CN109917053A - A method of amanita hemolysin in detection mushroom - Google Patents
A method of amanita hemolysin in detection mushroom Download PDFInfo
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Abstract
The invention discloses a kind of methods of amanita hemolysin in detection mushroom.This method comprises the following steps: (1) extracting mushroom fruitbody using methanol and obtain Toxic extraction liquid；(2) the Toxic extraction liquid is detected using HPLC, obtains the peak area of the absorption peak of amanita hemolysin；(3) the standard specimen preparing standard solution for using amanita hemolysin, then detects the standard solution using HPLC, obtains peak area when different sample volumes, and then obtain the equation of linear regression between the sample volume and peak area of the standard specimen of the amanita hemolysin；Step (2) is identical with the condition that HPLC in (3) is detected；(4) equation of linear regression that the peak area according to obtained in step (2) and step (3) obtain to get the amanita hemolysin into mushroom content.This emperor's son-in-law method can be used for the quantitative detection of amanita hemolysin in mushroom, be suitable for Shandong or even the multifarious feature of Mount Taishan poisonous mushroom species resource.
Description
Technical field
The present invention relates to a kind of methods of amanita hemolysin in detection mushroom, belong to field of detection of food safety.
Background technique
Mushroom belongs to macro fungi, including wild edible fungus and medicinal fungus, wherein a small number of is poisonous mushroom.Poisonous mushroom is containing difference
Type causes evil toxin (fourth of the twelve Earthly Branches morning mist, 1987).The sixties in 19th century, the mankind are for the first time from Amanita muscaria Amanita muscaria
(L.) discovery causes evil toxin (Schimedeberg et al, 1869).Poisonous mushroom is had reported at present containing nine big toxoids: goose cream
Toxic cyclic peptide, single methyl hydrazine toxin, cortina verticillium toxin, muscarine, Iibotenicacid, coprinin, psilocybin and dephosphorylation are naked
Lid mushroom element, GI irritation agent and Russula subnigricans toxin (Liu Jikai, 2004).Poisonous mushroom toxin intoxicating symptom is divided into stomach and intestine
Type, nervous system type, Hemolytic Type, liver damage type, breathing and the big poisoning symptom (fourth of the twelve Earthly Branches of circulatory failure type and hypericism type etc. 6
Morning mist, 2006).Amanitine is strong fatal type toxin, such poisonous mushroom toxin is primarily present in Amanita poisonous mushroom thallus,
Very little dosage (LD50It can 0.1mg/kg) cause people in death, higher than cyanide toxicity more than ten again (Wieland, 1986).Amanita fuliginea peptide
Toxoid has had now been found that 22 kinds (Wieland, 1986).Peptide toxin is cyclic peptide compounds, is formed according to its amino acid
It can be divided into three classes with structure: amanita hemolysin class, Phallus toxic peptides and amanita phalloides toxic peptides.
Amanita hemolysin is the structure of the carbocyclic skeleton of a kind of bicyclic octapeptide, and structural formula is as shown in Equation 1, and chemical property is steady
It is fixed, high temperature resistant, acid and alkali-resistance.Amanita hemolysin has very strong lethal, and action site is the nucleus of eukaryocyte
(Wieland, 1986).The toxoid has now been found that 9 kinds, and as known from Table 1, their amino acid composition difference is smaller, mainly
Difference is only that the different substituent group (being shown in Table 2) of its side chain.It is the molecular formula of several important amanita hemolysins individually below:
C39H54N10O14S(α-amanitin)、C39H53O15S (β-amanitin) and C39H54N10O13S (γ-amanitin)。
The amino acid of 1 amanita hemolysin toxin of table forms
The side-chain radical and toxicity * of 2 amanita hemolysin toxin of table
* document (Vetter, 1998) is derived from.
