CN113671076A - Detection method of amantadine compounds and triazine herbicides in algae - Google Patents
Detection method of amantadine compounds and triazine herbicides in algae Download PDFInfo
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Abstract
The invention belongs to the technical field of substance analysis, and provides a detection method of an amantadine compound and a triazine herbicide in algae. The method adopts acetonitrile, ethyl acetate, a formic acid-acetonitrile mixed solution with the formic acid volume concentration of 1% or a formic acid-ethyl acetate mixed solution with the formic acid volume concentration of 1% as an extracting agent to release the amantadine compounds and the triazine herbicides in the algae sample, then adopts a PSA adsorbent as an adsorbent to adsorb and remove impurities which greatly interfere with the amantadine compounds and the triazine herbicides in the extracting solution to the maximum extent, adopts the ultra-high liquid chromatography, the primary mass spectrum and the secondary mass spectrum to perform qualitative detection, can improve the qualitative accuracy, eliminates false positive and improves the detection accuracy; then, the quantitative analysis is carried out by utilizing the ultra-high liquid chromatography and the primary chromatography, the accurate quantification of the amantadine compounds and the triazine herbicides can be accurately carried out, and the detection sensitivity is higher.
Description
Technical Field
The invention relates to the technical field of substance analysis, in particular to a detection method of an amantadine compound and a triazine herbicide in algae.
Background
Because the amantadine drugs belong to antiviral drugs and the triazine herbicides belong to pesticide residues, and because the two drugs belong to different types of drugs, the existing detection method respectively detects the amantadine drugs and the triazine herbicides, the operation is more complicated, and experimenters need to carry out repeated treatment and then respectively carry out on-machine detection, so the cost is higher. In addition, the existing detection method is mainly aimed at the adamantanamine of animal origin and triazine herbicide in the environment, and no method for simultaneously detecting two types of medicines in algae exists at present.
Disclosure of Invention
In view of the above, the present invention aims to provide a method for detecting an amantadine compound and a triazine herbicide in algae. The detection method provided by the invention has the advantages that the pretreatment method is simple, the obtained upper machine solution can be used for simultaneously detecting the amantadine compounds and the triazine herbicides in the algae samples, and the detection sensitivity is high.
In order to achieve the above object, the present invention provides the following technical solutions:
the invention provides a detection method of an amantadine compound and a triazine herbicide in algae, which comprises the following steps:
mixing the algae sample with an extracting agent, and extracting to obtain an extracting solution;
mixing the extracting solution with an adsorbent, and purifying to obtain an upper machine solution;
performing qualitative analysis on the upper computer solution by adopting a super high liquid chromatogram, a primary mass spectrum and a secondary mass spectrum;
performing quantitative analysis on the upper computer solution by adopting a super high liquid chromatography and a primary mass spectrum to obtain the contents of the adamantanamine compounds and the triazine herbicides in the algae sample;
the extractant is acetonitrile, ethyl acetate, a formic acid-acetonitrile mixed solution with the volume concentration of formic acid of 1 percent or a formic acid-ethyl acetate mixed solution with the volume concentration of formic acid of 1 percent;
the adsorbent is a PSA adsorbent.
Preferably, the ratio of the amount of the algae sample to the amount of the extractant is 3 g: (15-30) mL.
Preferably, the extraction comprises performing vortex extraction and ultrasonic extraction in sequence; the rotational speed of vortex extraction is 1500-2500 r/min, and the time is 30 s.
Preferably, the power of ultrasonic extraction is 300-600W, the frequency is 30-50 Hz, and the time is 30 min.
Preferably, before mixing the extracting solution and the adsorbent, the method further comprises the steps of concentrating and redissolving the extracting solution; the redissolution reagent is an acetonitrile aqueous solution with the volume concentration of formic acid of 0.1%, and the volume ratio of acetonitrile to water in the acetonitrile aqueous solution is 6: 4.
preferably, the dosage ratio of the extracting solution to the adsorbent is 5 mL: (50-150) mg.
