CN109916992A - A kind of Mass Spectrometric Identification detection method of Wild Edible Fungi in Yunnan species - Google Patents
A kind of Mass Spectrometric Identification detection method of Wild Edible Fungi in Yunnan species Download PDFInfo
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- CN109916992A CN109916992A CN201910300856.3A CN201910300856A CN109916992A CN 109916992 A CN109916992 A CN 109916992A CN 201910300856 A CN201910300856 A CN 201910300856A CN 109916992 A CN109916992 A CN 109916992A
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- 241000894007 species Species 0.000 title claims abstract description 41
- 241000233866 Fungi Species 0.000 title claims abstract description 29
- 238000001514 detection method Methods 0.000 title claims abstract description 23
- 235000001674 Agaricus brunnescens Nutrition 0.000 claims abstract description 124
- BDAGIHXWWSANSR-UHFFFAOYSA-N methanoic acid Natural products OC=O BDAGIHXWWSANSR-UHFFFAOYSA-N 0.000 claims abstract description 58
- 238000000034 method Methods 0.000 claims abstract description 51
- 239000010414 supernatant solution Substances 0.000 claims abstract description 33
- OSWFIVFLDKOXQC-UHFFFAOYSA-N 4-(3-methoxyphenyl)aniline Chemical compound COC1=CC=CC(C=2C=CC(N)=CC=2)=C1 OSWFIVFLDKOXQC-UHFFFAOYSA-N 0.000 claims abstract description 29
- 235000019253 formic acid Nutrition 0.000 claims abstract description 29
- 230000010355 oscillation Effects 0.000 claims abstract description 22
- WEVYAHXRMPXWCK-UHFFFAOYSA-N Acetonitrile Chemical compound CC#N WEVYAHXRMPXWCK-UHFFFAOYSA-N 0.000 claims description 48
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 claims description 12
- 238000005119 centrifugation Methods 0.000 claims description 8
- 239000007788 liquid Substances 0.000 claims description 8
- 239000000203 mixture Substances 0.000 claims description 8
- 229910052757 nitrogen Inorganic materials 0.000 claims description 6
- 238000001819 mass spectrum Methods 0.000 claims description 3
- 238000012360 testing method Methods 0.000 abstract description 8
- 239000000523 sample Substances 0.000 description 37
- 108020004414 DNA Proteins 0.000 description 10
- 239000006228 supernatant Substances 0.000 description 6
- 241000894006 Bacteria Species 0.000 description 5
- 208000005374 Poisoning Diseases 0.000 description 5
- 235000013305 food Nutrition 0.000 description 5
- 231100000572 poisoning Toxicity 0.000 description 5
- 230000000607 poisoning effect Effects 0.000 description 5
- 108090000623 proteins and genes Proteins 0.000 description 5
- 239000000243 solution Substances 0.000 description 5
- 241000121220 Tricholoma matsutake Species 0.000 description 4
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- 108091028043 Nucleic acid sequence Proteins 0.000 description 1
- 238000001237 Raman spectrum Methods 0.000 description 1
- 241000222562 Suillus Species 0.000 description 1
- 150000001413 amino acids Chemical class 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 238000012455 bioassay technique Methods 0.000 description 1
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Abstract
The present invention relates to a kind of Mass Spectrometric Identification detection methods of Wild Edible Fungi in Yunnan species, and the method includes the steps of, and step (1) grinds wild mushroom mushroom；Step (2) takes the wild fungus of crushing to be packed into centrifuge tube, and formic acid is added and mixes well；Step (3) oscillation shakes up, and is centrifuged, taking supernatant solution is the sample prepared；Step (4) sample-taking spot is to target center；Step (5) carries out target practice identification using Mass Spectrometric Identification system.Method of the invention substantially reduces the detection time of Wild Edible Fungi in Yunnan species identification, and testing result is stablized, will quick and simplicity carry out wild mushroom species identification in the season of a large amount of fruiting listings of wild mushroom.