In order to quickly and effectively detect whether poisonous mushroom contains amatoxin, the dry rear dense salt of paper fresh mushroom pressure juice trace is had developed
Sour hanging drop development process quickly and effectively detects the presence or absence of fresh mushroom amatoxin (Wieland, 1949), and Wieland is used again later
1% cinnamic acid methanol solution, in the presence of concentrated hydrochloric acid, amatoxin shows darkviolet, and Phallus phallotoxins shows yellowish-brown, is afterwards in sky-blue；
These methods were improved again later, improvement is advanced optimized and establishes the conventional detection side that poisonous mushroom mainly poisons ingredient
Method.Beuthler&Marderosin in 1981 uses high performance liquid chromatography (HPLC) to detect Amanita fuliginea peptide toxin for the first time.With
Pastorelllo etc. (1982) also detects α-amanita hemolysin using HPLC method afterwards.Silva in 1983 attempts to be detected with HPLC method
Eat α-amanita hemolysin and β-amanita hemolysin in malicious human body.HPLC method is because having the advantages that quick, sensitive, the quilt in mushroom Mycotoxin identification
It is widely used (Enjalbert et al, 1993,1999).In the 1990s, Chinese scholar Zhang Zhiguang uses HPLC method first
The Amanita fuliginea type amanita hemolysin of the provinces such as Hunan, Hubei is tested and analyzed, detection has found East Asia endemic species and sociales-ash
Decorative pattern In The Studies On Toxins of Amanita content is high.In addition, Chen Zuohong (2003), Hu Jingsong (2003) and Bao Haiying (1999) et al. also use HPLC
Method has detected the content of toxins of other a variety of Amanita fuligineas, the overall merit Amanita fuliginea in China torrid zone and refrigerant latitudes poisonous mushroom
Content of toxins and its variation dynamic provide feasible technical method to eat poisonous mushroom state of an illness quick diagnosis and treatment in time by mistake.
Summary of the invention
The object of the present invention is to provide a kind of methods of amanita hemolysin in detection mushroom, can be used for amanita hemolysin in mushroom
Quantitative detection is suitable for Shandong or even the multifarious feature of Mount Taishan poisonous mushroom species resource.
The method of amanita hemolysin, includes the following steps: in detection mushroom provided by the present invention
(1) mushroom fruitbody is extracted using methanol and obtains Toxic extraction liquid；
(2) the Toxic extraction liquid is detected using HPLC, obtains the peak area of the absorption peak of amanita hemolysin；
(3) the standard specimen preparing standard solution for using amanita hemolysin, then detects the standard solution using HPLC, obtains not
With peak area when sample volume, and then obtain the linear regression side between the sample volume and peak area of the standard specimen of the amanita hemolysin
Journey；
Step (2) is identical with the condition that HPLC in (3) is detected；
(4) equation of linear regression that the peak area according to obtained in step (2) and step (3) obtain is to get into mushroom
The content of amanita hemolysin.
In above-mentioned method, in step (1), the methanol extract the step of it is as follows:
After the dry mushroom fruitbody is crushed, extracted using methanol aqueous solution；Supernatant is taken after centrifugation.
Silt sundries is rejected, in thermostatic drying chamber after choosing the complete fructification of individual of acquisition if dry fructification
70 DEG C drying to constant weight, is then crushed；
If new fresh sporophore, the complete fructification of individual for choosing acquisition is crushed.
In above-mentioned method, the volumn concentration of the methanol aqueous solution can be 50%.
The dosage of the methanol aqueous solution are as follows: mushroom fruitbody described in 1g: 200mL methanol aqueous solution；
The condition of the extraction is as follows: temperature is 30~37 DEG C, and the time is 12~30h；
Precipitating after centrifugation extracts again, then merges supernatant.
In above-mentioned method, the method also includes handling the supernatant using petroleum ether degreasing；
Extracting solution after the ungrease treatment is concentrated, the filtering with microporous membrane after constant volume with 0.22 μm.