Preferably, the purification is carried out under the condition of vortex, the rotation speed of the vortex is 1500-2500 r/min, and the time is 0.5-1.5 min.
Preferably, the parameters of the ultra-high liquid chromatography include;
a chromatographic column: waters BEH C18，2.1mm×100mm 1.7μm；
Column temperature: 40-50 ℃;
mobile phase A: acetonitrile;
mobile phase B: an aqueous solution of formic acid at a concentration of 0.1% by volume;
flow rate: 0.25 mL/min;
sample introduction volume: 10 mu L of the solution;
the gradient elution procedure included:
-3.00～0min：5％A；
0.00～6min：5％A→95％A；
6～8min：95％A；
8～9min：95％A→5％A；
9～10min：5％A；
the parameters of the primary mass spectrum comprise:
scanning mode: scanning positive ions;
capillary temperature: 200-350 ℃;
ion source temperature: 110-450 ℃;
spraying voltage: 3000-4000V;
lens voltage: 50V;
sheath gas: n is a radical of2The flow rate is 35-60 arb;
auxiliary gas: n is a radical of2The flow rate is 0-15 arb;
scanning range m/z: 100 to 900;
resolution R: 70000;
AGC target：1×106；
maximum residence time: 100 ms;
the parameters of the secondary mass spectrum include:
scanning mode: scanning positive ions;
capillary temperature: 200-350 ℃;
ion source temperature: 110-450 ℃;
spraying voltage: 3000-4000V;
lens voltage: 50V;
sheath gas: n is a radical of2The flow rate is 35-60 arb;
auxiliary gas: n is a radical of2The flow rate is 0-15 arb;
resolution R: 17500-35000;
collision energy: 60V;
AGC target：2×105。
preferably, the amantadine-based compound comprises amantadine, memantine, and rimantadine; the triazine herbicide comprises atrazine, cyanazine, dimethomorph, hexazinone, metamitron, metribuzin, prometryn, sec-butyl, simazine, terbuthylazine and ametryn.
The invention provides a detection method of an amantadine compound and a triazine herbicide in algae, which comprises the following steps: mixing the algae sample with an extracting agent, and extracting to obtain an extracting solution; mixing the extracting solution with an adsorbent, and purifying to obtain an upper machine solution; performing qualitative analysis on the upper computer solution by adopting a super high liquid chromatogram, a primary mass spectrum and a secondary mass spectrum; performing quantitative analysis on the upper computer solution by adopting a super high liquid chromatography and a primary mass spectrum to obtain the contents of the adamantanamine compounds and the triazine herbicides in the algae sample; the extractant is acetonitrile, ethyl acetate, a formic acid-acetonitrile mixed solution with the volume concentration of formic acid of 1 percent or a formic acid-ethyl acetate mixed solution with the volume concentration of formic acid of 1 percent; the adsorbent is a PSA adsorbent.
The invention adopts acetonitrile, ethyl acetate, formic acid-acetonitrile mixed solution with the volume concentration of 1% of formic acid or formic acid-ethyl acetate mixed solution with the volume concentration of 1% of formic acid as an extracting agent, can release amantadine compounds and triazine herbicides in algae samples, and then adopts PSA adsorbent as adsorbent, can adsorb and remove impurities which greatly interfere with the amantadine compounds and the triazine herbicides in the extracting solution to the maximum extent; the qualitative detection is carried out by adopting the ultra-high liquid chromatography, the primary mass spectrum and the secondary mass spectrum, so that the qualitative accuracy can be improved, false positives can be eliminated, and the detection accuracy can be improved; and then, the quantitative analysis is carried out by utilizing the ultra-high liquid chromatography and the primary chromatography, so that the amantadine compound and the triazine herbicide can be accurately quantified, the simultaneous detection of the amantadine compound and the triazine herbicide in the solution to be detected is realized, and the detection sensitivity is higher.
