CN106619804A - Herba rabdosiae extract having antibacterial activity as well as preparation method and application of herba rabdosiae extract - Google Patents
Herba rabdosiae extract having antibacterial activity as well as preparation method and application of herba rabdosiae extract Download PDFInfo
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Abstract
The invention relates to a herba rabdosiae extract having an antibacterial activity as well as a preparation method and an application of the herba rabdosiae extract. The herba rabdosiae extract consists of the following ingredients: oridonin, 5,6,7-trimethoxy flavone, 2-methoxy-4-vinylphenol, eugenol, [alpha]-linolenic acid and androstenedione. The preparation method of the herba rabdosiae extract comprises the following steps: selecting and drying herba rabdosiae, crushing the dried herba rabdosiae by virtue of a traditional Chinese medicine pulverizer, and soaking the crushed herba rabdosiae in an alcohol water solution; conducting ultrasonic ethanol reflux extraction on a soaked herba rabdosiae powder mixture so as to obtain a filtrate; and decompressing and filtering the filtrate, and concentrating the processed filtrate by virtue of a rotary evaporator at 45 DEG C so as to obtain an extractum. The herba rabdosiae extract which has the antibacterial activity is obtained by drying the extractum in a vacuum environment in a freeze-drying mode. The preparation method provided by the invention is reasonable in process; the prepared herba rabdosiae extract has a quite strong inhibitory effect on a plurality of microorganisms; and the herba rabdosiae extract, as an antibacterial active ingredient of a feed additive, can be used for preventing bacterial pollution of feed.
Description
Technical field
The present invention relates to a kind of Rabdosia rubescens extract, specifically a kind of Rabdosia rubescens extract with antibacterial activity and
Its preparation method and application.
Background technology
Self-discovery antibiotic has after facilitation to growth of animal, and antibiotic feed additive has obtained widely should
With being made that huge contribution in animal husbandry.But with the abuse of antibiotic, antibiotic feed additive produces the pair of three aspects
Effect：（1）, make pathogen produce drug resistance；（2）, in body remain.Affect the immune effect of humans and animals；（3）, easily produce
The cross-infection of raw pathogenic bacteria.Scientists have proven to：The numerous disease of the mankind all with animal products in antibiotic, hormone and
The residual of medicine is relevant.With people's living standard, the raising of health perception, green products of domestic animal is increasingly subject to numerous people
Like.The safety issue of feed addictive, is focus of concern all the time.
1986. Sweden completely forbids the antibiotic used in animal and fowl fodder. and become first and forbidden to make using antibiotic
For the country of growth promoter.From in January, 2006, European Union has completely forbidden and has added antibiotic in animal feed.Subsequently, Japan
Livestock and poultry antibiotic-free has also been formulated in succession with the Asian countries such as Korea has raised specification.Before and after the 2008 Beijing Olympic Games, China exists
Biological feedstuff and livestock and poultry antibiotic-free cultivation aspect have also obtained the unprecedented attention of various circles of society.2015, China fired
First rifle of " feed is prohibited anti-", Lomefloxacin, Pefloxacin, Ofloxacin, 4 kinds of veterinary drugs of Norfloxacin start to completely forbid and exist
Used in animal, with the development of situation, future will have more antibiotic and be inhibited in animal feed addition and use.
Therefore, it is current cultivation industry urgent problem to develop safe and effective green feed additive.
Rabdosia rubescens is referred to as rabdosia rubesens, and scientific name is cracked rice fork, is Labiatae Rabdosia herbaceous plant.The activity of Rabdosia rubescens into
Divide and mainly include terpene substances（Diterpenes, triterpenes）And steroid, it is flavonoids, volatile oil, amino acids, polysaccharide, organic
Acid and other classes.
The content of the invention
The present invention seeks to the deficiency to solve above-mentioned technical problem, there is provided a kind of Rabdosia rubescens with antibacterial activity is extracted
Thing and its preparation method and application, has very strong inhibitory action to multiple-microorganism, can have as the antibacterial of feed addictive
Effect composition, prevents feed Bacterial Contamination.
