CN104605013A - Preparation method of thalictrum plant extract, thalictrum plant extract prepared according to preparation method, and application of thalictrum plant extract - Google Patents
Preparation method of thalictrum plant extract, thalictrum plant extract prepared according to preparation method, and application of thalictrum plant extract Download PDFInfo
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- CN104605013A CN104605013A CN201510002789.9A CN201510002789A CN104605013A CN 104605013 A CN104605013 A CN 104605013A CN 201510002789 A CN201510002789 A CN 201510002789A CN 104605013 A CN104605013 A CN 104605013A
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- thalictrum spp
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- 238000002360 preparation method Methods 0.000 title claims abstract description 39
- 239000000419 plant extract Substances 0.000 title abstract 9
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- 239000000284 extract Substances 0.000 claims description 123
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims description 98
- 241000675108 Citrus tangerina Species 0.000 claims description 45
- 238000000605 extraction Methods 0.000 claims description 32
- KRKNYBCHXYNGOX-UHFFFAOYSA-N citric acid Chemical compound OC(=O)CC(O)(C(O)=O)CC(O)=O KRKNYBCHXYNGOX-UHFFFAOYSA-N 0.000 claims description 24
- 239000000843 powder Substances 0.000 claims description 22
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- RIOXQFHNBCKOKP-UHFFFAOYSA-N benomyl Chemical compound C1=CC=C2N(C(=O)NCCCC)C(NC(=O)OC)=NC2=C1 RIOXQFHNBCKOKP-UHFFFAOYSA-N 0.000 description 3
- MITFXPHMIHQXPI-UHFFFAOYSA-N benzoxaprofen Natural products N=1C2=CC(C(C(O)=O)C)=CC=C2OC=1C1=CC=C(Cl)C=C1 MITFXPHMIHQXPI-UHFFFAOYSA-N 0.000 description 3
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- BCPFWSWROVXGQA-UHFFFAOYSA-N 2-(2,4,4-trimethylpentan-2-yl)guanidine Chemical compound CC(C)(C)CC(C)(C)N=C(N)N BCPFWSWROVXGQA-UHFFFAOYSA-N 0.000 description 2
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- GCLAFEGUXXHIFT-IWLDQSELSA-N Homoplantaginin Chemical compound C1=C2OC(C=3C=CC(O)=CC=3)=CC(=O)C2=C(O)C(OC)=C1O[C@@H]1O[C@H](CO)[C@@H](O)[C@H](O)[C@H]1O GCLAFEGUXXHIFT-IWLDQSELSA-N 0.000 description 1
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- JYAPBWCAZXNDAR-UHFFFAOYSA-N Oxyacanthine Natural products COc1cc2CCN(C)C3Cc4ccc(Oc5cccc(CC6N(C)CCc7cc(OC)c(OC)c(Oc(c1)c23)c67)c5)c(O)c4 JYAPBWCAZXNDAR-UHFFFAOYSA-N 0.000 description 1
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Landscapes
- Medicines Containing Plant Substances (AREA)
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
Abstract
The invention provides a preparation method of a thalictrum plant extract. The preparation method comprises ultrasonic-assisted ethanol extraction of thalictrum plant(s). Preferably, the thalictrum plant(s) are/is radix thalictri simplicis and/or radix et rhizoma thalictri. The invention further provides the thalictrum plant extract obtained according to the preparation method, and application of the thalictrum plant extract in preparation of a medicine composition used for inhibiting citrus postharvest pathogen(s). The thalictrum plant extract has a strong inhibitory effect on the citrus postharvest pathogen(s), and no chemical residue exists after the thalictrum plant extract is used for fresh citrus preservation, so that the thalictrum plant extract is harmless to health of human bodies, free from environmental pollution, and low in drug resistance causing possibility.
Description
Technical field
The invention belongs to fruit disease control and fresh-keeping field, be specifically related to a kind of to there is thalictrum spp extract of suppression oranges and tangerines postharvest pathogen effect and preparation method thereof and novelty teabag.
Background technology
Oranges and tangerines are the large trade fruit of the first in the world, are also the third-largest trade agricultural product.China's oranges and tangerines aboundresources, plant with a long history.Cultivated area and the output of current China oranges and tangerines all rank first in the world, and within 2013, oranges and tangerines cultivated area reaches more than 3,600 ten thousand mu, and total output is about more than 3,200 ten thousand tons, and Citrus Industry occupies very important status in China's agricultural economy.
Citrusfruit is organized delicate succulence and is rich in sugar, in the processes such as harvesting, storing and sale, mechanical damage easily occurs, and is subject to multiple infecting of disease fungus and produces postharvest disease after pericarp damage.The pathologic disease species that rear generation adopted by oranges and tangerines reaches more than 20 kinds, mainly contain penicilliosis that Italian mould (Penicillium italicum) causes and the green mould that Penicillium digitatum (Penicillium digitatum) causes, these diseases easily cause fruit rot to go bad, rotting rate is generally 10% ~ 30%, up to more than 40% ~ 50%, huge economic loss can be caused time serious.Other pathologic disease of oranges and tangerines also comprises black rot, sour rot, brown rot, brown Supreme Being maize ear rot, anthracnose and burnt maize ear rot etc.
