CN103940921A - Microcystic toxins liquid chromatogram-tandem mass spectrum detection method - Google Patents
Microcystic toxins liquid chromatogram-tandem mass spectrum detection method Download PDFInfo
- Publication number
- CN103940921A CN103940921A CN201410011060.3A CN201410011060A CN103940921A CN 103940921 A CN103940921 A CN 103940921A CN 201410011060 A CN201410011060 A CN 201410011060A CN 103940921 A CN103940921 A CN 103940921A
- Authority
- CN
- China
- Prior art keywords
- ion
- tandem mass
- formic acid
- liquid chromatography
- microcystin
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Granted
Links
Abstract
The invention discloses a microcystic toxins liquid chromatogram-tandem mass spectrum detection method, which comprises the following steps: 1)pre-treating a sample, 2)enriching and purifying; and 3)performing LC-MS/MS detection. The detection method has the advantages of simple operation and strong qualitative/quantitative ability, and can be a reliable method for detecting water quality and evaluating the safety risk of food.
Description
Technical field
The present invention relates to a kind of liquid chromatography-tandem mass of Microcystin, belong to instrument detection technique field.
Background technology
Algae health food be a kind of by algae one-celled plants powder, be mainly principal ingredient, add a small amount of excipient or with algae powder, be the health food that raw material is made merely.Main edible algae be take spirulina as representative at present.Spirulina contains rich in protein, mineral matter (calcium, iron, zinc, phosphorus, magnesium and iodine etc.), vitamin (B1, B2, B12, C, E, K etc.) and beta carotene, gamma-linoleic acid, chlorophyll and the various amino acid useful to health.
By manual method, cultivating spirulina is the major way of producing at present algae health food, and can toxigenic microcystic aeruginosa, Microcystis viridis, Hui Shi Microcystis aeruginosa and the algae that quivers, anabena etc. all belong to blue-green algae with spirulina.Theoretically, under strict condition of culture, can control the growth of producing malicious blue-green algae.But because the algae slurry of current domestic spirulina manufacturing enterprise is cultivated under open environment, results, in process of production, culturing pool and spirulina slurry all are very likely subject to producing the pollution of malicious blue-green algae, in results spirulina slurry, gathered in the crops and produced malicious blue-green algae, due to the wash-out of spirulina slurry can not be removed completely and produced malicious blue-green algae, and will pollution algae health food by producing Microcystin that malicious blue-green algae produces, thereby human health be caused to serious harm.
Microcystin be take liver as target organ, can impel the growth of tumour cell, is one of the strongest liver cancer promoter of having found up to now.The primary hepatic carcinoma incidence of disease of China some areas is high relevant with local resident's Long Term Contact Microcystin.Microcystin is class ring-type seven peptide compounds, finds that there is at present about 60 kinds of isomeride, and kind common in China's eutrophication water is MC-LR and MC-RR, and wherein MC-RR is the most common, and MC-LR toxicity is maximum.
At present also many about extraction and the detection method of Microcystin, also the method for useful liquid chromatography-tandem mass spectrometry detects, but in detection method before, the pre-treatment step Toxic extraction of sample is incomplete, and has other impurity to disturb.Especially for algae sample, current existing pre-treating method is very limited for the removal of pigment, causes in Mass Spectrometer Method, occurring comparatively significantly matrix effect, thereby reduces detection sensitivity and accuracy.In addition, because Microcystin has polypeptide class formation, in its molecular structure, there is hydroxyl and amino isopolarity group, very easily generate [M+H]
+with [M+2H]
2+ion.
Set up the detection method of Microcystin in a kind of algae health food of fast and reliable, to guaranteeing the security of algae health food, promote the technological innovation of manufacturing enterprise, improve domestic algae health food quality and class, the guarantee people's is healthy significant.
