CN105372353B - A kind of assay method of food glyphosate and its metabolin AminomethylphosphoniAcid Acid residual quantity - Google Patents
A kind of assay method of food glyphosate and its metabolin AminomethylphosphoniAcid Acid residual quantity Download PDFInfo
- Publication number
- CN105372353B CN105372353B CN201510973309.3A CN201510973309A CN105372353B CN 105372353 B CN105372353 B CN 105372353B CN 201510973309 A CN201510973309 A CN 201510973309A CN 105372353 B CN105372353 B CN 105372353B
- Authority
- CN
- China
- Prior art keywords
- glyphosate
- metabolin
- food
- aminomethylphosphoniacid acid
- residual quantity
- 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.)
- Expired - Fee Related
Links
Classifications
-
- G—PHYSICS
- G01—MEASURING; TESTING
- 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
-
- G—PHYSICS
- G01—MEASURING; TESTING
- 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
- G01N30/04—Preparation or injection of sample to be analysed
- G01N30/06—Preparation
-
- G—PHYSICS
- G01—MEASURING; TESTING
- 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
- G01N2030/022—Column chromatography characterised by the kind of separation mechanism
- G01N2030/027—Liquid chromatography
-
- G—PHYSICS
- G01—MEASURING; TESTING
- 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
- G01N30/04—Preparation or injection of sample to be analysed
- G01N30/06—Preparation
- G01N2030/062—Preparation extracting sample from raw material
Landscapes
- Physics & Mathematics (AREA)
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Analytical Chemistry (AREA)
- Biochemistry (AREA)
- General Health & Medical Sciences (AREA)
- General Physics & Mathematics (AREA)
- Immunology (AREA)
- Pathology (AREA)
- Other Investigation Or Analysis Of Materials By Electrical Means (AREA)
- Investigating Or Analysing Biological Materials (AREA)
Abstract
The present invention relates to the assay method of a kind of food glyphosate and its metabolin AminomethylphosphoniAcid Acid residual quantity, including standard working solution is prepared, food samples test solution is prepared, draw standard working solution regression curve and calculates food samples glyphosate and its residual quantity of metabolin AminomethylphosphoniAcid Acid.The method of the present invention can go out food glyphosate and its metabolin AminomethylphosphoniAcid Acid residual quantity with Accurate Determining, quantify accurately and reliably, and reappearance is good；Purified, will not be blocked using bag filter plus divinylbenzene solid phase extraction column, and the rate of recovery more than 80%, it is not required that correct recovery using expensive internal standard compound；Using the 2D posts of NH2P 50, using 5mmol/L ammonium acetate solution and acetonitrile as mobile phase, with good response and peak shape.
Description
Technical field
The present invention relates to a kind of detection method, especially a kind of food glyphosate and its metabolin AminomethylphosphoniAcid Acid residual
The assay method of amount.
Background technology
Glyphosate (glyphosate, GLY), chemical name N- ((phosphonomethyl)) amion acetic acid is in the world should at present
With the wide spectrum nonselective herbicide that most wide, output is maximum, but its irrational is excessively high residued in using can still result in it
In plant.Glyphosate and its major metabolite AminomethylphosphoniAcid Acid (aminomethylphosphonic acid, AMPA) have
The effect toxicity similar to organic phosphorus compound, mainly causes nervous system function to lose by suppressing the activity of CHE
Adjust, the case such as the water pollution vegetation deterioration as caused by glyphosate, livestock poisoning also increases year by year.
China《National food safety standard Pesticide MRL》(GB 2763-2014) is in August, 2014
Gone into effect from 1 day, its clear stipulaties：Glyphosate is that the residues of pesticides of cereal, oil plant and grease, sugar material, fruit, tealeaves etc. are strong
(must the examine) project of inspection, its residue limits in these products is between 0.05-5 milligrams per kilogram.With Chinese Main Trade state/
Residue limits requirement of the area to food glyphosate is China 1mg/kg, Hong-Kong 1mg/kg, U.S. 1mg/kg, Japan
1mg/kg。
Because glyphosate and its metabolin AminomethylphosphoniAcid Acid are highly polar compound, insoluble in most of organic solvent,
Both without UV absorption, but it is not volatile, therefore, generally required with gas-chromatography or high performance liquid chromatography detection by means of before post
Or post column derivatization, its complex steps, poor repeatability, Quality Control hardly possible；Or use the chromatography of ions, but when separating GLY due to
Its reservation is very strong, its separate must use strong eluent sodium carbonate, and other Common Anions retain it is weaker can not be to GLY and its
Its anion carries out analysis simultaneously, and is only limitted to the test to the more single matrix such as water.
Patent of invention CN 101646348B disclose a kind of test matrix glyphosate, N- acetyl glyphosates, N- acetyl
The method of the presence or content of AMPA or AminomethylphosphoniAcid Acid (AMPA) and its various metabolins, it is purified is consolidated using ion exchange
Phase extraction column, many samples such as banana, lemon, tea dust etc. can block solid-phase extraction column and cause to operate, and the rate of recovery is non-
It is often low, less than 5%, it is necessary to correct recovery with expensive internal standard compound.Patent of invention CN 103529148B disclose a kind of food
The detection method of middle herbicide glyphosate and its metabolin AminomethylphosphoniAcid Acid residual quantity, mainly by extract, absorption, elution, elution,
Five steps of qualitative and quantitative analysis composition, its glyphosate and aminomethyl phosphine to be remained in binary magnesium-aluminum hydrotalcite adsorbing and extracting liquid
Acid, then respectively with water and methanol elution binary magnesium-aluminum hydrotalcite, in addition it is also necessary to sodium carbonate liquor to adsorbing in hydrotalcite
Glyphosate and AminomethylphosphoniAcid Acid are desorbed, troublesome poeration.Its limitation is had based on the above method, a kind of quick, letter is set up
Victory detects that the method for food glyphosate and its metabolin is very necessary.