Drawings
FIG. 1 is a graph of the effect of different extractants on the extraction of amantadine-based compounds and triazine-based herbicides;
FIG. 2 is a graph of the effect of different adsorbents on the recovery of adamantanes and triazines herbicides;
FIG. 3 is an extraction chromatogram resolution chart of amantadine compounds and triazine herbicides (5. mu.g/L Waters BEH C)182.1 mm×100mm 1.7μm)；
FIG. 4 is an extraction chromatogram resolution chart of amantadine compounds and triazine herbicides (5. mu.g/L Waters BEH C)182.1 mm×100mm 1.7μm)；
Fig. 5 is a memantine ion full scan (second hadron ion 107.0855, NCE 60V);
fig. 6 is a full scan of rimantadine ion (second hadronate 81.0699, NCE 60V);
FIG. 7 is a screening parameter setting interface diagram.
Detailed Description
The invention provides a detection method of an amantadine compound and a triazine herbicide in algae, which comprises the following steps:
mixing the algae sample with an extracting agent, and extracting to obtain an extracting solution;
mixing the extracting solution with an adsorbent, and purifying to obtain an upper machine solution;
performing qualitative analysis on the upper computer solution by adopting a super high liquid chromatogram, a primary mass spectrum and a secondary mass spectrum;
and carrying out quantitative analysis on the upper computer solution by adopting a super-high liquid chromatography and a primary mass spectrum to obtain the contents of the adamantanamine compounds and the triazine herbicides in the algae sample.
In the present invention, the starting materials used in the present invention are preferably commercially available products unless otherwise specified.
The invention mixes the algae sample and the extractant to extract, and obtains the extracting solution.
The kind of the algae sample is not particularly limited in the present invention, as long as the obtained algae sample can be obtained by those skilled in the art. In the present invention, the algae sample is preferably prepared by including the steps of: homogenizing an algae raw material; the homogenization is preferably carried out in a homogenizer; the operation of the homogenization in the present invention is not particularly limited, and the homogenization operation known to those skilled in the art may be used.
In the invention, the extractant is acetonitrile, ethyl acetate, a formic acid-acetonitrile mixed solution with a formic acid volume concentration of 1% or a formic acid-ethyl acetate mixed solution with a formic acid volume concentration of 1%, and is preferably acetonitrile.
In the present invention, the ratio of the amount of the algae sample to the amount of the extractant is preferably 3 g: (15-30) mL, more preferably 3 g: (20-25) mL.
In the present invention, the extraction preferably comprises performing vortex extraction and ultrasonic extraction sequentially. In the invention, the rotation speed of the vortex extraction is preferably 1500-2500 r/min, and more preferably 2000 r/min; the time is preferably 30 s. In the present invention, the vortex extraction can mix and immerse the extractant and the algae sample sufficiently, and also provides enough air gaps and bubbles for the next ultrasonic extraction.
In the invention, the power of ultrasonic extraction is preferably 300-600W, and more preferably 400-500W; the frequency is preferably 30-50 Hz, and more preferably 40 Hz; the time is preferably 30 min. In the present invention, the ultrasonic extraction enables the extraction of the target substance.
After the extraction, the method preferably further comprises the step of carrying out first solid-liquid separation on the obtained extraction liquid, and taking the obtained supernatant as an extracting solution.
In the invention, the first solid-liquid separation mode is preferably centrifugal separation, and the rotating speed of the centrifugal separation is 8000-10000 r/min, and more preferably 9000 r/min; the time is preferably 10-15 min.
After the extracting solution is obtained, the extracting solution and the adsorbent are mixed and purified to obtain the upper machine solution.
