CN104292355B - A kind of extracting method of platycodon root polysaccharide - Google Patents
A kind of extracting method of platycodon root polysaccharide Download PDFInfo
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Abstract
The invention belongs to technical field of polysaccharide extraction, be specifically related to the extracting method of a kind of platycodon root polysaccharide.A kind of extracting method of platycodon root polysaccharide, step including following: select the Radix Platycodonis dried, 40 80 mesh sieves are crossed after pulverizing, obtain balloonflower powder, adding water in balloonflower powder and stir, balloonflower powder is 1:5 10 with the part by weight of water, soaks 8 16 hours in water, add cellulase degradation, add ficin and bromelain, enzymolysis under microwave condition, concentrate after enzymolysis and decolour, centrifugal, take supernatant concentration precipitate with ethanol, then be centrifuged, use ethanol purge precipitation, after lyophilization and pulverize, obtain platycodon root polysaccharide.The method using the present invention extracts polysaccharide from Radix Platycodonis, uses enzyme action condition gentle, uses various different enzyme by the fat in Radix Platycodonis, cellulase hydrolysis, by Polyose extraction therein out, improve the utilization rate of Radix Platycodonis, adopt and be obtained by the present invention polysaccharide yield height, and purity is the highest.
Description
Technical field
The invention belongs to technical field of polysaccharide extraction, be specifically related to the extracting method of a kind of platycodon root polysaccharide.
Background technology
Polysaccharide is the sugar chain combined by glycosidic bond, is to be formed by condensation, dehydration by more than at least 10 monosaccharide, is connected with glycosidic bond between structural units, is molecule huge structure and the glucide of complexity.Polysaccharide is extensive in distributed in nature, is present in animal cell membrane, plant cell wall, microorganism wall more, is one of four big base substances constituting vital movement.In recent years, along with the development of Celluar and Molecular Biology, scholars find that it has the effects such as antitumor, enhancing human body immunity, anti-inflammatory, anticoagulation, antiviral, defying age, blood sugar lowering.Since the sixties in 20th century, scholars' discovery polysaccharide successively has a lot of pharmacologically active, up to now, people have isolated more than 300 kind of polysaccharide compound from naturally-produced, wherein mostly important with the polysaccharide that extracts in plant, because vegetable polysaccharides has, function is many, toxic and side effects is little, the feature of high safety, causes the concern of numerous scholars, makes it the study hotspot of medicine and health care of food industry.
Platycodon root polysaccharide is a kind of inulin type polysaccharide, is the important indicator evaluating Radix Platycodonis edible quality.Modern pharmacology research finds that platycodon root polysaccharide has a variety of biological activity, as immunomodulating, expectorant, antiinflammatory, protect the liver, antitumor, antioxidation, defying age, enhancing human body immunity function and improve the effect such as insulin, therefore, for the research of platycodon root polysaccharide, will have the highest medical value.
The purity of Radix Platycodonis directly affects the physiological function of polysaccharide, and the extracting method having patent application disclosure platycodon root polysaccharide is as follows:
CN101830999 discloses the extracting method of a kind of platycodon root polysaccharide, its step is as follows: one, choose dry Radix Platycodonis rhizome, cross 60 mesh sieves after pulverizing, be separately added into Platycodon Root and aqueous solution with the ratio of certain solid-liquid ratio 1g:20mL-1 g:20mL, 0 soak 1 hour after to extract under certain microwave power after, 40-70 DEG C of water-bath, obtain the extract rich in platycodon root polysaccharide, carry out solid-liquid separation with horizontal spiral shell formula centrifuge with the speed of 1500-2000r/min, isolate extracting solution and residue；Two, with sevag method (chloroform: n-butyl alcohol=4:1) deproteinization repeatedly, bag filter is dialysed, 45-55 DEG C of vacuum rotary evaporator concentrated extracting solution；Three, in extracting solution, add the ethanol solution of times volume after measuring solution absorbance value with phend-sulphuric acid；Four, alcohol hypostasis is separated, obtains platycodon root polysaccharide powder with the vacuum lyophilization under the conditions of 0.4-0.7mbar, temperature-40～-50 DEG C of vacuum freeze-drying machine.Above-mentioned method extracts platycodon root polysaccharide, and its extraction ratio can reach 20-30%, extracts than common water-bath and has exceeded 15-20%, substantially increases the economic worth of the polysaccharide of Radix Platycodonis.