The content of the invention
Problem to be solved by this invention be overcome the shortcomings of prior art exist there is provided a kind of food glyphosate and its
The assay method of metabolin AminomethylphosphoniAcid Acid residual quantity.
Concrete scheme is as follows：
The assay method of a kind of food glyphosate and its metabolin AminomethylphosphoniAcid Acid residual quantity, comprises the following steps：(1)
Prepare standard working solution：With water as solvent, the standard work for being each configured to glyphosate and its metabolin AminomethylphosphoniAcid Acid is molten
Liquid；(2) food samples test solution is prepared：(a) sample specimens are weighed, quality is m, adds water in centrifuging homogeneous on high speed homogenization device, receive
Collect aqueous phase or upper phase, it is v that volume is settled to water；(b) test solution of constant volume in above-mentioned steps (a) is taken, bag filter is added
It is interior, it is placed in glass tube, adds ultra-pure water ultrasound, takes out in the raffinate given, injection divinylbenzene solid phase extraction column,
Discard efflux after efflux before divinylbenzene solid phase extraction column, collection；(c) graphite is added into the rear efflux of collection
Change carbon black, removing water colo(u)r in sample extraction solution as adsorbent disturbs, and is centrifuged after rotation nest concussion, takes supernatant, nitrogen
Dry, residue aqueous dissolution is blown to, poly tetrafluoroethylene is crossed, obtains food samples test solution, it is standby；(3) efficient liquid phase is used
The triple level Four bar GC-MSs of chromatogram-series connection, liquid chromatography-tandem matter is carried out to the standard working solution of step (1) respectively
Spectrum is determined：Concentration x using standard working solution glyphosate and its metabolin AminomethylphosphoniAcid Acid is measured as abscissa with corresponding
Quota ion peak area y be ordinate, draw standard working solution regression curve y=ax+b, calculate the oblique of regression curve
Rate a and intercept b；(4) the triple level Four bar GC-MSs of high performance liquid chromatography-series connection are used, the food samples of step (2) are tried
Liquid carries out liquid chromatography tandom mass spectrometry determination, and the linear equation y=ax+b according to obtained in (3) extrapolates food samples examination
The concentration c of liquid glyphosate and its metabolin AminomethylphosphoniAcid Acid0, and then calculate food samples glyphosate and its metabolin ammonia first
The residual quantity c, c=c of base phosphonic acids0×v/m。
Its further technical scheme is, the color of the triple level Four bar GC-MSs of described high performance liquid chromatography-series connection
Spectrum post is NH2P-502D posts.Its further technical scheme is that described adsorbent Graphon addition is 12-32mg,
40-60 μm of particle diameter, ProElut fillers.Its further technical scheme is, the described triple level Four of high performance liquid chromatography-series connection
The column Mobile phase pH of bar GC-MS is 10-11.Its further technical scheme is, described high performance liquid chromatography-
The mobile phase of triple level Four bar GC-MSs of connecting contains ammonium acetate solution.Its further technical scheme is, described
The mobile phase of the triple level Four bar GC-MSs of high performance liquid chromatography-series connection is the mixed solution of ammonium acetate solution and acetonitrile.
Its further technical scheme is that the concentration of described ammonium acetate solution is 5mmol/L.Its further technical scheme is,
Described food includes cereal, oil plant and grease, sugar material, fruit and tealeaves.
Because glyphosate and its metabolin AminomethylphosphoniAcid Acid are dissolved in water but are insoluble in most organic solvent, this method
A variety of organic solvents and water are compared to glyphosate and its extraction effect of metabolin AminomethylphosphoniAcid Acid, methanol：Water (1:1), second
Nitrile：Water (1:1), pure methanol, pure acetonitrile, pure water find that acetonitrile and methanol dissolve the organic impurities in a large amount of samples, interference is very
Greatly, and the rate of recovery be less than 30%, finally determine water be used as extractant.The present invention need to the greatest extent may be used to reduce matrix effect to sample
Can purification.This method compare dispersive solid-phase extraction method, MAX pillars, amino pillar, HLB pillars, MCX, MAX columns in series and thoroughly
The purification method that analysis bag+divinylbenzene solid phase extraction column is used in conjunction, as a result as shown in Figure 1.
From figure 1 it appears that dispersive solid-phase extraction method there is no the rate of recovery to both, its reason is to add
PSA has adsorption effect to glyphosate and its metabolin AminomethylphosphoniAcid Acid.Amino pillar and HLB pillars are to both rate of recovery
It is relatively low, no more than 50%；And when only with a MAX pillar, AminomethylphosphoniAcid Acid is substantially more than 50% for the glyphosate rate of recovery
Do not reclaim, reason is that clean-up effect is not good, and sample generates very strong substrate inhibition, makes the response of two kinds of materials relatively low；And
The method of MCX, MAX series connection pillar is to the rate of recovery of glyphosate and its metabolin AminomethylphosphoniAcid Acid more than 60%, and purification is imitated
Fruit is general, but food substrate such as banana, lemon, tea dust SPE posts easily cause pillar blocking, extract solution is solid by series connection
Mutually extraction pillar is substantially impossible；And the method rate of recovery that bag filter is used in conjunction with divinylbenzene solid phase extraction column exceedes
80%.Therefore the method for this method selection bag filter and divinylbenzene solid phase extraction column is purified to sample.Wherein, thoroughly
Analysis bag effect be take out sample extraction solution in big molecular impurity chaff interference, divinylbenzene solid phase extraction column be in order to
Remove and extract hydrophobic compound interference in solution.