Description
Technical field
The invention belongs to bio-assay technique field, the Mass Spectrometric Identification of especially a kind of suitable Wild Edible Fungi in Yunnan species is detected
Method and technology field.
Background technique
More than global Known Edible Mushrooms 2000, China accounts for 978 kinds, and the Yunnan Province as Kingdom of Animals and Plants good reputation up to has
882 kinds, type accounts for 90% or more of the total type of national wild mushroom.16, Yunnan Province ground (state), city have the distribution of wild mushroom,
About 500,000 tons of its natural production, height ranks first in the country, and the wild edible fungus in 70% or more the whole nation is that is produced from Yunnan Province.
Wild edible fungus delicious flavour, superior in taste, strong flavor are unique, amino acid is abundant, fatty low, dietary fiber
It is high, minerals and vitamins are abundant, the natural food with food therapy effect, deep to be liked by masses.The wild mushroom in Yunnan is not only
Domestic many provinces are sold to, numerous countries such as the U.S., Japan, South Korea, European Union, Philippine, Malaysia is also exported to, is earned for China
Take a large amount of foreign exchange earnings.Since the 21th century, Yunnan Provincial Party committee, provincial government are using edible mushroom as plateau strong industry, characteristic
Modern agriculture industry vigorously supports its development of promotion in turn.Yunnan mushroom industry has become peasant in Plateau Characteristic agricultural and increases
Receipts get rich, the outstanding specialty industries with broad based growth potentiality.
However, wild mushroom is many kinds of, kind (complexion) is similar, always exists identification unclear the case where eating poisoning by mistake.It is wild
The poisoning of raw bacterium has become that China is common and the higher sitotoxismus reason of lethality and each year Yunnan Province's food safety risk
Hidden danger most distinct issues.The event that Yunnan Province has edible wild bacterium to be poisoned every year, What is more for up to 40 people's
Collective poisoning matters of aggravation.
Rapid verification, which has toadstool, especially mode of appearance to be broken the wild of ring, toadstool, and ascertains whether by wild
Food accident etc. caused by bacterium is poisoned is required to carry out species identification to wild mushroom.In addition, most wild edible fungus are current
It is unable to artificial growth, belongs to pure natural biological species resource, is precious food materials or medicinal material that the Nature vouchsafes the mankind.Object is with dilute
To be expensive, selling price remains high throughout the year, such as 800 yuan/kilogram of matsutake, 1200/ kilogram of bacterium of drying up, in great number economic interests
It drives down, is easy to appear the case where bad retailer is adulterated, mix, kind substitution wild mushroom.When expanding domestic and international trade, also need
The official to carry out identity examination to wild mushroom species proves, but currently, quickly reflects without reference to wild mushroom species both at home and abroad
Other standard method.
Applicant it is very long by 3 years experimental studies have found that, existing detection method realizes the species identification of Wild Edible Fungi in Yunnan
There are numerous defects, is described below.
Method 1: the method for the DNA sequencing in the method for species identification is the species mirror of vast research institutions and research institution
Unusual research means, principle is to be compared according to DNA sequence dna information with the database of gene bank, to obtain object
Kind information.Can this technology be extracted by DNA, and can the linear mass of DNA meet requirement of experiment, and can DNA mass, which reach, wants
It asks, whether the database of gene bank has the influence of the problems such as data information of this species.Through inquiring, gene bank data
Library only has matsutake and a kind of whole genome sequence information of Suillus albidipes (Peck) Sing, and DNA bar code has much, but whether exhaustive Yunnan is all
The type of wild mushroom is apt to belong to unknown.At present from extract DNA to obtain analysis data at least need 8 hours, in addition, detection at
This is very expensive, and about 300 yuan/sample, and detect gene sequencer price used and be up to 850,000 RMB or more, such great number
Instrument and the not general detection class laboratory of use cost can have.