In above-mentioned method, in step (2) and (3), the condition of the HPLC detection is as follows:
Highly effective liquid phase chromatographic system is Shimadzu LC-20A；
Analytical column is YWG C18,5 μm of inverse analysis columns；
Diode array detector is SPD-M20A；
Detection wavelength is 295nm；
Analysis uses LCsolution software；
Eluent is made of A liquid and B liquid, pH value 5.0；Wherein, the acetonitrile and 90% that A liquid is 10% by volume content
0.02mol/L ammonium acetate aqueous solution mixed liquor, B liquid is the 0.02mol/L of the acetonitrile that volume content is 24% and 76%
Ammonium acetate aqueous solution mixed liquor；
Gradient (for linear gradient elution) are as follows: the volume content of 0~15min, A liquid is adjusted to 95%, B liquid by 100%
Volume content be adjusted to 5% by 0%；The volume content of 15~35min, A liquid by 95% be adjusted to the volume content of 20%, B liquid by
5% is adjusted to 80%, and keeps 5min；The volume content of 40~45min, A liquid by 20% be adjusted to the volume content of 0%, B liquid by
80% is adjusted to 100%, and keeps 5min；The volume content that the volume content of 50~55min, A liquid modulates 100%, B liquid by 0%
It is adjusted to 0% by 100%, and keeps 10min；
Flow rate of mobile phase is 1.0mL/min；
Sample volume is 20 μ L.
In above-mentioned method, in step (3), sample introduction can be carried out according to the amount of 3 μ g, 5 μ g, 10 μ g, 15 μ g, 20 μ g；With face
Product, external standard method obtain the peak area data of the gradient sample volume of each toxin respectively, acquire each standard specimen toxin with minimum square law
The equation of linear regression of sample volume and peak area.
The present invention has tested and analyzed the fructification toxin situation of the fatal goose cream in Mount Taishan using HPLC method, the results showed that Mount Taishan
More than 10 toxic substances such as amanita hemolysin and Phallus phallotoxins of the fatal goose ointment-like medicine for oral or plastering use entity containing strong toxic action, and fructification individual
Between Peptide toxin content notable difference, Peptide toxin content be 1336~3627 μ g/g dry weights.Furthermore, it was confirmed that fructification is prosperous
Peak period Peptide toxin content highest, immature flower bud phase are taken second place, and the ripening and senscence phase is minimum.By analyzing HPLC detection data, discovery is caused
New 157 μ g/g of fresh sporophore amanita hemolysin content average out to, the 21 μ g/g of fructification content average out to boiled that life goose cream does not boil,
The former is higher than 7.5 times of the latter, provides important scientific reference frame for the treatment of poisoning patient.
Detailed description of the invention
Fig. 1 is the aspect graph of fatal goose cream different stages of growth fructification, wherein Fig. 1 (a) is the aspect graph of button phase,
Fig. 1 (b) is the aspect graph for growing animated period, and Fig. 1 (c) is the aspect graph in maturity period.
Fig. 2 is fatal goose ointment-like medicine for oral or plastering use entity Raw toxin (Fig. 2 (a)) and α-amanita hemolysin standard specimen HPLC map (Fig. 2 (b)).
Fig. 3 is the ultraviolet spectrogram of amanita hemolysin.
Fig. 4 is the ultraviolet spectrogram of Phallus phallotoxins.
Fig. 5 be the fatal goose ointment-like medicine for oral or plastering use entity different stages of growth toxin in Mount Taishan and α-amanita hemolysin standard specimen HPLC map,
In, Fig. 5 (a) is the HPLC map of button phase, and Fig. 5 (b) is the HPLC map for growing animated period, and Fig. 5 (c) is the maturity period
HPLC map, Fig. 5 (d) are the HPLC map of amanita hemolysin standard specimen.
Fig. 6 is the Raw toxin and α-amanita hemolysin HPLC figure of fatal goose cream Different Individual, wherein Fig. 6 (a)-Fig. 6 (c)
For the HPLC map of different samples, Fig. 6 (d) is α-amanita hemolysin standard specimen HPLC map.