Summary of the invention
Technical matters to be solved by this invention is: the liquid chromatography-tandem mass that a kind of easy and simple to handle, Microcystin that qualitative, quantitative ability is stronger is provided
For solving the problems of the technologies described above, the technical solution used in the present invention is:
A liquid chromatography-tandem mass for Microcystin, comprises the following steps:
(1) sample pre-treatments
Accurately take through the algae health products 1.0g pulverizing, homogeneous is crossed, with 10mL60% methanol aqueous solution, extract toxin, 200rpm shaking table concussion 60min, more ultrasonic processing 30min, 20 ℃ of following centrifugal 10min of 8000rpm, get supernatant;
With 2mol/L formic acid, regulate supernatant pH value to 4.0, standing 3h can be observed and in solution, occurs a large amount of flocculent deposits, 20 ℃ of following centrifugal 10min of 8000rpm, obtain light yellow supernatant or water white transparency liquid, in solution, add weak aqua ammonia that pH value is adjusted to 7.0, add subsequently ultrapure water to be accurately settled to 30mL, now obtain comparatively pure crude extract;
(2) enrichment and purification
First with 10mL methyl alcohol and 10mL ultrapure water activation C18 solid-phase extraction column, reactivation process flow control is at 1-2 drop/sec, get 15mL crude extract injection C18 solid-phase extraction column and carry out enrichment, flow control is at 1 drop/sec, then more respectively with each 10mL drip washing of 20% and 30% methanol aqueous solution C18 solid-phase extraction column, to remove impurity, finally with 5mL70% methanol aqueous solution (containing 0.05% formic acid) wash-out Microcystin, nitrogen blows near dry, with 50% acetonitrile solution, be settled to 0.5mL, treat that liquid chromatography-tandem mass spectrometry (LC-MS/MS) detects;
Solid-phase extraction column can play good enrichment, clean-up effect, the multiplex methanol-eluted fractions of bibliographical information, and the recovery is better.In the present invention, find that the MC-LR recovery is better as eluent in the situation that usining pure methyl alcohol, and the recovery of MC-RR is extremely low.Add the formic acid of 0.05% concentration in methyl alcohol after, again carry out recovery experiment, find that the recovery of MC-RR is significantly improved.
(3) LC-MS/MS detects
Liquid-phase condition Waters Atlantis dC18 chromatographic column (2.1mm * 150mm, 5 μ m), 30 ℃ of column temperatures; Sample size 20 μ L; Mobile phase A is water (containing 0.05% formic acid, 5mmol/L ammonium formate), and B is 95% acetonitrile (containing 0.05% formic acid, 5mmol/L ammonium formate); Condition of gradient elution is in Table 1:
Table 1 condition of gradient elution
Mass spectrum condition ionization mode is electro-spray ionization holotype (ESI+); Multiple-reaction monitoring pattern (MRM); Collision atmospheric pressure (CAD): 8; Gas curtain atmospheric pressure (CUR): 15; Electron spray voltage (IS): 5000V; Ion source temperature (TEM): 600 ℃.
Multiple-reaction monitoring ion pair and the mass spectrum correlation parameter of compound are as shown in table 2.
The multiple-reaction monitoring ion pair of table 2 compound and mass spectrum correlation parameter table
* be quota ion
When liquid chromatography-mass spectrography detects, due to the impact of matrix effect, cause the effect of target compound generation ion enhancer or inhibitor, do not adopt and obtain matrix blank solution preparation gradient hybrid standard product solution (0 containing target determinand, 0.1,0.2,0.5,1.0,2.0,5.0 μ g/kg).Condition is measured according to the above analysis, with peak area to concentration linear regression.After need to diluting, the sample that exceeds the range of linearity redeterminates.
Table 3 typical curve and linear relationship
In sample, the retention time of Microcystin chromatographic peak is compared with the retention time of respective standard chromatographic peak, and variation range should be within ± 2.5%.The signal to noise ratio (S/N ratio) at the reconstruct chromatography of ions peak of the qualitative ion of Microcystin should be more than or equal to 3(S/N >=3), the signal to noise ratio (S/N ratio) at the reconstruct chromatography of ions peak of quota ion should be more than or equal to 10(S/N >=10).The detection lower bound of MC-LR and MC-RR is all 0.1 μ g/kg.