It is noted that patent of invention CN200880010069 purification samples use Ion Exchange Solid Phase extraction column, very
Multi-example can block solid-phase extraction column such as banana, lemon, tea dust and cause to operate, and the rate of recovery is very low is less than
5% with expensive internal standard compound, it is necessary to correct recovery.The present invention adds divinylbenzene solid phase extraction column to carry out using dialysis membrane
Purification, will not be blocked, and the rate of recovery more than 80%, it is not necessary to correct recovery using expensive internal standard compound.
Graphon can be very good to remove the interference of water colo(u)r in sample extraction solution as adsorbent, its
Addition is 12-32mg, and 40-60 μm of particle diameter, the effect using ProElut fillers is preferable.
Due to the highly polar and water solubility of glyphosate and its metabolin AminomethylphosphoniAcid Acid, it is extremely difficult on common reverse-phase chromatographic column
Retain, thus the present invention compare two kinds can be to glyphosate and its hydrophilic separation chromatography post of metabolin AminomethylphosphoniAcid Acid：HILIC colors
Spectrum post (amphion chromatographic column) and amino chromatographic column post are separated, and wherein HILIC hydrophilic chromatographics post is that one kind can retain
With the chromatographic column of separating polar-ionic compound, polarity-ionic compound does not retain when organic Phase Proportion is high in some,
And polarity is weak or non-polar compound is then not preserved.Therefore, HILIC posts can only be under the relatively low precondition of organic phase
The reservation of polarity-ionic compound is solved the problems, such as, and anti-reverse-phase chromatographic column has reverse-phase chromatography mechanism and normal-phase chromatography simultaneously
Mechanism, in this case, can just be changed between reverse-phase chromatography and positive level as long as changing mobile phase, can be not required to
Water removal is gone to can be carried out normal-phase chromatography operation completely from mobile phase；Pillar will not be also produced using the elution of 100% aqueous phase
Avalanche；The releveling of gradient elution can be just completed within one minute.These advantages are compared with HILIC pillars,
Hydrone can fix phase surface formation moisture film in HILIC, and in anti-anti-phase fixed phase surface, because its strong-hydrophobicity is without shape
Into moisture film.Therefore, it need not be cleaned for anti-reversed-phase column as HILIC posts with water.
In view of these advantages of anti-reverse-phase chromatographic column, using NH2P-502D posts, compared for different mobile phases and chromatogram are divided
From and sensitivity.Different PH influence of the mobile phase to chromatographic isolation is compared for first, is as a result shown, pH=10-11 mobile phases
Effect to glyphosate and its metabolin is best；Next compared for the influence of ammoniacal liquor and ammonium acetate solution to chromatographic isolation, knot
Fruit shows that the chromatogram peak type of ammonium acetate solution more conforms to the requirement of chromatography；Finally, it compared for various concentrations ammonium acetate
Influence of the aqueous solution to chromatographic isolation, as a result shows, the reservation with the increase glyphosate and its metabolin of acetic acid ammonium concentration increases
By force, it is but on a declining curve more than after 5mmol/L, therefore mobile phase is used for A 5mmol/L ammonium acetate solutions, B acetonitriles, flowing
Phase gradient elution program is shown in Table 1.LC-MS instrument sets up assay method, with good response and peak shape.
Beneficial effect：(1) method of the invention can go out food glyphosate and its metabolin aminomethyl phosphine with Accurate Determining
Sour residual quantity, is quantified accurately and reliably, and reappearance is good；(2) divinylbenzene solid phase extraction column is added to carry out using bag filter
Purification, will not be blocked, and the rate of recovery more than 80%, it is not required that correct recovery using expensive internal standard compound；(3) use
NH2P-502D posts, using 5mmol/L ammonium acetate solution and acetonitrile as mobile phase, with good response and peak shape.