The present invention is for the solution technical scheme that adopted of above-mentioned technical problem：
A kind of Rabdosia rubescens extract with antibacterial activity, its composition includes Oridonin, 5,6,7 trimethoxy flavones, 2-
Methoxyl group -4- phenol ethylenes, eugenol, alpha-linolenic acid and androstenedione, each Ingredient percent content is：Oridonin
10.17%, 5,6,7 trimethoxy flavones 16.46%, 2- methoxyl group -4- phenol ethylenes 1.95%, eugenol 0.83%, alpha-linolenic acid
0.67%, androstenedione 2.79%；Its to staphylococcus aureus, vibrio harveyi, salmonella typhimurium, Aeromonas hydrophila and
Pasteurella multocida has bacteriostatic activity.
Rabdosia rubescens extract with antibacterial activity can prevent feed bacterium as the antibacterial effective ingredient of feed addictive
Contact scar.
The preparation method of the Rabdosia rubescens extract with antibacterial activity, comprises the following steps：
（1）Rabdosia rubescens aerial part is selected, oven for drying is put into, bar and leaf are separated, the leaf Jing Chinese medicines selected are crushed
Machine is crushed, and is crossed 60 mesh sieves and is put into vacuum glass drier, and Rabdosia rubescens powder after being dried is standby；
（2）Rabdosia rubescens powder after the drying that above-mentioned steps are prepared is weighed, the volume fraction for adding its quality 6-10 times is 95% alcohol
The aqueous solution, soaks 3-5 days, daily stirring 3-4 time, Rabdosia rubescens powder mixture after being soaked；Rabdosia rubescens powder mixing after soaking
Thing is added in reflux, and reflux is put into supersonic generator carries out ultrasonic alcohol reflux extraction, Extracting temperature
40 DEG C, the min of each extraction time 30, the W of ultrasonic power 150, extract 2 times, merge filtrate twice, it is standby；
（3）Above-mentioned steps gained filtrate is entered in Buchner funnel with glass bar drainage, pumping switch pump is opened, starts suction filtration, mistake
Filtering flask adapter is first taken out after having filtered, aspiration pump is closed afterwards, 45 DEG C of Rotary Evaporators concentrations of filtrate Jing after suction filtration are obtained into medicinal extract,
It is standby；
（4）Medicinal extract is carried out after Components identification, to be dried by vacuum freeze drying, medicinal extract enters after pulverizer is to drying
Row is crushed, that is, prepare the Rabdosia rubescens extract with antibacterial activity.
Step（4）Described in vacuum freeze drying step be：Freezing 3-4h drops to 40 DEG C of ﹣ to temperature, vacuumizes, finally etc.
Temperature reaches 14-16 DEG C, is dried 48h.
Step（4）Medicinal extract is crushed to 130-160 mesh after middle drying.
Step（4）Middle employing gas chromatography mass spectrometry method carries out Components identification to medicinal extract, and Rabdosia rubescens extract is diluted into 1mg/ml,
Chromatographic condition：300 DEG C of injector temperature, temperature programming：300 DEG C are risen to by 80 DEG C keep 8min, 10 DEG C/min of heating rate；Carrier gas
For high-purity helium, flow velocity is 1ml/min；Split ratio 5:1, the μ L of sample size 1；EI ionization spectrum 70eV.
Beneficial effect is：
1st, Rabdosia rubescens extract of the present invention is to staphylococcus aureus, pasteurella multocida, Aeromonas hydrophila, vibrio harveyi
There is very strong inhibitory action etc. common easy pathogenic bacteria.It is that one kind can suppress to be harmful to micro- life as the Rabdosia rubescens of " integration of drinking and medicinal herbs "
The good source of thing, the plant resources to developing China greatly facilitates effect；Feed addition is prepared with Rabdosia rubescens extract
Agent, can be added in feed to prevent bacterial infection green, safe and pollution-free as natural feed addictive, meet current
The requirement in livestock breed aquatics nonreactive epoch, can be added to prevention bird bacterial infection disease in feed as feed addictive.