Method of the present invention is easy and simple to handle, qualitative, quantitative ability is stronger, can detect the reliable method of algae health products quality and assessment food safety risk.This method, by regulating the means such as pH to remove pigment and floccus, reduces the matrix effect of sample on mass spectrum greatly; In mass spectrum mrm parameter, this method adopts MC-LR to be with doubly charged [M+2H]
2+498 molions are as parent ion, than using and be with unicharged [M+H] in other patented claims
+995 molions have higher detection sensitivity as mothers and sons.
Accompanying drawing explanation
Below in conjunction with accompanying drawing, the specific embodiment of the present invention is described in further detail.
Fig. 1 be MC-LR's [M+H]
+fragment ion spectrogram for parent ion generation.
Fig. 2 be MC-LR's [M+2H]
2+fragment ion spectrogram for parent ion generation.
Fig. 3 is that MC-LR is with [M+H]
+ion, as in parent ion situation, is determined two pairs of ion pairs, 995.8/135.1 and 995.8/509.5.
Fig. 4 is that MC-LR is with [M+2H]
2+ion, as in parent ion situation, is determined two couples of ion pair 498.4/135.1 and 498.4/861.5.
Fig. 5 is the second order ms figure of MC-LR.
Fig. 6 is the second order ms figure of MC-RR.
Fig. 7 is MC-LR and MC-RR total ion current figure.
Embodiment
The selection of embodiment 1 parent ion
Existing MC-LR Mass Spectrometer Method method adopts [M+H] more
+ion is as parent ion, and the present invention has compared [M+H] of MC-LR
+with [M+2H]
2+the response of secondary fragmention, result shows [M+2H] of MC-LR
2+the secondary fragment ion that ion can meet with a response stronger as parent ion, the present invention finally selects [M+2H]
2+ion is as MC-LR parent ion, and its Mass Spectrometer Method sensitivity is apparently higher than existing detection method.[M+2H] of the two positive charges of parent ion select tape of MC-LR
2+ion.
Fig. 1 be MC-LR's [M+H]
+for the fragment ion spectrogram that parent ion produces, [M+2H] that Fig. 2 is MC-LR
2+fragment ion spectrogram for parent ion produces again carries out the scanning of secondary fragment after mass spectrum parameter is optimized, and finds [M+2H] of MC-LR
2+the secondary fragment ion that ion can meet with a response stronger as parent ion.
Fig. 3 is that MC-LR is with [M+H]
+ion, as in parent ion situation, is determined two pairs of ion pairs, 995.8/135.1 and 995.8/509.5; Fig. 4 is that MC-LR is with [M+2H]
2+ion, as in parent ion situation, is determined two couples of ion pair 498.4/135.1 and 498.4/861.5.Above two class ion pairs are carried out respectively after MRM optimization and ion gun optimization, and the MC-LR standard items to same concentrations under optimal conditions separately carry out liquid chromatography-tandem mass spectrometry detection, by the comparative analysis to mass spectrogram, find that MC-LR is with [M+2H]
2+ion has stronger response as two pairs of ion pairs determining in parent ion situation, and sensitivity is higher.
Embodiment 2 sample pre-treatments conditions
Sample pre-treatments
The present invention be take algae sample as basis, and the matrix of algae sample is more complicated than the matrix of water sample, and pre-treatment is more complicated.
Accurately take through the algae health products 1.0g pulverizing, homogeneous is crossed, with 10mL60% methanol aqueous solution, extract toxin, 200rpm shaking table concussion 60min, more ultrasonic processing 30min, 20 ℃ of following centrifugal 10min of 8000rpm, get supernatant.
With 2mol/L formic acid, regulate supernatant pH value to 4.0, standing 3h can be observed and in solution, occurs a large amount of flocculent deposits, 20 ℃ of following centrifugal 10min of 8000rpm, obtain light yellow supernatant or water white transparency liquid, in solution, add weak aqua ammonia that pH value is adjusted to 7.0, add subsequently ultrapure water to be accurately settled to 30mL, now obtain comparatively pure crude extract.
