phosphoric acid,diluted, in controlled fermentation process, pH value remains on 5;
Control of additive raw material: when the content of reducing sugar in fermentation liquid is down to 3mg/ml, start to add supplemented medium, feed supplement amount is to maintain fermentation liquid content of reducing sugar as 4mg/ml;
Put tank standard: the old and feeble self-dissolving of 40% thalline.
Described fermentation medium consists of: maltodextrin 100g, Semen Maydis powder 50g, soybean cake powder 25g, Herba speranskiae tuberculatae 10g, Radix Scutellariae 20g, yeast powder 6g, Semen Maydis pulp 3g, ammonium sulfate 2g, dipotassium hydrogen phosphate 1g, potassium dihydrogen phosphate 1g, sodium citrate 3g, defoamer 0.1g, pure water l000mL, 6,121 ℃ of sterilizing 20min of pH value;
Described supplemented medium weight consists of: maltodextrin 25%, and Semen Maydis powder 15%, Semen Glycines powder 20%, insufficient section pure water is supplied, 6,121 ℃ of sterilizing 30min of pH value.
The concocting method of described fermentation medium is:
Accurately take in proportion raw material, pure water in raw material, Semen Maydis powder, soybean cake powder are dropped in material-compound tank, regulate pH value 6, add midrange thermal stable amylase (3u/g Semen Maydis powder) and alpha-amylase (30u/g Semen Maydis powder), ℃ insulation 15min of warming while stirring to 70 simultaneously, is then slowly warming up to 90 ℃ of insulation 30min and liquefies, finally add other raw material, stir, adjust initial pH6,121 ℃ of sterilizing 30min are standby.
From fermentation liquid, extracting antibacterial substance method is organic solvent precipitation method:
First by fermentation liquid high speed centrifugation, 2 ℃ of temperature, rotating speed is 7000 revs/min, centrifugal ten minutes, retains supernatant; In supernatant, add equal-volume organic solvent (methanol: ethanol=1:3), fully dissolve and be placed in there-necked flask, 50 degrees Celsius of water-bath control temperature, mixing speed is 40rpm, the time is 4 hours, collects organic facies, adopts vacuum distillation method to remove organic solvent.55 ℃ of vapo(u)rizing temperatures, distill 20 minutes, and organic solvent is removed.Collecting precipitation material and get final product.
The preparation method of described bactericidal liquid is as follows: by proportionally mix homogeneously of above-mentioned several materials, fill, sterilization packaging.
Embodiment 2:
The parts by weight of bactericidal liquid are composed as follows:
3 parts of Rhizoma Gastrodiae powders, 8 parts of Fructus Corni extracts, 6 parts of Herba speranskiae tuberculatae extracts, 25 parts of fermentation of bacillus subtilis liquid extracts, 9 parts of Radix Scutellariae extracts, 8 parts of the concentrated hydrochloric acid of containing hydrogen chloride 36%, 3 parts, iodine tincture, 21 parts, water.
Embodiment 4: experiment effect
Example: treatment psoriasis
Zhai Tan township, Songyang in Zhejiang Province county in 2007 spends the Zhou Yufa in end of the bridge village, 28 years old age.The state of an illness of observing, left side item neck connects near shoulder, approximately has the surface of 10 square centimeters, has the dry scurf of silvery white to come off, and the time has had 8 years.Itching in the inside, extremely feels bad.One scratch just know subcutaneous inflamed.Right side item neck approximately has the surface of more than 10 square centimeter near connecting shoulder, with left side difference, is but a slice black, tears a small amount of casting skin, subcutaneous also inflamed, can't bear to stand.It is all invalid for many years that patient mentions commune hospital, township, county hospital is sought medical advice.