For a long time, people mainly adopt chemical synthesis material to carry out fresh-keeping as antistaling agent to oranges and tangerines, and the chemical bactericide that different times uses is different.Borax, sorbic acid and its esters etc. are the oranges and tangerines anticorrisive agents used the earliest; The benzimidazole germicide comprising benomyl (Benomyl), thiophanate (Thiophanate), thiophanate methyl (Thiophanatemethyl), carbendazim (Carbendazim), probenazole (Thiabendazole) etc. is the second generation oranges and tangerines anticorrisive agent of the application seventies; The third generation oranges and tangerines anticorrisive agent of current use comprises imazalil (Imazalil), Guanoctine (Guazatine), Prochloraz (Prochloraz) etc.These chemical bactericides can kill or suppress the growth of pathogenic microorganism, have good corrosion-resistanting fresh-keeping effect.Because chemical bactericide can cause, medicament is residual, contaminated environment, and Long-Time Service causes oranges and tangerines to adopt the reasons such as rear germ develops immunity to drugs making the safety in utilization of chemical bactericide day by day obtain the concern of people.In addition, along with science and technology development, detect improving and the further investigation of chemical bactericide toxicity of analysis means, find that some chemical preservatives have the possibility of carcinogenic or potential carcinogenic, mutagenesis, teratogenesis.Some chemical bactericides have been forbidden by developed countries such as America and Europes or have limited use at present, and as benomyl and Guanoctine series bactericidal agent etc., oneself is prohibited use or restriction use in part fruit products by EPA.
In recent years, people start the research and development turning to natural plant antistaling agent.The advantages such as natural plant antistaling agent has efficiently, low toxicity, low-residual, pollution-free, use safety, adopt its process oranges and tangerines to carry out anti-corrosive fresh-keeping and will be subject to the popular welcome of consumer.In addition, medicine based on herbs is by multicomponent, and Mutiple Targets plays bacteriostasis, not easily develops immunity to drugs.Therefore, be necessary to find from natural plants, exploitation natural plant source antiseptic agent carries out oranges and tangerines fresh-keeping to reduce dependence to chemical agent.
Thalictrum is a genus under Ranunculaceae, is herbaceos perennial, is distributed in Southwestern China more, as to view and admire or medicinal.This platymiscium of China about have 29 kinds can hyoscine, many kinds all have heat-clearing, it is wet to control, sweating, only dysentery, control the effects such as hot eyes.Wherein, SHUIHUANGLIAN is the root of thalictrum spp T.simplex (Thalictrumsimplex L.var.brevipes Hara), and with all herbal medicine, cold in nature, bitter, can be used for jaundice, ascites, the diseases such as difficult urination; Washing eye is decocted in external application can control eye conjunctivitis.Horse hair is linked as the root and rhizome of thalictrum spp spun gold manyleaf meadowure rhizome and root (Thalictrum glandulosissimum), leafy grass of meadow rue (Thalictrum foliolosum DC.), Zhaotong grass of meadow rue (Thalictrum glandulosissimum var.chaotungense) and plateau grass of meadow rue (Thalictrumcultratum Wall.T.deciternatum Boivin) etc., cold in nature, bitter.Manyleaf meadowure rhizome and root has effect of heat-clearing and damp-drying drug, purging intense heat and detonicating, cures mainly diarrhea due to damp-heat, dysentery jaundice, cold, fever, measles, carbuncle swell the diseases such as sore furuncle, red eye, swell pain.
Chemical composition and pharmacology activity research prove, alkaloid is the main active of thalictrum spp, comprise the alkaloids compositions such as Tang Song Xining, Tang Songximingding, oxyacanthine, thalcimine, Homoplantaginin, there is the effects such as antiviral, Tumor suppression, immunological regulation.(Lee is holy each, Su Yuqing, etc.; The progress of alkaloids chemical composition and pharmacological action in thalictrum spp, " modern medicines and clinical ", 2014,20 (3): 312-321).Bibliographical information thalictrum spp also has effect (Deng Xiaodong, the Yang Zhao for the treatment of acute bacillary dysentery; The routine clinical analysis of thalictrum spp treatment bacillary dysentery 30, " contemporary Chinese medical journal ", 1996,6 (8): 65; Tian Huayong; The routine observation of curative effect of thalictrum spp treatment acute bacillary dysentery 300, " Hunan medical science ", 1986,3 (1): 20-21).The people such as Wang Haikun report bacteriostasis (Wang Haikun, the Li Shuyi of manyleaf meadowure rhizome and root to six kinds of pathogens such as staphylococcus aureus, epidermis coccus, beta hemolytic streptococcus, Escherichia coli, shigella dysenteriae, Pseudomonas aeruginosas; The antibacterial action of manyleaf meadowure rhizome and root, " China National folk medicine magazine ", 1999,8 (5): 289-290,310).The Chinese invention patent application of denomination of invention " manyleaf meadowure rhizome and root extract is preparing the application in antineoplastic " (publication number CN101306094A), discloses manyleaf meadowure rhizome and root water or extractive with organic solvent is preparing the application in antineoplastic.
Chinese invention patent application " a kind of method (publication number CN102516257A) extracting thalcimine from hard thalictrum spp ", discloses the method adopting supercritical carbon dioxide extracting to extract thalcimine from hard thalictrum spp.