2nd, the extraction process of Rabdosia rubescens extract of the present invention, has selected suitable organic solvent to be extracted, such that it is able to
Effectively antimicrobial component is extracted.What is taken is that ultrasonic ultrasonic wave added alcohol reflux is extracted, and is carried with common alcohol reflux
Take and compare, extraction efficiency is higher.Use vacuum freeze dryer to be dried, with conventional drying, be spray-dried, dry
Etc. comparing, vacuum freeze drying is carried out at low temperature, and the nutrient content of material, shape can be kept also to keep being basically unchanged；Freeze
The moisture content of dry material can reach very low degree, facilitate long term storage, long-distance transport；Due to carrying out in the state of freezing
It is dried, therefore the volume of product, shape are almost unchanged, maintain original structure, and concentration phenomena will not occur；Material is in low pressure
Lower drying, make oxidizable composition in material will not oxidation deterioration, while because of hypobaric hypoxia, can sterilize or suppress some bacteriums
Vigor.
Specific embodiment
A kind of Rabdosia rubescens extract with antibacterial activity, its composition includes that Oridonin, 5,6,7 trimethoxies are yellow
Ketone, 2- methoxyl group -4- phenol ethylenes, eugenol, alpha-linolenic acid and androstenedione, each Ingredient percent content is：Winter insults
Careless A prime 10.17%, 5,6,7 trimethoxy flavones 16.46%, 2- methoxyl group -4- phenol ethylenes 1.95%, eugenol 0.83%, α -
Leukotrienes 0.67%, androstenedione 2.79%；It is to staphylococcus aureus, vibrio harveyi, salmonella typhimurium, thermophilic aqueous vapor list
Born of the same parents bacterium and pasteurella multocida have bacteriostatic activity.
Rabdosia rubescens extract with antibacterial activity can prevent feed bacterium as the antibacterial effective ingredient of feed addictive
Contact scar.
The preparation method of the Rabdosia rubescens extract with antibacterial activity, comprises the following steps：
（1）Rabdosia rubescens aerial part is selected, oven for drying is put into, bar and leaf are separated, the leaf Jing Chinese medicines selected are crushed
Machine is crushed, and is crossed 60 mesh sieves and is put into vacuum glass drier, and Rabdosia rubescens powder after being dried is standby；
（2）Rabdosia rubescens powder after the drying that above-mentioned steps are prepared is weighed, the volume fraction for adding its quality 6-10 times is 95% alcohol
The aqueous solution, soaks 3-5 days, daily stirring 3-4 time, Rabdosia rubescens powder mixture after being soaked；Rabdosia rubescens powder mixing after soaking
Thing is added in reflux, and reflux is put into supersonic generator carries out ultrasonic alcohol reflux extraction, Extracting temperature
40 DEG C, the min of each extraction time 30, the W of ultrasonic power 150, extract 2 times, merge filtrate twice, it is standby；
（5）Above-mentioned steps gained filtrate is entered in Buchner funnel with glass bar drainage, pumping switch pump is opened, starts suction filtration, mistake
Filtering flask adapter is first taken out after having filtered, aspiration pump is closed afterwards, 45 DEG C of Rotary Evaporators concentrations of filtrate Jing after suction filtration are obtained into medicinal extract,
It is standby；
（4）Medicinal extract is carried out after Components identification, to be dried by vacuum freeze drying, medicinal extract enters after pulverizer is to drying
Row is crushed, that is, prepare the Rabdosia rubescens extract with antibacterial activity.
Step（4）Described in vacuum freeze drying step be：Freezing 3-4h drops to 40 DEG C of ﹣ to temperature, vacuumizes, finally etc.
Temperature reaches 14-16 DEG C, is dried 48h.
Step（4）Medicinal extract is crushed to 130-160 mesh after middle drying.
Step（4）Middle employing gas chromatography mass spectrometry method carries out Components identification to medicinal extract, and Rabdosia rubescens extract is diluted into 1mg/ml,
Chromatographic condition：300 DEG C of injector temperature, temperature programming：300 DEG C are risen to by 80 DEG C keep 8min, 10 DEG C/min of heating rate；Carrier gas
For high-purity helium, flow velocity is 1ml/min；Split ratio 5:1, the μ L of sample size 1；EI ionization spectrum 70eV.
