CN108982689A - A kind of method of two kinds of rice song toxin in quick detection paddy - Google Patents
A kind of method of two kinds of rice song toxin in quick detection paddy Download PDFInfo
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- CN108982689A CN108982689A CN201810726387.7A CN201810726387A CN108982689A CN 108982689 A CN108982689 A CN 108982689A CN 201810726387 A CN201810726387 A CN 201810726387A CN 108982689 A CN108982689 A CN 108982689A
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- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
- G01N30/02—Column chromatography
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- G—PHYSICS
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- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
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Abstract
The invention discloses a kind of methods of two kinds of rice song toxin in quickly detection paddy, include the following steps: to choose and crushes rice sample, and it is placed in rice sample is crushed in centrifuge tube, after Plus acidic aqueous solution high-speed homogenization extracts, high speed centrifugation processing, take supernatant, Plus acidic aqueous solution high-speed homogenization homogeneous extracts residue again, high speed centrifugation again, merge supernatant in concentrate bottle, after concentration, it is to be clean, by treated, supernatant is placed in C18 solid-phase extraction column, successively use methanol, ultrapure water activation, supernatant methanol and water mixed solution elution, concentration is close dry, then methanol and formic acid water mixed solution constant volume are utilized again, filter membrane, purified rice sample is carried out ultra performance liquid chromatography Tandem Mass Spectrometry Analysis, and the object of the present invention is to provide a kind of simple, quickly, it is economical, it is sensitive, safety.
Description
Technical field
The present invention relates to a kind of methods of two kinds of rice song toxin in detection technique field, more particularly to quickly detection paddy.
Background technique
Rice originates in China, have 7,000 year many years plantation history, China be in the world maximum Rice Production state and
Country of consumption, rice are one of national important staple food grains.China's rice region is vast, and in addition to Qinghai Province, there is rice cultivation in remaining each province.Rice
Bent disease is that a kind of fungal disease as caused by green solani infestation Rice Panicle also causes after infecting in addition to endangering itself grain
Increase disease fringe sterile grain rate, mass of 1000 kernel reduces, and causes the underproduction in poor harvest, directly affects rice yield and quality, the bent poison of the rice of generation
Element is to the toxic effect of people and animals, and the content highest and activity of semilate rice song toxin A is maximum, followed by rice song toxin B.Current
Detection method is mainly to be combined method, liquid chromatogram high-resolution matter by liquid chromatography, liquid chromatogram matter after organic reagent extracts
Spectrometry etc. tests and analyzes rice song toxin, current method is long there are the pre-treatment time, reagent consumption is big, poor repeatability,
Equipment component valuableness is unfavorable for the shortcomings that implementing.For this purpose, we have proposed two kinds of rice song toxin in a kind of quickly detection paddy
Method.
Summary of the invention
The invention proposes a kind of methods of two kinds of rice song toxin in quickly detection paddy, to solve in above-mentioned background technique
The problem of proposition.