Diagnosis: belong to serious symptom psoriasis.1. first with the anti-cleanout fluid that spreads anti-recurrence, clean once.2. because patient is young and vigorous, holding capacity is strong, and case history is also longer.In order to cure the disease at a high speed, clean every 10 minutes, at once by this high-grade bactericidal liquid to 5-7 seconds of affected part wiping.Be worth reminding: must be very careful while getting it filled cotton, first with tweezers, clamp 1 cubic centimetre of big or small cotton balls, open bottleneck, insert in medicament, be then pulled to bottleneck and rub slightly pressure, humidity is as alcohol swab.First build bottleneck, then to affected part wiping.The speed that speed is walked about with second hand is as the criterion.Interval 2 days in dispenser for the second time.By that analogy, left neck affected part treatment 4 times, right neck affected part is also treated 4 times, 8 times altogether.Several years, Zhou Yufa cervical region was safe and sound, did manual work at county town, Songyang always.
Adopt the method product of the present invention successively to cure this type of patient of 100 many cases.
Claims (8)
1. bactericidal liquid, parts by weight are composed as follows: Rhizoma Gastrodiae powder 3-6 part, Fructus Corni extract 5-10 part, Herba speranskiae tuberculatae extract 6-12 part, fermentation of bacillus subtilis liquid extract 15-25 part, Radix Scutellariae extract 1-9 part, concentrated hydrochloric acid 8-13 part of containing hydrogen chloride 36%-38%, iodine tincture 1-3 part, water 9-25 part;
Described fermentation of bacillus subtilis liquid extract preparation method is as follows:
Described bacterial strain is specially bacillus subtilis (Bacillus subtilis) CGMCC7926.
The method of extracting antibacterial substance from fermentation liquid is organic solvent precipitation method:
First by fermentation liquid high speed centrifugation, temperature 0-4 ℃, rotating speed is 7000 revs/min, centrifugal ten minutes, retains supernatant; In supernatant, add equal-volume organic solvent (methanol: ethanol=1:3), fully dissolve and be placed in there-necked flask, 50 degrees Celsius of water-bath control temperature, mixing speed is 40rpm, the time is 4 hours, collects organic facies, adopts vacuum distillation method to remove organic solvent.Vapo(u)rizing temperature 45-65 ℃, distills 20 minutes, and organic solvent is removed.Collecting precipitation material and get final product.
2. bactericidal liquid according to claim 1, is characterized in that described Herba speranskiae tuberculatae extract preparation method is as follows:
Take Herba speranskiae tuberculatae; Being crushed to particle diameter is below 2 millimeters, then in container, evenly mix and add the water of 3-6 times of weight, control temperature 70 C-90 ℃ and keep 2-4h, add the ethanol of 2-3 times of weight of mixed material and the mixture of propanol, control temperature to 60 ℃-78 ℃ of maintenance 3-4h, filter; Filtrate vacuum concentration postlyophilization obtains Herba speranskiae tuberculatae extract.
3. bactericidal liquid according to claim 1, is characterized in that the preparation method of described Fructus Corni extract is as follows:
Powder of Fructus Corni is broken to particle diameter to be 2 millimeters and to be placed in below container, add the water of 3-6 times of weight, control 40 ℃-60 ℃ of temperature and keep 2-4h, control temperature to 65 ℃-75 ℃ of maintenance 3-5h, add the ethanol of 2-3 times of weight of mixed material and the mixture of methanol, control temperature to 30 ℃-40 ℃ of maintenance 3-8h, filter; Filtrate vacuum concentration postlyophilization obtains Fructus Corni extract.
The mass ratio of described ethanol and methanol is 1:2, concentration of alcohol 85-95%.Methanol concentration 100%.
4. bactericidal liquid according to claim 1, is characterized in that described Radix Scutellariae extract preparation method is as follows:
Take the Radix Astragali; Being crushed to particle diameter is below 2 millimeters, then be placed in the water that container adds 3-6 times of weight, control 75 ℃～80 ℃ of temperature and keep 2～4h, then be cooled to 45-60 ℃, add mixing enzyme preparation to carry out enzymolysis, with newborn acid for adjusting pH value be 5.5-6.8, enzymolysis 2-4h, finally add the ethanol of mixed material 0.5-3 times weight and the mixture of propanol, control temperature to 60 ℃～78 ℃ and keep 3～4h, filter; Filtrate vacuum concentration postlyophilization obtains Radix Astragali extract.