Embodiment 1
With Rabdosia rubescens as raw material, drying, crushing, the extraction of ultrasonic wave added alcohol reflux, reduced pressure concentration, vacuum freeze drying are obtained
A kind of Rabdosia rubescens extract with antibacterial activity, its preparation method is as follows：
（1）Rabdosia rubescens leaf is selected, Henan Province Jiyuan City is picked up from, Jing medicinal herb grinders are crushed after drying, and excessively 60 mesh sieves are put into dry
It is standby in dry device.
（2）Above-mentioned Rabdosia rubescens 20g is weighed, 95% alcohol of 6 times of Rabdosia rubescens quality is added, is soaked 3 days, 4 are stirred daily
It is secondary.Reflux is put into supersonic generator carries out ultrasonic alcohol reflux extraction, 40 DEG C of Extracting temperature, each extraction time
30 min, the W of ultrasonic power 150, extract 2 times, merge filtrate twice.
（3）Filtrate is reduced pressure, suction filtration, and 45 DEG C of Rotary Evaporators concentrations of Jing are obtained medicinal extract.
（4）Medicinal extract is dried by vacuum freeze drying, carries out being crushed to 150 mesh by pulverizer after being dried, i.e.,
Obtain a kind of Rabdosia rubescens extract with antibacterial activity.
（5）Components identification is carried out to Rabdosia rubescens extract using gas chromatography mass spectrometry method, Rabdosia rubescens extract is diluted into 1mg/
Ml, chromatographic condition：300 DEG C of injector temperature, temperature programming：300 DEG C are risen to by 80 DEG C keep 8min, 10 DEG C/min of heating rate；
Carrier gas is high-purity helium, and flow velocity is 1ml/min；Split ratio 5:1, the μ L of sample size 1；EI ionization spectrum 70eV.
Embodiment 2
With Rabdosia rubescens as raw material, drying, crushing, the extraction of ultrasonic wave added alcohol reflux, reduced pressure concentration, vacuum freeze drying are obtained
A kind of Rabdosia rubescens extract with antibacterial activity, its preparation method is as follows：
（1）Rabdosia rubescens leaf is selected, Henan Province Jiyuan City is picked up from, Jing medicinal herb grinders are crushed after drying, and excessively 60 mesh sieves are put into dry
It is standby in dry device.
（2）Above-mentioned Rabdosia rubescens 20g is weighed, 95% alcohol of 8 times of Rabdosia rubescens quality is added, is soaked 5 days, 3 are stirred daily
It is secondary.Reflux is put into supersonic generator carries out ultrasonic alcohol reflux extraction, 40 DEG C of Extracting temperature, each extraction time
30 min, the W of ultrasonic power 150, extract 2 times, merge filtrate twice.
（3）Filtrate is reduced pressure, suction filtration, and 45 DEG C of Rotary Evaporators concentrations of Jing are obtained medicinal extract.
（4）Medicinal extract is dried by vacuum freeze drying, carries out being crushed to 150 mesh by pulverizer after being dried, i.e.,
Obtain a kind of Rabdosia rubescens extract with antibacterial activity.
（5）Components identification is carried out to Rabdosia rubescens extract using gas chromatography mass spectrometry method, Rabdosia rubescens extract is diluted into 1mg/
Ml, chromatographic condition：300 DEG C of injector temperature, temperature programming：300 DEG C are risen to by 80 DEG C keep 8min, 10 DEG C/min of heating rate；
Carrier gas is high-purity helium, and flow velocity is 1ml/min；Split ratio 5:1, the μ L of sample size 1；EI ionization spectrum 70eV.
Embodiment 3
With Rabdosia rubescens as raw material, drying, crushing, the extraction of ultrasonic wave added alcohol reflux, reduced pressure concentration, vacuum freeze drying are obtained
A kind of Rabdosia rubescens extract with antibacterial activity, its preparation method is as follows：
（1）Rabdosia rubescens leaf is selected, Henan Province Jiyuan City is picked up from, Jing medicinal herb grinders are crushed after drying, and excessively 60 mesh sieves are put into dry
It is standby in dry device.