Enrichment and purification
First with 10mL methyl alcohol and 10mL ultrapure water activation C18 solid-phase extraction column, reactivation process flow control is at 1-2 drop/sec, get 15mL crude extract injection C18 solid-phase extraction column and carry out enrichment, flow control is at 1 drop/sec, then more respectively with each 10mL drip washing of 20% and 30% methanol aqueous solution C18 solid-phase extraction column, to remove impurity, finally with 5mL70% methanol aqueous solution (containing 0.05% formic acid) wash-out Microcystin, nitrogen blows near dry, with 50% acetonitrile solution, be settled to 0.5mL, treat that liquid chromatography-tandem mass spectrometry (LC-MS/MS) detects.
This method, by regulating the means such as pH to remove pigment and floccus, reduces the matrix effect of sample on mass spectrum greatly.
Determining of embodiment 3LC-MS/MS testing conditions
As shown in Fig. 5-7, according to relevant regulations in CAC and No. 657/2002/EEC resolution of EU, selecting two pairs of ions to carry out MRM monitoring can meet, the simultaneously selection of daughter ion mainly considers that wherein parent ion and daughter ion are chosen according to the mass spectrogram of every kind of compound and architectural characteristic, and at actual sample, analyzes mesostroma and disturb less.Determine the molion of various materials, then the molion of various compounds of take is respectively parent ion, its daughter ion is carried out to full scan and choose that abundance is strong, to disturb two pairs of less daughter ions be qualitative ion, as shown in Figure 1-2, the ion flow graph of final two pairs of ions determining as shown in Figure 3 for MC-LR and MC-RR second order ms figure.Finally with multiple-reaction monitoring (MRM) positive ion mode, optimize various mass spectrum parameters.
LC-MS/MS testing conditions:
Liquid-phase condition Waters Atlantis dC18 chromatographic column (2.1mm * 150mm, 5 μ m), 30 ℃ of column temperatures; Sample size 20 μ L; Mobile phase A is water (containing 0.05% formic acid, 5mmol/L ammonium formate), and B is 95% acetonitrile (containing 0.05% formic acid, 5mmol/L ammonium formate); Gradient elution;
Mass spectrum condition ionization mode is electro-spray ionization holotype (ESI+); Multiple-reaction monitoring pattern (MRM); Collision atmospheric pressure (CAD): 8; Gas curtain atmospheric pressure (CUR): 15; Electron spray voltage (IS): 5000V; Ion source temperature (TEM): 600 ℃.
Embodiment 4 recovery experiments
(1) sample adds:
Get blank algae health products sample and carry out the interpolation of three levels, the interpolation level of MC-LR and MC-RR toxin is all 0.1 μ g/kg, 1.0 μ g/kg, 5.0 μ g/kg, each level do 6 parallel.
(2) sample pre-treatments
Accurately take the algae health products 1.0g that has added toxin, with 10mL60% methanol aqueous solution, extract toxin, 200rpm shaking table concussion 60min, more ultrasonic processing 30min, 20 ℃ of following centrifugal 10min of 8000rpm, get supernatant.
With 2mol/L formic acid, regulate supernatant pH value to 4.0, standing 3h, 20 ℃ of following centrifugal 10min of 8000rpm, add weak aqua ammonia that pH value is adjusted to 7.0, add subsequently ultrapure water to be accurately settled to 30mL.
(3) enrichment and purification
First with 10mL methyl alcohol and 10mL ultrapure water activation C18 solid-phase extraction column, reactivation process flow control is at 1-2 drop/sec, get 15mL crude extract injection C18 solid-phase extraction column and carry out enrichment, flow control is at 1 drop/sec, then more respectively with each 10mL drip washing of 20% and 30% methanol aqueous solution C18 solid-phase extraction column, finally with 5mL70% methanol aqueous solution (containing 0.05% formic acid) wash-out Microcystin, nitrogen blows near dry, with 50% acetonitrile solution, be settled to 0.5mL, treat that liquid chromatography-tandem mass spectrometry (LC-MS/MS) detects.