Described mixing enzyme preparation addition is the 5-10% of mixed material gross weight.
The parts by weight of described mixing enzyme preparation consist of: endo-beta-glucanase 10-20 part, outer 1,4 beta-glucanase 10-20 part, beta-glucosidase 10-15 part, xylanase 15-20 part, beta amylase 10-15 part, neutral protease 10-15 part.
5. bactericidal liquid according to claim 1, is characterized in that the preparation method of described bactericidal liquid is as follows:
By proportionally mix homogeneously of above-mentioned several materials, fill, sterilization packaging.
6. bactericidal liquid according to claim 1, is characterized in that the parts by weight of described bactericidal liquid are composed as follows:
5 parts of Rhizoma Gastrodiae powders, 8 parts of Fructus Corni extracts, 10 parts of Herba speranskiae tuberculatae extracts, 20 parts of fermentation of bacillus subtilis liquid extracts, 5 parts of Radix Scutellariae extracts, 10 parts of the concentrated hydrochloric acid of containing hydrogen chloride 36%-38%, 2 parts, iodine tincture, 20 parts, water.
7. according to bactericidal liquid described in claim 1 or 6, it is characterized in that described fermentation of bacillus subtilis liquid extract preparation method is as follows:
Bacterial strain CGMCC7926;
(1) actication of culture
The slant strains of intact bacillus subtilis is inoculated in to slant medium, cultivates 30h for 40 ℃ and carry out actication of culture, so activate 2-3 time;
Described slant medium consists of: Carnis Bovis seu Bubali cream 6g, sodium chloride 10g, peptone 15g, glucose 3g, agar 18g, distilled water l000mL, 6,121 ℃ of sterilizing 20min of pH value;
(2) liquid seeds amplification culture
1. first order seed is cultivated: the slant strains 1-2 articulating after step (1) activation is entered in 500 ml shake flasks to 100 milliliters of culture medium loading amounts, 180 revs/min of rotary shaking tables, 40 ℃ of cultivation temperature, incubation time 12h;
2. secondary seed is cultivated: by first order seed, according in 500 milliliters of secondary seed shaking flasks of inoculum concentration access of 10%, condition of culture is identical with first order seed;
3. three grades of seed culture: secondary seed is accessed in 5000 milliliters of three grades of seed shaking flasks to 1000 milliliters of culture medium loading amounts, 100 revs/min of rotary shaking tables, 40 ℃ of cultivation temperature, incubation time 12h with 10% inoculum concentration;
4. first class seed pot is cultivated: the first class seed pot by three grades of seeds take 10% inoculum concentration access total measurement (volume) as 150L, fermentation medium loading amount 100L, 40 ℃ of cultivation temperature, mixing speed 300rpm, ventilation (V/V) 1:1, tank pressure 0.05Mpa, incubation time 15h;
Described one-level, secondary, three grades of seed culture medium weight consist of:
Yeast powder 0.3%, glucose 1.5%, peptone 0.3%, Carnis Bovis seu Bubali cream 0.5%, dipotassium hydrogen phosphate 0.8%, calcium sulfate 1g, magnesium chloride 2g, sodium citrate 1g, insufficient section pure water is supplied, 5,121 ℃ of sterilizing 30min of pH value.
Described seed tank culture basic weight amount consists of:
Maltodextrin 10%, yeast powder 0.4%, peptone 0.3%, Semen Maydis pulp 0.4%, dipotassium hydrogen phosphate 1.0%, magnesium sulfate 0.08%, sodium citrate 0.3%, insufficient section pure water is supplied, 6,121 ℃ of sterilizing 30min of pH value.