（2）Above-mentioned Rabdosia rubescens 20g is weighed, 95% alcohol of 10 times of Rabdosia rubescens quality is added, is soaked 4 days, 4 are stirred daily
It is secondary.Reflux is put into supersonic generator carries out ultrasonic alcohol reflux extraction, 40 DEG C of Extracting temperature, each extraction time
30 min, the W of ultrasonic power 150, extract 2 times, merge filtrate twice.
（3）Filtrate is reduced pressure, suction filtration, and 45 DEG C of Rotary Evaporators concentrations of Jing are obtained medicinal extract.
（4）Medicinal extract is dried by vacuum freeze drying, carries out being crushed to 150 mesh by pulverizer after being dried, i.e.,
Obtain a kind of Rabdosia rubescens extract with antibacterial activity.
（5）Components identification is carried out to Rabdosia rubescens extract using gas chromatography mass spectrometry method, Rabdosia rubescens extract is diluted into 1mg/
Ml, chromatographic condition：300 DEG C of injector temperature, temperature programming：300 DEG C are risen to by 80 DEG C keep 8min, 10 DEG C/min of heating rate；
Carrier gas is high-purity helium, and flow velocity is 1ml/min；Split ratio 5:1, the μ L of sample size 1；EI ionization spectrum 70eV.
Pharmacological test example：The bacteriostasis of Rabdosia rubescens extract
1 test material
1.1 materials and reagent
Rabdosia rubescens extract, peptone, sodium chloride, soya peptone, agar powder, distilled water
1.2 for examination bacterial classification
Staphylococcus aureus（S.aureus）, University Of Science and Technology Of He'nan's food and Biological Engineering College's experiment teaching center preservation.
Pasteurella multocida（Pasteurella multocida）, Aeromonas hydrophila（Aeromonas
hydrophila）, vibrio harveyi（Vibrio harveyi）, salmonella typhimurium（Salmonella typhimurium）,
Purchased from Chinese microorganism strain collection, Jing after inclined-plane culture activation, 4 DEG C of Refrigerator stores are standby.
2 test methods
2.1 bacteriostatic tests
（1）Prepare containing bacterium culture medium
TSA culture mediums：Peptone 15g, soya peptone 5g, sodium chloride 5g, 15 ~ 20g of agar, water 1000ml.121 DEG C of sterilizing 20min.
TSB culture mediums：Peptone 15g, soya peptone 5g, sodium chloride 5g, water 1000ml.121 DEG C of sterilizing 20min.
Sterilized straw draws the bacteria suspension 1mL prepared with TSB culture mediums and adds 100mL to melt and be incubated in a water bath
（48～50 DEG C）TSA culture mediums in, make bacterial content reach 1 × 106Cfu/mL, shakes up stand-by.
（2）Bacteriostatic test
By it is above-mentioned prepare pour into sterile petri dish containing bacterium culture medium, stand cooling, wait its solidification after, with aseptic nipper will
Sterilizing Oxford cup is vertically placed on surface containing bacterium culture medium, presses lightly on（Each flat board puts 4 Oxford cups）.Drawn with pipettor
100 μ L Rabdosia rubescens extract solution, are added in the cup aperture of Oxford, while doing blank with methyl alcohol.Then, culture dish is put into
In incubator, after 37 DEG C of culture 15h of bacterium, after 30 DEG C of culture 48h of fungi antibacterial circle diameter is measured.
3 results and analysis
The fungistatic effect of Rabdosia rubescens extract such as table 1.
The fungistatic effect of the Rabdosia rubescens extract of table 1
As shown in Table 1, Rabdosia rubescens extract is to staphylococcus aureus（S.aureus）, pasteurella multocida（P.
multocida）, Aeromonas hydrophila（A. hydrophila）, vibrio harveyi（V. harveyi）And salmonella typhimurium
（S. typhimurium）With certain inhibitory action, wherein, staphylococcus aureus（S.aureus）And vibrio harveyi（V.
harveyi）More than 20mm is reached to the antibacterial circle diameter of Rabdosia rubescens chloroform extract, the antibacterial sensitivity of many antibacterials is reached
Standard.