In the present invention, before the extracting solution is mixed with the adsorbent, it is preferable that the extracting solution is concentrated and redissolved. In the invention, the concentration mode is preferably nitrogen blowing, and the temperature of the nitrogen blowing is preferably 40 ℃; the time for the concentration is not particularly limited in the present invention, as long as the concentration can be performed without an excessive solvent. In the present invention, the redissolving agent is preferably an acetonitrile aqueous solution having a formic acid volume concentration of 0.1%, and the volume ratio of acetonitrile to water in the acetonitrile aqueous solution is preferably 6: 4. in the present invention, the volume ratio of the extraction solution to the redissolved reagent is preferably 5: 1.
in the present invention, the adsorbent is a PSA adsorbent. In the present invention, the particle size of the adsorbent is preferably 50 to 75 μm.
In the present invention, the amount ratio of the extraction liquid to the adsorbent is preferably 5 mL: (50-150) mg, more preferably 5 mL: (75-125) mg, more preferably 5 mL: 100 mg.
In the invention, the purification is preferably carried out under the condition of vortex, and the rotation speed of the vortex is preferably 1500-2500 r/min, and is further preferably 2000 r/min; the time is preferably 0.5 to 1.5min, and more preferably 1.0 min.
After purification, the invention preferably also performs second solid-liquid separation on the obtained purified feed liquid, and the obtained supernatant is filtered to obtain the upper machine solution. In the invention, the second solid-liquid separation mode is preferably centrifugation, the rotation speed of the centrifugation is preferably 5000-10000 r/min, and the time is preferably 10-15 min. In the present invention, the filtration membrane for filtration is preferably a 0.22 μm PVDF filtration membrane.
After the upper computer solution is obtained, the invention adopts the ultra-high liquid chromatography, the primary mass spectrum and the secondary mass spectrum to carry out qualitative analysis on the upper computer solution.
In the present invention, the parameters of the ultra-high liquid chromatography include
The chromatographic column is preferably: waters BEH C18，2.1mm×100mm 1.7μm；
The column temperature is preferably: 40-50 ℃, and further preferably 45 ℃;
the mobile phase a is preferably: acetonitrile;
the mobile phase B is preferably: an aqueous solution of formic acid at a concentration of 0.1% by volume;
the flow rates are preferably: 0.25 mL/min;
the sample introduction volume is preferably: 10 mu L of the solution;
the gradient elution procedure is preferably as shown in table 1.
TABLE 1 gradient elution procedure
In the present invention, the parameters of the primary mass spectrum include:
the scan pattern is preferably: positive ion scanning, data dependent scanning (Full MS/dd-MS)2)；
The capillary temperature is preferably: 200 to 350 ℃, and more preferably 250 to 300 ℃;
the ion source temperature is preferably: 110 to 450 ℃, more preferably 150 to 400 ℃, and even more preferably 200 to 300 ℃;
the spray voltage is preferably: 3000-4000V, more preferably 3500V;
the lens voltage is preferably: 50V;
the sheath gas is preferably: n is a radical of2The flow rate is preferably 35 to 60arb, and more preferably 35 to 60arb40 to 50 arb;
the auxiliary gas is preferably: n is a radical of2The flow rate is preferably 0 to 15arb, and more preferably 5 to 10 arb;
the scanning range m/z is preferably: 100 to 900;
the resolution R is preferably: 70000;
the AGC target is preferably: 1X 106；
The maximum residence time is preferably: 100 ms.
In the present invention, the parameters of the secondary mass spectrum include:
the scan pattern is preferably: positive ion scanning, data dependent scanning (Full MS/dd-MS)2)；
The capillary temperature is preferably: 200 to 350 ℃, and more preferably 250 to 300 ℃;
the ion source temperature is preferably: 110 to 450 ℃, and more preferably 150 to 400 ℃;
the spray voltage is preferably: 3000-4000V, more preferably 3500V;
the lens voltage is preferably: 50V;
the sheath gas is preferably: n is a radical of2The flow rate is preferably 35-60 arb, and more preferably 40-50 arb;
the auxiliary gas is preferably: n is a radical of2The flow rate is preferably 0 to 15arb, and more preferably 5 to 10 arb;
the resolution R is preferably: 17500-35000;
the collision energy is preferably: 60V;
the AGC target is preferably: 2X 105；
The maximum residence time is preferably: 50-100 ms.