Fig. 7 is the HPLC map of the Raw toxin of fatal goose ointment-like medicine for oral or plastering use entity fresh goods, wherein Fig. 7 (a) is not boil new fresh sporophore
HPLC map, Fig. 7 (b) is the HPLC map for boiling 1h sample.
Specific embodiment
Experimental method used in following embodiments is conventional method unless otherwise specified.
The materials, reagents and the like used in the following examples is commercially available unless otherwise specified.
One, the fatal poisonous mushroom fructification of material-
Fatal goose ointment-like medicine for oral or plastering use entity was collected in the wind such as the electrodeless mausoleum in Mount Taishan, fan precipice, Hou Shiwu, bamboo grove temple in 2017~2018 years
Scenic spot, and correctly identified by document and its type specimen control.
The sample of different stages of growth Peptide toxin test experience selects: pressing fatal goose cream different stages of growth fructification shape
In three periods of the notable difference of state feature point, as shown in Figure 1, button period: the non-parachute-opening of fructification, interior teleoblema have；It is raw
Long animated period: interior mycoderm is only deposited in incomplete parachute-opening；The ripening and senscence phase: the complete parachute-opening of fructification, cap is open and flat or upwarps, inside and outside
Mycoderm all without.The individual difference between differences of environmental factors and same growth phase sample different strains to reduce different samples
Different, experimental material selects multiple complete drying fructifications of same when and where acquisition, each growth phase sample is equal
The repetition of 3 Different Individuals is set.
Two, method
1, fatal poisonous mushroom poisons composition detection
(1) Toxic extraction method
(1-1) does the Extracted toxin of fructification
Using methanol extraction, concrete operation step: the complete fatal goose ointment-like medicine for oral or plastering use entity sample of individual of acquisition is chosen, is picked
Except silt sundries, in thermostatic drying chamber 70 DEG C drying to constant weight.It is milled to pulverizer powdered, accurately weighs sample
3 repetitions are arranged in 1.0g.20mL 50% (v/v) methanol solution is added, sets shaking table oscillation, 130g extracts 12h at 37 DEG C.
4000g is centrifuged 10 minutes, and supernatant is taken to be stored in 4 DEG C of refrigerators, and precipitating repeats extracting once, takes supernatant and previous conjunction
And.With the petroleum ether (30~60 DEG C) of same volume, degreasing is twice.Extracting solution is concentrated in Rotary Evaporators, and ultrapure water is added to be settled to
10mL.With 0.22 μm of filtering with microporous membrane, Toxic extraction liquid is obtained.
The Extracted toxin of (1-2) new fresh sporophore
Using methanol extraction, concrete operation step: the complete fresh fatal goose ointment-like medicine for oral or plastering use entity sample of individual of acquisition is chosen
Product, each individual averagely cut two halves, then respectively weigh 70g, and 3 repetitions are arranged in every part of sample, and a copy of it is ground with pulverizer
Be broken into it is powdered, then be added 100mL 50% (v/v) methanol aqueous solution carry out Toxic extraction according to the above method.Another adds
1h is boiled in 1000mL boiling, and the methanol-water of 100mL 50% (v/v) is added then by after the fructification sample comminution boiled in filtering
Solution carries out Toxic extraction according to the above method.Fructification will not be boiled and the Toxic extraction liquid for the fructification acquisition for boiling 1h adds
Volume petroleum ether (30~60 DEG C) degreasing twice, is concentrated in Rotary Evaporators, ultrapure water is added to be settled to 10mL.