(4) LC-MS/MS detects
Liquid-phase condition Waters Atlantis dC18 chromatographic column (2.1mm * 150mm, 5 μ m), 30 ℃ of column temperatures; Sample size 20 μ L; Mobile phase A is water (containing 0.05% formic acid, 5mmol/L ammonium formate), and B is 95% acetonitrile (containing 0.05% formic acid, 5mmol/L ammonium formate); Condition of gradient elution is in Table 1:
Table 1 condition of gradient elution
Mass spectrum condition ionization mode is electro-spray ionization holotype (ESI+); Multiple-reaction monitoring pattern (MRM); Collision atmospheric pressure (CAD): 8; Gas curtain atmospheric pressure (CUR): 15; Electron spray voltage (IS): 5000V; Ion source temperature (TEM): 600 ℃.
Multiple-reaction monitoring ion pair and the mass spectrum correlation parameter of compound are as shown in table 2.
The multiple-reaction monitoring ion pair of table 2 compound and mass spectrum correlation parameter table
* be quota ion
Experimental result demonstration, the recovery of this experimental technique is 91.4%～99.6%, the coefficient of variation is 1.2%～6.3%.
Embodiment 5
2 kinds of health-care spinulina foods producing in 2 domestic enterprises of supermarket stochastic buying, a kind of is tablet, a kind of is capsule.Tablet samples adopts homogenizer to pulverize homogeneous, and capsule product removes outside capsule, gets internal helicoid algae powder.Utilize this method to carry out the detection of Microcystin to 2 kinds of products.
(1) sample pre-treatments
Accurately take Powdered spirulina sample 1.0g, with 10mL60% methanol aqueous solution, extract toxin, 200rpm shaking table concussion 60min, more ultrasonic processing 30min, 20 ℃ of following centrifugal 10min of 8000rpm, get supernatant.
With 2mol/L formic acid, regulate supernatant pH value to 4.0, standing 3h, 20 ℃ of following centrifugal 10min of 8000rpm, add weak aqua ammonia that pH value is adjusted to 7.0, add subsequently ultrapure water to be accurately settled to 30mL.
Get blank spirulina powder and add recovery experiment, MC-LR, MC-RR toxin add water product and are all 1.0 μ g/kg, and pre-treating method is identical with sample method.
(2) enrichment and purification
First with 10mL methyl alcohol and 10mL ultrapure water activation C18 solid-phase extraction column, reactivation process flow control is at 1-2 drop/sec, get 15mL crude extract injection C18 solid-phase extraction column and carry out enrichment, flow control is at 1 drop/sec, then more respectively with each 10mL drip washing of 20% and 30% methanol aqueous solution C18 solid-phase extraction column, finally with 5mL70% methanol aqueous solution (containing 0.05% formic acid) wash-out Microcystin, nitrogen blows near dry, with 50% acetonitrile solution, be settled to 0.5mL, treat that liquid chromatography-tandem mass spectrometry (LC-MS/MS) detects.
(3) LC-MS/MS detects
Liquid-phase condition Waters Atlantis dC18 chromatographic column (2.1mm * 150mm, 5 μ m), 30 ℃ of column temperatures; Sample size 20 μ L; Mobile phase A is water (containing 0.05% formic acid, 5mmol/L ammonium formate), and B is 95% acetonitrile (containing 0.05% formic acid, 5mmol/L ammonium formate); Condition of gradient elution sees the following form:
Table condition of gradient elution
Mass spectrum condition ionization mode is electro-spray ionization holotype (ESI+); Multiple-reaction monitoring pattern (MRM); Collision atmospheric pressure (CAD): 8; Gas curtain atmospheric pressure (CUR): 15; Electron spray voltage (IS): 5000V; Ion source temperature (TEM): 600 ℃.
Multiple-reaction monitoring ion pair and the mass spectrum correlation parameter of compound are as shown in the table.