Analyze above-mentioned literature search content, the purposes finding thalictrum spp extract is mainly antitumor and suppress some infection pathogens; Report is had no to the bacteriostasis of plant pathogenic fungi, is especially suppressing the system research in oranges and tangerines postharvest pathogenic fungi to have no report.Based on above-mentioned research background, inventor has carried out the preparation of thalictrum spp extract and prepared product thereof and has suppressed oranges and tangerines to adopt the research of rear mould germ, black rot and sour germ.
Summary of the invention
The object of the invention is for the deficiencies in the prior art, a kind of preparation method of thalictrum spp extract and the thalictrum spp extract novelty teabag of preparation thereof are provided.The present invention is with natural Chinese herbal medicine for raw material, and the thalictrum spp extract of preparation to the fungistatic effect of oranges and tangerines postharvest pathogenic fungi obviously and without chemical residual, to human body without health hazard, environmentally safe, imply that good application prospect.
For solving the problems of the technologies described above, the present invention adopts following technical scheme:
A preparation method for thalictrum spp extract, adopts ultrasonic assistant to extract, take ethanol as Extraction solvent, comprise the steps:
The mass percent concentration adding thalictrum spp weight 10 ~ 40 times in described thalictrum spp is the ethanol of 40% ~ 100%, it is 40 ~ 60 DEG C at Extracting temperature, extraction time is 30min ~ 120min, and supersonic frequency is extract under the condition of 30 ~ 40KHz; Filter, reclaim ethanol, concentrated, to obtain final product.
In preparation method of the present invention, preferably, the weight of ethanol is 10 ~ 30 times of described thalictrum spp weight; Be more preferably 20 ~ 30 times.
Preferably, the mass percent concentration of ethanol is preferably 40% ~ 70%; Be more preferably 50% ~ 60%.
Preferably, ultrasonic assistant extraction time is preferably 60 ~ 120min, is more preferably 60 ~ 90min.
Preferably, the temperature that ultrasonic assistant extracts is 45 DEG C.
Also preferred, before alcohol extract, described thalictrum spp 40 ~ 50 DEG C oven dry, crosses 40 mesh sieves after pulverizing, gets minus sieve powder and carry out ultrasonic assistant extraction.
As a preferred detailed description of the invention, the invention provides a kind of preparation method of thalictrum spp extract, comprise the steps:
I. by thalictrum spp 40 ~ 50 DEG C oven dry, cross 40 mesh sieves after pulverizing, get minus sieve powder;
II. the mass percent concentration adding described thalictrum spp weight 20 ~ 30 times in the powder that step I obtains be 40% ~ 70% ethanol carry out ultrasonic assistant extraction, Extracting temperature is 40 DEG C ~ 60 DEG C, extraction time is 60min ~ 120min, and supersonic frequency is 30 ~ 40KHz; Filter, obtain alcohol extract;
III. the alcohol extract that obtains of Step II, decompression recycling ethanol, to without alcohol taste, to obtain final product.
As a preferred detailed description of the invention, the invention provides a kind of preparation method of thalictrum spp extract, comprise the steps:
I. by thalictrum spp 40 ~ 50 DEG C oven dry, cross 40 mesh sieves after pulverizing, get minus sieve powder;
II. the mass percent concentration adding described thalictrum spp weight 20 ~ 30 times in the powder that step I obtains be 50% ~ 60% ethanol carry out ultrasonic assistant extraction, Extracting temperature is 45 DEG C, extraction time is 60 ~ 90min, and supersonic frequency is 30 ~ 40KHz; Filter, obtain alcohol extract;
III. the alcohol extract that obtains of Step II, decompression recycling ethanol, to without alcohol taste, to obtain final product.
Preferably, thalictrum spp of the present invention is selected from SHUIHUANGLIAN and/or manyleaf meadowure rhizome and root.
As one preferred embodiment, the invention provides a kind of preparation method of SHUIHUANGLIAN extract, comprise the steps:
I. by SHUIHUANGLIAN 40 ~ 50 DEG C oven dry, cross 40 mesh sieves after pulverizing, get minus sieve powder;
II. the mass percent concentration adding described SHUIHUANGLIAN weight 20 times in the powder that step I obtains be 60% ethanol carry out ultrasonic assistant extraction, Extracting temperature is 45 DEG C, and extraction time is 60min, and supersonic frequency is 30 ~ 40KHz; Filter, obtain alcohol extract;
III. the alcohol extract that obtains of Step II, decompression recycling ethanol, to without alcohol taste, to obtain final product.
As another preferred implementation method, the invention provides a kind of preparation method of manyleaf meadowure rhizome and root extract, specifically comprise the steps:
I. by manyleaf meadowure rhizome and root 40 ~ 50 DEG C oven dry, cross 40 mesh sieves after pulverizing, get minus sieve powder;
II. the mass percent concentration adding described manyleaf meadowure rhizome and root weight 30 times in the powder that step I obtains be 50% ethanol carry out ultrasonic assistant extraction, Extracting temperature is 45 DEG C, and extraction time is 90min, and supersonic frequency is 30 ~ 40KHz; Filter, obtain alcohol extract;
III. the alcohol extract that obtains of Step II, decompression recycling ethanol, to without alcohol taste, to obtain final product.
Another object of the present invention, is the thalictrum spp extract providing above-mentioned preparation method to prepare; Particularly SHUIHUANGLIAN extract and manyleaf meadowure rhizome and root extract.
