CN102525901B - Animal codonopsis pilosula polysaccharide oral solution and preparation method thereof - Google Patents
Animal codonopsis pilosula polysaccharide oral solution and preparation method thereof Download PDFInfo
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Abstract
The invention relates to animal codonopsis pilosula polysaccharide oral solution. A preparation method of the animal codonopsis pilosula polysaccharide oral solution comprises the steps of smashing codonopsis pilosula, degreasing and decolorizing, adding 8 times to 12 times of water, extracting for 1 hour to 3 hours 1 time to 3 times, performing enzymolysis with amylase, concentrating ethanol precipitation, adding papain of 18 kilograms to 48 kilograms for two hours, adding a Sevag agent with the volume of 18% to 30% volume of filter liquid to fully oscillate, centrifuging to obtain supernate, concentrating the supernate to 2/3 of crude drug and finally diluting, distributing and packing. The animal codonopsis pilosula polysaccharide oral solution is a novel drug form of traditional Chinese veterinary medicine with codonopsis pilosula polysaccharide as a raw material, uses reasonable and convenient extraction refining process to effectively remove impurities, greatly improves content of active ingredient, is not prone to sedimentation, is favorable of industrial batch production, has the advantages of being small in using dosage, high in biological utilization degree and convenient to take, and can enhance organism immunity, anti-cancer effect, anti-oxidation effect, anti-stress effect, antibiosis and antiviral effect of livestock.
Description
Technical field
The present invention relates to a kind of modern herbal medicine compound preparation, specifically related to Codonopsis pilosula polysaccharide oral liquid.
Background technology
Radix Codonopsis is Campanulaceae Radix Codonopsis, codonopsis pilosula var. modesta or the dry root of radix codonpsis tangshen, with tonifying middle-Jiao and Qi, the effect of strengthening spleen and tonifying lung, cures mainly spleen and lung weakness, shortness of breath palpitations, anorexia and loose stool, virtual asthma cough.It is distributed mainly on the ground such as northeast, North China and Shaanxi, Ningxia, Gansu, Qinghai, Henan, Sichuan, Yunnan, Tibet, and resource relative abundance fully excavates its practical value, with good Social benefit and economic benefit.In animal health-care product, it can be used for livestock and poultry weakness of the spleen and the stomach, and few food diarrhoea, deficiency syndrome of the lung cough and asthma, body is tired powerless, and the deficiency of vital energy is hung down de-.
The main component of Radix Codonopsis has polysaccharide, tangshenoside, sterol constituents, triterpenes components, alkaloid, lactone constituents, legumin class.Codonopsis pilosula polysaccharide is made up of important monose and monosaccharide derivatives, and wherein monose is mainly pentose and hexose, the xylose of such as pentose, arabinose, glucose, galactolipin, fructose of hexose etc.；Monosaccharide derivatives are mainly sugar alcohol such as mannitol, saccharic acid such as galacturonic acid etc..Modern pharmacological research shows that Codonopsis pilosula polysaccharide is one of active ingredient of Radix Codonopsis, with blood glucose is adjusted, and promotes hematopoietic function, and step-down, anti anoxia, endurance strengthens immunity of organisms, a variety of effects such as the contraction of regulation stomach and antiulcer.
Asiabell preparation develops still few in recent years, mainly based on the exploitation of compound preparation, such as pill, powder, there is that dose is big, the low problem of active ingredient availability.
Having by main ingredient preparation of Radix Codonopsis dissipated in shenling baizhu powder, reason at present, four gentleman dissipate etc..These powders exist based on spice and mix uneven, dosage is difficult the defects such as grasp, it is impossible to function well as therapeutic action.Such as：Strong livestock and poultry material feeding is more, and often material feeding is few for ill livestock and poultry, or due to age, kind, the high low factor influence collection capacity of temperature, causes dosage to be difficult grasp.But to colony's disease prevention and cure effect preferably, particularly in ill livestock and poultry, often material feeding is few for Codonopsis pilosula polysaccharide oral liquid（Not even material feeding）In the case of, can be with colony's drinking water administration, this enough medicine that can be ingested to guarantee ill domestic animal fowl is very good.