Method 2: the round pcr identified for species includes regular-PCR technology and real-time fluorescence PCR, is also research institutions
The technological means that biological species identify is used for testing agency.Can DNA be also depended on extract, and can the linear mass of DNA
Meet requirement of experiment, can DNA mass reach the influence of requirement.And the specificity of primer or probe, accuracy need system
The process of property groped and optimized, and the method system of each species needs to be established one by one.From DNA is extracted to being divided
Analysis data at least about need the time of 3 hours, about 500 yuan/sample of testing cost.
Method 3: the foundation of genetic chip needs to be integrated into high-throughput detection means on the basis of PCR establishes mature.
The time of 8 hours, about 500 yuan/sample of testing cost are at least about needed to analysis data are obtained from DNA is extracted.
Method 4: the identification of Raman spectrum and near infrared spectrum depends on size, the shape of sample, for being cut into silk, being cut into
Block, the sample of frost, freeze-drying sample, must all establish a large amount of database can be only used for identifying, at present such lossless mirror
It is fixed no for biological sample, and the substance characteristics of each of the wild edible fungus in Yunnan are unclear, so can not use
The method is rapidly performed by species identification.
Applicant is in 3 years it was found that, above-mentioned detection method is not able to satisfy Wild Edible Fungi in Yunnan species Rapid identification
Needs, predominantly two aspects: 1) for Wild Edible Fungi in Yunnan when being exported to foreign countries through trade, the states such as Malaysia, Philippine are at present
It is required that prove that the requirement becomes new international trade rule gradually with the official of the subsidiary related wild mushroom species identification of goods, because
This China is also researching and developing relevant national standard to meet the needs of the following international market；2) in terms of the people's livelihood: domestic or province
Interior citizen is badly in need of carrying out species identification to wild mushroom after eating wild mushroom poisoning by mistake, and then hospital could suit the medicine to the illness and make a definite diagnosis, and is solved
Poison treatment, up to the very long waiting of a few houres, sacrifice be citizen life and health interests, or even the cost to be given one's life for.
Therefore it is badly in need of controlling the species identification time of wild mushroom within half an hour, thus to meet, save the doctor of poisoner's life
It treats and needs.
Summary of the invention
The present invention is precisely in order to the defect that solves the above problems, provides a kind of Mass Spectrometric Identification detection side of Wild Edible Fungi in Yunnan species
Method.
The present invention adopts the following technical scheme that realization.
A kind of Mass Spectrometric Identification detection method of Wild Edible Fungi in Yunnan species, the method includes the steps of by the present invention, step (1)
Wild mushroom mushroom is ground；Step (2) takes the wild fungus of crushing to be packed into centrifuge tube, and formic acid is added and mixes well；Step
Suddenly (3) oscillation shakes up, and is centrifuged, taking supernatant solution is the sample prepared；Step (4) sample-taking spot is to target center；Step (5)
Target practice identification is carried out using Mass Spectrometric Identification system.
It is further that the method includes the steps of by the present invention, and step (1) grinds wild mushroom mushroom；Step (2)
It takes the wild mushroom mushroom of crushing to be packed into centrifuge tube, formic acid is added and acetonitrile mixes well；Step (3) oscillation shakes up, and is centrifuged, takes
Supernatant solution is the sample prepared；Step 4) sample-taking spot is dried to target center；Step (5) uses Mass Spectrometric Identification system
Carry out identification target practice identification of practicing shooting.
It is further that the method includes the steps of by the present invention, and step (1) grinds wild mushroom mushroom；Step (2)
It takes the wild mushroom mushroom of crushing to be packed into centrifuge tube, formic acid is added and acetonitrile mixes well；Step (3) oscillation shakes up, and is centrifuged, takes
Supernatant solution is the sample prepared；Step (4) sample-taking spot is dried to target center；Supernatant solution is added dropwise in step (5) again,
It dries again, then reuses Mass Spectrometric Identification system and carry out target practice identification.