A further object of the invention, is to provide thalictrum spp or thalictrum spp extract for the preparation of the purposes in the pharmaceutical composition of control oranges and tangerines postharvest pathogen and/or the agent of oranges and tangerines post-harvest fresh-keeping; Preferably, described thalictrum spp extract is prepared by preparation method of the present invention.
Described oranges and tangerines postharvest pathogen is selected from Italian mould (Penicillium italicum), one or more in citric acid maize ear rot bacterium (Geotrichum candidum) and oranges and tangerines black rot (Alternaria citri).
Thus, the invention provides a kind of pharmaceutical composition preventing and treating oranges and tangerines postharvest pathogen, comprise the thalictrum spp extract that preparation method of the present invention prepares.
In addition, the present invention also provides the agent of a kind of oranges and tangerines post-harvest fresh-keeping, comprises the described thalictrum spp extract that preparation method of the present invention prepares.
Pharmaceutical composition of the present invention or the agent of oranges and tangerines post-harvest fresh-keeping, except described thalictrum spp extract, acceptable auxiliary material can also be comprised, such as be selected from the film forming agent such as carboxymethyl cellulose, shitosan, be selected from the cosolvent of glycerine, sucrose ester, tween etc., be selected from the anticorrisive agent of citric acid, calcium propionate, potassium sorbate or Sodium Benzoate etc., and suitable solvent, as water.
Ultrasonic wave has " cavitation effect ", the characteristic such as " mechanical oscillation " and " fuel factor ", " cavitation effect " can produce several thousand atmospheric pressures instantaneously, the micro-bubble of Extraction medium is compressed, bursts, the broken cell membrane extracting raw material, accelerate the stripping of natural medicinal component, " mechanical oscillation " and " fuel factor " further enhance the diffusion of stripping composition, make medicinal component reach concentration balance fast in Extraction medium, greatly can shorten extraction cost.And Extracting temperature is lower, avoids the destruction of long-time high-temperature water decoction to thalictrum spp chemical composition, improve the quality of products.
The extract that the present invention obtains for raw material with natural thalictrum spp, has very strong suppression Italy mould, the effect, particularly SHUIHUANGLIAN extract of oranges and tangerines black rot and sour germ and manyleaf meadowure rhizome and root extract.Pharmaceutical composition and the citrus preservative containing thalictrum spp extract of the present invention, without chemical residual, to human body without health hazard, environmentally safe, has good application prospect.
Accompanying drawing explanation
The photo display of Fig. 1 be in test example 1 thalictrum spp extract to the fungistatic effect of oranges and tangerines postharvest pathogen, arrow instruction be the edge of bacterial restrain; Wherein Figure 1A-1C shows the SHUIHUANGLIAN extract fungistatic effect to Italian mould, oranges and tangerines black rot and sour germ, and Fig. 1 D-1F shows the fungistatic effect of manyleaf meadowure rhizome and root extract to Italian mould, oranges and tangerines black rot and sour germ.
The photo display of Fig. 2 be that in test example 1, thalictrum spp extract inoculates the biocontrol effect of Italian mould to south, Jiangxi navel orange; Wherein Fig. 2 A shows SHUIHUANGLIAN extract inoculates Italian mould biocontrol effect to south, Jiangxi navel orange, and Fig. 2 B shows manyleaf meadowure rhizome and root extract inoculates Italian mould biocontrol effect to south, Jiangxi navel orange.
Detailed description of the invention
By concrete example given below, the invention will be further described, but it can not be interpreted as limitation of the invention.
Medicinal material, reagent, biomaterial that following embodiment is used, if no special instructions, all derive from commercial sources.
Embodiment 1 one kinds of SHUIHUANGLIAN extracts
Described SHUIHUANGLIAN extract is prepared as follows:
I. by SHUIHUANGLIAN 40 DEG C oven dry, cross 40 mesh sieves after pulverizing, get minus sieve powder;
II. the weight percent concentration adding described SHUIHUANGLIAN weight 20 times in the powder that step I obtains be 60% ethanol carry out ultrasonic assistant extraction, Extracting temperature is 45 DEG C, and extraction time is 1 hour, and supersonic frequency is 30KHz; Filter, obtain alcohol extract;
III. the alcohol extract that obtains of Step II, decompression recycling ethanol, to without alcohol taste, obtains described SHUIHUANGLIAN extract.
Embodiment 2 one kinds of manyleaf meadowure rhizome and root extracts
Prepare as follows:
I. by manyleaf meadowure rhizome and root 40 DEG C oven dry, cross 40 mesh sieves after pulverizing, get minus sieve powder;
II. the weight percent concentration adding described manyleaf meadowure rhizome and root weight 30 times in the powder that step I obtains be 50% ethanol carry out ultrasonic assistant extraction, Extracting temperature is 45 DEG C, and extraction time is 1 hour, and supersonic frequency is 30KHz; Filter, obtain alcohol extract;
III. the alcohol extract that obtains of Step II, decompression recycling ethanol to without alcohol taste, i.e. described manyleaf meadowure rhizome and root extract.
Test example 1 thalictrum spp extract is to the bacteriostasis of oranges and tangerines postharvest pathogenic fungi
Experiment material:
1. thalictrum spp extract:
SHUIHUANGLIAN extract prepared by embodiment 1, is mixed with distilled water the solution that concentration is 1g medicinal material/mL;
Manyleaf meadowure rhizome and root extract prepared by embodiment 2, is mixed with distilled water the solution that concentration is 1g medicinal material/mL.