But in the prior art, traditional water extraction and alcohol precipitation method can not efficiently extracting polysaccharide composition, and a large amount of impurity are mixed with product, thus need to repeatedly be extracted, this not only influences production efficiency, increases production cost, and also product efficacy is had a greatly reduced quality.
The content of the invention
Have purity high it is an object of the invention to provide one kind, bioavilability is high, easy to use, and the rational Codonopsis pilosula polysaccharide oral liquid of production technology and preparation method thereof.
Much starch, water-solubility protein and oil-soluble impurities are mixed with Codonopsis pilosula polysaccharide product after conventional water extract-alcohol precipitation, these impurity not only influence the color and lusters of all kinds of Codonopsis pilosula polysaccharide preparations, character, stability, product purity is also reduced, so as to have impact on the drug effect and production efficiency of product.
To solve the above problems, the present invention is adopted the following technical scheme that：
A kind of Codonopsis pilosula polysaccharide oral liquid, is made up of following raw material proportioning：Radix Codonopsis 1800-2400g, Sodium Benzoate 1-5g.
Above-mentioned Codonopsis pilosula polysaccharide oral liquid, is made by following steps：
1st, pre-process：Precise codonopsis pilosula, cut-out is crushed；
2nd, petroleum ether takes off ester：By the codonopsis pilosula after processing under the conditions of 60-90 DEG C, petroleum ether refluxing extraction 1-2h is measured with 5-12 times, filters, takes filter residue；
3rd, alcohol reflux decolouring removal of impurities：Filter residue is measured into ethanol with 5-12 times and extracts 1-2h, the dregs of a decoction are obtained after filtering；
4th, water extraction：The dregs of a decoction add 8-12 times to measure decocting to boil 1-3 times, each 1-3 hours, collecting decoction, filtration；
5th, except starch：20-40g amylase water enzyme digestion 1-2h is added, react rear boiling water and has gone out enzyme 5min, room temperature is cooled to, 3000-5000r/min centrifuges 5min, takes supernatant；
6th, concentrate：Supernatant concentration to crude drug amount 2/3 (V/W)；
7th, alcohol precipitation：Concentrate adds 85%-95% ethanol, makes alcohol content up to 80%, leaching sediment is washed with ethanol；
8th, removing protein：18-48g papains are added in filtrate, digest 2h, the Sevag reagents added after the Sevag reagents or adjustment ratio of 15%-25% filtrate volumes fully vibrate 30min, stratification to water layer and organic layer intersection is precipitated without white protein, 12-20min is centrifuged with 3000-5000r/min rotating speed, supernatant is taken；
9th, dilution packing：It is dissolved in water and is diluted to nearly 1000ml, add Sodium Benzoate, adjustment total amount stirs evenly to 1000ml, dispenses, produce.
Wherein, Sevag reagents are by chloroform：Amylalcohol=5.5-4:1 proportional arrangement is formed.
Embodiment
With reference to specific embodiment, the present invention is described further, to help understanding present disclosure.
Embodiment 1：
A kind of Codonopsis pilosula polysaccharide oral liquid, is made up of following raw material proportioning：Radix Codonopsis 2000g, Sodium Benzoate 3g.
Above-mentioned Codonopsis pilosula polysaccharide oral liquid, is made by following steps：
1st, pre-process：Precise codonopsis pilosula 2000g, cut-out is crushed；
2nd, petroleum ether takes off ester：By the codonopsis pilosula after processing under the conditions of 80 DEG C, with 6 times of medicinal material weight petroleum ether refluxing extraction 1h, filtration takes filter residue；
3rd, alcohol reflux decolouring removal of impurities：Filter residue is extracted into 1h with 10 times of filter residue w ethanols, the dregs of a decoction are obtained after filtering；
4th, water extraction：The dregs of a decoction add 8 times of dregs of a decoction weight waters to decoct 2 times, first time 2h, second of 1.5h, collecting decoction, filtration；
5th, except starch：24g amylase water enzyme digestion 1h is added, react rear boiling water and has gone out enzyme 5min, room temperature is cooled to, 3500r/min centrifuges 5min, takes supernatant；
6th, concentrate：Supernatant concentration to crude drug amount 2/3 (V/W)；
7th, alcohol precipitation：Concentrate adds 95% ethanol, makes alcohol content up to 80%, leaching sediment is washed with ethanol；
8th, removing protein：40g papains are added in filtrate, 2h is digested, the Sevag reagents for adding 16% filtrate volume amount fully vibrate 30min, and stratification to water layer is precipitated with organic layer intersection without white protein, is centrifuged 15min with 4000r/min rotating speed, is taken supernatant；
9th, dilution packing：It is dissolved in water and is diluted to nearly 1000ml, add 3g Sodium Benzoates, adjustment total amount stirs evenly to 1000ml, dispenses, produce.