It is further that the method includes the steps of by the present invention, and step (1) grinds wild mushroom mushroom；Step (2)
It takes the wild mushroom mushroom 100-5000mg of crushing to be packed into centrifuge tube, 70% formic acid of 10-5000 μ L concentration and 20-3000 μ L is added
Acetonitrile mixes well；Step (3) oscillation shakes up 5-20min, and revolving speed is no less than 10000r/min centrifugation 1min or more, takes supernatant molten
Liquid is the sample prepared；Step (4) takes 1-5 μ L sample point to target center, dries；1-10 μ L supernatant is added dropwise in step (5) again
Solution, then dry, it then reuses Mass Spectrometric Identification system and carries out target practice identification.
It is further that the method includes the steps of by the present invention, and step (1) grinds wild mushroom mushroom；Step (2)
It takes the wild mushroom mushroom 200-500mg of crushing to be packed into centrifuge tube, 70% formic acid of 10-500 μ L concentration and 20-600 μ L second is added
Nitrile mixes well；Step (3) oscillation shakes up 10-15min, and revolving speed is no less than 12000r/min centrifugation 1.5min or more, takes supernatant
Solution is the sample prepared；Step (4) takes 1-3 μ L sample point to target center, dries；1-5 μ L supernatant is added dropwise in step (5) again
Solution, then dry, it then reuses Mass Spectrometric Identification system and carries out target practice identification.
Further be, in this method step (2) of the present invention the volume ratio of added 70% formic acid of concentration and acetonitrile be 1:1~
1.2。
It is further that the volume ratio of added 70% formic acid of concentration and acetonitrile is 1:1 in this method step (2) of the present invention.
It is further that the method includes the steps of by the present invention, and step (1) grinds wild mushroom mushroom；Step (2)
It takes the wild mushroom mushroom 200mg of crushing to be packed into centrifuge tube, 50 μ L concentration, 70% formic acid is added and 50 μ L acetonitriles mix well；Step
Suddenly (3) oscillation shakes up 10min, and revolving speed 12000r/min is centrifuged 2min, and taking supernatant solution is the sample prepared；Step (4) takes 1
μ L sample point is dried to target center；1 μ L supernatant solution is added dropwise in step (5) again, then dries, and then reuses Mass Spectrometric Identification system
System carries out target practice identification.
It is further that step (1) of the present invention grinds specifically, wild mushroom mushroom is under liquid nitrogen frozen state,
Up to fine-powdered.
It is further that step (1) of the present invention grinds it specifically, wild mushroom mushroom is under liquid nitrogen frozen state
It is preceding first to be blended with tissue grinder instrument.
Mass Spectrometric Identification system set forth in the present invention is Matrix-assisted laser desorption ionization identification systems,
The system is described as https: //baike.baidu.com/item/%E5%9F%BA%E8%B4%A8%E8%BE%
85%E5%8A%A9%E6%BF%80%E5%85%89%E8%A7%A3%E5%90%B8%E7%94%B5 %
E7%A6%BB%E9%A3%9E%E8%A1%8C%E6%97%B6%E9%97%B4%E8%B4%A8%E8 %
B0%B1/3833495? fr=aladdin.
Beneficial effects of the present invention are, in conjunction with method and Matrix-assisted laser desorption ionization of the invention
Identification systems (instrument) detect the proteins and peptides in wild mushroom, compare the species specific mass-spectrogram of wild mushroom object, thus real
The purpose of existing precise Identification wild mushroom.The method requires considerably less, 10 minutes/sample of pre-treatment step, mass spectral analysis 2 to sample size
Minute/sample, total time are only 12 minutes；And testing cost is only 20 yuan/sample.Method of the invention substantially reduces Yunnan Wild
The detection time of bacterium species identification, and testing result is stablized, it, will be quickly and easy in the season of a large amount of fruiting listings of wild mushroom
Carry out wild mushroom species identification.
The present invention is further explained with reference to the accompanying drawings and detailed description.