After the upper computer solution is obtained, the invention adopts the ultra-high liquid chromatography and the primary mass spectrum to carry out quantitative analysis on the upper computer solution, and the contents of the adamantanamine compounds and the triazine herbicides in the algae sample are obtained.
In the present invention, the parameters of the hplc and the primary mass spectrum are preferably the same as those of the above technical solution, and are not described herein again.
In the invention, the chromatographic peaks of the amantadine compounds and the triazine herbicides are preferably obtained by carrying out quantitative analysis on the upper computer solution by adopting an ultra-high liquid chromatography and a primary mass spectrum, and the contents of the adamantanamine compounds and the triazine herbicides in the algae sample are obtained based on the standard curves of the amantadine compounds and the triazine herbicides.
The standard curves of the amantadine compounds and the triazine herbicides are not particularly limited, and a standard curve establishing method well known to those skilled in the art can be adopted. In the present invention, when the standard curve of each amantadine compound and triazine herbicide is established, the scanning is preferably performed by using parameters of high performance liquid chromatography and primary mass spectrometry.
In the present invention, the amantadine-based compounds include amantadine, memantine, and rimantadine; the triazine herbicide comprises atrazine, cyanazine, dimethomorph, hexazinone, metamitron, metribuzin, prometryn, sec-butyl, simazine, terbuthylazine and ametryn.
The following examples are provided to illustrate the detection method of the adamantanamine compounds and triazine herbicides in algae according to the present invention, but they should not be construed as limiting the scope of the present invention.
Example 1
Sample pretreatment
Weighing 3.0g (+ -0.01 g) of algae sample to be detected, homogenizing by a homogenizer, cleaning a tool bit by 15mL of acetonitrile, adding the obtained product into the sample, performing ultrasonic extraction for 30min after vortex for 30s at 2000r/min, and centrifuging for 10min at 8000 r/min; and (2) putting 5mL of supernatant into a 15mL centrifuge tube, drying the supernatant by using nitrogen at 40 ℃, adding 1mL of acetonitrile aqueous solution containing 0.1 vol% formic acid (the volume ratio of acetonitrile to water in the acetonitrile aqueous solution is 6: 4) and 1mL of 50mg of PSA adsorbent (the particle size is 50-75 mu m), carrying out vortex at 2000r/min for 1min, centrifuging at 5000r/min for 10min, taking the supernatant, and filtering the supernatant through a 0.22 mu m PVDF filter membrane to obtain the solution to be detected.
The parameters of ultra-high liquid chromatography include:
liquid phase type: dionex UltiMate 3000 type ultra-high performance liquid phase system
A chromatographic column: waters BEH C18(2.1mm×100mm 1.7μm)；
Column temperature: 45 ℃;
mobile phase A: acetonitrile; mobile phase B: 0.1 vol% formic acid water, flow rate: 0.250mL/min, injection volume: 10 mu L of the solution;
the gradient elution procedure is shown in table 1.
The parameters of the primary mass spectrum include:
the mass spectrum model is as follows: thermo Q active Focus
Scanning mode: scanning positive ions; data dependent scanning (Full MS/dd-MS)2)；
Capillary temperature: 350 ℃;
ion source temperature: 350 ℃;
spraying voltage: 3500V;
lens voltage: 50V;
sheath gas: n is a radical of2The flow rate is 40 arb;
auxiliary gas: n is a radical of2The flow rate is 10 arb;
scanning range m/z: 100 to 900;
resolution R: 70000;
AGC target：1×106；
maximum residence time: 100 ms;
parameters of the secondary mass spectrum include:
the mass spectrum model is as follows: thermo Q active Focus;
scanning mode: positive ion scanning, data dependent scanning (Full MS/dd-MS)2)；
Capillary temperature: 350 ℃;
ion source temperature: 350 ℃;
spraying voltage: 3500V;
lens voltage: 50V;
sheath gas: n is a radical of2The flow rate is 40 arb;
auxiliary gas: n is a radical of2The flow rate is 10 arb;
resolution R: 17500-35000;
collision energy: 60V;
AGC target：2×105；
maximum residence time: 50-100 ms;
other parameters are shown in table 2.
