CN103142742A - Method for preparing oral solution of poplar flower - Google Patents
Method for preparing oral solution of poplar flower Download PDFInfo
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- CN103142742A CN103142742A CN2013100918574A CN201310091857A CN103142742A CN 103142742 A CN103142742 A CN 103142742A CN 2013100918574 A CN2013100918574 A CN 2013100918574A CN 201310091857 A CN201310091857 A CN 201310091857A CN 103142742 A CN103142742 A CN 103142742A
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Abstract
The invention discloses a method for preparing an oral solution of the poplar flower and belongs to the technical field of veterinary medicaments. The method comprises the following steps of: (1) putting the poplar flower into an extraction tank, adding water into the poplar flower, stirring, and increasing the temperature to 45-50 DEG C, wherein the water is 4-6 times of the mass of the poplar flower; (2) adding cellulose into the extraction tank, adjusting the pH value of the materials to 5.0-5.5, and hydrolyzing under the action of the cellulose for 90-120 minutes at the temperature of 45-50 DEG C, wherein the mass of the cellulose is 3-7% of that of the poplar flower; and (3) filtering a solution of enzymolysis, and collecting a filtrate to obtain the poplar-flower oral solution. The poplar flower is extracted by adopting the technology of hydrolyzing under the action of the cellulose, and a novel process for extracting the poplar flower is developed, thus the time spent in extracting the poplar flower is shortened, the heat consumption is reduced, the amount of water used in the process is reduced, and the purposes of saving energy, reducing emission, improving the work efficiency and the extraction efficiency are achieved. The method has good social and economic benefits.
Description
Technical field
The present invention relates to the veterinary drug technical field, especially a kind of preparation method of poplar flower oral liquid.
Background technology
Flower of poplar is Salicaceous Plants Canada willow populus canadensis Mench. inflorescence, is rich in the compositions such as flavone, alkaloid, organic acid, vitamin, phenols, has the effect of strengthening the spleen and nourishing the stomach, removing dampness dysentery relieving, relieving diarrhea with astringents, heat-clearing and toxic substances removing.Poplar flower oral liquid is recorded in 2010 editions " Chinese veterinary pharmacopoeias " two ones, and is clinical multiplex in pig, sheep, fowl dysentery.Modern pharmacology studies show that flower of poplar has the effects such as antibiotic, antipyretic and anti-inflammatory, dysentery relieving antidiarrheal.The tradition poplar flower oral liquid is to be prepared from by immersion, heated and boiled, decoction, the method such as concentrated, and the process time is long, and thermal energy consumption is high, process water consumption is large, extraction ratio is low.1 ton of oral liquid of every production needs more than tens hour before soaking, decoct, concentrate, precipitate, be filled into packing; Decoction, the concentrated hectic fever steam 3-5 ton that needs; Medical material decocts 2 times, needs to consume 8 tons of purified water (needing 40 tons by drinking water standard), concentrated 10 tons of the waters that need; The effective ingredient flavones content is up to 0.132%.
Middle the effective elements of the medicine stores in the framework that is cell wall cellulose formation mostly, in being wrapped in by cellulose, extracts its effective ingredient, and the cellulose framework that must first break its cell wall fully is dissolved out effective ingredient.Be used for Chinese medicine extraction research more be cellulase, cellulose is to be connected with Isosorbide-5-Nitrae-β glucoside bond by β-D-Glucose, can destroy β-D-Glucose key with cellulase degradation, makes plant cell wall destruction, is conducive to the extraction to effective ingredient.In recent years, the report of domestic employing cellulase extraction Effective Component of Chinese Medicine constantly occurs.The introductions such as Lv Weiming employing enzyme hydrolysis method extracts the separation baicalin and reaches 75.67% from Radix Scutellariae, yield is 2.46%.Someone extracts total flavones from Radix Puerariae under the effect of cellulase, yield has improved 13%.In the Radix Notoginseng leaching process, destroy cell wall with the cellulase degradation effect, the total soap of Radix Notoginseng former times extraction ratio improves 23.5%.In addition, at first Shanghai Chinese medicine one factory uses enzyme process and has successfully prepared SHENGMAI YIN KOUFUYE.Have with traditional extraction process comparison cellulase solution and improve yield, energy savings, reduce the solvent use amount etc. advantage, adopt the report of cellulase extraction flower of poplar but so far there are no.