Brief description of the drawings
Fig. 1 is Extraction solvent rate of recovery investigation figure of the present invention；
Fig. 2 is the triple level Four bars of high performance liquid chromatography-series connection of glyphosate of the present invention and its metabolin AminomethylphosphoniAcid Acid
GC-MS multiple-reaction monitoring chromatogram；
Fig. 3 is the working curve of glyphosate and AminomethylphosphoniAcid Acid；
Fig. 4 a are the glyphosate quota ion extraction chromatography figures of blank tealeaves；
Fig. 4 b are the quota ion extraction chromatography figures of blank tealeaves；
Fig. 5 a are the glyphosate quota ion extraction chromatography figures of blank citrus；
Fig. 5 b are the quota ion extraction chromatography figures of blank citrus；
Fig. 6 a are the glyphosate quota ion extraction chromatography figures of blank wheat；
Fig. 6 b are the quota ion extraction chromatography figures of blank wheat；
Fig. 7 a are the glyphosate quota ion extraction chromatography figures of blank apple；
Fig. 7 b are the quota ion extraction chromatography figures of blank apple；
Fig. 8 a are the glyphosate quota ion extraction chromatography figures of blank corn；
Fig. 8 b are the quota ion extraction chromatography figures of blank corn；
Fig. 9 a are the glyphosate quota ion extraction chromatography figures of blank peach；
Fig. 9 b are the AminomethylphosphoniAcid Acid quota ion extraction chromatography figures of blank peach；
Figure 10 a are the glyphosate quota ion extraction chromatography figures of blank paddy；
Figure 10 b are the AminomethylphosphoniAcid Acid quota ion extraction chromatography figures of blank paddy；
Figure 11 a are the glyphosate quota ion extraction chromatography figures of blank grape；
Figure 11 b are the AminomethylphosphoniAcid Acid quota ion extraction chromatography figures of blank grape；
Figure 12 a are the glyphosate quota ion extraction chromatography figures of blank watermelon；
Figure 12 b are the AminomethylphosphoniAcid Acid quota ion extraction chromatography figures of blank watermelon；
Figure 13 a are the glyphosate quota ion extraction chromatography figures of blank banana；
Figure 13 b are the AminomethylphosphoniAcid Acid quota ion extraction chromatography figures of blank banana；
Figure 14 a are the glyphosate quota ion extraction chromatography figures of blank soybean；
Figure 14 b are the AminomethylphosphoniAcid Acid quota ion extraction chromatography figures of blank soybean；
Figure 15 a are the glyphosate quota ion extraction chromatography figures of blank sugarcane；
Figure 15 b are the AminomethylphosphoniAcid Acid quota ion extraction chromatography figures of blank sugarcane；
Figure 16 a are the glyphosate quota ion extraction chromatography figures of blank baked barley tea；
Figure 16 b are the AminomethylphosphoniAcid Acid quota ion extraction chromatography figures of blank baked barley tea；
Figure 17 a are the glyphosate quota ion extraction chromatography figures of blank cottonseed oil；
Figure 17 b are the AminomethylphosphoniAcid Acid quota ion extraction chromatography figures of blank cottonseed oil；
Figure 18 a are the glyphosate quota ion extraction chromatography figures of tealeaves matrix mark-on；
Figure 18 b are the AminomethylphosphoniAcid Acid quota ion extraction chromatography figures of tealeaves matrix mark-on；
Figure 19 a are the glyphosate quota ion extraction chromatography figures of citrus matrix mark-on；
Figure 19 b are the AminomethylphosphoniAcid Acid quota ion extraction chromatography figures of citrus matrix mark-on；
Figure 20 a are the glyphosate quota ion extraction chromatography figures of wheat matrix mark-on；
Figure 20 b are the AminomethylphosphoniAcid Acid quota ion extraction chromatography figures of wheat matrix mark-on；
Figure 21 a are the glyphosate quota ion extraction chromatography figures of apple matrix mark-on；
Figure 21 b are the AminomethylphosphoniAcid Acid quota ion extraction chromatography figures of apple matrix mark-on；
Figure 22 a are the glyphosate quota ion extraction chromatography figures of corn-base mark-on；
Figure 22 b are the AminomethylphosphoniAcid Acid quota ion extraction chromatography figures of corn-base mark-on；
Figure 23 a are the glyphosate quota ion extraction chromatography figures of peach matrix mark-on；
Figure 23 b are the AminomethylphosphoniAcid Acid quota ion extraction chromatography figures of peach matrix mark-on；
Figure 24 a are the glyphosate quota ion extraction chromatography figures of paddy matrix mark-on；
Figure 24 b are the AminomethylphosphoniAcid Acid quota ion extraction chromatography figures of paddy matrix mark-on；
Figure 25 a are the glyphosate quota ion extraction chromatography figures of grape matrix mark-on；
Figure 25 b are the AminomethylphosphoniAcid Acid quota ion extraction chromatography figures of grape matrix mark-on；
Figure 26 a are the glyphosate quota ion extraction chromatography figures of watermelon matrix mark-on；
Figure 26 b are the AminomethylphosphoniAcid Acid quota ion extraction chromatography figures of watermelon matrix mark-on；
Figure 27 a are the glyphosate quota ion extraction chromatography figures of banana matrix mark-on；
Figure 27 b are the AminomethylphosphoniAcid Acid quota ion extraction chromatography figures of banana matrix mark-on；
Figure 28 a are the glyphosate quota ion extraction chromatography figures of soybean substrate mark-on；
Figure 28 b are the AminomethylphosphoniAcid Acid quota ion extraction chromatography figures of soybean substrate mark-on；
Figure 29 a are the glyphosate quota ion extraction chromatography figures of sugarcane matrix mark-on；
Figure 29 b are the AminomethylphosphoniAcid Acid quota ion extraction chromatography figures of sugarcane matrix mark-on；
Figure 30 a are the glyphosate quota ion extraction chromatography figures of baked barley tea matrix mark-on；
Figure 30 b are the quota ion extraction chromatography figures of baked barley tea matrix mark-on；
Figure 31 a are the glyphosate quota ion extraction chromatography figures of cottonseed oil matrix mark-on；
Figure 31 b are the AminomethylphosphoniAcid Acid quota ion extraction chromatography figures of cottonseed oil matrix mark-on.
From Fig. 4-Figure 31, each width figure includes two figures, is respectively designated as a, b, wherein a figures are determining for sample glyphosate
The ion extraction chromatogram is measured, b figures are the quota ion extraction chromatography figure of AminomethylphosphoniAcid Acid in sample.
Embodiment
Technical solution of the present invention is further elaborated with reference to the accompanying drawings and examples.Unreceipted specific skill in embodiment
Art or condition person, are carried out according to the technology or condition described by document in the art or according to product description.Examination used
Agent or the unreceipted production firm person of instrument, being can be by the conventional products of acquisition purchased in market.