The invention proposes a kind of methods of two kinds of rice song toxin in quickly detection paddy, include the following steps:
S1: the preparation of standard sample solution: choosing rice song toxin A and rice song toxin B, respectively with ultrapure water by rice song toxin
A and rice song toxin B be diluted to mass concentration be 0.005mg/L, 0.01mg/L, 0.05mg/L, 0.1mg/L, 0.5mg/L,
The standard solution of 1.0mg/L；
S2: it the extraction of sample: chooses and crushes rice sample, and be placed in rice sample is crushed in centrifuge tube, Plus acidic water
After solution high-speed homogenization extracts, high speed centrifugation processing takes supernatant, and Plus acidic aqueous solution high-speed homogenization homogeneous extracts residue again,
High speed centrifugation again merges supernatant in concentrate bottle, to be clean after concentration；
S3: the purification of sample: by treated in S2, supernatant is placed in C18 solid-phase extraction column, successively with methanol, ultrapure
Water activation, supernatant methanol and water mixed solution elution, are concentrated and closely do, and are then mixed again using methanol with formic acid water molten
Liquid constant volume, filter membrane are to be measured after the completion；
S4: rice sample purified in S3 is subjected to ultra performance liquid chromatography Tandem Mass Spectrometry Analysis, specific step is as follows；
(1), a chromatographic column is chosen, and injects A, B mixed flow phase thereto, and A phase is specially water, the water includes
There is formic acid, B phase is specially hplc grade methanol, and A, B mixed flow are mutually with gradient elution mode, by the flow velocity of 0.1-0.4ml/min
Into in chromatographic column, analysis time 5 minutes, it can guarantee that object and impurity are sufficiently separated；
(2), after the completion of above-mentioned, it is placed in again in triple level Four rod-type mass spectrographs, sample is bombarded with stream of energetic electrons,
So that the sample is lost molecular ion and fragment ion that electronics becomes positively charged, passes through atomizer, dry gas and collision gas
Mass spectrum optimization processing is carried out to molecular ion, and atomizer air-flow is 1.5-3L/min, dry gas air-flow is 15-20L/min, is touched
Hitting gas is 200-230Kpa；
S5: in addition a certain concentration standard solution to the blank rice sample crushed, addition recycling sample is obtained, will be added
Recycling sample, sample to be tested are handled sample according to S2, S3 and S4, with prepared in S1 0.005mg/L,
Six concentration of 0.01mg/L, 0.05mg/L, 0.1mg/L, 0.5mg/L, 1.0mg/L are standard working curve, then carry out LC-
MSMS respectively obtains corresponding peak area, brings addition recycling sample and sample to be tested peak area into standard working curve side
Journey, calculates the TIANZHU XINGNAO Capsul and the coefficient of variation of rice song toxin A and rice song toxin B, and obtains the toxin in sample to be tested and contain
Amount completes the detection of paddy semilate rice song toxin.
Preferably, it is 10-20g that rice sample amount is weighed in S2, is separately added into 20-30ml acidic aqueous solution twice.
Preferably, the acidic aqueous solution pH=3-5 of sample extraction is then adjusted using formic acid if being not located at the region
Section.
Preferably, in S2 in centrifugal treating, high-speed homogenization revolving speed is 10000-12000rpm, and the time is 2min.
Preferably, the volume ratio of the methanol in S3 and water is 5:5-8:2.
Preferably, in carrying out ultra performance liquid chromatography Tandem Mass Spectrometry Analysis, the content of formic acid is 0.1- in the water of A phase
0.2%.
Preferably, the column temperature of the chromatographic column is 30-40 degrees Celsius.
Preferably, the chromatographic column is specially Shimadzu Shim-pack XR-C8,2.0 × 100mm of specification, and 1.6 μm of partial size,
10 μ l of sample volume.
The method of two kinds of rice song toxin, beneficial effect are in a kind of quickly detection paddy proposed by the present invention: the paddy
The detection method of semilate rice song toxin pass through the HCl dropwise water of sample after, extracted using high speed homogenization, can analyze simultaneously rice song toxin A with
Rice song toxin B, method time-consuming is short, and extraction does not consume organic reagent, reduces environmental pollution, purification selection less expensive
C18 solid phase extraction column, saves testing cost, and favorable reproducibility improves work efficiency.The object of the present invention is to provide a kind of letters
It is single, quick, economical, sensitive, safe, it can be used for analyzing and monitoring the detection side of the rice song toxin A and rice song toxin B in rice
Method.
Specific embodiment
It is next combined with specific embodiments below that the present invention will be further described.