Summary of the invention
The invention provides a kind of preparation method of poplar flower oral liquid, utilize the cellulase hydrolysis technology to extract flower of poplar, developed a kind of new technology of extracting flower of poplar, greatly shorten extraction time, reduced thermal energy consumption, reduced process water consumption, reach energy-saving and emission-reduction, increased work efficiency and the purpose of extraction ratio, had good society and economic benefit.
A kind of preparation method of poplar flower oral liquid comprises the steps:
(1) flower of poplar is put into extraction pot, add flower of poplar quality 4-6 water doubly, stir and be warming up to 45-50 ℃;
(2) add the cellulase of flower of poplar quality 3-7% in extraction pot, regulate pH value to 5.0-5.5,45-50 ℃ of lower enzymolysis 90-120min;
(3) enzymolysis solution is filtered, collect filtrate, namely get poplar flower oral liquid.
Regulate pH value with the lactic acid solution of 10-50% or the sodium hydroxide solution of 10-50% in step (2).
The quality of step (2) cellulase is 4% of flower of poplar quality.
In step (2), enzymolysis time is 120min.
Its usage and dosage is: horse, cattle 50～100ml; Sheep, pig 10～20ml; Rabbit, fowl 1～2ml.
Single factors such as enzyme addition, hydrolysis temperature, enzymolysis time, solvent pH value affect test to the flavone extraction ratio:
1, cellulase activity detects
Cellulase is a kind of important compound bio-enzyme, mainly is comprised of circumscribed 1,4 beta-glucanase, Endo-β-glucanase and beta-glucosidase etc., also has the xylanase of very high vigor.
Cellulose and hemicellulose have consisted of the plant cell wall framework, and the flower of poplar effective ingredient is present in the framework that cellulose and hemicellulose formation are arranged.Cellulase has specificity, the cellulase decomposition of cellulose, and xylanase decomposes hemicellulose, and glucanase continues to resolve into glucose with cellulose and hemicellulose catabolite.Flower of poplar cell wall under cellulase and xylanase effect fully decomposes and discharges the flower of poplar effective ingredient.
Adopt spectrophotometry cellulase and Xylanase activity.
Detection fibers element enzyme cellulase 〉=30000U/g, xylanase 〉=100000U/g, 1,4 beta-glucanase 〉=120000U/g.
2, standard solution preparation and Specification Curve of Increasing
2.1 the preparation of control substance of Rutin solution:
Data shows that flavone detects take rutin as standard reference material.
Accurately take control substance of Rutin 10mg, be placed in the 50ml volumetric flask, add 30% ethanol 30ml, ultrasonic concussion makes it dissolving, is cooled to room temperature, and is diluted to scale, shakes up to such an extent that concentration is the control substance of Rutin solution of 200 μ g/ml, and cold preservation is standby.
2.2 the selection of absorbing wavelength:
Reference substance is measured determining of wavelength: get reference substance solution 1ml, undertaken by the method under the standard curve item, in the interscan of 400～700nm wave-length coverage, result is absorption maximum at 500nm wavelength place.Determining of sample determination wavelength: the ethanol extract 1ml of sample thief, undertaken by the method under the standard curve item, in the interscan of 400～700nm wave-length coverage, result is absorption maximum at 500nm wavelength place.
2.3 Specification Curve of Increasing:
Precision takes control substance of Rutin solution 0,1.0,2.0,3.0 4.0,5.0ml is respectively to the 25ml volumetric flask, add respectively 5% sodium nitrite solution 0.3ml, shake up, place 6min, add 10% aluminum nitrate solution 0.3ml, place 6min, add 1mol sodium hydroxide solution 4ml, use respectively 30% ethanol standardize solution, shake up, place 15min, put in cuvette, the place measures absorbance at the 500nm wavelength.Take absorbance A as vertical coordinate, concentration C is abscissa, drawing standard curve (see figure 1).Through returning statistics, get the standard curve equation: A=0.0486+0.0002 R
2=0.9996.Concentration in 0-400 μ g/ml scope R square reach 0.9996, linear relationship is good.