2. strains tested: Italian mould (Penicillium italicum, ACCC30399) is from Chinese agriculture Culture Collection (ACCC); Citric acid maize ear rot bacterium (Geotrichum candidum) and oranges and tangerines black rot (Alternaria citri) laboratory are separated from south, the Jiangxi sick fruit of navel orange and obtain.
Experimental technique:
1, agar diffusion method measures thalictrum spp extract to the bacteriostasis of Italian mould
Get and be inoculated in test tube slant culture medium for examination bacterium, cultivate 2-3d for 30 DEG C, will wash in triangular flask for examination bacterium PDB nutrient solution by spore in test-tube culture medium, shake up, bead be broken up, and makes spore suspension, counts, make bacterial concentration reach 10 with blood counting chamber
6-10
7cfu/mL is for subsequent use.
Draw above-mentioned pityrosporion ovale suspension 5mL to be mixed into and to be cooled to about 50 DEG C PDA culture mediums, preparation band bacterium culture medium.Then place aseptic Oxford cup, respectively at adding thalictrum spp extract solution described in 200 μ L in the cup of Oxford, being placed in 28 DEG C of insulating boxs and cultivating 48h, right-angled intersection method measures the size of inhibition zone.Replication three times, calculates inhibition zone diameter mean value.
2, growth rate method measures the thalictrum spp extract bacteriostasis to citric acid maize ear rot bacterium and black rot
Under aseptic condition, with pure PDA culture medium for contrast, the each colony edge bacterium block for examination bacterium got and activated is beaten with diameter 6mm card punch, be transferred to containing in the PDA culture medium of described thalictrum spp extract and control medium respectively, each process in triplicate, and is placed in 28 DEG C of constant incubators and cultivates, and records each colony growth situation after 5d, and measure colony diameter by right-angled intersection method, calculate inhibiting rate.
Inhibiting rate (%)=100 × (contrast colony diameter-process colony diameter)/contrast colony diameter
3, doubling dilution measures minimum inhibitory concentration (MIC) and the MBC (MFC) of thalictrum spp extract
By ten concentration gradients between described thalictrum spp extract solution doubling dilution to 1.95mg medicinal material/mL ~ 1000mg medicinal material/mL, then the thalictrum spp extract solution of 10mL variable concentrations is drawn in the PDA culture medium do not melted containing 90mL, pour into after shaking up in flat board, make pastille culture medium, after culture medium solidifying, evenly be applied on pastille PDA culture medium by Italian mould pityrosporion ovale suspension with spread plate, each concentration in triplicate.And blank group (culture medium does not inoculate bacterium), bacterium liquid control group (culture medium inoculated bacterium but not containing thalictrum spp alcohol extract) are set.The incubator that test group and control group are placed in 28 DEG C is cultivated 2d, in test group with the culture dish of asepsis growth minimum containing concentration be its MIC, being continued to cultivate after 7d by the culture dish of all asepsis growths and observe, is its MFC with the culture dish of asepsis growth minimum containing concentration.
4, thalictrum spp extract is to the biocontrol effect of south, Jiangxi navel orange
By south, Jiangxi Navel Orange Fruits with after the process of 2%NaClO solution disinfection, cleaner with running water, dry for subsequent use.Stab a diameter 4mm with transfer needle at fruit waist, the wound of degree of depth 3mm, inoculate thalictrum spp extract and the 20 μ L 1 × 10 of 20 μ L variable concentrations with micropipettor
6cFU/mL Italy mould spore suspension, test point 3 process: 1. citrusfruit inoculates Italian mould spore suspension, as test blank; 2. Italian mould spore suspension is inoculated after thalictrum spp extract-treated citrusfruit wound; 3. Prochloraz positive controls.
Fruit is placed on pallet, puts bag film, and each process 9 fruits, repeat 3 times.Keep relative humidity about 95%, after 27 DEG C of cultivation 7d, investigation citrusfruit lesion diameter, according to following formulae discovery prevention effect.
Prevention effect (%)=100 × (control group lesion diameter-processed group lesion diameter)/control group lesion diameter
Experimental result
1, thalictrum spp extract is to the inhibitory action for examination pathogen
Result is see table 1 and Fig. 1.SHUIHUANGLIAN and manyleaf meadowure rhizome and root extract have significant bacteriostasis (respectively see Figure 1A and 1D) to Italian mould under the concentration of 1g medicinal material/mL, the size of the antibacterial circle diameter of SHUIHUANGLIAN is 47.4 ± 1.1mm, and the size of the antibacterial circle diameter of manyleaf meadowure rhizome and root is 53.1 ± 0.7mm.SHUIHUANGLIAN and the inhibiting rate of manyleaf meadowure rhizome and root extract to oranges and tangerines black rot and citric acid maize ear rot bacterium all reached for 100% (respectively see Figure 1B, Fig. 1 C, Fig. 1 E and Fig. 1 F).Simultaneously with the Prochlorazs of 1000 times of dilutions for positive control, find that the bacteriostasis of SHUIHUANGLIAN and manyleaf meadowure rhizome and root extract is suitable with 1000 times of Prochlorazs diluted.