Embodiment 1
The invention proposes a kind of methods of two kinds of rice song toxin in quickly detection paddy, include the following steps:
S1: the preparation of standard sample solution: choosing rice song toxin A and rice song toxin B, respectively with ultrapure water by rice song toxin
A and rice song toxin B be diluted to mass concentration be 0.005mg/L, 0.01mg/L, 0.05mg/L, 0.1mg/L, 0.5mg/L,
The standard solution of 1.0mg/L；
S2: it the extraction of sample: chooses and crushes rice sample, and be placed in rice sample is crushed in centrifuge tube, Plus acidic water
After solution high-speed homogenization extracts, high speed centrifugation processing takes supernatant, and Plus acidic aqueous solution high-speed homogenization homogeneous extracts residue again,
High speed centrifugation again merges supernatant in concentrate bottle, to be clean after concentration；
S3: the purification of sample: by treated in S2, supernatant is placed in C18 solid-phase extraction column, successively with methanol, ultrapure
Water activation, supernatant methanol and water mixed solution elution, are concentrated and closely do, and are then mixed again using methanol with formic acid water molten
Liquid constant volume, filter membrane are to be measured after the completion；
S4: rice sample purified in S3 is subjected to ultra performance liquid chromatography Tandem Mass Spectrometry Analysis, specific step is as follows；
(1), a chromatographic column is chosen, and injects A, B mixed flow phase thereto, and A phase is specially water, the water includes
There is formic acid, B phase is specially hplc grade methanol, and A, B mixed flow are mutually with gradient elution mode, by the flow velocity of 0.1-0.4ml/min
Into in chromatographic column, analysis time 5 minutes, it can guarantee that object and impurity are sufficiently separated；
Gradient elution
(2), after the completion of above-mentioned, it is placed in again in triple level Four rod-type mass spectrographs, sample is bombarded with stream of energetic electrons,
So that the sample is lost molecular ion and fragment ion that electronics becomes positively charged, passes through atomizer, dry gas and collision gas
Mass spectrum optimization processing is carried out to molecular ion, and atomizer air-flow is 1.5L/min, dry gas air-flow is 15L/min, collision gas
For 200Kpa；Specifically it see the table below:
Mass spectrum optimal conditions table
A: quota ion；B: qualitative ion
S5: in addition a certain concentration standard solution to the blank rice sample crushed, addition recycling sample is obtained, will be added
Recycling sample, sample to be tested are handled sample according to S2, S3 and S4, with prepared in S1 0.005mg/L,
Six concentration of 0.01mg/L, 0.05mg/L, 0.1mg/L, 0.5mg/L, 1.0mg/L are standard working curve, then carry out LC-
MSMS respectively obtains corresponding peak area, brings addition recycling sample and sample to be tested peak area into standard working curve side
Journey, calculates the TIANZHU XINGNAO Capsul and the coefficient of variation of rice song toxin A and rice song toxin B, and obtains the toxin in sample to be tested and contain
Amount completes the detection of paddy semilate rice song toxin.
It is 10-20g that rice sample amount is weighed in S2, is separately added into 20-30ml acidic aqueous solution twice.
The acidic aqueous solution pH=3-5 of sample extraction is then adjusted using formic acid if being not located at the region.
In S2 in centrifugal treating, high-speed homogenization revolving speed is 10000rpm, and the time is 2min.
The volume ratio of methanol and water in S3 is 5:5-8:2.
In carrying out ultra performance liquid chromatography Tandem Mass Spectrometry Analysis, the content of formic acid is 0.1-0.2% in the water of A phase.
The column temperature of the chromatographic column is 30 degrees Celsius.