TABLE 2 database information for amantadine-based compounds and triazine-based herbicide drugs (mass number deviation range 5ppm)
Example 2
Selection of the extractant
An extract was obtained by replacing the extractant acetonitrile in example 1 with 1 vol% acetonitrile formate, methanol, 1 vol% methanol formate, ethyl acetate, and 1 vol% ethyl acetate formate.
Calculating the extraction rate in the extracting solution by using the formula 1:
in equation 1:
instrument detection values: adding a standard substance solution into the blank matrix, and performing pretreatment, and then loading a machine to obtain a detection result of mu g/L;
blank sample matrix addition value: value of standard solution added to blank matrix sample, μ g/L.
The extraction rate of each of the obtained extractants is shown in FIG. 1. As can be seen from fig. 1: the extraction efficiency of acetonitrile is similar to the effects of 1 vol% acetonitrile formate, ethyl acetate and 1 vol% ethyl formate, which shows that acetonitrile, 1 vol% acetonitrile formate, ethyl acetate and 1 vol% ethyl formate can be used as the extracting agent.
Example 3
Selection of adsorbents
The PSA adsorbent of example 1 was driedIs replaced by C18Adsorbent, recovery calculated using equation 2:
in equation 2:
and (3) a computer-operated detection result: pretreating a blank matrix sample, fixing the volume by using a standard solution, purifying by using an adsorbent, and then loading the sample on a machine to obtain a detection result of mu g/L;
matrix-calibrated value: after the blank matrix sample is pretreated, the volume of the blank matrix sample is determined by using a standard solution with a certain concentration, and the volume of the blank matrix sample is mu g/L.
The effect of different adsorbents on the recovery of adamantanes and triazines herbicides is shown in figure 2. As can be seen from fig. 2: c18The adsorbent has a certain adsorption on part of herbicide medicaments, so that the recovery rate is reduced, therefore, the invention mainly adopts PSA adsorbent as a purification means, C18Adsorbent purification is used as an aid only when processing biological samples with relatively high oil content.
Example 4
Selection of reconstitution reagents
Since the flow rate of the mobile phase of the high performance liquid chromatography is only 0.25mL/min and the sample injection volume is 10 μ L, the peak shape of the compound paired with the organic phase of the redissolution reagent is large, and in this embodiment, the volume ratio of acetonitrile to water is 1: 1 as a redissolution reagent; adjusting according to the peak shape condition of each peak, and finally determining that the acetonitrile water solution with the formic acid concentration of 0.1 vol% is adopted (the volume ratio of acetonitrile to water in the acetonitrile water solution is 6: 4), wherein the peak shape and the response value of amantadine, rimantadine and memantine can be effectively improved by adding 0.1% formic acid.
Example 5
Working conditions of the apparatus
Using Waters BEH C18(2.1 mm. times.100 mm 1.7 μm) standard data were collected to create an mzVault spectrum library and a database.