Multiple-reaction monitoring ion pair and the mass spectrum correlation parameter table of table compound
* be quota ion
(4) testing result
The recovery of method is: MC-LR is that 92.5%, MC-RR is 97.1%.
After the testing result of sample is proofreaied and correct by the recovery, obtain final testing result: in tablet health-care spinulina food, MC-LR content is 0.52 μ g/kg, MC-RR content is 12.6 μ g/kg, and in capsule health-care spinulina food, MC-LR content is 0.23 μ g/kg, and MC-RR content is 27.1 μ g/kg.
Claims (4)
1. a liquid chromatography-tandem mass for Microcystin, is characterized in that, comprises the following steps:
(1) sample pre-treatments
Accurately take through algae health products 1.0 g that pulverize, homogeneous is crossed, with 10 mL 60 % methanol aqueous solutions, extract toxin, 200 rpm shaking tables shake 60 min, more ultrasonic processing 30 min, and 20 ℃ of following centrifugal 10 min of 8000 rpm, get supernatant;
Regulate supernatant pH value to 4.0, standing 3 h can be observed and in solution, occur a large amount of flocculent deposits, 20 ℃ of following centrifugal 10 min of 8000 rpm, obtain light yellow supernatant or water white transparency liquid, in solution, add weak aqua ammonia that pH value is adjusted to 6.8-7.2, add subsequently ultrapure water to be accurately settled to 30 mL, now obtain comparatively pure crude extract;
(2) enrichment and purification
First with 10 mL methyl alcohol and 10 mL ultrapure waters activation C18 solid-phase extraction column, reactivation process flow control is at 1-2 drop/sec, get 15 mL crude extracts injection C18 solid-phase extraction columns and carry out enrichment, flow control is at 1 drop/sec, then more respectively with 20 % and each 10 mL drip washing C18 solid-phase extraction columns of 30 % methanol aqueous solutions, to remove impurity, finally with 5 mL 70 % methanol aqueous solutions (containing 0.05 % formic acid) wash-out Microcystins, nitrogen blows near dry, with 50 % acetonitrile solutions, be settled to 0.5 mL, treat that liquid chromatography-tandem mass spectrometry (LC-MS/MS) detects;
(3) LC-MS/MS detects
Liquid-phase condition Waters Atlantis dC18 chromatographic column (2.1 mm * 150 mm, 5 μ m), 30 ℃ of column temperatures; Sample size 20 μ L; Mobile phase A is water (containing 0.05% formic acid, 5 mmol/L ammonium formates), and B is 95 % acetonitriles (containing 0.05% formic acid, 5 mmol/L ammonium formates); Gradient elution;
Mass spectrum condition ionization mode is electro-spray ionization holotype (ESI+); Multiple-reaction monitoring pattern (MRM); Collision atmospheric pressure (CAD): 8; Gas curtain atmospheric pressure (CUR): 15; Electron spray voltage (IS): 5000 V; Ion source temperature (TEM): 600 ℃.
2. the liquid chromatography-tandem mass of Microcystin according to claim 1, is characterized in that, in step (1) solution, adds weak aqua ammonia that pH value is adjusted to 7.0.
3. the liquid chromatography-tandem mass of Microcystin according to claim 1, is characterized in that, in step (1), with 2 mol/L formic acid, regulates supernatant pH value to 4.0.