Wherein, Sevag reagents are by chloroform：Amylalcohol=5:1 proportional arrangement is formed.
Embodiment 2
A kind of Codonopsis pilosula polysaccharide oral liquid, is made up of following raw material proportioning：Radix Codonopsis 2200g, Sodium Benzoate 3g.
Above-mentioned Codonopsis pilosula polysaccharide oral liquid, is made by following steps：
1st, pre-process：Precise codonopsis pilosula 2200g, cut-out is crushed；
2nd, petroleum ether takes off ester：By the codonopsis pilosula after processing under the conditions of 85 DEG C, with 8 times of medicinal material weight petroleum ether refluxing extraction 1.5h, filtration takes filter residue；
3rd, alcohol reflux decolouring removal of impurities：Filter residue is extracted into 2h with 10 times of filter residue w ethanols, the dregs of a decoction are obtained after filtering；
4th, water extraction：The dregs of a decoction add 8 times of dregs of a decoction weight waters to decoct 2 times, first time 3h, second of 1.5h, collecting decoction, filtration；
5th, except starch：28g amylase water enzyme digestion 1.5h is added, react rear boiling water and has gone out enzyme 5min, room temperature is cooled to, 4000r/min centrifuges 5min, takes supernatant；
6th, concentrate：Supernatant concentration to crude drug amount 2/3 (V/W)；
7th, alcohol precipitation：Concentrate adds 95% ethanol, makes alcohol content up to 80%, leaching sediment is washed with ethanol；
8th, removing protein：30g papain is added in filtrate, 2h is digested, the Sevag reagents for adding 20% filtrate volume amount fully vibrate 30min, and stratification to water layer is precipitated with organic layer intersection without white protein, is centrifuged 15min with 5000r/min rotating speed, is taken supernatant；
9th, dilution packing：It is dissolved in water and is diluted to nearly 1000ml, add 3g Sodium Benzoates, adjustment total amount stirs evenly to 1000ml, dispenses, produce.
Wherein, Sevag reagents are by chloroform：Amylalcohol=4:1 proportional arrangement is formed.
Embodiment 3：
A kind of Codonopsis pilosula polysaccharide oral liquid, is made up of following raw material proportioning：Radix Codonopsis 1800g, Sodium Benzoate 3g.
Above-mentioned Codonopsis pilosula polysaccharide oral liquid, is made by following steps：
1st, pre-process：Precise codonopsis pilosula 1800g, cut-out is crushed；
2nd, water extraction：Plus 10 times of medicinal material weight waters are decocted 2 times, first time 3h, second of 2h, collecting decoction, filtration；
3rd, except starch：20g amylase water enzyme digestion 1h is added, react rear boiling water and has gone out enzyme 5min, room temperature is cooled to, 3000r/min centrifuges 5min, takes supernatant；
4th, concentrate：Supernatant concentration to crude drug amount 2/3 (V/W)；
5th, alcohol precipitation：Concentrate adds 95% ethanol, makes alcohol content up to 80%, leaching sediment is washed with ethanol；
6th, removing protein：22g papain is added in filtrate, 2h is digested, the Sevag reagents for adding 15% filtrate volume amount fully vibrate 30min, and stratification to water layer is precipitated with organic layer intersection without white protein, is centrifuged 15min with 3500r/min rotating speed, is taken supernatant；
7th, dilution packing：It is dissolved in water and is diluted to nearly 1000ml, add 3g Sodium Benzoates, adjustment total amount stirs evenly to 1000ml, dispenses, produce.