In full scan mode, mass spectrometry as resolution increasesThe accurate molecular weight of the analyte can be accurately extracted, otherwise, when the resolution is reduced, the interferent with the same fragment as the target substance can be extracted, and a false positive result is caused. However, sensitivity generally decreases as resolution increases. It is therefore necessary to select the best resolution based on the sensitivity of all analyte substances. Data-dependent scanning secondary mass spectrometry (Full MS/dd-MS2) based on high resolution mass spectrometry changes are made to the mass spectrometry parameters in order to obtain the best instrument response. Multiple experimental result comparisons show that when the MS scanning resolution is 70000 and the MS/MS resolution is 17500, the resolution and the separation degree are better, the obtained extraction chromatograms of the amantadine compounds and the herbicides are shown in figures 3 and 4, wherein figure 3 is an extraction chromatogram resolution chart of the amantadine compounds and the triazine herbicides (5 mu g/L Waters BEH C)182.1mm × 100mm 1.7 μm); FIG. 4 is an extraction chromatogram resolution chart of amantadine compounds and triazine herbicides (5. mu.g/L Waters BEH C)182.1mm × 100mm 1.7 μm); FIGS. 3 and 4 show the extraction chromatograms of amantadine and triazine herbicides (5. mu.g/L Waters BEH C18 2.1mm×100mm 1.7μm)。
Example 6
Determination of isomers of rimantadine and memantine
Since rimantadine and memantine are isomers, the peak times on their respective chromatograms are comparable, as shown in fig. 5 and 6. However, since the difference in the ratio of the kurtosis of the daughter ions is large, only the kurtosis of the second hadron ion can be used as a criterion.
Example 7
Screening parameter settings
Since the molecular weights of the target compounds of the research are very small, particularly amantadine, rimantadine and memantine, and the parent ions of the compounds have a lot of peaks, the instrument is used for performing Full MS-ddMS2During scanning, because a secondary graph is easily missed due to the problems of scanning speed and ion response intensity, when screening parameter setting is performed, the parameter setting shown in fig. 7 is adopted, and specifically: peaks, m/z, Threshold Override: 100,000; S/N RatioThreshold: 500.0, Mass Tolerance: 5.0 ppm; RetentionTime, Identify, Window Override (Sec): 30, of a nitrogen-containing gas; fragment Ions, Confirm, Ignore if Not Defined; min. # of Fragmens: 1; intensity Threshold 10,000; mass Tolerance: 5.0 ppm; MS Order: MS 2; isotropic Pattern, Confirm, Fit Threshold (%): 75; allowed Mass development (ppm): 20; allowed Intensity development (%): 15; library Search, Confirm, Library Search Type: mzVault. The method comprises the steps of firstly screening and confirming essential conditions of parent ions and peak output time through software, then, carrying out non-essential conditions on the other conditions, and further confirming the generated suspected result through a secondary mass spectrum result so as to finally carry out qualitative and quantitative determination.
Example 8
Method linearity, recovery, precision and detection limit
Preparing a standard solution of amantadine, rimantadine, memantine and triazine herbicide medicaments by using an acetonitrile aqueous solution containing 0.1 vol% of formic acid (the volume ratio of acetonitrile to water in the acetonitrile aqueous solution is 6: 4), wherein the concentration range is 1-100 ng/mL, and determining the linear range in QE; the standard solution was tested by ultra-high liquid chromatography and primary mass spectrometry, and the resulting standard curve is shown in table 3.
In this experiment, the recovery rate was shown in Table 3, where the addition concentration of the recovered sample was 10. mu.g/kg, and the results of the recovery rate were measured in 3 replicates.
The quantitative limit results are shown in table 3.
TABLE 3 Linear range of adamantanamine and triazine herbicide drug in algae and recovery, precision and detection limits at different addition concentrations (n ═ 6)
As can be seen from table 3: amantadine compounds and triazinesThe linear coefficient of the standard curve of the herbicide drug can reach R2>0.99, the limit of quantitation is 1. mu.g/kg, since the method of the present invention is a method for quantifying parent ions. The experiment carries out 3 times of parallel measurement on the added and recovered samples, when the added concentration is 10 mug/kg, the recovery rate is between 66.6% and 77.0%, the standard deviation is between 1.76% and 9.21%, the recovery rate is stable, and the experiment requirements are met.
The foregoing is only a preferred embodiment of the present invention, and it should be noted that, for those skilled in the art, various modifications and decorations can be made without departing from the principle of the present invention, and these modifications and decorations should also be regarded as the protection scope of the present invention.