Table 1 thalictrum spp extract is to the bacteriostasis of Italian mould
2, thalictrum spp alcohol extract suppresses MIC and MFC of Italian mould
SHUIHUANGLIAN is respectively 6.25mg medicinal material/mL and 50mg medicinal material/mL to Italian mould minimum inhibitory concentration (MIC) and MBC (MFC), MIC and MFC of manyleaf meadowure rhizome and root extract to Italian mould is respectively 6.25mg medicinal material/mL and 25mg medicinal material/mL.
3, thalictrum spp alcohol extract is to the biocontrol effect of south, Jiangxi navel orange
Result is see Fig. 2 and table 2.
Table 2 thalictrum spp alcohol extract is to the biocontrol effect of south, Jiangxi navel orange
The SHUIHUANGLIAN extract of 100mg medicinal material/mL and manyleaf meadowure rhizome and root extract are respectively 56.86% and 68.62% to south, the Jiangxi navel orange disease-controlling effect that Italian mould causes.Simultaneously with 500 times of dilutions and 1000 times of Prochlorazs diluted for positive control, find the Prochloraz strong (37.5%) of the prevention effect of two kinds of extracts of above-mentioned concentration than 1000 times of dilutions, the effect of SHUIHUANGLIAN extract is weaker than the prevention effect of the Prochloraz (68.1%) of 500 times of dilutions, and the effect of manyleaf meadowure rhizome and root is suitable with it.Show as linear relationship between the logarithm value of thalictrum spp extract concentrations and prevention effect, the linear equation of SHUIHUANGLIAN extract is Y=7.0159+1.8797X, as calculated EC
50for 84.6mg/mL; The linear equation of manyleaf meadowure rhizome and root extract is Y=6.7202+1.5029X, as calculated EC
50for 45.8mg/mL.
Be linked as the thalictrum spp of representative with SHUIHUANGLIAN and horse hair, its extract is adopted rear multiple pathogen to oranges and tangerines and is had very strong inhibitory action, effectively can prevent and treat the microbial oranges and tangerines postharvest disease of these cause of diseases.
The agent of embodiment 3 one kinds of oranges and tangerines post-harvest fresh-keepings
Prescription: SHUIHUANGLIAN extract 100mg prepared by embodiment 1, carboxymethyl cellulose 10g, potassium citrate 0.6g, sucrose ester 5g, glycerine 5g, calcium propionate 10g, surplus is distilled water, makes 1000mL altogether.
Preparation method: the carboxymethyl cellulose taking recipe quantity, adds the swelling 8h of appropriate distilled water, then adds distilled water stirring preparation cmc soln.Then add citric acid, sucrose ester, glycerine, calcium propionate and SHUIHUANGLIAN extract successively, with distilled water diluting to 1000mL, stir, to obtain final product.
The agent of embodiment 4 one kinds of oranges and tangerines post-harvest fresh-keepings
Prescription: manyleaf meadowure rhizome and root extract 100mg prepared by embodiment 2, carboxymethyl cellulose 10g, potassium citrate 0.6g, sucrose ester 5g, glycerine 5g, calcium propionate 10g, surplus is distilled water, makes 1000mL altogether.
Preparation method: the carboxymethyl cellulose taking recipe quantity, adds the swelling 8h of appropriate distilled water, then adds distilled water stirring preparation cmc soln.Then add citric acid, sucrose ester, glycerine, calcium propionate and manyleaf meadowure rhizome and root extract successively, with distilled water diluting to 1000mL, stir, to obtain final product.
Embodiment 5-19 SHUIHUANGLIAN extract
Prepare by the following method:
Step I ~ III and embodiment 1 with, but in Step II except Extracting temperature, each technological parameter is all different from embodiment 1, and the technological parameter of each embodiment refers to table 3.
The process conditions of the ultrasonic extraction of table 3 embodiment 5-19
Test example 2 SHUIHUANGLIAN extract is to the bacteriostasis of oranges and tangerines postharvest pathogenic fungi
Experiment material:
1. supplying reagent thing: SHUIHUANGLIAN extract prepared by embodiment 5-19, is the solution of 1g medicinal material/mL with distilled water compound concentration.
2. strains tested: Italian mould (Penicillium italicum, ACCC30399).
Experimental technique:
1, agar diffusion method measures SHUIHUANGLIAN extract to the bacteriostasis of Italian mould
With test example 1.
Experimental result: in table 4.
Table 4 SHUIHUANGLIAN extract is to the bacteriostasis of Italian mould
| Embodiment | Antibacterial circle diameter | Embodiment | Antibacterial circle diameter | Embodiment | Antibacterial circle diameter |
| 5 | 36.17±0.55 | 10 | 40.17±0.55 | 15 | 47.67±0.52 |
| 6 | 38.83±0.41 | 11 | 37.50±0.41 | 16 | 43.27±0.75 |
| 7 | 37.00±0.98 | 12 | 36.50±0.98 | 17 | 40.17±0.82 |
| 8 | 34.17±0.52 | 13 | 44.17±0.52 | 18 | 38.67±0.52 |
| 9 | 39.00±0.52 | 14 | 45.83±0.63 | 19 | 37.00±0.52 |
Result of the test shows, the extracting factor described in embodiment of the present invention 5-19 extracts the SHUIHUANGLIAN extract obtained fungistatic effect, but the strongest with the ethanol extract effect of embodiment 15, is secondly embodiment 14,13 and 16.