Embodiment 1：Prepare standard working solution
Standard working solution is the standard work for three kinds of concentration being equipped by glyphosate and its metabolin AminomethylphosphoniAcid Acid
Solution, specific preparation method is：(1) glyphosate and its each 10mg of metabolin AminomethylphosphoniAcid Acid standard items accurately are weighed, it is fixed with water
Hold to 10mL, be configured to the Standard Stock solutions that concentration is 1mg/mL；(2) standard items stock solution is drawn, is diluted with water, respectively
Interstitial fluid in the standard that concentration is 10 μ g/mL is configured to, less than 4 DEG C lucifuges are stored refrigerated；(3) draw in the middle of a certain amount of standard
Liquid, 0.01,0.02,0.05,0.1,0.2,0.5 μ g/mL series of standards working solutions is diluted to the aqueous solution, less than 4 respectively
DEG C lucifuge is stored refrigerated.
Embodiment 2：Use the triple level Four bar GC-MS analytical standard working solutions of high performance liquid chromatography-series connection
Parameter set by the triple level Four bar GC-MSs of high performance liquid chromatography-series connection should ensure that tested during chromatographic determination
Component can be efficiently separated with other components, such as following parameter：
(1) liquid-phase condition：
1) chromatographic column：NH2P-502D posts, long 150mm, internal diameter 2.0mm, quite 5 μm of particle diameter or person；
2) mobile phase：A：5mmol/L ammonium acetate solutions (ammoniacal liquor adjusts pH=10-11), B：Acetonitrile solution；Flow velocity：
0.25mL/min, gradient elution program is shown in Table 1；
3) column temperature：35℃；4) sample size：10μL.
The eluent gradient elution program table of table 1
| Time (min) | Mobile phase A (%) | Mobile phase B (%) |
| 0.00 | 20.0 | 80.0 |
| 2.00 | 20.0 | 80.0 |
| 2.01 | 80.0 | 20.0 |
| 8.00 | 80.0 | 20.0 |
| 8.01 | 20.0 | 80.0 |
| 12.00 | 20.0 | 80.0 |
(2) Mass Spectrometry Conditions：
The multiple-reaction monitoring parameter list of table 2
Fig. 2 is the triple level Four bars of high performance liquid chromatography-series connection of glyphosate of the present invention and its metabolin AminomethylphosphoniAcid Acid
GC-MS multiple-reaction monitoring chromatogram：It is with secondary from top to bottom：Glyphosate (167.8/63.0,167.8/81.0,167.8/
150.0), AminomethylphosphoniAcid Acid (110.0/63.0,110.0/79.0,110.0/81.0).
As seen from Figure 2, under the conditions of described in embodiment 2, glyphosate and aminomethyl glyphosate obtain retaining very well
And separation, and the sensitivity of glyphosate and the detection of aminomethyl glyphosate is very high.
Embodiment 3：Draw standard working solution regression curve
Using the triple level Four bar GC-MSs of high performance liquid chromatography-series connection, the series of standards in embodiment 1 is worked
Solution carries out liquid chromatography tandom mass spectrometry determination respectively, and instrument parameter setting is according to progress listed in embodiment 2, then
Concentration x using standard working solution glyphosate and its metabolin AminomethylphosphoniAcid Acid as abscissa, with it is corresponding measure quantify
Ion peak areas y is ordinate, draws standard working solution regression curve, sees Fig. 3.From figure 3, it can be seen that solution glyphosate
And its concentration and the corresponding quota ion peak area measured of metabolin AminomethylphosphoniAcid Acid are linear correlations.
According to the straight line drawn, the slope and intercept of straight line, linearly dependent coefficient R are tried to achieve2Be 0.9998, obtain with
Lower linear equation：Glyphosate：Y=242.47x+3601 AminomethylphosphoniAcid Acids y=172.4x+92.275
Embodiment 4：Prepare food samples test solution
(1) for fluid sample, Cereals class and fruits and vegetables sample：Sample 4g is weighed, in 50mL centrifuge tubes, 6mL is added
Water, in homogeneous 3min on (15000r/min) homogenizer at a high speed, 5min is centrifuged with 15000r/min, aqueous phase is collected, uses water constant volume
To 10mL.For tealeaves class sample：Sample 2g is weighed, in 50mL centrifuge tubes, 10mL water is added, at a high speed (15000r/min)
Homogeneous 3min on homogenizer, centrifuges 5min with 15000r/min by centrifuge tube, collects upper liquid, 10mL is settled to water.(2) it is accurate
Standby bag filter：Cut to 10-20cm segments, boiled 10 minutes with water with scissors, secondary water is cleaned, preserved into ethanol；Before use
Cleaned only with secondary water afterwards.(3) divinylbenzene solid phase extraction column (1.0cc posts) is prepared：Use preceding 5mL methanol, 5mL water
Activation, stands half an hour；Cleaned and preserved using rear methanol.(4) sample extracting solution of above-mentioned steps (1) is taken to add in step (2)
In the bag filter, it is placed in by 1:1 ratio is taken out, from above-mentioned glass tube in the 50mL glass tubes of ultra-pure water after ultrasonic 1h
Middle taking-up 5mL raffinates, with divinylbenzene solid phase extraction column described in 5mL syringes implantation step (3), discard divinyl
2mL effluxes before benzene solid phase extraction column, 3mL effluxes after being collected with 5mL centrifuge tubes；Add 20mg Graphon adsorbents
(40-60 μm, ProElut fillers), rotation nest concussion 2min centrifuges 5min after 15000r/min, takes 2mL supernatants to cleaning
Nitrogen is blown to dry, residue 0.5mL aqueous dissolution at 60 DEG C in glass tube, crosses polytetrafluoroethylene (PTFE) (PTFE) film.