Summary of the invention
In order to solve above-mentioned technical problem, the invention provides the extracting method of the higher platycodon root polysaccharide of a kind of yield.
The extracting method of platycodon root polysaccharide of the present invention is to be realized by following technical scheme:
A kind of extracting method of platycodon root polysaccharide, the step including following:
Select the Radix Platycodonis dried, cross 40-80 mesh sieve after pulverizing, obtain balloonflower powder, add water in balloonflower powder and stir, balloonflower powder is 1:5-10 with the part by weight of water, soaking 8-16 hour in water, add cellulase, the addition of cellulase is 0.5-0.9%, hydrolysis temperature 45 DEG C, pH value 5, above-mentioned raw material is placed in Microwave Extraction Equipment extraction, the power of microwave is 400-600W, and frequency is 800MHZ, enzymolysis 0.1-0.5h；
Add ficin and bromelain, the addition of ficin is 0.2-0.6%, the addition of bromelain is 0.2-0.5%, hydrolysis temperature 45 DEG C, pH value 5.5-6.5, be placed in Microwave Extraction Equipment extraction by above-mentioned raw material, and the power of microwave is 400-600W, frequency is 800MHZ, enzymolysis 0.1-0.5h, after enzymolysis at 100-105 DEG C enzyme denaturing 5-10min, obtain enzymolysis solution；
Above-mentioned enzymolysis solution is placed in vacuum concentrator concentration, and decompression cryoconcentration, to enzymolysis solution to the 1/3 of original volume, obtains Radix Platycodonis crude polysaccharides concentrated solution；
Adding activated carbon in the concentrated solution of Radix Platycodonis crude polysaccharides, described activated carbon accounts for the 2-6% of Radix Platycodonis crude polysaccharides concentrated solution weight, stirs 0.2-0.5h, leaches activated carbon；
Centrifugal 10-20 min, centrifugal rotational speed is 4500 rpm, is collected respectively with precipitation by supernatant；
Take supernatant liquid filtering to remove slag, the filtrate obtained after filtration concentrates at 60-70 DEG C, being concentrated into its concentration is 60-70%, concentrated solution adds 95% ethanol precipitation in 1:4 ratio and stands 10-20 hour, is centrifuged 10-20 min, centrifugal rotational speed is 4500 rpm, collects precipitation and repeatedly cleans 2-3 time with ethanol, after lyophilization to moisture is 4-6%, pulverize at-1-4 DEG C again, obtain soluble polysaccharide；
Above-mentioned enzyme concentration is enzyme and accounts for the percentage by weight of the Radix Platycodonis after micronizing.
Preferably, Radix Platycodonis is 1:9 with the part by weight of water；
Preferably, the consumption of ficin is 0.45%；
The addition of bromelain is 0.4%.
Preferably, the extracting method of a kind of platycodon root polysaccharide, the method includes following step:
Select the Radix Platycodonis dried, cross 60 mesh sieves after pulverizing, obtain balloonflower powder, add water in balloonflower powder and stir, balloonflower powder is 1:7 with the part by weight of water, soaking 10 hours in water, add cellulase, the addition of cellulase is 0.8%, hydrolysis temperature 45 DEG C, pH value 5, above-mentioned raw material is placed in Microwave Extraction Equipment extraction, the power of microwave is 400-600W, and frequency is 800MHZ, enzymolysis 0.1-0.5h；
Add ficin and bromelain, the addition of ficin is 0.45%, the addition of bromelain is 0.4%, hydrolysis temperature 45 DEG C, pH value 6.0, be placed in Microwave Extraction Equipment extraction by above-mentioned raw material, and the power of microwave is 500W, frequency is 800MHZ, enzymolysis 0.3h, after enzymolysis at 100 DEG C enzyme denaturing 6min, obtain enzymolysis solution；
Above-mentioned enzymolysis solution is placed in vacuum concentrator concentration, and decompression cryoconcentration, to enzymolysis solution to the 1/3 of original volume, obtains Radix Platycodonis crude polysaccharides concentrated solution；
Adding activated carbon in the concentrated solution of Radix Platycodonis crude polysaccharides, described activated carbon accounts for the 4% of Radix Platycodonis crude polysaccharides concentrated solution weight, stirs 0.3h, leaches activated carbon；
Centrifugal 15 min, centrifugal rotational speed is 4500 rpm, is collected respectively with precipitation by supernatant；
Take supernatant liquid filtering to remove slag, the filtrate obtained after filtration concentrates at 65 DEG C, being concentrated into its concentration is 65%, concentrated solution adds 95% ethanol precipitation in 1:4 ratio and stands 10-20 hour, is centrifuged 15 min, centrifugal rotational speed is 4500 rpm, collects precipitation and repeatedly cleans 2-3 time with ethanol, after lyophilization to moisture is 5%, pulverize at-1-4 DEG C again, obtain soluble polysaccharide；
Above-mentioned enzyme concentration is enzyme and accounts for the percentage by weight of the Radix Platycodonis after micronizing.