Wherein, Sevag reagents are by chloroform：Amylalcohol=4.5:1 proportional arrangement is formed.
Embodiment 4
A kind of Codonopsis pilosula polysaccharide oral liquid, is made up of following raw material proportioning：Radix Codonopsis 2000g, Sodium Benzoate 3g.
Above-mentioned Codonopsis pilosula polysaccharide oral liquid, is made by following steps：
1st, pre-process：Precise codonopsis pilosula 2000g, cut-out is crushed；
2nd, petroleum ether takes off ester：By the codonopsis pilosula after processing under the conditions of 80 DEG C, with 6 times of medicinal material weight petroleum ether refluxing extraction 1.5h, filtration takes filter residue；
3rd, alcohol reflux decolouring removal of impurities：Filter residue is extracted into 1.5h with 10 times of filter residue w ethanols, the dregs of a decoction are obtained after filtering；
4th, water extraction：The dregs of a decoction add 8 times of dregs of a decoction weight waters to decoct 2 times, first time 2h, second of 1.5h, collecting decoction, filtration；
5th, except starch：30g amylase water enzyme digestion 2h is added, react rear boiling water and has gone out enzyme 5min, room temperature is cooled to, 3000r/min centrifuges 5min, takes supernatant；
6th, alcohol precipitation：Concentrate adds 95% ethanol, makes alcohol content up to 80%, leaching sediment is washed with ethanol；
7th, concentrate：Supernatant concentration to crude drug amount 2/3 (V/W)；
8th, dilution packing：It is dissolved in water and is diluted to nearly 1000ml, add 3g Sodium Benzoates, adjustment total amount stirs evenly to 1000ml, dispenses, produce.
Embodiment 5
A kind of Codonopsis pilosula polysaccharide oral liquid, is made up of following raw material proportioning：Radix Codonopsis 2000g, Sodium Benzoate 3g.
Above-mentioned Codonopsis pilosula polysaccharide oral liquid, is made by following steps：
1st, pre-process：Precise codonopsis pilosula 2000g, cut-out is crushed；
2nd, petroleum ether takes off ester：By the codonopsis pilosula after processing under the conditions of 80 DEG C, with 6 times of medicinal material weight petroleum ether refluxing extraction 1.5h, filtration takes filter residue；
3rd, alcohol reflux decolouring removal of impurities：Filter residue is extracted into 1.5h with 10 times of filter residue w ethanols, the dregs of a decoction are obtained after filtering；
4th, water extraction：The dregs of a decoction add 8 times of dregs of a decoction weight waters to decoct 2 times, first time 2h, second of 1.5h, collecting decoction, filtration；
5th, concentrate：Supernatant concentration to crude drug amount 2/3 (V/W)；
6th, alcohol precipitation：Concentrate adds 95% ethanol, makes alcohol content up to 80%, leaching sediment is washed with ethanol；
7th, removing protein：20g papain is added in filtrate, 2h is digested, the Sevag reagents for adding 20% filtrate volume amount fully vibrate 30min, and stratification to water layer is precipitated with organic layer intersection without white protein, is centrifuged 15min with 4000r/min rotating speed, is taken supernatant；
8th, dilution packing：It is dissolved in water and is diluted to nearly 1000ml, add 3g Sodium Benzoates, adjustment total amount stirs evenly to 1000ml, dispenses, produce.
Wherein, Sevag reagents are by chloroform：Amylalcohol=5.5:1 proportional arrangement is formed.
Comparative example
A kind of Codonopsis pilosula polysaccharide oral liquid, is made up of following raw material proportioning：Radix Codonopsis 2000g, Sodium Benzoate 3g.