3, single-factor influence test
3.1 the impact (A factor) of cellulase addition on flavone extraction ratio in poplar flower oral liquid
Get 7 parts of flower of poplar, every part of 100g, the cellulase addition is respectively 2g, 3g, 4g, 5g, 6g, 7 g, 8 g, hydrolysis temperature is controlled at 45 ℃, enzymolysis pH value and is controlled under 5.0 conditions, enzymolysis time is controlled at 90 min, collect enzymolysis solution, detect general flavone content according to the assay method that detects general flavone content in poplar flower oral liquid, the cellulase addition is seen Fig. 2 to the impact of flavone extraction ratio in poplar flower oral liquid.
as shown in Figure 2, enzyme addition single-factor influence result of the test shows, get the 100g flower of poplar and prepare poplar flower oral liquid, along with the enzyme addition increases, in oral liquid, the flavone extraction ratio raises, flavone extraction ratio in sample when enzyme adds 3g to, significantly raise than the extraction ratio that adds 2g, when adding 4-8g to, the flavone extraction ratio improves always, adding the 7g amplitude that the flavone extraction ratio improves when above to diminishes, the many factors such as flavone extraction ratio in comprehensive production cost and flower of poplar, get the 100g flower of poplar and prepare oral liquid, add the 3-7g cellulase more suitable, be preferably 4g.
3.2 the impact (B factor) of hydrolysis temperature on flavone extraction ratio in poplar flower oral liquid
Get 5 parts of flower of poplar, every part of 100g, the cellulase addition is 4g, hydrolysis temperature is respectively 35 ℃, 40 ℃, 45 ℃, 50 ℃, 55 ℃, the enzymolysis pH value is controlled under the conditions such as 5.0, and enzymolysis time is controlled at 90 min, collects enzymolysis solution, detect general flavone content according to the assay method that detects general flavone content in poplar flower oral liquid, hydrolysis temperature is seen Fig. 3 to the impact of flavone extraction ratio in poplar flower oral liquid.
As shown in Figure 3, under same enzyme addition, enzymolysis time and enzymolysis pH condition, hydrolysis temperature single-factor influence result of the test shows, hydrolysis temperature is from 35 ℃ to 45 ℃, be elevated to 0.154% with flavone extraction ratio in temperature rising poplar flower oral liquid by 0.102%, the hydrolysis temperature extraction ratio 0.144% 50 ℃ time that raises, extraction ratio drops to 0.016% when hydrolysis temperature reaches 55 ℃, in view of this, hydrolysis temperature impact in the cellulose enzyme process extracts is larger, and the temperature of suitable extraction is between 45 ℃ to 50 ℃.
The impact (C factor) of enzymolysis time on flavone extraction ratio in poplar flower oral liquid
Get flower of poplar 500g, be divided into 5 parts, every part of 100g, cellulase addition are 4g, 45 ℃ of hydrolysis temperatures, enzymolysis time is respectively 30min, 60 min, 90 min, 120 min, 150 min, the enzymolysis pH value is controlled at 5.0, and enzymolysis time arrives, and collects enzymolysis solution, detect general flavone content according to the assay method that detects general flavone content in poplar flower oral liquid, enzymolysis time is seen Fig. 4 to the impact of flavone extraction ratio in poplar flower oral liquid.
As shown in Figure 4, under same enzyme addition, hydrolysis temperature and enzymolysis pH condition, enzymolysis time single-factor influence result of the test shows: begin to take a sample from the enzymolysis 30min time, extending extraction ratio with enzymolysis time improves constantly, extraction ratio is substantially constant after enzymolysis time reaches 120min, and the extraction ratio of enzymolysis 150min and 120min is basically identical.Therefore, from single factor experiment, enzymolysis time is more suitable at 90-120min, is preferably 120min.