2, HPLC determination condition and parameter
Highly effective liquid phase chromatographic system is Shimadzu LC-20A, and analytical column is 250mm × 4.6mm, YWG C18 (blood testing point
Analysis column is Hypersil ODS2), 5 μm of inverse analysis columns, diode array detector SPD-M20A, Detection wavelength 295nm,
Analysis uses LCsolution software.Eluent is mixed by two liquid of A, B, A liquid: 10% acetonitrile, 90%0.02mol/L ammonium acetate；B
Liquid: 24% acetonitrile, 76%0.02mol/L ammonium acetate are adjusted to pH5.0 with glacial acetic acid.Gradient are as follows: 0 → 15min, A liquid
100% → 95%, B liquid 0 → 5%；15 → 35min, A liquid 95% → 20%, B liquid 5% → 80%, and keep 5min；40→
45min, A liquid 20% → 0%, B liquid 80% → 100%, and keep 5min；50 → 55min, A liquid 0% → 100%, B liquid
100% → 0%, and keep 10min.Flow rate of mobile phase is 1.0mL/min, and sample volume is 20 μ L.
3, the acquisition of linearity of regression equation
Standard specimen toxin is taken accurately to be configured to the solution that concentration is 1mg/mL, by 3 μ g, 5 μ g, 10 μ g, 15 μ g, 20 μ g sample introductions,
The peak area data for obtaining the gradient sample volume of each toxin respectively with area, external standard method acquire each standard specimen poison with minimum square law
The sample volume of element and the equation of linear regression of peak area.
Three, result and analysis
1, fatal poisonous mushroom causes evil toxin
(1-1) toxin standard specimen equation of linear regression
The equation of linear regression of Mount Taishan fatal amanita hemolysin standard specimen sample amount and peak area is obtained by calculating are as follows: y=
657000x+366000, r2=0.999391 (x: sample volume/μ g；Y: peak area；R: related coefficient).It can from related coefficient
Out, the correlation of sample volume and peak area is quite high.When measuring sample, can be led to according to the elution peak area of its HPLC figure
Cross the quantitative analysis that above-mentioned equation of linear regression carries out toxin.
(1-2) fatal goose ointment-like medicine for oral or plastering use entity Peptide toxin HPLC analysis
The fatal goose ointment-like medicine for oral or plastering use entity toxin crude extract HPLC finger-print in Mount Taishan, it is more apparent there are about 10 as shown in Fig. 2 (a)
Toxic substance eluting peak, wherein content is higher 6~7 kinds of toxin, amanita hemolysin content highest.It is accordingly washed by HPLC
The retention time at de- peak is compared with standard specimen (Fig. 2 (b)), and the equation of linear regression pass between elution peak area and content of toxins
System show that the content of amanita hemolysin in sample is 2736 μ g/g dry weights.The ultraviolet spectrogram of toxin elution peak is analyzed it is found that being examined
The toxin measured is mainly made of two major classes: one kind is the amanita hemolysin (Fig. 3) that obtained the maximum absorption is 303 nm, and another kind of is most
The Phallus phallotoxins (Fig. 4) that big absorption value is 290nm.By table 3 it is found that the former includes 2,3, No. 6 three toxin peaks, the latter includes
Five toxin peaks that number is 4,7,8,9,10.The eluting peak size of more identical ultraviolet absorption value is it is found that three kinds of goose cream
The content of phallotoxins is answered are as follows: No. 3 peak (amanita hemolysin) content highests, followed by No. 2 peaks, minimum are No. 6 peaks.Another five kinds of Phallus phallotoxins
Content height then successively are as follows: 7,8,4,10, No. 9 peaks.