Above-mentioned Codonopsis pilosula polysaccharide oral liquid, is made by following steps：
1st, pre-process：Precise codonopsis pilosula 2000g, cut-out is crushed；
2nd, water extraction：The dregs of a decoction add 8 times of dregs of a decoction weight waters to decoct 2 times, first time 2h, second of 1.5h, collecting decoction, filtration；
3rd, concentrate：Supernatant concentration to crude drug amount 2/3 (V/W)；
4th, alcohol precipitation：Concentrate adds 95% ethanol, makes alcohol content up to 80%, leaching sediment is washed with ethanol；
5th, dilution packing：It is dissolved in water and is diluted to nearly 1000ml, add 3g Sodium Benzoates, adjustment total amount stirs evenly to 1000ml, dispenses, produce.
Test example
The Codonopsis pilosula polysaccharide COMPARISON OF CHARACTERS of test example 1
Codonopsis pilosula polysaccharide made from choosing embodiment 1-5 and comparative example, puts and dried in vacuum drying chamber to constant mass, observe Codonopsis pilosula polysaccharide character, the results are shown in Table 1：
The Codonopsis pilosula polysaccharide COMPARISON OF CHARACTERS of table 1
| Color | Character | |
| Embodiment 1 | Canescence | Pulverulent solids |
| Embodiment 2 | Canescence | Pulverulent solids |
| Embodiment 3 | Brown color | Chip solid |
| Embodiment 4 | Canescence | Pulverulent solids |
| Embodiment 5 | Canescence | Pulverulent solids |
| Comparative example | Brown color | Chip solid |
As seen from the above table, ester is taken off without petroleum ether, the Radix Codonopsis sample of ethanol decolorization removing impurities, color is deeper, is brown color, wherein containing more impurity, and character is chip solid.
The albumen clearance of test example 2 compares
Determine after embodiment 1-5 and comparative example alcohol precipitation protein content a in supernatant, and protein content b during last dilution respectively using Coomassie Brilliant Blue, with formula albumen clearance=（Protein content after before processing protein content-processing）/ before processing protein content % is calculated, and the results are shown in Table 2：
Table 2 takes the quantity of fluid to be measured of equal volume, and albumen clearance is compared as follows：
| a | b | Albumen clearance（%） | |
| Embodiment 1 | 25.5 milligrams | 1.3 milligrams | 94.9 |
| Embodiment 2 | 27.1 milligrams | 2.2 milligrams | 91.9 |
| Embodiment 3 | 23.8 milligrams | 1.7 milligrams | 92.9 |
| Embodiment 4 | 24.9 milligrams | 18.5 milligrams | 25.7 |
| Embodiment 5 | 26.2 milligrams | 2.4 milligrams | 90.8 |
| Comparative example | 25.3 milligrams | 20.8 milligrams | 17.8 |
Compare through albumen clearance, it is seen that first through papain enzymolysis protein, then through Sevag agent treatments, can effectively remove the protein in Codonopsis pilosula polysaccharide, it is ensured that the purity of product.And adjusted Sevag ratio of reagents is 5:When 1, removing protein rate is optimal.
The accelerated test of test example 3
By Codonopsis pilosula polysaccharide oral liquid made from embodiment 1-5 and comparative example, be respectively placed in 40 DEG C of temperature, the environment of relative humidity 75% preserve 6 months, 3 months during testing, 6 the end of month it is separately sampled once, its precipitation capacity and clarity are determined respectively, 3 are the results are shown in Table：
Table 3 takes the quantity of fluid to be measured of equal volume, and precipitation capacity and clarity are compared as follows：
Accelerated test process, not plus except the embodiment that starch is handled starts slightly to precipitate for three month, shows as muddiness on the 6th month, does not possess stability, and comparative example began to muddiness occurred at three month.And embodiment 1-4 is stable in properties, clarify without precipitation.