3.4 the impact (D factor) of enzymolysis pH value on flavone extraction ratio in poplar flower oral liquid
Get flower of poplar 500g, be divided into 5 parts, every part of 100g, the cellulase addition is 4g, 45 ℃ of enzymolysis time 90 min of hydrolysis temperature, pH value is respectively 4.0,4.5,5.0,5.5,6.0, collects enzymolysis solution, detect general flavone content according to the assay method that detects general flavone content in poplar flower oral liquid, the enzymolysis pH value is seen Fig. 5 to the impact of flavone extraction ratio in poplar flower oral liquid.
As shown in Figure 5, under the condition of same enzyme addition, hydrolysis temperature and enzymolysis time, enzymolysis pH value single-factor influence result of the test shows: enzymolysis pH value flavone extraction ratio in 5.0,5.5 o'clock poplar flower oral liquids is the extraction ratio of 4.0,4.5,6.0 o'clock apparently higher than the enzymolysis pH value, and wherein the enzymolysis pH value is that to be starkly lower than pH value be 4.0 extraction ratio 0.082% for the extraction ratio 0.023% of 6.0 o'clock.In view of this pH value is that 6.0 pairs of cellulase destroy seriously, causes the cellulase inactivation, has affected hydrolysis result.Therefore, setting enzymolysis pH scope in cellulase solution extraction poplar flower oral liquid technique is 5.0-5.5.
4, orthogonal experiments:
Select L9(3
4) orthogonal array is take the flavone extraction ratio as performance assessment criteria, investigate the cellulase solution and extract enzyme addition factor A, hydrolysis temperature factor B in poplar flower oral liquid, enzymolysis time factor C, enzymolysis pH value factor D to the impact of poplar flower oral liquid extraction process, orthogonal experiments sees Table 1.
Table 1 L9(3
4) orthogonal array
By to L9(3
4) orthogonal experiments carries out range analysis, sequentially is followed successively by A affecting the impact on the flavone extraction ratio of the A factor enzyme addition of flavone extraction ratio, B factor hydrolysis temperature, C factor enzymolysis time and D factor enzymolysis pH value B C D.KA3 is almost consistent with kA2 in the A factor, and kA3, kA2 and KA1 difference are obvious aobvious; In the B factor, KB2 and KB3 difference are little, both and KB1 obvious difference; KC3 is apparently higher than kC1 and kC2 in the C factor; KD3 and kD2 are more approaching, apparently higher than KD1.
Comprehensive above factor influence degree and controlling factors ability, addition and the enzymolysis time of enzyme are easily controlled, and are not subject to other condition influence, and hydrolysis temperature and enzymolysis pH value are distributed with different in extracting solvent.Therefore, by quadrature L9(3
4) filtering out cellulase solution extraction flower of poplar process conditions for adding enzyme 3-7g, enzymolysis time 90-120min, hydrolysis temperature are controlled between 45-50 ℃, and the enzymolysis pH value is controlled between 5.0-5.5.
5, demonstration test
According to L9(34) the interpolation enzyme 3-7% that filters out of orthogonal test, enzymolysis time 90-120min, hydrolysis temperature is controlled between 45-50 ℃, the enzymolysis pH value is controlled at the process conditions between 5.0-5.5, carried out the proving test of continuous three batches at pilot plant, three batches of test temperatures are respectively 46 ℃, 48 ℃, 48 ℃, and pH is respectively 5.2,5.5,5.1, three batches of test products flavone extraction ratios are respectively 0.185%, 0.189%, 0.188%, and weighted average demonstration test extraction ratio is 0.187%.According to 2010 editions " Chinese veterinary pharmacopoeias " two poplar flower oral liquid quality standard checks, every assay is up to specification.
6, clinical efficacy:
The drug sensitive test result shows that cellulase solution extraction poplar flower oral liquid of the present invention has obvious fungistatic effect to the broiler escherichia coli, can be used for the treatment of broiler colibacillosis.