Detailed description of the invention
Fig. 1 is using the method for the present invention to adopting to the map of five, Yunnan Province not ferfas Mass Spectrometric Identification of region；
Fig. 2 is using the method for the present invention to adopting to the map of six, Yunnan Province not cepe Mass Spectrometric Identification of region；
Fig. 3 is using the method for the present invention to adopting to the map of four, Yunnan Province not matsutake Mass Spectrometric Identification of region.
Specific embodiment
A kind of Mass Spectrometric Identification detection method of Wild Edible Fungi in Yunnan species, the method includes the steps of by the present invention, step (1)
Wild mushroom mushroom is ground；Step (2) takes the wild fungus of crushing to be packed into centrifuge tube, and formic acid is added and mixes well；Step
Suddenly (3) oscillation shakes up, and is centrifuged, taking supernatant solution is the sample prepared；Step (4) sample-taking spot is to target center；Step (5)
Target practice identification is carried out using Mass Spectrometric Identification system.
It is further that the method includes the steps of by the present invention, and step (1) grinds wild mushroom mushroom；Step (2)
It takes the wild mushroom mushroom of crushing to be packed into centrifuge tube, formic acid is added and acetonitrile mixes well；Step (3) oscillation shakes up, and is centrifuged, takes
Supernatant solution is the sample prepared；Step 4) sample-taking spot is dried to target center；Step (5) uses Mass Spectrometric Identification system
Carry out identification target practice identification of practicing shooting.
It is further that the method includes the steps of by the present invention, and step (1) grinds wild mushroom mushroom；Step (2)
It takes the wild mushroom mushroom of crushing to be packed into centrifuge tube, formic acid is added and acetonitrile mixes well；Step (3) oscillation shakes up, and is centrifuged, takes
Supernatant solution is the sample prepared；Step (4) sample-taking spot is dried to target center；Supernatant solution is added dropwise in step (5) again,
It dries again, then reuses Mass Spectrometric Identification system and carry out target practice identification.
It is further that the method includes the steps of by the present invention, and step (1) grinds wild mushroom mushroom；Step (2)
It takes the wild mushroom mushroom 100-5000mg of crushing to be packed into centrifuge tube, 70% formic acid of 10-5000 μ L concentration and 20-3000 μ L is added
Acetonitrile mixes well；Step (3) oscillation shakes up 5-20min, and revolving speed is no less than 10000r/min centrifugation 1min or more, takes supernatant molten
Liquid is the sample prepared；Step (4) takes 1-5 μ L sample point to target center, dries；1-10 μ L supernatant is added dropwise in step (5) again
Solution, then dry, it then reuses Mass Spectrometric Identification system and carries out target practice identification.
It is further that the method includes the steps of by the present invention, and step (1) grinds wild mushroom mushroom；Step (2)
It takes the wild mushroom mushroom 200-500mg of crushing to be packed into centrifuge tube, 70% formic acid of 10-500 μ L concentration and 20-600 μ L second is added
Nitrile mixes well；Step (3) oscillation shakes up 10-15min, and revolving speed is no less than 12000r/min centrifugation 1.5min or more, takes supernatant
Solution is the sample prepared；Step (4) takes 1-3 μ L sample point to target center, dries；1-5 μ L supernatant is added dropwise in step (5) again
Solution, then dry, it then reuses Mass Spectrometric Identification system and carries out target practice identification.
Further be, in this method step (2) of the present invention the volume ratio of added 70% formic acid of concentration and acetonitrile be 1:1~
1.2。
It is further that the volume ratio of added 70% formic acid of concentration and acetonitrile is 1:1 in this method step (2) of the present invention.