The polyoses content of test example 4
Precision weighs obtained Codonopsis pilosula polysaccharide precipitation 3.0g before embodiment 1-5 and comparative example dilution, is dissolved in water and is transferred in 100ml volumetric flask, add water to scale, shakes up standby.Precision weighs the dry DEXTROSE ANHYDROUS reference substance to constant weight in right amount, adds water and solution of every 1ml containing 1mg is made, be used as reference substance.Sample 0.2ml is measured respectively, put in 10ml tool scale test tubes, according to reference substance solution preparation method, the phenol solution 1ml for plus 5%, then add the 5ml concentrated sulfuric acid, shake up, heating 15min taking-ups in 60 DEG C of water-bath are put to let cool, corresponding blank is the mensuration absorbance value at 490nm, then the quality containing DEXTROSE ANHYDROUS from reading sample solution on standard curve, and calculating is produced.It the results are shown in Table 4：
The polyoses content of table 4 compares
| Polysaccharide quality（g） | Content（%） | |
| Embodiment 1 | 2.87 | 95.67 |
| Embodiment 2 | 2.85 | 95.00 |
| Embodiment 3 | 2.41 | 80.33 |
| Embodiment 4 | 2.33 | 77.67 |
| Embodiment 5 | 2.64 | 88.00 |
| Comparative example | 1.96 | 65.33 |
As seen from the above table, comparative example polyoses content is substantially more relatively low than polyoses content in embodiment, and the polyoses content of embodiment 1 is up to 95.67%, it is seen that preparation method of the present invention can effectively reject impurity, extracts polysaccharide component in Radix Codonopsis.
Experimental example
The long term toxicity of experimental example 1 is studied
By rat 80, random point four groups, every group 20.Choose Codonopsis pilosula polysaccharide oral liquid made from embodiment 1, Codonopsis pilosula polysaccharide oral liquid decoction that heavy dose group concentration is 18g crude drugs/ml, middle dosage are that 12g crude drugs/ml, low dose are 6g crude drugs/ml, control group gives isometric(al) physiological saline, carries out hematology, blood biochemical analysis and main organs pathological examination to experimental rat respectively after two months.
As a result：Each group rat ordinary circumstance and the change without exception such as behavioral activity, excrement.Each administration group animal hematology, blood biochemical analysis and main organs pathological examination, have no significant change.As a result show, Codonopsis pilosula polysaccharide oral liquid safety non-toxic.
The influence of the Codonopsis pilosula polysaccharide Oral Liquid On Mice swimming time of experimental example 2
Method：Mouse 160, random point of 8 groups, every group 20, i.e. blank group, positive drug control group（Ginseng oral liquid）, Codonopsis pilosula polysaccharide oral liquid group made from embodiment 1（Including basic, normal, high three groups of dosage）, Codonopsis pilosula polysaccharide oral liquid group made from embodiment 4（Including basic, normal, high three groups of dosage）, each administration group continuous gavage is administered 5 days, 1 time a day, each 0.2ml/10g, the physiological saline of the capacity such as blank group gavage.Born a heavy burden in mouse tail, observation mouse, which is swum to power, to exhaust, and calculates the time.It the results are shown in Table 5：
The Codonopsis pilosula polysaccharide Oral Liquid On Mice swimming time of table 5 influence (
)
Compared with blank group：*P<0.05,**P<0.01
As seen from the above table, Codonopsis pilosula polysaccharide oral liquid can significantly extend mouse swimming time, and increase with dosage, and mouse swimming time increases.And Codonopsis pilosula polysaccharide oral liquid effect made from embodiment 1 is than embodiment 4 more preferably.
Influence of the polyoses oral liquid of experimental example 3 to mouse normobaric hypoxia
Method：Mouse 96, random point of 8 groups, every group 12, i.e. blank group, positive drug control group（Ginseng oral liquid）, Codonopsis pilosula polysaccharide oral liquid group made from embodiment 1（Including basic, normal, high three groups of dosage）, Codonopsis pilosula polysaccharide oral liquid group made from embodiment 4（Including basic, normal, high three groups of dosage）, each administration group continuous gavage is administered 5 days, 1 time a day, each 0.2ml/10g, the physiological saline of the capacity such as blank group gavage.Mouse is put into soda lime closed glass jar, observes and records the anoxia in mice death time.It the results are shown in Table 6：
The Radix Codonopsis the water extracted immersing paste stomach of table 6 to mouse normobaric hypoxia influence (
)
Compared with blank group：*P<0.05,**P<0.01
Shown by the above results, Codonopsis pilosula polysaccharide oral liquid can significantly extend mouse hypoxia endurance time, and increase with dosage, the extension of mouse hypoxia endurance time.The effect of the Codonopsis pilosula polysaccharide oral liquid of embodiment 4 is compared, the experimental mice hypoxia endurance time for eating Codonopsis pilosula polysaccharide oral liquid made from embodiment 1 is considerably longer.