6.1 poplar flower oral liquid is to the colibacillary drug sensitive test of broiler.Seminar adopts the pathological material of disease of the colibacillosis that the chicken house clinical onset be positioned at three regions makes a definite diagnosis to be numbered A, B, C, be numbered 1 take poplar flower oral liquid of the present invention as trial drug susceptibility sheet, take the former powder of florfenicol, the former powder of colistin sulfate, the former powder of polygynax, the former powder in amoxicillin as control drug (being numbered 2,3,4,5), carried out drug sensitivity test.Result of the test sees Table 2:
Table 2 drug sensitivity test result (unit: mm)
Shown by the result of the test of table 2: no matter whether medication was treated from three groups of chicken groups' of different geographical bacterial strain, and was all responsive to poplar flower oral liquid.
6.2 the clinical effect trial of poplar flower oral liquid control broiler colibacillosis
12600 plumages that project team raises for certain broiler breeding field, 15 ages in days, the colibacillosis of the clinical generation of Ross 308 Broiler has been carried out the clinical efficacy contrast test.This test is control drug with poplar flower oral liquid and 10% colistin sulfate soluble powder and 10% ofloxacin soluble powder, investigates cellulase hydrolysis poplar flower oral liquid clinical efficacy, and result of the test sees Table 3.
Table 3 poplar flower oral liquid clinical efficacy
Result of the test by table 3 shows: it is extremely remarkable than C group, D group difference that A group, B organize death rate, and there were significant differences in weightening finish, and feedstuff-meat ratio is without significant difference; The dead naughty number of A group, death rate, weightening finish, material meat test effect are organized significantly better than B, and single statistical data is without significant difference.
7, Economic and Efficiency Analysis
Through experimental analysis, adopt enzymolysis process to prepare poplar flower oral liquid, steam consumption descends 86.4%, and ample resources has been saved in using water wisely 53.5%.
45 tons of every production poplar flower oral liquids are 17.5 ten thousand yuan by price per ton, 787.5 ten thousand yuan of new output values, and after 97.058 ten thousand yuan of the newly-increased manufacturing cost of deduction and development expenses, newly-increased net earnings is 690.442 ten thousand yuan; Its input-output ratio is 1:8.11, and scientific and technological investment yield is that 1:7.11 popularization investment yield is 1:40.28.Namely in this project, 10000 yuan of the every increases of social investment can increase by 81100 yuan of outputs, can repay 71100 yuan of net earnings; Promote 10000 yuan of the every increases of specific investment cost, can make society obtain 402800 yuan of net earnings.Therefore, implement this project and obtained very significant society and economic benefit.
The beneficial effect that adopts technique scheme to produce is:
1. the present invention utilizes the cellulase hydrolysis technology to extract flower of poplar, developed a kind of new technology of extracting flower of poplar, shorten the process time, reduced thermal energy consumption, reduced process water consumption, reach energy-saving and emission-reduction, increased work efficiency and the purpose of extraction ratio, had good society and economic benefit.
2. the extraction ratio of flavone is brought up to more than 0.18%, improved more than 40% than traditional handicraft.
Description of drawings
The present invention is further detailed explanation below in conjunction with the drawings and specific embodiments.
Fig. 1 is the canonical plotting of control substance of Rutin;
Fig. 2 is that the addition of enzyme affects schematic diagram to the flavone extraction ratio;
Fig. 3 is that hydrolysis temperature affects schematic diagram to the flavone extraction ratio;
Fig. 4 is that enzymolysis time affects schematic diagram to the flavone extraction ratio;
Fig. 5 is that the enzymolysis pH value affects schematic diagram to the flavone extraction ratio.
The specific embodiment
The cellulase that uses in following examples is provided by Shandong Kang Yuan bio tech ltd.
(1) the 100g flower of poplar is put into extraction pot, add flower of poplar quality 400g water, stir and be warming up to 45 ℃;
(2) 5g cellulase in extraction pot is regulated pH value to 5.5,45 ℃ of lower enzymolysis 95min;
(3) enzymolysis solution is filtered, collect filtrate, get poplar flower oral liquid 105mL.
Detect general flavone content in the present embodiment according to the assay method that detects general flavone content in poplar flower oral liquid, the extraction ratio of total flavones is as calculated: 0.188%.According to 2010 editions " Chinese veterinary pharmacopoeias " two poplar flower oral liquid quality standard checks, every assay is up to specification.