Claims (9)
1. A detection method of an amantadine compound and a triazine herbicide in algae comprises the following steps:
mixing the algae sample with an extracting agent, and extracting to obtain an extracting solution;
mixing the extracting solution with an adsorbent, and purifying to obtain an upper machine solution;
performing qualitative analysis on the upper computer solution by adopting a super high liquid chromatogram, a primary mass spectrum and a secondary mass spectrum;
performing quantitative analysis on the upper computer solution by adopting a super high liquid chromatography and a primary mass spectrum to obtain the contents of the adamantanamine compounds and the triazine herbicides in the algae sample;
the extractant is acetonitrile, ethyl acetate, a formic acid-acetonitrile mixed solution with the volume concentration of formic acid of 1 percent or a formic acid-ethyl acetate mixed solution with the volume concentration of formic acid of 1 percent;
the adsorbent is a PSA adsorbent.
2. The method of claim 1, wherein the ratio of the amount of the algae sample to the amount of the extractant is 3 g: (15-30) mL.
3. The detection method according to claim 1 or 2, wherein the extraction comprises performing vortex extraction and ultrasonic extraction in sequence; the rotational speed of vortex extraction is 1500-2500 r/min, and the time is 30 s.
4. The detection method according to claim 3, wherein the power of the ultrasonic extraction is 300-600W, the frequency is 30-50 Hz, and the time is 30 min.
5. The detection method according to claim 1, wherein the step of concentrating and redissolving the extract before mixing the extract with the adsorbent; the redissolution reagent is an acetonitrile aqueous solution with the volume concentration of formic acid of 0.1%, and the volume ratio of acetonitrile to water in the acetonitrile aqueous solution is 6: 4.
6. the detection method according to claim 1, wherein the amount ratio of the extraction solution to the adsorbent is 5 mL: (50-150) mg.
7. The detection method according to claim 1 or 6, wherein the purification is performed under the condition of vortex, and the rotation speed of the vortex is 1500-2500 r/min for 0.5-1.5 min.
8. The detection method according to claim 1, wherein the parameters of the ultra-high liquid chromatography include;
a chromatographic column: WatersBEHC18，2.1mm×100mm1.7μm；
Column temperature: 40-50 ℃;
mobile phase A: acetonitrile;
mobile phase B: an aqueous solution of formic acid at a concentration of 0.1% by volume;
flow rate: 0.25 mL/min;
sample introduction volume: 10 mu L of the solution;
the gradient elution procedure included:
-3.00～0min：5％A；
0.00～6min：5％A→95％A；
6～8min：95％A；
8～9min：95％A→5％A；
9～10min：5％A；
the parameters of the primary mass spectrum comprise:
scanning mode: scanning positive ions;
capillary temperature: 200-350 ℃;
ion source temperature: 110-450 ℃;
spraying voltage: 3000-4000V;
lens voltage: 50V;
sheath gas: n is a radical of2The flow rate is 35-60 arb;
auxiliary gas: n is a radical of2The flow rate is 0-15 arb;
scanning range m/z: 100 to 900;
resolution R: 70000;
AGCtarget：1×106；
maximum residence time: 100 ms;
the parameters of the secondary mass spectrum include:
scanning mode: scanning positive ions;
capillary temperature: 200-350 ℃;
ion source temperature: 110-450 ℃;
spraying voltage: 3000-4000V;
lens voltage: 50V;
sheath gas: n is a radical of2The flow rate is 35-60 arb;
auxiliary gas: n is a radical of2The flow rate is 0-15 arb;
resolution R: 17500-35000;
collision energy: 60V;
AGCtarget：2×105。
9. the detection method according to claim 1, wherein the amantadine-based compound includes amantadine, memantine, and rimantadine; the triazine herbicide comprises atrazine, cyanazine, dimethomorph, hexazinone, metamitron, metribuzin, prometryn, sec-butyl, simazine, terbuthylazine and ametryn.
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