The influence of ability is cleaned up in phagocytosis of the Codonopsis pilosula polysaccharide Oral Liquid On Mice reticuloendothelial system of experimental example 4 to inertia carbon granule in blood flow
Method：Male mice 80 is taken, 8 groups, every group 10, i.e. blank group, positive drug control group are randomly divided into（Ginseng oral liquid）, Codonopsis pilosula polysaccharide oral liquid group made from embodiment 1（Including basic, normal, high three groups of dosage）, Codonopsis pilosula polysaccharide oral liquid group made from embodiment 4（Including basic, normal, high three groups of dosage）, each administration group gastric infusion, blank group gives the physiological saline of respective volume amount, and one time a day, 0.2ml/10g, continuous 4 days.Extraction mouse thymus and spleen are simultaneously weighed.Thymus gland or spleen index, phagocytic index K and phagocytosis factor alpha is calculated as follows.It the results are shown in Table 7-8.
C is carbon granule concentration in blood in formula, and T is the time, and W is body weight, and WLS is that liver and spleen closes weight.
The Codonopsis pilosula polysaccharide Oral Liquid On Mice immune organ weight of table 7 influence (
)
Compared with blank group：* P<0.05, * * P<0.01
The influence of the Codonopsis pilosula polysaccharide Oral Liquid On Mice reticuloendothelial system phagocytic function of table 8
Compared with blank group：* P<0.05, * * P<0.01
Shown by the above results, Codonopsis pilosula polysaccharide oral liquid administration group thymus index, spleen index, K values and α values substantially increase compared with blank group, it is seen that it can strengthen mouse reticuloendothelial system phagocytic function.Meanwhile, the effect of embodiment 1 is obvious compared with the drug effect of embodiment 4.
The influence that the Codonopsis pilosula polysaccharide oral liquid of experimental example 5 is generated to sheep red blood cell (SRBC) induced mice hemolytic antibody
Immune and administration：Mouse 80 is chosen, 8 groups, male and female half and half, every group 10, i.e. blank group, positive drug control group are randomly divided into（Ginseng oral liquid）, Codonopsis pilosula polysaccharide oral liquid group made from embodiment 1（Including basic, normal, high three groups of dosage）, Codonopsis pilosula polysaccharide oral liquid group made from embodiment 4（Including basic, normal, high three groups of dosage）, each administration group gastric infusion, blank group gives the physiological saline of respective volume amount, and one time a day, 0.2ml/10g, continuous 8 days.Each administration group is in 2 days pneumoretroperitoneum injection sheep erythrocyte suspension 0.2ml/ mouse of administration, and control group is immune with SRBC with method, and 6 days after immune, eye socket takes blood to carry out hemolysin test.It the results are shown in Table 9：
The hemolysin test method of table 9
| Group | Sample cell | Control tube |
| 500 times of dilute serums | 1ml | / |
| Physiological saline | / | 1ml |
| Sheep red blood cell (SRBC) dilution | 0.5ml | 0.5ml |
| 1：10 guinea pig serum dilutions | 1ml | 1ml |
Calculate：The half hemolytic value HC of every mouse samples is calculated by following equation50
The Codonopsis pilosula polysaccharide Oral Liquid On Mice hemolysin level of table 10 influence ( n=10）
Compared with blank group：*P<0.05,**P<0.01
Shown by the above results, the HC of each dosage group mouse50Control group is above, and amount increases and raised immediately.I.e. Codonopsis pilosula polysaccharide oral liquid has effects that to strengthen mouse humoral immune function.And the drug effect of embodiment 1 is better than embodiment 4.