(1) the 100g flower of poplar is put into extraction pot, add flower of poplar quality 500g water, stir and be warming up to 45-50 ℃;
(2) add the 3g cellulase in extraction pot, regulate pH value to 5.1,46 ℃ of lower enzymolysis 110min;
(3) enzymolysis solution is filtered, collect filtrate, get poplar flower oral liquid 120mL.
Detect general flavone content in the present embodiment according to the assay method that detects general flavone content in poplar flower oral liquid, the extraction ratio of total flavones is as calculated: 0.181%.According to 2010 editions " Chinese veterinary pharmacopoeias " two poplar flower oral liquid quality standard checks, every assay is up to specification.
(1) the 100g flower of poplar is put into extraction pot, add flower of poplar quality 600g water, stir and be warming up to 45-50 ℃;
(2) add the 7g cellulase in extraction pot, regulate pH value to 5.4,48 ℃ of lower enzymolysis 100min;
(3) enzymolysis solution is filtered, collect filtrate, get poplar flower oral liquid 130mL.
Detect general flavone content in the present embodiment according to the assay method that detects general flavone content in poplar flower oral liquid, the extraction ratio of total flavones is as calculated: 0.189%.According to 2010 editions " Chinese veterinary pharmacopoeias " two poplar flower oral liquid quality standard checks, every assay is up to specification.
(1) the 100g flower of poplar is put into extraction pot, add flower of poplar quality 450g water, stir and be warming up to 45-50 ℃;
(2) add the 4g cellulase in extraction pot, regulate pH value to 5.2,50 ℃ of lower enzymolysis 120min;
(3) enzymolysis solution is filtered, collect filtrate, get poplar flower oral liquid 108mL.
Detect general flavone content in the present embodiment according to the assay method that detects general flavone content in poplar flower oral liquid, the extraction ratio of total flavones is as calculated: 0.186%.According to 2010 editions " Chinese veterinary pharmacopoeias " two poplar flower oral liquid quality standard checks, every assay is up to specification.
(1) the 100g flower of poplar is put into extraction pot, add flower of poplar quality 500g water, stir and be warming up to 45-50 ℃;
(2) add the 6g cellulase in extraction pot, regulate pH value to 5.0,46 ℃ of lower enzymolysis 90min;
(3) enzymolysis solution is filtered, collect filtrate, get poplar flower oral liquid 120mL.
Detect general flavone content in the present embodiment according to the assay method that detects general flavone content in poplar flower oral liquid, the extraction ratio of total flavones is as calculated: 0.188%.According to 2010 editions " Chinese veterinary pharmacopoeias " two poplar flower oral liquid quality standard checks, every assay is up to specification.
Claims (5)
1. the preparation method of a poplar flower oral liquid, is characterized in that comprising the steps:
(1) flower of poplar is put into extraction pot, add flower of poplar quality 4-6 water doubly, stir and be warming up to 45-50 ℃;
(2) add the cellulase of flower of poplar quality 3-7% in extraction pot, regulate pH value to 5.0-5.5,45-50 ℃ of lower enzymolysis 90-120min;
(3) enzymolysis solution is filtered, collect filtrate, namely get poplar flower oral liquid.
2. the preparation method of poplar flower oral liquid according to claim 1 is characterized in that adding in described step (1) water of 4 times of flower of poplar quality.
3. the preparation method of poplar flower oral liquid according to claim 1, is characterized in that regulating pH value with the lactic acid solution of 10-50% or the sodium hydroxide solution of 10-50% in described step (2).
4. the preparation method of poplar flower oral liquid according to claim 1, is characterized in that the quality of described step (2) cellulase is 4% of flower of poplar quality.
5. the preparation method of poplar flower oral liquid according to claim 1, is characterized in that in described step (2), enzymolysis time is 120min.
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| RU2669612C2 (en) * | 2014-04-18 | 2018-10-12 | Таркетт Гдл | Polyurethane coating, which is cured using the actinic radiation for decorative lining coatings |
| CN110433219A (en) * | 2019-08-09 | 2019-11-12 | 江苏禾宝药业有限公司 | A kind of compound essence of veterinary oral liquid proposes technique |
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