4. the liquid chromatography-tandem mass of Microcystin according to claim 1, is characterized in that, in step (3) with [M+2H]
2+ion is as parent ion.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201410011060.3A CN103940921B (en) | 2014-01-09 | 2014-01-09 | A kind of liquid chromatography-tandem mass of Microcystin |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201410011060.3A CN103940921B (en) | 2014-01-09 | 2014-01-09 | A kind of liquid chromatography-tandem mass of Microcystin |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN103940921A true CN103940921A (en) | 2014-07-23 |
| CN103940921B CN103940921B (en) | 2015-09-30 |
Family
ID=51188675
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201410011060.3A Expired - Fee Related CN103940921B (en) | 2014-01-09 | 2014-01-09 | A kind of liquid chromatography-tandem mass of Microcystin |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN103940921B (en) |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106279359A (en) * | 2016-08-11 | 2017-01-04 | 中国水稻研究所 | A kind of method preparing five kinds of ustilaginoidea virens toxin |
| CN107389825A (en) * | 2017-08-11 | 2017-11-24 | 浙江省食品药品检验研究院 | The method that algae toxin in water is determined based on full-automatic on-line solid phase extraction ultra performance liquid chromatography linear ion hydrazine tandem mass spectrum |
| CN113447581A (en) * | 2021-06-23 | 2021-09-28 | 中国食品药品检定研究院 | Method for detecting microcystin in algae food and raw materials for producing algae food |
Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1401660A (en) * | 2002-09-24 | 2003-03-12 | 中国科学院水生生物研究所 | Method for extracting and purifying microcytis toxin |
| KR20110115681A (en) * | 2010-04-16 | 2011-10-24 | 강원대학교산학협력단 | Microcystin detection kit using paper chromatography and chemical luminescence and detection method thereof |
| CN103076420A (en) * | 2012-12-31 | 2013-05-01 | 江苏省环境监测中心 | Method for detecting multicomponent microcystins through ultra-high performance liquid chromatography/triple quadrupole tandem mass spectrometry |
| CN103163001A (en) * | 2013-03-27 | 2013-06-19 | 云南省环境监测中心站 | Method for extracting and purifying microcystic toxins LR and RR by taking cyanobacterial bloom in Dian Lake as raw material |
-
2014
- 2014-01-09 CN CN201410011060.3A patent/CN103940921B/en not_active Expired - Fee Related
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1401660A (en) * | 2002-09-24 | 2003-03-12 | 中国科学院水生生物研究所 | Method for extracting and purifying microcytis toxin |
| KR20110115681A (en) * | 2010-04-16 | 2011-10-24 | 강원대학교산학협력단 | Microcystin detection kit using paper chromatography and chemical luminescence and detection method thereof |
| CN103076420A (en) * | 2012-12-31 | 2013-05-01 | 江苏省环境监测中心 | Method for detecting multicomponent microcystins through ultra-high performance liquid chromatography/triple quadrupole tandem mass spectrometry |
| CN103163001A (en) * | 2013-03-27 | 2013-06-19 | 云南省环境监测中心站 | Method for extracting and purifying microcystic toxins LR and RR by taking cyanobacterial bloom in Dian Lake as raw material |
Non-Patent Citations (6)
| Title |
|---|
| PING TONG 等: "Analysis of microcystins by capillary zone electrophoresis coupling with electrospray ionization mass spectrometry", 《TALANTA》 * |
| SUSANN HILLER 等: "Rapid detection of cyanobacterial toxins in precursor ion mode by liquid chromatography tandem mass spectrometry", 《JOURNAL OF MASS SPECTROMETRY》 * |
| 张维昊 等: "天然水华蓝藻中微囊藻毒素的提取和净化研究", 《环境污染与防治》 * |
| 范赛 等: "水产品中微囊藻毒素检测方法的研究进展", 《色谱》 * |
| 郑雪琴 等: "高效液相色谱法测定蓝绿藻中微囊藻毒素", 《理化检验（化学分册）》 * |
| 陈立坚 等: "高效液相色谱-串联质谱法测定水产品中7种微囊藻毒素", 《现代食品科技》 * |
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106279359A (en) * | 2016-08-11 | 2017-01-04 | 中国水稻研究所 | A kind of method preparing five kinds of ustilaginoidea virens toxin |
| CN107389825A (en) * | 2017-08-11 | 2017-11-24 | 浙江省食品药品检验研究院 | The method that algae toxin in water is determined based on full-automatic on-line solid phase extraction ultra performance liquid chromatography linear ion hydrazine tandem mass spectrum |
| CN107389825B (en) * | 2017-08-11 | 2020-07-07 | 浙江省食品药品检验研究院 | Method for determining algae toxins in water based on full-automatic online solid-phase extraction-ultra-high performance liquid chromatography-linear ion trap tandem mass spectrometry |