The chromatographic column is specially Shimadzu Shim-pack XR-C8,2.0 × 100mm of specification, and 1.6 μm of partial size, sample volume 10
μl。
Embodiment 2
The invention proposes a kind of methods of two kinds of rice song toxin in quickly detection paddy, include the following steps:
S1: the preparation of standard sample solution: choosing rice song toxin A and rice song toxin B, respectively with ultrapure water by rice song toxin
A and rice song toxin B be diluted to mass concentration be 0.005mg/L, 0.01mg/L, 0.05mg/L, 0.1mg/L, 0.5mg/L,
The standard solution of 1.0mg/L；
S2: it the extraction of sample: chooses and crushes rice sample, and be placed in rice sample is crushed in centrifuge tube, Plus acidic water
After solution high-speed homogenization extracts, high speed centrifugation processing takes supernatant, and Plus acidic aqueous solution high-speed homogenization homogeneous extracts residue again,
High speed centrifugation again merges supernatant in concentrate bottle, to be clean after concentration；
S3: the purification of sample: by treated in S2, supernatant is placed in C18 solid-phase extraction column, successively with methanol, ultrapure
Water activation, supernatant methanol and water mixed solution elution, are concentrated and closely do, and are then mixed again using methanol with formic acid water molten
Liquid constant volume, filter membrane are to be measured after the completion；
S4: rice sample purified in S3 is subjected to ultra performance liquid chromatography Tandem Mass Spectrometry Analysis, specific step is as follows；
(1), a chromatographic column is chosen, and injects A, B mixed flow phase thereto, and A phase is specially water, the water includes
There is formic acid, B phase is specially hplc grade methanol, and A, B mixed flow are mutually with gradient elution mode, by the flow velocity of 0.1-0.4ml/min
Into in chromatographic column, analysis time 5 minutes, it can guarantee that object and impurity are sufficiently separated；
Gradient elution
(2), after the completion of above-mentioned, it is placed in again in triple level Four rod-type mass spectrographs, sample is bombarded with stream of energetic electrons,
So that the sample is lost molecular ion and fragment ion that electronics becomes positively charged, passes through atomizer, dry gas and collision gas
Mass spectrum optimization processing is carried out to molecular ion, and atomizer air-flow is 2L/min, dry gas air-flow is 16L/min, and collision gas is
210Kpa；
Mass spectrum optimal conditions table
A: quota ion；B: qualitative ion
S5: in addition a certain concentration standard solution to the blank rice sample crushed, addition recycling sample is obtained, will be added
Recycling sample, sample to be tested are handled sample according to S2, S3 and S4, with prepared in S1 0.005mg/L,
Six concentration of 0.01mg/L, 0.05mg/L, 0.1mg/L, 0.5mg/L, 1.0mg/L are standard working curve, then carry out LC-
MSMS respectively obtains corresponding peak area, brings addition recycling sample and sample to be tested peak area into standard working curve side
Journey, calculates the TIANZHU XINGNAO Capsul and the coefficient of variation of rice song toxin A and rice song toxin B, and obtains the toxin in sample to be tested and contain
Amount completes the detection of paddy semilate rice song toxin.
It is 10-20g that rice sample amount is weighed in S2, is separately added into 20-30ml acidic aqueous solution twice.
The acidic aqueous solution pH=3-5 of sample extraction is then adjusted using formic acid if being not located at the region.
In S2 in centrifugal treating, high-speed homogenization revolving speed is 10500rpm, and the time is 2min.
The volume ratio of methanol and water in S3 is 5:5-8:2.
In carrying out ultra performance liquid chromatography Tandem Mass Spectrometry Analysis, the content of formic acid is 0.1-0.2% in the water of A phase.
The column temperature of the chromatographic column is 33 degrees Celsius.