Embodiment 20-34 manyleaf meadowure rhizome and root extract
Prepare by the following method:
Step I ~ III and embodiment 2 with, but in Step II except Extracting temperature, other technological parameter is all different from embodiment 2, the concrete technological parameter of each embodiment is in table 5.
The process conditions of the ultrasonic extraction of table 5 embodiment 20-34
Test example 3 manyleaf meadowure rhizome and root extract is to the bacteriostasis of oranges and tangerines postharvest pathogenic fungi
Experiment material:
1. supply reagent thing: manyleaf meadowure rhizome and root extract prepared by embodiment 20-34, concentration is the solution of 1g medicinal material/mL.
2. strains tested: Italian mould (Penicillium italicum, ACCC30399).
Experimental technique:
1, agar diffusion method measures manyleaf meadowure rhizome and root extract to the bacteriostasis of Italian mould
With test example 1.
Experimental result: in table 6.
Table 6 manyleaf meadowure rhizome and root extract is to the bacteriostasis of Italian mould
| Embodiment | Antibacterial circle diameter | Embodiment | Antibacterial circle diameter | Embodiment | Antibacterial circle diameter |
| 20 | 38.33±0.89 | 25 | 41.17±1.03 | 30 | 53.83±0.89 |
| 21 | 40.67±0.82 | 26 | 43.50±1.05 | 31 | 53.17±0.82 |
| 22 | 41.00±0.98 | 27 | 41.33±1.03 | 32 | 47.67±0.75 |
| 23 | 41.67±1.03 | 28 | 52.67±0.75 | 33 | 45.00±0.89 |
| 24 | 40.50±0.89 | 29 | 55.50±1.36 | 34 | 41.67±0.75 |
Result of the test shows, the extracting factor described in embodiment of the present invention 20-34 extracts the manyleaf meadowure rhizome and root extract obtained fungistatic effect, but the strongest with the ethanol extract effect of embodiment 29, is secondly embodiment 30,31 and 28.
The result of compbined test example 2 and 3, optimizes a kind of preparation method of thalictrum spp extract, comprises the steps:
I. by thalictrum spp 40 ~ 50 DEG C oven dry, cross 40 mesh sieves after pulverizing, get minus sieve powder;
II. the mass percent concentration adding described thalictrum spp weight 20 ~ 30 times in the powder that step I obtains be 40% ~ 70% ethanol carry out ultrasonic assistant extraction, Extracting temperature is 40 DEG C ~ 60 DEG C, extraction time is 60min ~ 120min, and supersonic frequency is 30 ~ 40KHz; Filter, obtain alcohol extract;
III. the alcohol extract that obtains of Step II, decompression recycling ethanol, to without alcohol taste, to obtain final product.
The preparation method of the preferred thalictrum spp extract of the present invention, comprises the steps:
I. by thalictrum spp 40 ~ 50 DEG C oven dry, cross 40 mesh sieves after pulverizing, get minus sieve powder;
II. the mass percent concentration adding described thalictrum spp weight 20 ~ 30 times in the powder that step I obtains be 50% ~ 60% ethanol carry out ultrasonic assistant extraction, Extracting temperature is 45 DEG C, extraction time is 60min ~ 90min, and supersonic frequency is 30 ~ 40KHz; Filter, obtain alcohol extract;
III. the alcohol extract that obtains of Step II, decompression recycling ethanol, to without alcohol taste, to obtain final product.
In a word, the invention provides and a kind ofly rear multiple pathogen is adopted to oranges and tangerines there is very strong inhibiting thalictrum spp extract, effectively can prevent and treat the microbial oranges and tangerines postharvest disease of these cause of diseases.
Claims (10)
1. a preparation method for thalictrum spp extract, adopts ultrasonic assistant extractive technique, is Extraction solvent, comprises the steps: with ethanol
Be the ethanol of 40% ~ 100% by the mass percent concentration adding thalictrum spp weight 10 ~ 40 times in described thalictrum spp, it is 40 ~ 60 DEG C at Extracting temperature, extraction time is 30min ~ 120min, and supersonic frequency is extract under the condition of 30 ~ 40KHz; Filter, reclaim ethanol, concentrated, dry or moist, to obtain final product.
2. preparation method according to claim 1, is characterized in that: the weight of ethanol is described thalictrum spp weight 10 ~ 30 times, is preferably 20 ~ 30 times;
Preferably, the mass percent concentration of ethanol is 40% ~ 70%, is more preferably 50 ~ 60%;
Preferably, extraction time is 60 ~ 120min, is more preferably 60 ~ 90min;
Preferably, the temperature that ultrasonic assistant extracts is 45 DEG C.
3. preparation method according to claim 1 and 2, is characterized in that: before alcohol extract, described thalictrum spp 40 ~ 50 DEG C oven dry, crosses 40 mesh sieves, get minus sieve powder and carry out ultrasonic assistant extraction after pulverizing.