Constituent analysis test example：Gc-mss
1 test method
（1）Rabdosia rubescens aerial part is selected, Henan Province Jiyuan City is picked up from, oven for drying is put into, bar and leaf are separated, will be selected
Leaf Jing medicinal herb grinders crush, cross 60 mesh sieves be put into it is standby in vacuum glass drier.
（2）Above-mentioned Rabdosia rubescens is weighed, 95% alcohol is added, Rabdosia rubescens is 1 with 95% alcohol ratio:10（g/ml）, immersion 3 ~ 5
My god, stir 3 ~ 4 times daily.Reflux is put into supersonic generator carries out ultrasonic alcohol reflux extraction, Extracting temperature 40
DEG C, the min of each extraction time 30, the W of ultrasonic power 150, extract 2 times, merge filtrate twice.
（3）Filtrate is entered in Buchner funnel with glass bar drainage, pumping switch pump is opened, starts suction filtration, after having filtered
Filtering flask adapter is first taken out, aspiration pump is closed afterwards, 45 DEG C of Rotary Evaporators concentrations of filtrate Jing after suction filtration are obtained into medicinal extract.
（4）Using liquid-liquid extraction method, extract medicinal extract is extracted successively with petroleum ether, chloroform, to Rabdosia rubescens
Extract is purified.
（5）Components identification is carried out to its bacteriostatic active ingredients using gas chromatography mass spectrometry method, Rabdosia rubescens extract chloroform is mutually dilute
It is interpreted into 1mg/ml, chromatographic condition：300 DEG C of injector temperature, temperature programming：300 DEG C of holding 8min, heating rate 10 are risen to by 80 DEG C
℃/min；Carrier gas is high-purity helium, and flow velocity is 1ml/min；Split ratio 5:1, the μ L of sample size 1；EI ionization spectrum 70eV.
2 test results and analysis
The bacteriostatic active ingredients of Rabdosia rubescens extract are primarily present in chloroform phase, and main component has Oridonin, 5,6,7 front threes
Epoxide flavones, 2- methoxyl group -4- phenol ethylenes, eugenol, alpha-linolenic acid, androstenedione.Each component content is：Oridonin
10.17%；5,6,7 trimethoxy flavones 16.46%；2- methoxyl group -4- phenol ethylenes 1.95%；Eugenol 0.83%；Alpha-linolenic acid
0.67%, androstenedione 2.79%.
Claims (6)
1. a kind of Rabdosia rubescens extract with antibacterial activity, it is characterised in that：Its composition include Oridonin, 5,6,7 3
Methoxy flavone, 2- methoxyl group -4- phenol ethylenes, eugenol, alpha-linolenic acid and androstenedione, each Ingredient percent content
For：The trimethoxy flavone 16.46% of Oridonin 10.17%, 5,6,7,2- methoxyl group -4- phenol ethylenes 1.95%, eugenol
0.83%, alpha-linolenic acid 0.67%, androstenedione 2.79%；Its to staphylococcus aureus, vibrio harveyi, salmonella typhimurium,
Aeromonas hydrophila and pasteurella multocida have bacteriostatic activity.
2. the application of the Rabdosia rubescens extract with antibacterial activity as claimed in claim 1, it is characterised in that：As feed addition
The antibacterial effective ingredient of agent, prevents feed Bacterial Contamination.
3. there is as claimed in claim 1 the preparation method of the Rabdosia rubescens extract of antibacterial activity, it is characterised in that：Including following
Step：
（1）Rabdosia rubescens aerial part is selected, oven for drying is put into, bar and leaf are separated, the leaf Jing Chinese medicines selected are crushed
Machine is crushed, and is crossed 60 mesh sieves and is put into vacuum glass drier, and Rabdosia rubescens powder after being dried is standby；
（2）Rabdosia rubescens powder after the drying that above-mentioned steps are prepared is weighed, the volume fraction for adding its quality 6-10 times is 95% alcohol
The aqueous solution, soaks 3-5 days, daily stirring 3-4 time, Rabdosia rubescens powder mixture after being soaked；Rabdosia rubescens powder mixing after soaking
Thing is added in reflux, and reflux is put into supersonic generator carries out ultrasonic alcohol reflux extraction, Extracting temperature
40 DEG C, the min of each extraction time 30, the W of ultrasonic power 150, extract 2 times, merge filtrate twice, it is standby；
（3）Above-mentioned steps gained filtrate is entered in Buchner funnel with glass bar drainage, pumping switch pump is opened, starts suction filtration, mistake
Filtering flask adapter is first taken out after having filtered, aspiration pump is closed afterwards, 45 DEG C of Rotary Evaporators concentrations of filtrate Jing after suction filtration are obtained into medicinal extract,
It is standby；
（4）Medicinal extract is carried out after Components identification, to be dried by vacuum freeze drying, medicinal extract enters after pulverizer is to drying
Row is crushed, that is, prepare the Rabdosia rubescens extract with antibacterial activity.