| CN113447581A (en) * | 2021-06-23 | 2021-09-28 | 中国食品药品检定研究院 | Method for detecting microcystin in algae food and raw materials for producing algae food |
Also Published As
| Publication number | Publication date |
|---|---|
| CN103940921B (en) | 2015-09-30 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Nardi et al. | Soil–root cross‐talking: The role of humic substances | |
| Magalhães et al. | Evaluation of the composition of street cocaine seized in two regions of Brazil | |
| Corte et al. | Assessment of safety and efficiency of nitrogen organic fertilizers from animal‐based protein hydrolysates—a laboratory multidisciplinary approach | |
| Zhao et al. | Vortex-assisted dispersive liquid-liquid microextraction for the analysis of major Aspergillus and Penicillium mycotoxins in rice wine by liquid chromatography-tandem mass spectrometry | |
| CN105651899B (en) | Method for detecting aflatoxin with high sensitivity and application thereof | |
| CN102200530B (en) | Method for detecting 33 kinds of narcotic drugs in urine by liquid chromatography-tandem mass spectrometry | |
| Llorente-Mirandes et al. | Inorganic arsenic determination in food: a review of analytical proposals and quality assessment over the last six years | |
| Kong et al. | Analysis of fumonisins B 1 and B 2 in spices and aromatic and medicinal herbs by HPLC-FLD with on-line post-column derivatization and positive confirmation by LC-MS/MS | |
| Tsednee et al. | A sensitive LC‐ESI‐Q‐TOF‐MS method reveals novel phytosiderophores and phytosiderophore–iron complexes in barley | |
| Hu et al. | Profiling of triacylglycerols in plant oils by high-performance liquid chromatography–atmosphere pressure chemical ionization mass spectrometry using a novel mixed-mode column | |
| CN104698114A (en) | Method for detecting 205 kinds of pesticide residues in rice | |
| CN103940921B (en) | A kind of liquid chromatography-tandem mass of Microcystin | |
| Cai et al. | Dispersive solid-phase extraction followed by high-performance liquid chromatography/tandem mass spectrometry for the determination of ricinine in cooking oil | |
| CN102519930A (en) | Method for rapidly determining selenium content in plant sample | |
| Li et al. | Determination of 35 free amino acids in tea using ultra-performance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry | |
| CN103278594A (en) | Universal rapid detection method for micromolecule poisonous and harmful materials in powdery food | |
| CN105319313B (en) | Liquid chromatogram-tandem mass spectrum detection method of toxoflavin | |
| Mélançon et al. | Impact of ocean acidification on phytoplankton assemblage, growth, and DMS production following Fe-dust additions in the NE Pacific high-nutrient, low-chlorophyll waters | |
| Kim et al. | Metabolomics-based optimal koji fermentation for tyrosinase inhibition supplemented with Astragalus Radix | |
| Amaral et al. | Evaluation of sample preparation procedures and krypton as an interference standard probe for arsenic speciation by HPLC-ICP-QMS | |
| Bartók et al. | Identification of unknown isomers of fumonisin B5 mycotoxin in a Fusarium verticillioides culture by high-performance liquid chromatography/electrospray ionization time-of-flight and ion trap mass spectrometry | |
| US11474088B2 (en) | Method for determining origin of amino acid | |
| Krüger et al. | Development of an analytical method for the unambiguous structure elucidation of cyclic peptides with special appliance for hepatotoxic desmethylated microcystins | |
| CN106324169B (en) | The SPE gas-chromatography tandem mass spectrum detection method of acid amide fungicides in grape wine | |
| CN104977383A (en) | Method for rapidly and quantitatively detecting microcystins in spirulina food |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C14 | Grant of patent or utility model | ||
| GR01 | Patent grant | ||
| CF01 | Termination of patent right due to non-payment of annual fee |
Granted publication date: 20150930 Termination date: 20210109 |
|
| CF01 | Termination of patent right due to non-payment of annual fee |