It is further that the method includes the steps of by the present invention, and step (1) grinds wild mushroom mushroom；Step (2)
It takes the wild mushroom mushroom 200mg of crushing to be packed into centrifuge tube, 50 μ L concentration, 70% formic acid is added and 50 μ L acetonitriles mix well；Step
Suddenly (3) oscillation shakes up 10min, and revolving speed 12000r/min is centrifuged 2min, and taking supernatant solution is the sample prepared；Step (4) takes 1
μ L sample point is dried to target center；1 μ L supernatant solution is added dropwise in step (5) again, then dries, and then reuses Mass Spectrometric Identification system
System carries out target practice identification.
It is further that the method includes the steps of by the present invention, and step (1) grinds wild mushroom mushroom；Step (2)
It takes the wild mushroom mushroom 100mg of crushing to be packed into centrifuge tube, 10 μ L concentration, 70% formic acid is added and 20 μ L acetonitriles mix well；Step
Suddenly (3) oscillation shakes up 5min, and revolving speed 10000r/min is centrifuged 3min, and taking supernatant solution is the sample prepared；Step (4) takes 5 μ
L sample point is dried to target center；5 μ L supernatant solutions are added dropwise in step (5) again, then dry, and then reuse Mass Spectrometric Identification system
Carry out target practice identification.
It is further that the method includes the steps of by the present invention, and step (1) grinds wild mushroom mushroom；Step (2)
It takes the wild mushroom mushroom 600mg of crushing to be packed into centrifuge tube, 100 μ L concentration, 70% formic acid is added and 300 μ L acetonitriles mix well；
Step (3) oscillation shakes up 15min, and revolving speed 13000r/min is centrifuged 5min, and taking supernatant solution is the sample prepared；Step (4)
It takes 5 μ L sample points to target center, dries；10 μ L supernatant solutions are added dropwise in step (5) again, then dry, and then reuse mass spectrum mirror
Determine system and carries out target practice identification.
It is further that the method includes the steps of by the present invention, and step (1) grinds wild mushroom mushroom；Step (2)
It takes the wild mushroom mushroom 300mg of crushing to be packed into centrifuge tube, 200 μ L concentration, 70% formic acid is added and 200 μ L acetonitriles mix well；
Step (3) oscillation shakes up 15min, and revolving speed 13000r/min is centrifuged 5min, and taking supernatant solution is the sample prepared；Step (4)
It takes 5 μ L sample points to target center, dries；10 μ L supernatant solutions are added dropwise in step (5) again, then dry, and then reuse mass spectrum mirror
Determine system and carries out target practice identification.
It is further that the method includes the steps of by the present invention, and step (1) grinds wild mushroom mushroom；Step (2)
It takes the wild mushroom mushroom 500mg of crushing to be packed into centrifuge tube, 500 μ L concentration, 70% formic acid is added and 600 μ L acetonitriles mix well；
Step (3) oscillation shakes up 20min, and revolving speed 14000r/min is centrifuged 5min, and taking supernatant solution is the sample prepared；Step (4)
It takes 3 μ L sample points to target center, dries；5 μ L supernatant solutions are added dropwise in step (5) again, then dry, and then reuse Mass Spectrometric Identification
System carries out target practice identification.
It is further that step (1) of the present invention grinds specifically, wild mushroom mushroom is under liquid nitrogen frozen state,
Up to fine-powdered.
It is further that step (1) of the present invention grinds it specifically, wild mushroom mushroom is under liquid nitrogen frozen state
It is preceding first to be blended with tissue grinder instrument.
The background of the method for the present invention can be applied: the wild mushroom that certain dining room will shred (into strips or fourth is blocky) carries out general
Stir-fry and eat, poisoning occurs after having eaten the wild mushroom for customer, then will poisoning person sponging on an aristocrat while be sent to hospital, by dining room work
Quickly the wild mushroom raw material shredded is sent to testing agency as personnel's motor of riding, testing agency using method of the invention only
Need Rapid identification can be completed in more than 10 minutes, the hospital that the patient that then tells by telephone send, hospital be connected to tell by telephone after clearly
Chu know in institute wild mushroom why species, and select and targetedly detoxify scheme, to save the life of poisoner.This is applicable in back
Scape is only one of particular situation, and similar background is very more.