In the method for the present invention, freezing and pulverizing principle freezing and pulverizing is " black brittleness " utilizing material under low temperature state, and i.e. material is along with the reduction of temperature, its hardness and fragility increase, and plasticity and toughness reduce, at a certain temperature, just can be pulverized by a power the least.The material of chilled pulverizing, its granularity can reach the degree of " superfine ", therefore can produce " superfine food ".
" black brittleness " of material is closely-related with the phenomenon of a kind of referred to as glass transition.So-called glass transition refers to that amorphous polymer there will be the change of mechanical property when variations in temperature, forms rubbery state and two kinds of physical states of glassy state originally；And temperature changing process can produce by rubbery state to the transformation of glassy state.When rubbery state, the toughness of material is big, and deformability is strong；And when glassy state, material hardness and fragility are big, deformability is the least.Glass transition phenomenon non-polymer institute are peculiar in fact, and food and agricultural product there will be Glass Transition equally.But, because the composition complicated structure of food and agricultural product, so its glass transition is more complex, as there may be multistage Glass Transition and devitrification transition phenomenon.Usually, our material by rubbery state to glassy transition time required temperature be called glass transition temperature.According to above-mentioned rubbery state and the character of glassy state, it is believed that the glass transition temperature of material correspond to " brittle temperature " of material.
Therefore, the freezing and pulverizing principle of food and agricultural product is exactly: first makes low-temperature material be chilled to below glass transition temperature or brittle temperature, then is pulverized with pulverizer.During food and agricultural product fast cooling, the uneven contraction in internal each position can be caused to produce internal stress, under the effect of this stress, the internal weak part of material produces micro-crack and also causes the adhesion of interior tissue to reduce.Just make underbead crack expand rapidly at outside low-force and crush.
Freezing crusher system is during crushing material, its low-temperature receiver forms a closed circuit circulatory system, makes the energy be fully used, and saves energy consumption: the sink temperature pulverized can be down to negative 196 degree, brittle point temperature according to material, its temperature adjustable in crushing process, selects most preferably to pulverize temperature, reduces energy consumption: smashing fineness can reach 10-700 mesh, the fineness such as even up to micron μ: use liquid nitrogen as abrasive media, realize pulverizing at ultralow temperature, material explosion-proof, anti-oxidation wait resultant effect.
Freezing and pulverizing is applicable to polysaccharide, because polysaccharide easily produces bonding, blocking and the problem such as qualitative change when ambient ground, and effect and inefficient.
Therefore, using the polysaccharide that extraction is obtained by the method for freezing and pulverizing in the present invention, the quality maintaining polysaccharide is unaffected.
The beneficial effects of the present invention is, use enzyme action condition gentle, use various different enzyme by the fat in Radix Platycodonis, cellulase hydrolysis, from Radix Platycodonis, extract polysaccharide simultaneously, utilizing Radix Platycodonis to greatest extent, and adopt the polysaccharide being obtained by the present invention, its yield and purity are high.
Detailed description of the invention
Below in conjunction with specific embodiment, the present invention is further described, in order to those skilled in the art knows more about the present invention, but and is not so limited the present invention.
Following detection method, if no special instructions, uses phend-sulphuric acid to survey the content of polysaccharide.