Embodiment 5：Use the triple level Four bar GC-MS analysis food samples of high performance liquid chromatography-series connection
Food samples test solution, including tealeaves, citrus, wheat, apple, corn, peach, paddy, Portugal are prepared according to embodiment 4
Grape, watermelon, banana, soybean, sugarcane, baked barley tea, cottonseed oil.Each based food includes two kinds of samples, and one kind is free from glyphosate
And its blank food samples of metabolin AminomethylphosphoniAcid Acid, one kind be to blank food samples addition concentration be 0.05mg/kg
The mark-on food samples of glyphosate and its metabolin AminomethylphosphoniAcid Acid.It is equal to 10 by signal to noise ratio (S/N) to calculate, this method is quantified
Lower limit is that 0.05mg/kg can meet limitation requirement.
Using with the triple level Four bar GC-MSs of high performance liquid chromatography-series connection, according to the instrument gone out given in embodiment 2
Parameter, analyzes prepared blank food samples and mark-on food samples respectively, obtains corresponding chromatography of ions figure, sees Fig. 4-figure
31, each width figure includes two figures, is respectively designated as a, b, wherein a figures are the quota ion extraction chromatography of sample glyphosate
Figure, b figures are the quota ion extraction chromatography figure of AminomethylphosphoniAcid Acid in sample, and such as Fig. 4 is that the quota ion of blank tealeaves is extracted
Chromatogram, then Fig. 4 a are the glyphosate quota ion extraction chromatography figures of blank tealeaves, and Fig. 4 b are the AminomethylphosphoniAcid Acids of blank tealeaves
Quota ion extraction chromatography figure, figure below is similarly.For simplicity, it is as follows to figure Unify legislation below：Fig. 5 is blank mandarin orange
The quota ion extraction chromatography figure of tangerine, Fig. 6 is the quota ion extraction chromatography figure of blank wheat, and Fig. 7 is quantifying for blank apple
The ion extraction chromatogram, Fig. 8 is the quota ion extraction chromatography figure of blank corn, and Fig. 9 is that the quota ion of blank peach extracts color
Spectrogram, Figure 10 is the quota ion extraction chromatography figure of blank paddy, and Figure 11 is the quota ion extraction chromatography figure of blank grape, figure
12 be the quota ion extraction chromatography figure of blank watermelon, and Figure 13 is the quota ion extraction chromatography figure of blank banana, and Figure 14 is empty
The quota ion extraction chromatography figure of white soybean, Figure 15 is the quota ion extraction chromatography figure of blank sugarcane, and Figure 16 is blank barley
The quota ion extraction chromatography figure of tea, Figure 17 is the quota ion extraction chromatography figure of blank cottonseed oil, and Figure 18 is that tealeaves matrix adds
Target quota ion extraction chromatography figure, Figure 19 is the quota ion extraction chromatography figure of citrus matrix mark-on, and Figure 20 is wheat matrix
The quota ion extraction chromatography figure of mark-on, Figure 21 is the quota ion extraction chromatography figure of apple matrix mark-on, and Figure 22 is corn-based
The quota ion extraction chromatography figure of matter mark-on, Figure 23 is the quota ion extraction chromatography figure of peach matrix mark-on, and Figure 24 is paddy base
The quota ion extraction chromatography figure of matter mark-on, Figure 25 is the quota ion extraction chromatography figure of grape matrix mark-on, and Figure 26 is watermelon
The quota ion extraction chromatography figure of matrix mark-on, Figure 27 is the quota ion extraction chromatography figure of banana matrix mark-on, and Figure 28 is big
The quota ion extraction chromatography figure of beans matrix mark-on, Figure 29 is the quota ion extraction chromatography figure of sugarcane matrix mark-on, Tu30Shi
The quota ion extraction chromatography figure of baked barley tea matrix mark-on, Figure 31 is the quota ion extraction chromatography figure of cottonseed oil matrix mark-on.
From Fig. 4-Figure 31 can be seen that blank food samples in do not contain the spy of glyphosate and its metabolin AminomethylphosphoniAcid Acid
Peak is levied, mark-on food samples go out peak position and do not have impurity peaks interference measurement in target compound appearance, show the special of method
Property can meet detection tealeaves, citrus, wheat, apple, corn, peach, paddy, grape, watermelon, banana, soybean, sugarcane, barley
The requirement of tea and a series of food of cottonseed oil.
Embodiment 6：Survey the precision and the rate of recovery of this method
The precision and the rate of recovery of this method are detected using additive process：With what is remained without glyphosate and AminomethylphosphoniAcid Acid
Tealeaves, citrus, wheat, apple, corn, peach, paddy, grape, watermelon, banana, soybean, sugarcane, baked barley tea, cottonseed oil are blank
Sample substrate, three concentration levels of addition (are higher than the linear upper limit, detected by being diluted in level of linearity), Mei Genong
Degree level carries out six repetitions and tested, and the rate of recovery and precision for measuring GLY and AMPA collect and be shown in Table 3.Here the rate of recovery refers to
Recovery of standard addition, computational methods are：Recovery of standard addition=(mark-on Specimen Determination value-Specimen Determination value) ÷ mark-on amount × 100%
From table 3 it can be seen that the average recovery rate of sample is in 71.9-92.2%, relative standard deviation is in 2.29-
7.68%, meet the requirement of residue detection, the testing result of this method is credible.
Although embodiments of the invention have been shown and described above, it is to be understood that above-described embodiment is example
Property, it is impossible to limitation of the present invention is interpreted as, one of ordinary skill in the art is not departing from the principle and objective of the present invention
In the case of above-described embodiment can be changed within the scope of the invention, change, replace and modification.