4. there is as claimed in claim 3 the preparation method of the Rabdosia rubescens extract of antibacterial activity, it is characterised in that：Step（4）
Described in vacuum freeze drying step be：Freezing 3-4h drops to 40 DEG C of ﹣ to temperature, vacuumizes, and last equitemperature reaches 14-16
DEG C, it is dried 48h.
5. there is as claimed in claim 3 the preparation method of the Rabdosia rubescens extract of antibacterial activity, it is characterised in that：Step（4）
Medicinal extract is crushed to 130-160 mesh after middle drying.
6. there is as claimed in claim 3 the preparation method of the Rabdosia rubescens extract of antibacterial activity, it is characterised in that：Step（4）
Middle employing gas chromatography mass spectrometry method carries out Components identification to medicinal extract, and Rabdosia rubescens extract is diluted into 1mg/ml, chromatographic condition：Sample introduction temperature
300 DEG C of degree, temperature programming：300 DEG C are risen to by 80 DEG C keep 8min, 10 DEG C/min of heating rate；Carrier gas is high-purity helium, flow velocity
For 1ml/min；Split ratio 5:1, the μ L of sample size 1；EI ionization spectrum 70eV.
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107646940A (en) * | 2017-11-07 | 2018-02-02 | 安徽鸿泰食品有限公司 | A kind of preparation method of nutrition and health care purple potato hawthorn bread |
| CN107667939A (en) * | 2017-11-17 | 2018-02-09 | 金陵科技学院 | Use the method for the bacteriostatic activity of Procambius clarkii checking plant extraction liquid bacterium |
| CN112772927A (en) * | 2020-12-31 | 2021-05-11 | 重庆第二师范学院 | Probiotic freeze-dried powder |
| CN113754486A (en) * | 2021-09-01 | 2021-12-07 | 安徽省司尔特肥业股份有限公司 | Special organic-inorganic compound fertilizer for peanuts and preparation method thereof |
Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1486712A (en) * | 2003-08-09 | 2004-04-07 | 北海国发海洋生物产业股份有限公司 | Extractive of Chinese medicinal material rabdosia as antitumor medicine and its prepn process |
-
2016
- 2016-09-30 CN CN201610868646.0A patent/CN106619804A/en active Pending
Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1486712A (en) * | 2003-08-09 | 2004-04-07 | 北海国发海洋生物产业股份有限公司 | Extractive of Chinese medicinal material rabdosia as antitumor medicine and its prepn process |
Non-Patent Citations (1)
| Title |
|---|
| 汪伦记等: "超声辅助乙醇回流提取冬凌草抑菌活性成分的工艺优化", 《食品科技》 * |
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107646940A (en) * | 2017-11-07 | 2018-02-02 | 安徽鸿泰食品有限公司 | A kind of preparation method of nutrition and health care purple potato hawthorn bread |
| CN107667939A (en) * | 2017-11-17 | 2018-02-09 | 金陵科技学院 | Use the method for the bacteriostatic activity of Procambius clarkii checking plant extraction liquid bacterium |
| CN112772927A (en) * | 2020-12-31 | 2021-05-11 | 重庆第二师范学院 | Probiotic freeze-dried powder |
| CN113754486A (en) * | 2021-09-01 | 2021-12-07 | 安徽省司尔特肥业股份有限公司 | Special organic-inorganic compound fertilizer for peanuts and preparation method thereof |
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