Embodiment 1
A kind of extracting method of platycodon root polysaccharide, the method includes following step:
Select the Radix Platycodonis dried, cross 60 mesh sieves after pulverizing, obtain balloonflower powder, add water in balloonflower powder and stir, balloonflower powder is 1:7 with the part by weight of water, soaking 10 hours in water, add cellulase, the addition of cellulase is 0.8%, hydrolysis temperature 45 DEG C, pH value 5, above-mentioned raw material is placed in Microwave Extraction Equipment extraction, the power of microwave is 400-600W, and frequency is 800MHZ, enzymolysis 0.1-0.5h；
Add ficin and bromelain, the addition of ficin is 0.45%, the addition of bromelain is 0.4%, hydrolysis temperature 45 DEG C, pH value 6.0, be placed in Microwave Extraction Equipment extraction by above-mentioned raw material, and the power of microwave is 500W, frequency is 800MHZ, enzymolysis 0.3h, after enzymolysis at 100 DEG C enzyme denaturing 6min, obtain enzymolysis solution；
Above-mentioned enzymolysis solution is placed in vacuum concentrator concentration, and decompression cryoconcentration, to enzymolysis solution to the 1/3 of original volume, obtains Radix Platycodonis crude polysaccharides concentrated solution；
Adding activated carbon in the concentrated solution of Radix Platycodonis crude polysaccharides, activated carbon accounts for the 4% of Radix Platycodonis crude polysaccharides concentrated solution weight, stirs 0.3h, leaches activated carbon；
Centrifugal 15 min, centrifugal rotational speed is 4500 rpm, is collected respectively with precipitation by supernatant；
Take supernatant liquid filtering to remove slag, the filtrate obtained after filtration concentrates at 65 DEG C, being concentrated into its concentration is 65%, concentrated solution adds 95% ethanol precipitation in 1:4 ratio and stands 10-20 hour, is centrifuged 15 min, centrifugal rotational speed is 4500 rpm, collects precipitation and repeatedly cleans 2-3 time with ethanol, after lyophilization to moisture is 5%, pulverize at-1-4 DEG C again, obtain soluble polysaccharide；
Above-mentioned enzyme concentration is enzyme and accounts for the percentage by weight of the Radix Platycodonis after micronizing.
The extraction ratio of polysaccharide is: 36.44%, and purity is 94.64%.
Embodiment 2
A kind of extracting method of platycodon root polysaccharide, the method includes following step:
Select the Radix Platycodonis dried, cross 40 mesh sieves after pulverizing, obtain balloonflower powder, add water in balloonflower powder and stir, balloonflower powder is 1:5 with the part by weight of water, soaking 8 hours in water, add cellulase, the addition of cellulase is 0.5%, hydrolysis temperature 45 DEG C, pH value 5, above-mentioned raw material is placed in Microwave Extraction Equipment extraction, the power of microwave is 400-600W, and frequency is 800MHZ, enzymolysis 0.1-0.5h；
Add ficin and bromelain, the addition of ficin is 0.2%, the addition of bromelain is 0.2%, hydrolysis temperature 45 DEG C, pH value 5.5, be placed in Microwave Extraction Equipment extraction by above-mentioned raw material, and the power of microwave is 400W, frequency is 800MHZ, enzymolysis 0.3h, after enzymolysis at 100 DEG C enzyme denaturing 5min, obtain enzymolysis solution；
Above-mentioned enzymolysis solution is placed in vacuum concentrator concentration, and decompression cryoconcentration, to enzymolysis solution to the 1/3 of original volume, obtains Radix Platycodonis crude polysaccharides concentrated solution；
Adding activated carbon in the concentrated solution of Radix Platycodonis crude polysaccharides, activated carbon accounts for the 4% of Radix Platycodonis crude polysaccharides concentrated solution weight, stirs 0.3h, leaches activated carbon；
Centrifugal 15 min, centrifugal rotational speed is 4500 rpm, is collected respectively with precipitation by supernatant；
Take supernatant liquid filtering to remove slag, the filtrate obtained after filtration concentrates at 65 DEG C, being concentrated into its concentration is 65%, concentrated solution adds 95% ethanol precipitation in 1:4 ratio and stands 10 hours, is centrifuged 15 min, centrifugal rotational speed is 4500 rpm, collects precipitation and repeatedly cleans 2-3 time with ethanol, after lyophilization to moisture is 5%, pulverize at-1-4 DEG C again, obtain soluble polysaccharide；
Above-mentioned enzyme concentration is enzyme and accounts for the percentage by weight of the Radix Platycodonis after micronizing.
The extraction ratio of polysaccharide is: 36.11%, and purity is 93.15%.