Fig. 1, Fig. 2 are seen, shown in Fig. 3, although its ferfas, cepe, matsutake are what Yunnan Province's different sources acquired
Species, but after being analyzed by the method for the invention, the map wave crest of same species is consistent, therefore establishes numerous differences
After the mass-spectrogram database of wild mushroom, it is only necessary to wild mushroom species to be identified be operated one time by means of the present invention, so
Database comparison is carried out afterwards, can be realized quickly, the purpose of precise Identification.Actual implementation of the present invention applies open country used in mode
Raw strain is not only above-mentioned three kinds, due to the limitation of length, cannot enumerate, illustrate hereby.
Above-described is only specific embodiments of the present invention, and well known common sense does not describe excessively herein in scheme.It answers
When pointing out, above-described embodiment is not limit the invention in any way, for those skilled in the art, all using equivalent
The technical solution that the mode of replacement or equivalent transformation obtains is fallen within the scope of protection of the present invention.This application claims protection model
The content of its claim should be subject to by enclosing, and the records such as specific embodiment in specification can be used for explaining claim
Content.
Claims (10)
1. a kind of Mass Spectrometric Identification detection method of Wild Edible Fungi in Yunnan species, which is characterized in that the method includes the steps of, step
(1) wild mushroom mushroom is ground；Step (2) takes the wild fungus of crushing to be packed into centrifuge tube, and formic acid is added and mixes well；
Step (3) oscillation shakes up, and is centrifuged, taking supernatant solution is the sample prepared；Step (4) sample-taking spot is to target center；Step
(5) target practice identification is carried out using Mass Spectrometric Identification system.
2. a kind of Mass Spectrometric Identification detection method of Wild Edible Fungi in Yunnan species according to claim 1, which is characterized in that the party
Method comprises the steps of that step (1) grinds wild mushroom mushroom；Step (2) takes the wild mushroom mushroom of crushing to be packed into centrifugation
In pipe, formic acid is added and acetonitrile mixes well；Step (3) oscillation shakes up, and is centrifuged, taking supernatant solution is the sample prepared；Step
Rapid 4) sample-taking spot is dried to target center；Step (5) carries out identification target practice identification of practicing shooting using Mass Spectrometric Identification system.
3. a kind of Mass Spectrometric Identification detection method of Wild Edible Fungi in Yunnan species according to claim 2, which is characterized in that the party
Method comprises the steps of that step (1) grinds wild mushroom mushroom；Step (2) takes the wild mushroom mushroom of crushing to be packed into centrifugation
In pipe, formic acid is added and acetonitrile mixes well；Step (3) oscillation shakes up, and is centrifuged, taking supernatant solution is the sample prepared；Step
Suddenly (4) sample-taking spot is dried to target center；Supernatant solution is added dropwise in step (5) again, then dries, and then reuses Mass Spectrometric Identification
System carries out target practice identification.
4. a kind of Mass Spectrometric Identification detection method of Wild Edible Fungi in Yunnan species according to claim 3, which is characterized in that the party
Method comprises the steps of that step (1) grinds wild mushroom mushroom；Step (2) takes the wild mushroom mushroom 100- of crushing
5000mg is packed into centrifuge tube, and 70% formic acid of 10-5000 μ L concentration is added and 20-3000 μ L acetonitrile mixes well；Step (3) vibration
It swings and shakes up 5-20min, revolving speed is no less than 10000r/min centrifugation 1min or more, and taking supernatant solution is the sample prepared；Step
(4) it takes 1-5 μ L sample point to target center, dries；1-10 μ L supernatant solution is added dropwise in step (5) again, then dries, and then reuses
Mass Spectrometric Identification system carries out target practice identification.