This method of table 3 adds concentration and rate of recovery range table (n=6)
Claims (9)
1. the assay method of a kind of food glyphosate and its metabolin AminomethylphosphoniAcid Acid residual quantity, it is characterised in that：Including with
Lower step：
(1) standard working solution is prepared：With water as solvent, the standard of glyphosate and its metabolin AminomethylphosphoniAcid Acid is configured to respectively
Working solution；
(2) food samples test solution is prepared：
(a) sample specimens are weighed, quality is m, adds water in centrifuging homogeneous on high speed homogenization device, collect upper phase, use water constant volume
It is v to volume；
(b) test solution of constant volume in above-mentioned steps (a) is taken, adds in bag filter, is placed in glass tube, ultra-pure water ultrasound is added, takes
Go out in the raffinate given, injection divinylbenzene solid phase extraction column, discard and flowed out before divinylbenzene solid phase extraction column
Efflux after liquid, collection；
(c) Graphon is added into the rear efflux of collection, water-soluble color in sample extraction solution is removed as adsorbent
Element interference, centrifuges after rotation nest concussion, takes supernatant, nitrogen is blown to dry, residue aqueous dissolution, crosses poly tetrafluoroethylene, obtains
Food samples test solution, it is standby；
(3) the triple level Four bar GC-MSs of high performance liquid chromatography-series connection are used, the standard working solution of step (1) is distinguished
Carry out liquid chromatography tandom mass spectrometry determination：Using the concentration x of standard working solution glyphosate and its metabolin AminomethylphosphoniAcid Acid as
Abscissa, using the corresponding quota ion peak area y measured as ordinate, draws standard working solution regression curve y=ax+b,
Calculate the slope a and intercept b of regression curve；
(4) the triple level Four bar GC-MSs of high performance liquid chromatography-series connection are used, the food samples test solution of step (2) is carried out
Liquid chromatography tandom mass spectrometry determination, the linear equation y=ax+b according to obtained in (3) extrapolates food samples test solution medium-height grass
The concentration c of sweet phosphine and its metabolin AminomethylphosphoniAcid Acid0, and then calculate food samples glyphosate and its metabolin AminomethylphosphoniAcid Acid
Residual quantity c, c=c0×v/m。
2. the measure side of a kind of food glyphosate according to claim 1 and its metabolin AminomethylphosphoniAcid Acid residual quantity
Method, it is characterised in that：The chromatographic column of the triple level Four bar GC-MSs of described high performance liquid chromatography-series connection is NH2P-50 2D
Post.
3. the measure side of a kind of food glyphosate according to claim 1 and its metabolin AminomethylphosphoniAcid Acid residual quantity
Method, it is characterised in that：Described adsorbent Graphon addition is 12-32mg, 40-60 μm of particle diameter, ProElut fillers.
4. the measure side of a kind of food glyphosate according to claim 1 and its metabolin AminomethylphosphoniAcid Acid residual quantity
Method, it is characterised in that：The column Mobile phase pH of the triple level Four bar GC-MSs of described high performance liquid chromatography-series connection is
10-11。
5. the measure side of a kind of food glyphosate according to claim 1 and its metabolin AminomethylphosphoniAcid Acid residual quantity
Method, it is characterised in that：The mobile phase of the triple level Four bar GC-MSs of described high performance liquid chromatography-series connection contains ammonium acetate water
Solution.
6. the measure side of a kind of food glyphosate according to claim 1 and its metabolin AminomethylphosphoniAcid Acid residual quantity
Method, it is characterised in that：The mobile phase of the triple level Four bar GC-MSs of described high performance liquid chromatography-series connection is that ammonium acetate is water-soluble
The mixed solution of liquid and acetonitrile.
7. the measure of a kind of food glyphosate and its metabolin AminomethylphosphoniAcid Acid residual quantity according to claim 5 or 6
Method, it is characterised in that：The concentration of described ammonium acetate solution is 5mmol/L.
8. a kind of food glyphosate as claimed in any of claims 1 to 6 and its metabolin AminomethylphosphoniAcid Acid are residual
The assay method of allowance, it is characterised in that：Described food includes cereal, oil plant, grease, sugar material, fruit and tealeaves.