Embodiment 3
A kind of extracting method of platycodon root polysaccharide, the method includes following step:
Select the Radix Platycodonis dried, cross 80 mesh sieves after pulverizing, obtain balloonflower powder, add water in balloonflower powder and stir, balloonflower powder is 1:10 with the part by weight of water, soaking 16 hours in water, add cellulase, the addition of cellulase is 0.9%, hydrolysis temperature 45 DEG C, pH value 5, above-mentioned raw material is placed in Microwave Extraction Equipment extraction, the power of microwave is 400-600W, and frequency is 800MHZ, enzymolysis 0.1-0.5h；
Add ficin and bromelain, the addition of ficin is 0.6%, the addition of bromelain is 0.5%, hydrolysis temperature 45 DEG C, pH value 6.5, be placed in Microwave Extraction Equipment extraction by above-mentioned raw material, and the power of microwave is 500W, frequency is 800MHZ, enzymolysis 0.3h, after enzymolysis at 100 DEG C enzyme denaturing 6min, obtain enzymolysis solution；
Above-mentioned enzymolysis solution is placed in vacuum concentrator concentration, and decompression cryoconcentration, to enzymolysis solution to the 1/3 of original volume, obtains Radix Platycodonis crude polysaccharides concentrated solution；
Adding activated carbon in the concentrated solution of Radix Platycodonis crude polysaccharides, activated carbon accounts for the 4% of Radix Platycodonis crude polysaccharides concentrated solution weight, stirs 0.3h, leaches activated carbon；
Centrifugal 15 min, centrifugal rotational speed is 4500 rpm, is collected respectively with precipitation by supernatant；
Take supernatant liquid filtering to remove slag, the filtrate obtained after filtration concentrates at 65 DEG C, being concentrated into its concentration is 65%, concentrated solution adds 95% ethanol precipitation in 1:4 ratio and stands 20 hours, is centrifuged 15 min, centrifugal rotational speed is 4500 rpm, collects precipitation and repeatedly cleans 2-3 time with ethanol, after lyophilization to moisture is about 5%, pulverize at-1-4 DEG C again, obtain soluble polysaccharide；
Above-mentioned enzyme concentration is enzyme and accounts for the percentage by weight of the Radix Platycodonis after micronizing.
The extraction ratio of polysaccharide is: 36.28%, and purity is 92.94%.
Claims (4)
1. an extracting method for platycodon root polysaccharide, the step including following:
Select the Radix Platycodonis dried, cross 40-80 mesh sieve after pulverizing, obtain balloonflower powder, add water in balloonflower powder and stir, balloonflower powder is 1:5-10 with the part by weight of water, soaking 8-16 hour in water, add cellulase, the addition of cellulase is 0.5-0.9%, hydrolysis temperature 45 DEG C, pH value 5, above-mentioned raw material is placed in Microwave Extraction Equipment extraction, the power of microwave is 400-600W, and frequency is 800MHz, enzymolysis 0.1-0.5h；
Add ficin and bromelain, the addition of ficin is 0.2-0.6%, the addition of bromelain is 0.2-0.5%, hydrolysis temperature 45 DEG C, pH value 5.5-6.5, be placed in Microwave Extraction Equipment extraction by above-mentioned raw material, and the power of microwave is 400-600W, frequency is 800MHz, enzymolysis 0.1-0.5h, after enzymolysis at 100-105 DEG C enzyme denaturing 5-10min, obtain enzymolysis solution；
Above-mentioned enzymolysis solution is placed in vacuum concentrator concentration, and decompression cryoconcentration, to enzymolysis solution to the 1/3 of original volume, obtains Radix Platycodonis crude polysaccharides concentrated solution；
Adding activated carbon in the concentrated solution of Radix Platycodonis crude polysaccharides, described activated carbon accounts for the 2-6% of Radix Platycodonis crude polysaccharides concentrated solution weight, stirs 0.2-0.5h, leaches activated carbon；
Centrifugal 10-20 min, centrifugal rotational speed is 4500 rpm, is collected respectively with precipitation by supernatant；
Take supernatant liquid filtering to remove slag, the filtrate obtained after filtration concentrates at 60-70 DEG C, being concentrated into its concentration is 60-70%, concentrated solution adds 95% ethanol precipitation in 1:4 ratio and stands 10-20 hour, is centrifuged 10-20 min, centrifugal rotational speed is 4500 rpm, collects precipitation and repeatedly cleans 2-3 time with ethanol, after lyophilization to moisture is 4-6%, pulverize at-1-4 DEG C again, obtain soluble polysaccharide；
Above-mentioned enzyme concentration is enzyme and accounts for the percentage by weight of the Radix Platycodonis after pulverizing.