Claims (2)
1. a kind of Codonopsis pilosula polysaccharide veterinary oral liquid, it is characterised in that be made up of following raw material proportioning：Radix Codonopsis 1800-2400g, Sodium Benzoate 1-5g, are made by following steps：
A, pretreatment：Precise codonopsis pilosula, cut-out is crushed；
B, petroleum ether take off ester：By the codonopsis pilosula after processing under the conditions of 60-90 DEG C, petroleum ether refluxing extraction 1-2h is measured with 5-12 times, filters, takes filter residue；
C, alcohol reflux decolouring removal of impurities：Filter residue is measured into ethanol with 5-12 times and extracts 1-2h, the dregs of a decoction are obtained after filtering；
D, water extraction：The dregs of a decoction add 8-12 times to measure decocting to boil 1-3 times, each 1-3 hours, collecting decoction, filtration；
E, except starch：20-40g amylase water enzyme digestion 1-2h is added, react rear boiling water and has gone out enzyme 5min, room temperature is cooled to, 3000-5000r/min centrifuges 5min, takes supernatant；
F, concentration：Supernatant volume is concentrated into the 2/3 of crude drug quality；
G, alcohol precipitation：Concentrate adds 85%-95% ethanol, makes alcohol content up to 80%, leaching sediment is washed with ethanol；
H, removing protein：18-48g papains are added in filtrate, 2h is digested, the Sevag reagents for adding 15%-25% filtrate volumes fully vibrate 30min, and stratification to water layer is precipitated with organic layer intersection without white protein, 12-20min is centrifuged with 3000-5000r/min rotating speed, supernatant is taken；
I, dilution packing：It is dissolved in water and is diluted to nearly 1000ml, add Sodium Benzoate, adjustment total amount stirs evenly to 1000ml, dispenses, produce；
Wherein, Sevag reagents are by chloroform：Amylalcohol=5.5-4:1 proportional arrangement is formed.
2. Codonopsis pilosula polysaccharide veterinary oral liquid according to claim 1, it is characterised in that be made by following steps：
A, pretreatment：Precise codonopsis pilosula 2000g, cut-out is crushed；
B, petroleum ether take off ester：By the codonopsis pilosula after processing under the conditions of 80 DEG C, with 6 times of medicinal material weight petroleum ether refluxing extraction 1h, filtration takes filter residue；
C, alcohol reflux decolouring removal of impurities：Filter residue is extracted into 1h with 10 times of filter residue w ethanols, the dregs of a decoction are obtained after filtering；
D, water extraction：The dregs of a decoction add 8 times of dregs of a decoction weight waters to decoct 2 times, first time 2h, second of 1.5h, collecting decoction, filtration；
E, except starch：24g amylase water enzyme digestion 1h is added, react rear boiling water and has gone out enzyme 5min, room temperature is cooled to, 3500r/min centrifuges 5min, takes supernatant；
F, concentration：Supernatant volume is concentrated into the 2/3 of crude drug quality；
G, alcohol precipitation：Concentrate adds 95% ethanol, makes alcohol content up to 80%, leaching sediment is washed with ethanol；
H, removing protein：40g papains are added in filtrate, 2h is digested, the Sevag reagents for adding 16% filtrate volume amount fully vibrate 30min, and stratification to water layer is precipitated with organic layer intersection without white protein, is centrifuged 15min with 4000r/min rotating speed, is taken supernatant；
I, dilution packing：It is dissolved in water and is diluted to nearly 1000ml, add 3g Sodium Benzoates, adjustment total amount stirs evenly to 1000ml, dispenses, produce；
Wherein, Sevag reagents are by chloroform：Amylalcohol=5:1 proportional arrangement is formed.
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| JP7460998B2 (en) * | 2017-05-18 | 2024-04-03 | 共栄化学工業株式会社 | Skin preparations |
| CN108606990A (en) * | 2018-06-01 | 2018-10-02 | 山西正来制药有限公司 | A kind of preparation method of RADIX CODONOPSIS PILOSULAE from Shanxi of China's oral solution |
| CN109021135A (en) * | 2018-07-27 | 2018-12-18 | 四川农业大学 | A kind of codonopsis pilosula var. modesta polysaccharide formulation and preparation method thereof |
| CN109350597A (en) * | 2018-09-28 | 2019-02-19 | 四川农业大学 | Codonopsis pilosula var. modesta polysaccharide/synanthrin oral liquid formulations and preparation method |
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