9. the measure side of a kind of food glyphosate according to claim 7 and its metabolin AminomethylphosphoniAcid Acid residual quantity
Method, it is characterised in that：Described food includes cereal, oil plant, grease, sugar material, fruit and tealeaves.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201510973309.3A CN105372353B (en) | 2015-12-22 | 2015-12-22 | A kind of assay method of food glyphosate and its metabolin AminomethylphosphoniAcid Acid residual quantity |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201510973309.3A CN105372353B (en) | 2015-12-22 | 2015-12-22 | A kind of assay method of food glyphosate and its metabolin AminomethylphosphoniAcid Acid residual quantity |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN105372353A CN105372353A (en) | 2016-03-02 |
| CN105372353B true CN105372353B (en) | 2017-09-05 |
Family
ID=55374735
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201510973309.3A Expired - Fee Related CN105372353B (en) | 2015-12-22 | 2015-12-22 | A kind of assay method of food glyphosate and its metabolin AminomethylphosphoniAcid Acid residual quantity |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN105372353B (en) |
Families Citing this family (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107576732B (en) * | 2016-07-04 | 2020-10-23 | 烟台杰科检测服务有限公司 | Method for determining glyphosate, aminomethylphosphonic acid and glufosinate in food |
| CN106153770B (en) * | 2016-07-22 | 2019-01-04 | 浙江省海洋水产研究所 | A kind of Solid Phase Extraction of aquatic products glyphosate-liquid chromatography-mass spectrography detection method |
| CN107843664A (en) * | 2017-10-30 | 2018-03-27 | 诺安实力可商品检验（青岛）有限公司 | The assay method of glufosinate-ammonium, glyphosate and its metabolin aminomethyl phosphonic acid residual quantity in tealeaves |
| CN108680663B (en) * | 2018-04-26 | 2020-10-30 | 广东省生物工程研究所（广州甘蔗糖业研究所） | Detection method of glyphosate and metabolite thereof in sugarcane |
| CN111487330B (en) * | 2019-01-29 | 2022-11-18 | 上海安谱实验科技股份有限公司 | Detection method for glyphosate and metabolites thereof in various matrixes |
| CN110441448A (en) * | 2019-09-12 | 2019-11-12 | 广东石油化工学院 | Method that is a kind of while measuring soil glyphosate and AminomethylphosphoniAcid Acid residual content |
| CN114965840A (en) * | 2021-02-24 | 2022-08-30 | 公安部物证鉴定中心 | Method for detecting glyphosate, glufosinate-ammonium and metabolites in organism liquid |
Family Cites Families (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US8003398B2 (en) * | 2007-03-27 | 2011-08-23 | E.I. De Pont De Nemours And Company | Methods and compositions for detecting glyphosate and metabolites thereof |
| CN103529148B (en) * | 2013-10-21 | 2014-11-05 | 福建出入境检验检疫局检验检疫技术中心 | Method for detecting residual quantity of glyphosate and aminomethyl phosphonic acid metabolite of glyphosate in food |
| CN104181257B (en) * | 2014-08-29 | 2016-04-13 | 福建中烟工业有限责任公司 | Extraction and purification methods and the composition therefor of mould clever residues of pesticides is disliked in tobacco |
-
2015
- 2015-12-22 CN CN201510973309.3A patent/CN105372353B/en not_active Expired - Fee Related
Also Published As
| Publication number | Publication date |
|---|---|
| CN105372353A (en) | 2016-03-02 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN105372353B (en) | A kind of assay method of food glyphosate and its metabolin AminomethylphosphoniAcid Acid residual quantity | |
| Solfrizzo et al. | Liquid chromatographic determination of Alternaria toxins in carrots | |
| CN105606757B (en) | Sweetener and preservative method for measuring simultaneously in flavouring essence for tobacco | |
| Perret et al. | Validation of a method for the determination of multiclass pesticide residues in fruit juices by liquid chromatography/tandem mass spectrometry after extraction by matrix solid-phase dispersion | |
| CN105758947B (en) | It is a kind of while measure the method for glufosinate-ammonium and its metabolite residue amount in food | |
| Wang et al. | Simultaneous determination of six plant growth regulators in fruits using high performance liquid chromatography based on solid-phase extraction and cleanup with a novel mixed-mode functionalized calixarene sorbent | |
| Abbaspour et al. | Monitoring of nine pesticides in different cereal flour samples with high performance liquid chromatography-diode array detection | |
| Hiemstra et al. | Fully automated solid-phase extraction cleanup and on-line liquid chromatographic determination of benzimidazole fungicides in fruit and vegetables | |
| CN109884207A (en) | A method of quick and precisely analyzing polyphenol content in rapeseed oil | |
| Gilvydis et al. | Ion-pairing liquid chromatographic determination of benzimidazole fungicides in foods | |
| AU2010280754B2 (en) | Method of preparation of samples for analysis and cartridge therefor | |
| CN113030331B (en) | Method for detecting chlorantraniliprole in plant | |
| Walker et al. | Determination of the Fusarium mycotoxins nivalenol, deoxynivalenol, 3-acetyldeoxynivalenol, and 15-0-acetyl-4-deoxynivalenol in contaminated whole wheat flour by liquid chromatography with diode array detection and gas chromatography with electron capture detection | |
| Qin et al. | Multi-residue method for determination of selected neonicotinoid insecticides in traditional Chinese medicine using modified dispersive solid-phase extraction combined with ultra-performance liquid chromatography tandem mass spectrometry | |
| CN113156034B (en) | Method for rapidly detecting various coffee flavor substances | |
| Wang et al. | Determination of agrochemical residues in aquatic vegetables by solid-phase extraction combined with HPLC spectrometry analyses | |
| CN103217498A (en) | Method for detecting dicyandiamide in milk powder with LC-MS (liquid chromatography/mass spectrometry) and sample preparation method | |
| CN107102078B (en) | A kind of method of aflatoxin B1 in measurement Gardenia Yellow | |
| CN113533608B (en) | Method suitable for rapidly detecting aflatoxin in large-batch edible oil samples | |
| Vázquez et al. | Application of coupled-column liquid chromatography combined with post-column photochemically induced fluorimetry derivatization and fluorescence detection to the determination of pyrethroid insecticides in vegetable samples | |
| CN109142589B (en) | Method for determining tetramycin residual quantity in soil by using high performance liquid chromatograph | |
| CN111175408A (en) | High performance liquid chromatography method for determining soluble sugar in tomato fruits | |
| CN110208405A (en) | A kind of remaining method of dinotefuran on detection rice | |
| CN103439424A (en) | Method for efficiently purifying nitro-methylene type neonicotinoid insecticides in plants | |
| Lawal et al. | The QuEChERS-dSPE Ionic-Liquid-Based Dispersive Liquid–Liquid Microextraction Coupled With High-Performance Liquid Chromatography–Tandem Mass Spectrometry for the Determination of Multiple Pesticide Residues in Red Syzygium samarangense Fruits |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant | ||
| CF01 | Termination of patent right due to non-payment of annual fee |
Granted publication date: 20170905 Termination date: 20211222 |
|
| CF01 | Termination of patent right due to non-payment of annual fee |