4. a preparation method for thalictrum spp extract, adopts ultrasonic assistant extractive technique, is Extraction solvent, comprises the steps: with ethanol
I. by thalictrum spp 40 ~ 50 DEG C oven dry, cross 40 mesh sieves after pulverizing, get minus sieve powder;
II. the mass percent concentration adding described thalictrum spp weight 20 ~ 30 times in the powder that step I obtains be 40% ~ 70% ethanol carry out ultrasonic assistant extraction, Extracting temperature is 40 DEG C ~ 60 DEG C, extraction time is 60min ~ 120min, and supersonic frequency is 30 ~ 40KHz; Filter, obtain alcohol extract;
III. the alcohol extract that obtains of Step II, decompression recycling ethanol, to without alcohol taste, to obtain final product;
Preferred technical scheme is:
I. by thalictrum spp 40 ~ 50 DEG C oven dry, cross 40 mesh sieves after pulverizing, get minus sieve powder;
II. the mass percent concentration adding described thalictrum spp weight 20 ~ 30 times in the powder that step I obtains be 50% ~ 60% ethanol carry out ultrasonic assistant extraction, Extracting temperature is 45 DEG C, extraction time is 60 ~ 90min, and supersonic frequency is 30 ~ 40KHz; Filter, obtain alcohol extract;
III. the alcohol extract that obtains of Step II, decompression recycling ethanol, to without alcohol taste, to obtain final product.
5. preparation method according to any one of claim 1 to 4, is characterized in that: described thalictrum spp is selected from SHUIHUANGLIAN and/or manyleaf meadowure rhizome and root.
6. the thalictrum spp extract that the preparation method according to any one of claim 1 to 5 prepares.
7. thalictrum spp or thalictrum spp extract are for the preparation of the purposes in the pharmaceutical composition of control oranges and tangerines postharvest pathogen and/or the agent of oranges and tangerines post-harvest fresh-keeping; Preferably, described thalictrum spp extract is prepared by the preparation method according to any one of claim 1 to 5.
8. purposes according to claim 7, it is characterized in that, described oranges and tangerines postharvest pathogen is selected from Italian mould (Penicillium italicum), one or more in citric acid maize ear rot bacterium (Geotrichum candidum) and oranges and tangerines black rot (Alternaria citri).
9. prevent and treat a pharmaceutical composition for oranges and tangerines postharvest pathogen, comprise thalictrum spp extract according to claim 6.
10. an oranges and tangerines post-harvest fresh-keeping agent, comprises thalictrum spp extract according to claim 6.
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| CN105496872A (en) * | 2015-12-25 | 2016-04-20 | 肇庆迪彩日化科技有限公司 | Sun-blocking composition and daily chemical product with sun-blocking effect |
| CN105613723A (en) * | 2016-03-04 | 2016-06-01 | 珠海真绿色技术有限公司 | Fruit and vegetable preservative and preparation method thereof |
| CN106106550A (en) * | 2016-07-13 | 2016-11-16 | 彭水县渝宏清脆李种植股份合作社 | A kind of compositions preventing and treating blue or green crisp plum red leaf spot |
| KR20190052256A (en) * | 2017-11-08 | 2019-05-16 | 대한민국(농촌진흥청장) | Antimicrobial composition comprising Thalictrum uchiyamai extracts and fractions for cladobotryum mycophilum and uses thereof |
| KR20190052254A (en) * | 2017-11-08 | 2019-05-16 | 대한민국(농촌진흥청장) | Antimicrobial composition comprising Thalictrum uchiyamai extracts and fractions for Blue Mold, and uses thereof |
| CN113200989A (en) * | 2021-05-18 | 2021-08-03 | 云南民族大学 | Preparation method and application of chromone alkaloid compound |
| CN113234084A (en) * | 2021-05-18 | 2021-08-10 | 云南民族大学 | Compound with anti-rotavirus activity in agrimony and thalictrum aquilegifolium and preparation method thereof |
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| CN105496872A (en) * | 2015-12-25 | 2016-04-20 | 肇庆迪彩日化科技有限公司 | Sun-blocking composition and daily chemical product with sun-blocking effect |
| CN105496872B (en) * | 2015-12-25 | 2019-09-13 | 肇庆迪彩日化科技有限公司 | A kind of sunscreen composition and the daily chemical product with sun-proof function |
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| CN106106550A (en) * | 2016-07-13 | 2016-11-16 | 彭水县渝宏清脆李种植股份合作社 | A kind of compositions preventing and treating blue or green crisp plum red leaf spot |
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| KR20190052254A (en) * | 2017-11-08 | 2019-05-16 | 대한민국(농촌진흥청장) | Antimicrobial composition comprising Thalictrum uchiyamai extracts and fractions for Blue Mold, and uses thereof |
| KR102069118B1 (en) * | 2017-11-08 | 2020-01-22 | 대한민국 | Antimicrobial composition comprising Thalictrum uchiyamai extracts and fractions for Blue Mold, and uses thereof |
| KR102069114B1 (en) * | 2017-11-08 | 2020-01-22 | 대한민국 | Antimicrobial composition comprising Thalictrum uchiyamai extracts and fractions for cladobotryum mycophilum and uses thereof |
| CN113200989A (en) * | 2021-05-18 | 2021-08-03 | 云南民族大学 | Preparation method and application of chromone alkaloid compound |
| CN113234084A (en) * | 2021-05-18 | 2021-08-10 | 云南民族大学 | Compound with anti-rotavirus activity in agrimony and thalictrum aquilegifolium and preparation method thereof |
| CN113234084B (en) * | 2021-05-18 | 2023-05-05 | 云南民族大学 | Compound with anti-rotavirus activity in Meadowrue of Crane celebration and preparation method thereof |
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