Described seed tank fermentation liquid cell concentration is 7.0x108/ml;
(3) ferment tank
First class seed pot fermentation liquid in step (2) is accessed to fermentation tank, 36 ℃ of cultivation temperature, mixing speed 300r/m, ventilation (V/V) 1:2, incubation time 36h with 6% inoculum concentration.
Dissolved oxygen control: by adjusting speed of agitator and ventilation, control dissolved oxygen 20%;
PH controls: by mending ammonia or phosphoric acid,diluted, in controlled fermentation process, pH value remains on 5;
Control of additive raw material: when the content of reducing sugar in fermentation liquid is down to 3mg/ml, start to add supplemented medium, feed supplement amount is to maintain fermentation liquid content of reducing sugar as 4mg/ml;
Put tank standard: the old and feeble self-dissolving of 40% thalline.
From fermentation liquid, extracting antibacterial substance method is organic solvent precipitation method:
First by fermentation liquid high speed centrifugation, 2 ℃ of temperature, rotating speed is 7000 revs/min, centrifugal ten minutes, retains supernatant; In supernatant, add equal-volume organic solvent (methanol: ethanol=1:3), fully dissolve and be placed in there-necked flask, 50 degrees Celsius of water-bath control temperature, mixing speed is 40rpm, the time is 4 hours, collects organic facies, adopts vacuum distillation method to remove organic solvent.55 ℃ of vapo(u)rizing temperatures, distill 20 minutes, and organic solvent is removed.Collecting precipitation material and get final product.
8. bactericidal liquid according to claim 1, is characterized in that the parts by weight of described bactericidal liquid are composed as follows:
3 parts of Rhizoma Gastrodiae powders, 8 parts of Fructus Corni extracts, 6 parts of Herba speranskiae tuberculatae extracts, 25 parts of fermentation of bacillus subtilis liquid extracts, 9 parts of Radix Scutellariae extracts, 8 parts of the concentrated hydrochloric acid of containing hydrogen chloride 36%, 3 parts, iodine tincture, 21 parts, water.
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201310741752.9A CN103735873A (en) | 2013-12-28 | 2013-12-28 | Sterilizing liquid |
Applications Claiming Priority (1)
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| CN201310741752.9A CN103735873A (en) | 2013-12-28 | 2013-12-28 | Sterilizing liquid |
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105146103A (en) * | 2015-09-17 | 2015-12-16 | 天津中天精科科技有限公司 | Feed additive |
| CN105192309A (en) * | 2015-09-17 | 2015-12-30 | 天津中天精科科技有限公司 | Composite feed additive |
| CN108659964A (en) * | 2018-07-17 | 2018-10-16 | 合肥东恒锐电子科技有限公司 | A kind of cleaning spraying of keyboard degerming disinfection of deoiling |
| CN108721336A (en) * | 2018-06-15 | 2018-11-02 | 江苏远山生物技术有限公司 | The preparation method and application of bacillus coagulans and bacillus licheniformis cocktail spray |
Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103610939A (en) * | 2013-11-19 | 2014-03-05 | 宁夏天地经纬电力设备工程有限公司 | Sterilization liquid |
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2013
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Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103610939A (en) * | 2013-11-19 | 2014-03-05 | 宁夏天地经纬电力设备工程有限公司 | Sterilization liquid |
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105146103A (en) * | 2015-09-17 | 2015-12-16 | 天津中天精科科技有限公司 | Feed additive |
| CN105192309A (en) * | 2015-09-17 | 2015-12-30 | 天津中天精科科技有限公司 | Composite feed additive |
| CN108721336A (en) * | 2018-06-15 | 2018-11-02 | 江苏远山生物技术有限公司 | The preparation method and application of bacillus coagulans and bacillus licheniformis cocktail spray |
| CN108659964A (en) * | 2018-07-17 | 2018-10-16 | 合肥东恒锐电子科技有限公司 | A kind of cleaning spraying of keyboard degerming disinfection of deoiling |
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Application publication date: 20140423 |