The fatal goose ointment-like medicine for oral or plastering use entity Raw toxin HPLC atlas analysis of table 3
The Peptide toxin of (1-3) fructification different stages of growth changes
The Toxic extraction liquid of the fatal goose ointment-like medicine for oral or plastering use entity different stages of growth in same place, same time acquisition is carried out
HPLC detection, as a result as shown in figure 5, display: fatal goose ointment-like medicine for oral or plastering use entity button phase, the goose cream for growing animated period and ripening and senscence phase
Phallotoxins concentration distinguishes 1725.9,2226.8 and 1462.7 μ g/g dry weights.This show amanita hemolysin during sporophore growth, poison
Highest cellulose content is growth animated period, and followed by button phase, the content of ripening and senscence phase is minimum.As a result it is also shown that growth is prosperous
The content of amanita hemolysin is higher by 34% and 22% than ripening and senscence phase and button phase respectively in flower phase fructification, and the button phase should
The content of toxin is then higher than the ripening and senscence phase by 15%.Therefore, by this experiment it is found that the fatal goose cream in Mount Taishan is grown in fructification difference
In the stage, the content of primary toxins amanita hemolysin has more apparent variation, and its changing rule is in normal distribution substantially.Compare 3
The sample of a different stages of growth, in the toxin peak area difference of same retention time, the content of other primary toxins of deducibility
Changing rule and amanita hemolysin it is almost the same.
Individual difference of (1-4) the Peptide toxin content in fructification and its with correlation locality
To it is same locality, the content of toxins of the fatal goose ointment-like medicine for oral or plastering use entity Different Individual of identical growth phase carry out detection hair
Existing, there are significant individual difference, concentration range 1336- in fatal goose ointment-like medicine for oral or plastering use entity for the content of α-amanitin
3627 μ g/g dry weights, as shown in Figure 6.Compare 3 different samples in the toxin peak area difference of same retention time it is found that other
There is also notable differences between Different Individual for the content of primary toxins, and the changes of contents of these toxin is with uniformity.
It is adopted respectively in the electrodeless mausoleum in Mount Taishan, rear stone depressed place and three, bamboo grove temple different acquisition using HPLC method detection same time
The fatal goose ointment-like medicine for oral or plastering use entity content of toxins of the same growth phase collected, the results show that three amanita hemolysins locality contain
Amount average value is respectively: 2398,2417 and 2425 μ g/g dry weight, the amanita hemolysin content of this with showing different acquisition sample is without bright
Significant difference is different.Therefore illustrate, the content of toxins of the fatal goose ointment-like medicine for oral or plastering use entity in Mount Taishan and locality being not significantly related to property.
The analysis of Peptide toxin after (1-4) new fresh sporophore and its boiling
In order to carry out the quantitative analysis of content of toxins difference to the fructification after fresh fatal Amanita fuliginea and its boiling, this
Experiment removes water on the basis of avoiding the individual difference of content of toxins, to fatal goose cream fresh sample and its after adding boiling to boil 1 hour
Fructification use methanol method to extract Raw toxin simultaneously, and carry out HPLC detection, as shown in fig. 7, display, the fatal fresh son of goose cream is real
The amanita hemolysin content of toxins of body is 145~152 μ g/g fresh weights, and average value is 148 μ g/g fresh weights, and the fructification toxin boiled
Content is 13~34 μ g/g fresh weights, and average value is 21 μ g/g fresh weights.Thus the amanita hemolysin content for boiling sample was not boiled only
The 13% of sample.Other toxin peak areas of two samples of comparative analysis also indicate that the new fresh sporophore that fatal goose cream does not boil is malicious
Cellulose content is significantly larger than the fructification boiled.Therefore after the fatal new fresh sporophore of goose cream boils 1h with boiling, 87% toxin can be from
It is dissolved out in fructification.Amatoxin steady chemical structure, high temperature resistant, toxin is substantially without degradation during boiling.
The present invention has tested and analyzed the fructification toxin situation of the fatal goose cream in Mount Taishan using HPLC method, and as a result Mount Taishan is fatal
More than 10 toxic substances such as amanita hemolysin and Phallus phallotoxins of the goose ointment-like medicine for oral or plastering use entity containing strong toxic action, and peptide between fructification individual
Toxoid content notable difference, Peptide toxin content are 1336~3627 μ g/g dry weights.Furthermore, it was confirmed that fructification animated period
Peptide toxin content highest, immature flower bud phase are taken second place, and the ripening and senscence phase is minimum.