The chromatographic column is specially Shimadzu Shim-pack XR-C8,2.0 × 100mm of specification, and 1.6 μm of partial size, sample volume 10
μl。
Embodiment 3
The invention proposes a kind of methods of two kinds of rice song toxin in quickly detection paddy, include the following steps:
S1: the preparation of standard sample solution: choosing rice song toxin A and rice song toxin B, respectively with ultrapure water by rice song toxin
A and rice song toxin B be diluted to mass concentration be 0.005mg/L, 0.01mg/L, 0.05mg/L, 0.1mg/L, 0.5mg/L,
The standard solution of 1.0mg/L；
S2: it the extraction of sample: chooses and crushes rice sample, and be placed in rice sample is crushed in centrifuge tube, Plus acidic water
After solution high-speed homogenization extracts, high speed centrifugation processing takes supernatant, and Plus acidic aqueous solution high-speed homogenization homogeneous extracts residue again,
High speed centrifugation again merges supernatant in concentrate bottle, to be clean after concentration；
S3: the purification of sample: by treated in S2, supernatant is placed in C18 solid-phase extraction column, successively with methanol, ultrapure
Water activation, supernatant methanol and water mixed solution elution, are concentrated and closely do, and are then mixed again using methanol with formic acid water molten
Liquid constant volume, filter membrane are to be measured after the completion；
S4: rice sample purified in S3 is subjected to ultra performance liquid chromatography Tandem Mass Spectrometry Analysis, specific step is as follows；
(1), a chromatographic column is chosen, and injects A, B mixed flow phase thereto, and A phase is specially water, the water includes
There is formic acid, B phase is specially hplc grade methanol, and A, B mixed flow are mutually with gradient elution mode, by the flow velocity of 0.1-0.4ml/min
Into in chromatographic column, analysis time 5 minutes, it can guarantee that object and impurity are sufficiently separated；
Gradient elution
(2), after the completion of above-mentioned, it is placed in again in triple level Four rod-type mass spectrographs, sample is bombarded with stream of energetic electrons,
So that the sample is lost molecular ion and fragment ion that electronics becomes positively charged, passes through atomizer, dry gas and collision gas
Mass spectrum optimization processing is carried out to molecular ion, and atomizer air-flow is 2.5L/min, dry gas air-flow is 18L/min, collision gas
For 220Kpa；Specifically it see the table below:
Mass spectrum optimal conditions table
A: quota ion；B: qualitative ion
S5: in addition a certain concentration standard solution to the blank rice sample crushed, addition recycling sample is obtained, will be added
Recycling sample, sample to be tested are handled sample according to S2, S3 and S4, with prepared in S1 0.005mg/L,
Six concentration of 0.01mg/L, 0.05mg/L, 0.1mg/L, 0.5mg/L, 1.0mg/L are standard working curve, then carry out LC-
MSMS respectively obtains corresponding peak area, brings addition recycling sample and sample to be tested peak area into standard working curve side
Journey, calculates the TIANZHU XINGNAO Capsul and the coefficient of variation of rice song toxin A and rice song toxin B, and obtains the toxin in sample to be tested and contain
Amount completes the detection of paddy semilate rice song toxin.
It is 10-20g that rice sample amount is weighed in S2, is separately added into 20-30ml acidic aqueous solution twice.
The acidic aqueous solution pH=3-5 of sample extraction is then adjusted using formic acid if being not located at the region.
In S2 in centrifugal treating, high-speed homogenization revolving speed is 11000rpm, and the time is 2min.
The volume ratio of methanol and water in S3 is 5:5-8:2.
In carrying out ultra performance liquid chromatography Tandem Mass Spectrometry Analysis, the content of formic acid is 0.1-0.2% in the water of A phase.
The column temperature of the chromatographic column is 37 degrees Celsius.
The chromatographic column is specially Shimadzu Shim-pack XR-C8,2.0 × 100mm of specification, and 1.6 μm of partial size, sample volume 10
μl。
Embodiment 4
The invention proposes a kind of methods of two kinds of rice song toxin in quickly detection paddy, include the following steps:
S1: the preparation of standard sample solution: choosing rice song toxin A and rice song toxin B, respectively with ultrapure water by rice song toxin
A and rice song toxin B be diluted to mass concentration be 0.005mg/L, 0.01mg/L, 0.05mg/L, 0.1mg/L, 0.5mg/L,
The standard solution of 1.0mg/L；
S2: it the extraction of sample: chooses and crushes rice sample, and be placed in rice sample is crushed in centrifuge tube, Plus acidic water
After solution high-speed homogenization extracts, high speed centrifugation processing takes supernatant, and Plus acidic aqueous solution high-speed homogenization homogeneous extracts residue again,
High speed centrifugation again merges supernatant in concentrate bottle, to be clean after concentration；
S3: the purification of sample: by treated in S2, supernatant is placed in C18 solid-phase extraction column, successively with methanol, ultrapure
Water activation, supernatant methanol and water mixed solution elution, are concentrated and closely do, and are then mixed again using methanol with formic acid water molten
Liquid constant volume, filter membrane are to be measured after the completion；
S4: rice sample purified in S3 is subjected to ultra performance liquid chromatography Tandem Mass Spectrometry Analysis, specific step is as follows；
(1), a chromatographic column is chosen, and injects A, B mixed flow phase thereto, and A phase is specially water, the water includes
There is formic acid, B phase is specially hplc grade methanol, and A, B mixed flow are mutually with gradient elution mode, by the flow velocity of 0.1-0.4ml/min
Into in chromatographic column, analysis time 5 minutes, it can guarantee that object and impurity are sufficiently separated；
Gradient elution
(2), after the completion of above-mentioned, it is placed in again in triple level Four rod-type mass spectrographs, sample is bombarded with stream of energetic electrons,
So that the sample is lost molecular ion and fragment ion that electronics becomes positively charged, passes through atomizer, dry gas and collision gas
Mass spectrum optimization processing is carried out to molecular ion, and atomizer air-flow is 3L/min, dry gas air-flow is 20L/min, and collision gas is
230Kpa；Specifically it see the table below:
Mass spectrum optimal conditions table
A: quota ion；B: qualitative ion
S5: in addition a certain concentration standard solution to the blank rice sample crushed, addition recycling sample is obtained, will be added
Recycling sample, sample to be tested are handled sample according to S2, S3 and S4, with prepared in S1 0.005mg/L,
Six concentration of 0.01mg/L, 0.05mg/L, 0.1mg/L, 0.5mg/L, 1.0mg/L are standard working curve, then carry out LC-
MSMS respectively obtains corresponding peak area, brings addition recycling sample and sample to be tested peak area into standard working curve side
Journey, calculates the TIANZHU XINGNAO Capsul and the coefficient of variation of rice song toxin A and rice song toxin B, and obtains the toxin in sample to be tested and contain
Amount completes the detection of paddy semilate rice song toxin.
It is 10-20g that rice sample amount is weighed in S2, is separately added into 20-30ml acidic aqueous solution twice.
The acidic aqueous solution pH=3-5 of sample extraction is then adjusted using formic acid if being not located at the region.
In S2 in centrifugal treating, high-speed homogenization revolving speed is 12000rpm, and the time is 2min.
The volume ratio of methanol and water in S3 is 5:5-8:2.
In carrying out ultra performance liquid chromatography Tandem Mass Spectrometry Analysis, the content of formic acid is 0.1-0.2% in the water of A phase.
The column temperature of the chromatographic column is 40 degrees Celsius.
The chromatographic column is specially Shimadzu Shim-pack XR-C8,2.0 × 100mm of specification, and 1.6 μm of partial size, sample volume 10
μl。
The foregoing is only a preferred embodiment of the present invention, but scope of protection of the present invention is not limited thereto,
Anyone skilled in the art in the technical scope disclosed by the present invention, according to the technique and scheme of the present invention and its
Inventive concept is subject to equivalent substitution or change, should be covered by the protection scope of the present invention.