The extracting method of a kind of platycodon root polysaccharide the most as claimed in claim 1, it is characterised in that the consumption of described ficin is 0.45%.
The extracting method of a kind of platycodon root polysaccharide the most as claimed in claim 1, it is characterised in that the addition of described bromelain is 0.4%.
4. the extracting method of a kind of platycodon root polysaccharide as according to any one of claim 1-3, it is characterised in that described method includes following step:
Select the Radix Platycodonis dried, cross 60 mesh sieves after pulverizing, obtain balloonflower powder, add water in balloonflower powder and stir, balloonflower powder is 1:7 with the part by weight of water, soaking 10 hours in water, add cellulase, the addition of cellulase is 0.8%, hydrolysis temperature 45 DEG C, pH value 5, above-mentioned raw material is placed in Microwave Extraction Equipment extraction, the power of microwave is 400-600W, and frequency is 800MHz, enzymolysis 0.1-0.5h；
Add ficin and bromelain, the addition of ficin is 0.45%, the addition of bromelain is 0.4%, hydrolysis temperature 45 DEG C, pH value 6.0, be placed in Microwave Extraction Equipment extraction by above-mentioned raw material, and the power of microwave is 500W, frequency is 800MHz, enzymolysis 0.3h, after enzymolysis at 100 DEG C enzyme denaturing 6min, obtain enzymolysis solution；
Above-mentioned enzymolysis solution is placed in vacuum concentrator concentration, and decompression cryoconcentration, to enzymolysis solution to the 1/3 of original volume, obtains Radix Platycodonis crude polysaccharides concentrated solution；
Adding activated carbon in the concentrated solution of Radix Platycodonis crude polysaccharides, described activated carbon accounts for the 4% of Radix Platycodonis crude polysaccharides concentrated solution weight, stirs 0.3h, leaches activated carbon；
Centrifugal 15 min, centrifugal rotational speed is 4500 rpm, is collected respectively with precipitation by supernatant；
Take supernatant liquid filtering to remove slag, the filtrate obtained after filtration concentrates at 65 DEG C, being concentrated into its concentration is 65%, concentrated solution adds 95% ethanol precipitation in 1:4 ratio and stands 10-20 hour, is centrifuged 15 min, centrifugal rotational speed is 4500 rpm, collects precipitation and repeatedly cleans 2-3 time with ethanol, after lyophilization to moisture is 5%, pulverize at-1-4 DEG C again, obtain soluble polysaccharide；
Above-mentioned enzyme concentration is enzyme and accounts for the percentage by weight of the Radix Platycodonis after pulverizing.
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| KR101976998B1 (en) * | 2018-10-18 | 2019-05-10 | 한국수목원관리원 | The method for preparing a Platycodon grandiflorum extract using a high pressure enzyme treatment and a composition having antioxidant or α-glucosidase inhibiting activity |
| CN114409821B (en) * | 2022-02-08 | 2023-04-07 | 合肥工业大学 | Platycodon grandiflorum polysaccharide extraction method based on micro-thermal explosion wall breaking |
| CN114478821B (en) * | 2022-03-01 | 2023-04-25 | 安徽中医药大学 | Radix Platycodi polysaccharide capable of improving constitution of mice after induction of high-fat diet and application thereof |
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| CN101830999A (en) * | 2010-05-21 | 2010-09-15 | 王振宇 | Method for extracting platycodon root polysaccharide |
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Effective date of registration: 20191205 Address after: 225300 Taizhou City, Taizhou City, Jiangsu Province, China Medical Chengkou, west of Tailu Road, east of Lujia Road, 0008 blocks, east of G78 No. 1 to 4 floors Patentee after: Jiangsu Kanglinbei Medical Devices Co., Ltd. Address before: 225300 Taihe Road 10, hailing Industrial Park, Jiangsu, Taizhou Patentee before: Jiangsu Huaneng Pharmaceuticals Co., Ltd. |