By analyzing HPLC detection data, the new fresh sporophore amanita hemolysin content average out to that fatal goose cream does not boil is found
157 μ g/g, the 21 μ g/g of fructification content average out to boiled, the former is higher than 7.5 times of the latter, provides for the treatment of poisoning patient
Important scientific reference frame.
Claims (5)
1. the method for amanita hemolysin, includes the following steps: in a kind of detection mushroom
(1) mushroom fruitbody is extracted using methanol and obtains Toxic extraction liquid；
(2) the Toxic extraction liquid is detected using HPLC, obtains the peak area of the absorption peak of amanita hemolysin；
(3) the standard specimen preparing standard solution for using amanita hemolysin, then detects the standard solution using HPLC, obtain it is different into
Peak area when sample amount, and then obtain the equation of linear regression between the sample volume and peak area of the standard specimen of the amanita hemolysin；
Step (2) is identical with the condition that HPLC in (3) is detected；
(4) equation of linear regression that the peak area according to obtained in step (2) and step (3) obtain is to get the goose cream into mushroom
The content of phallotoxins.
2. according to the method described in claim 1, it is characterized by: in step (1), the step of methanol extracts, is as follows:
After the dry mushroom fruitbody is crushed, extracted using methanol aqueous solution；Supernatant is taken after centrifugation.
3. according to the method described in claim 2, it is characterized by: the volumn concentration of the methanol aqueous solution is 50%；
The dosage of the methanol aqueous solution are as follows: mushroom fruitbody described in 1g: 200mL methanol aqueous solution；
The condition of the extraction is as follows: temperature is 30~37 DEG C, and the time is 12~30h.
4. according to the method in claim 2 or 3, it is characterised in that: the method also includes being handled using petroleum ether degreasing
The step of supernatant；
Extracting solution after the ungrease treatment is concentrated, the filtering with microporous membrane after constant volume with 0.22 μm.
5. method according to any of claims 1-4, it is characterised in that: in step (2) and (3), the HPLC inspection
The condition of survey is as follows:
Analytical column is YWG C18,5 μm of inverse analysis columns；
Diode array detector is SPD-M20A；
Detection wavelength is 295nm；
Eluent is made of A liquid and B liquid, pH value 5.0；Wherein, the acetonitrile and 90% that A liquid is 10% by volume content
The mixed liquor of the ammonium acetate aqueous solution of 0.02mol/L, B liquid are the 0.02mol/L of the acetonitrile that volume content is 24% and 76%
The mixed liquor of ammonium acetate aqueous solution；
Gradient are as follows: the volume content that the volume content of 0~15min, A liquid is adjusted to 95%, B liquid by 100% is adjusted to by 0%
5%；The volume content that the volume content of 15~35min, A liquid is adjusted to 20%, B liquid by 95% is adjusted to 80% by 5%, and keeps
5min；The volume content that the volume content of 40~45min, A liquid is adjusted to 0%, B liquid by 20% is adjusted to 100% by 80%, and keeps
5min；The volume content of 50~55min, A liquid is adjusted to 0% by 100% by the volume content of 0% modulation 100%, B liquid, and keeps
10min；
Flow rate of mobile phase is 1.0mL/min；
Sample volume is 20 μ L.
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| CN114878708A (en) * | 2022-04-29 | 2022-08-09 | 福建省疾病预防控制中心(福建省健康教育促进中心、福建省卫生检验检测中心) | Method for detecting 5 amanitin toxins, dephosphorized naked cap mushroom essence and toad tryptamine |
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN114878708A (en) * | 2022-04-29 | 2022-08-09 | 福建省疾病预防控制中心(福建省健康教育促进中心、福建省卫生检验检测中心) | Method for detecting 5 amanitin toxins, dephosphorized naked cap mushroom essence and toad tryptamine |
| CN114878708B (en) * | 2022-04-29 | 2023-11-07 | 福建省疾病预防控制中心(福建省健康教育促进中心、福建省卫生检验检测中心) | Method for detecting 5 amanita peptides, dephosphorized nupharin and bufadienolide |
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