5. a kind of Mass Spectrometric Identification detection method of Wild Edible Fungi in Yunnan species according to claim 4, which is characterized in that the party
Method comprises the steps of that step (1) grinds wild mushroom mushroom；Step (2) takes the wild mushroom mushroom 200-500mg of crushing
It is packed into centrifuge tube, 70% formic acid of 10-500 μ L concentration is added and 20-600 μ L acetonitrile mixes well；Step (3) oscillation shakes up
10-15min, revolving speed are no less than 12000r/min centrifugation 1.5min or more, and taking supernatant solution is the sample prepared；Step (4)
It takes 1-3 μ L sample point to target center, dries；1-5 μ L supernatant solution is added dropwise in step (5) again, then dries, and then reuses mass spectrum
Identification systems carry out target practice identification.
6. a kind of Mass Spectrometric Identification detection method of Wild Edible Fungi in Yunnan species according to claim 4 or 5, which is characterized in that
The volume ratio of added 70% formic acid of concentration and acetonitrile is 1:1~1.2 in this method step (2).
7. a kind of Mass Spectrometric Identification detection method of Wild Edible Fungi in Yunnan species according to claim 4 or 5, which is characterized in that
The volume ratio of added 70% formic acid of concentration and acetonitrile is 1:1 in this method step (2).
8. a kind of Mass Spectrometric Identification detection method of Wild Edible Fungi in Yunnan species according to claim 5, which is characterized in that the party
Method comprises the steps of that step (1) grinds wild mushroom mushroom；Step (2) takes the wild mushroom mushroom 200mg of crushing to be packed into
In centrifuge tube, 50 μ L concentration, 70% formic acid is added and 50 μ L acetonitriles mix well；Step (3) oscillation shakes up 10min, revolving speed
12000r/min is centrifuged 2min, and taking supernatant solution is the sample prepared；Step (4) takes 1 μ L sample point to target center, dries in the air
It is dry；1 μ L supernatant solution is added dropwise in step (5) again, then dries, and then reuses Mass Spectrometric Identification system and carries out target practice identification.
9. a kind of Mass Spectrometric Identification detection method of Wild Edible Fungi in Yunnan species described according to claim 1 or 2 or 3 or 4 or 5 or 8,
It is characterized in that, step (1) is ground specifically, wild mushroom mushroom is under liquid nitrogen frozen state, fine-powdered is reached.
10. a kind of Mass Spectrometric Identification detection method of Wild Edible Fungi in Yunnan species according to claim 9, which is characterized in that step
Suddenly (1) is first blended with tissue grinder instrument before specifically, wild mushroom mushroom is under liquid nitrogen frozen state and is ground.
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Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102608278A (en) * | 2012-02-25 | 2012-07-25 | 云南农业大学 | Method for simultaneously detecting contents of various heavy metal elements in edible mushrooms |
| CN109324007A (en) * | 2018-11-13 | 2019-02-12 | 辽宁省微生物科学研究院 | A kind of edible fungus species degeneration detection kit and detection method |
-
2019
- 2019-04-15 CN CN201910300856.3A patent/CN109916992A/en active Pending
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102608278A (en) * | 2012-02-25 | 2012-07-25 | 云南农业大学 | Method for simultaneously detecting contents of various heavy metal elements in edible mushrooms |
| CN109324007A (en) * | 2018-11-13 | 2019-02-12 | 辽宁省微生物科学研究院 | A kind of edible fungus species degeneration detection kit and detection method |
Non-Patent Citations (3)
| Title |
|---|
| CAROLE CASSAGNE 等: "Mould Routine Identification in the Clinical Laboratory by Matrix-Assisted Laser Desorption Ionization Time-Of-Flight Mass Spectrometry", 《PLOS ONE》 * |
| RYOTA SUGAWARA 等: "Rapid and reliable species identification of wild mushrooms by matrix assisted laser desorption/ionization time of flight mass spectrometry (MALDI-TOF MS)", 《ANALYTICA CHIMICA ACTA》 * |
| 王冲 等: "MALDI-TOF MS技术在临床病原真菌鉴定中的应用进展", 《中国真菌学杂质》 * |
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