Claims (8)
1. a kind of method of two kinds of rice song toxin in quickly detection paddy, which comprises the following steps:
S1: standard sample solution preparation: choose rice song toxin A and rice song toxin B, respectively with ultrapure water by rice song toxin A with
And it is 0.005mg/L, 0.01mg/L, 0.05mg/L, 0.1mg/L, 0.5mg/L, 1.0mg/ that rice song toxin B, which is diluted to mass concentration,
The standard solution of L；
S2: it the extraction of sample: chooses and crushes rice sample, and be placed in rice sample is crushed in centrifuge tube, Plus acidic aqueous solution
After high-speed homogenization extracts, high speed centrifugation processing takes supernatant, and Plus acidic aqueous solution high-speed homogenization homogeneous extracts residue again, again
High speed centrifugation merges supernatant in concentrate bottle, to be clean after concentration；
S3: the purification of sample: by treated in S2, supernatant is placed in C18 solid-phase extraction column, successively living with methanol, ultrapure water
Change, supernatant methanol and water mixed solution elution are concentrated and closely do, then fixed using methanol and formic acid water mixed solution again
Hold, filter membrane is to be measured after the completion；
S4: rice sample purified in S3 is subjected to ultra performance liquid chromatography Tandem Mass Spectrometry Analysis, specific step is as follows；
(1), a chromatographic column is chosen, and injects A, B mixed flow phase thereto, and A phase is specially water, contains first in the water
Acid, B phase are specially hplc grade methanol, and A, B mixed flow mutually with gradient elution mode, are entered by the flow velocity of 0.1-0.4ml/min
In chromatographic column, analysis time 5 minutes, it can guarantee that object and impurity are sufficiently separated；
(2), after the completion of above-mentioned, it is placed in again in triple level Four rod-type mass spectrographs, sample is bombarded with stream of energetic electrons, makes this
Sample loses the molecular ion and fragment ion that electronics becomes positively charged, by atomizer, dry gas and collision gas to point
Daughter ion carries out mass spectrum optimization processing, and atomizer air-flow is 1.5-3L/min, and dry gas air-flow is 15-20L/min, collision gas
For 200-230Kpa；
S5: in addition a certain concentration standard solution to the blank rice sample crushed, addition recycling sample is obtained, addition is recycled
Sample, sample to be tested are handled sample according to S2, S3 and S4, with prepared in S1 0.005mg/L, 0.01mg/L,
Six concentration of 0.05mg/L, 0.1mg/L, 0.5mg/L, 1.0mg/L are standard working curve, then carry out LC-MS MS analysis point
Corresponding peak area is not obtained, brings addition recycling sample and sample to be tested peak area into standard working curve equation, it is bent to calculate rice
The TIANZHU XINGNAO Capsul and the coefficient of variation of toxin A and rice song toxin B, and obtain the content of toxins in sample to be tested, that is, complete rice
The detection of paddy semilate rice song toxin.
2. the method for two kinds of rice song toxin in a kind of quickly detection paddy according to claim 1, it is characterised in that: in S2
In weigh rice sample amount be 10-20g, be separately added into 20-30ml acidic aqueous solution twice.
3. the method for two kinds of rice song toxin in a kind of quickly detection paddy according to claim 1, it is characterised in that: sample
The acidic aqueous solution pH=3-5 of extraction is then adjusted using formic acid if being not located at the region.
4. the method for two kinds of rice song toxin in a kind of quickly detection paddy according to claim 1, it is characterised in that: in S2
In middle centrifugal treating, high-speed homogenization revolving speed is 10000-12000rpm, and the time is 2min.
5. the method for two kinds of rice song toxin in a kind of quickly detection paddy according to claim 1, it is characterised in that: in S3
In methanol and water volume ratio be 5:5-8:2.
6. the method for two kinds of rice song toxin in a kind of quickly detection paddy according to claim 1, it is characterised in that: into
In row ultra performance liquid chromatography Tandem Mass Spectrometry Analysis, the content of formic acid is 0.1-0.2% in the water of A phase.
7. the method for two kinds of rice song toxin in a kind of quickly detection paddy according to claim 1, it is characterised in that: described
The column temperature of chromatographic column is 30-40 degrees Celsius.
8. the method for two kinds of rice song toxin in a kind of quickly detection paddy according to claim 1, it is characterised in that: described
Chromatographic column is specially Shimadzu Shim-pack XR-C8,2.0 × 100mm of specification, and 1.6 μm of partial size, 10 μ l of sample volume.
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111272521A (en) * | 2020-03-24 | 2020-06-12 | 中国水稻研究所 | Pretreatment device and method for extracting and purifying ustilaginoidin A |
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| CN112649523A (en) * | 2020-12-01 | 2021-04-13 | 华中农业大学 | Method for detecting ustilagin A or ustilagin B in food |
| CN112697898A (en) * | 2020-12-04 | 2021-04-23 | 华中农业大学 | Method for rapidly determining content of ustilagin A in urine or cell culture medium by liquid chromatography-mass spectrometry |
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