CN108387655A - A method of for detecting persticide residue in Radix Ophiopogonis - Google Patents
A method of for detecting persticide residue in Radix Ophiopogonis Download PDFInfo
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- CN108387655A CN108387655A CN201810142175.4A CN201810142175A CN108387655A CN 108387655 A CN108387655 A CN 108387655A CN 201810142175 A CN201810142175 A CN 201810142175A CN 108387655 A CN108387655 A CN 108387655A
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- G—PHYSICS
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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
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
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- 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
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- G01N30/04—Preparation or injection of sample to be analysed
- G01N30/06—Preparation
- G01N30/14—Preparation by elimination of some components
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- 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
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- G01N30/26—Conditioning of the fluid carrier; Flow patterns
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- G01N30/02—Column chromatography
- G01N30/04—Preparation or injection of sample to be analysed
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Abstract
The present invention relates to a kind of methods for detecting persticide residue in Radix Ophiopogonis, include the following steps：(1), Radix Ophiopogonis to be measured is weighed, is handled using QuEChERS pretreatment technologies, the first test solution is made；(2), ribose acid lactone and appropriate sorbierite are weighed, acetonitrile and water dissolution are added, as analysis protectant；(3), the first test solution is measured, is blown to nitrogen and is closely done, add acetone solution, and analysis protectant is added, mixing, as the second test solution；(4), agricultural chemical reference substance is weighed, respectively plus the first reference substance solution is made in acetonitrile；The first reference substance solution is measured, respectively plus the second reference substance solution is made in acetone；(5), measuring method；(6), the content of pesticide in first test solution and second test solution is calculated by external standard method；This method has the characteristics that repeated good, accuracy height, use cost are low, quick, effective, easy can be measured to the pesticide residue in Radix Ophiopogonis.
Description
Technical field
The present invention relates to Chinese medicine security technology areas, and in particular to a kind of side for detecting persticide residue in Radix Ophiopogonis
Method.
Background technology
It is more prevalent with Chinese medicine artificial growth, in planting process using pesticide residue caused by pesticide by
People's common concern.Radix Ophiopogonis is China's Chinese medicine general types, and the main place of production is Sichuan and Zhejiang, by carrying out the place of production to two producing regions
Investigation finds that Radix Ophiopogonis has the case where using pesticide in planting process, includes mainly herbicide and insecticide, in addition Radix Ophiopogonis
Plantation is carried out by the way of crop rotation, and crop rotation floristics is more, can also be used and be arrived pesticide.
After the growth period of Radix Ophiopogonis applied pesticide, pesticide can remain in Radix Ophiopogonis, can not only reduce the drug effect of Radix Ophiopogonis,
The also health of entail dangers to eater；Therefore it is necessary to be detected to the persticide residue in Radix Ophiopogonis, to ensure the matter of Radix Ophiopogonis
Amount safety；However in the prior art, since the type of pesticide is more, the residual condition of pesticide is complicated in Radix Ophiopogonis, there are no it is simple,
Effectively method can comprehensively detect the persticide residue in Radix Ophiopogonis, to provide guarantor for the quality safety of Radix Ophiopogonis
Barrier.
Invention content
It is an object of the invention to improve the deficiency in the presence of the prior art, provide a kind of for detecting pesticide in Radix Ophiopogonis
The method of residual quantity has the characteristics that repeatability is good, accuracy is high, use cost is low, can be quick, effective, easy to wheat
Pesticide residue in winter is measured, and reliable data monitoring is provided for the quality safety of Radix Ophiopogonis.
The technical scheme is that：
A method of for detecting persticide residue in Radix Ophiopogonis, including step (5) and step (6), wherein
Step (5), precision measures the first test solution and the first reference substance solution is appropriate, respectively sample introduction, using liquid phase
Chromatograph-mass spectrometer coupling method is measured, and records chromatogram；
Precision measures the second test solution and the second reference substance solution is appropriate, respectively sample introduction, using gas chromatography-mass spectrum
Combination method is measured, and records chromatogram；
Wherein, the chromatographic condition of the Liquid Chromatography-Mass Spectrometry includes：Using octadecylsilane chemically bonded silica
Column, column temperature is 45 DEG C, using 0.1% formic acid solution containing 5mmol/L ammonium formates as mobile phase A, to contain 5mmol/L ammonium formates
And 0.1% formic acid methanol solution be Mobile phase B, flow velocity 0.2ml/min, according to the form below carry out gradient elution；
Step (6) calculates containing for pesticide in first test solution and second test solution by external standard method
Amount.
Preferably in scheme, the chromatographic column of the Liquid Chromatography-Mass Spectrometry is CORTECSTM UPLC C18,
2.1mm × 150mm, 1.6 μm.
Further, the chromatographic condition of the Liquid Chromatography-Mass Spectrometry further includes：The column temperature of chromatographic column is 45 DEG C,
Sample size is 2 μ l, and the flow velocity of the mobile phase A and B are 0.2ml/min.
Preferably in scheme, the chromatographic condition of the gas chromatography-mass spectrometry includes：Using elastic quartz capillary tube
Chromatographic column, column flow rate are wire velocity control pattern, initial flow rate 1.3ml/min；Using Splitless injecting samples, injector temperature is
240℃；Temperature program is：60 DEG C of initial temperature is kept for 1 minute, first rises to 120 DEG C with 30 DEG C/min, then with 10 DEG C/min liters
To 160 DEG C, 230 DEG C then is risen to 2 DEG C/min, then 300 DEG C are risen to 15 DEG C/min, 6min is kept, finally with 20 DEG C/min
320 DEG C are risen to, 3min is kept.
In further scheme, the Mass Spectrometry Conditions of the Liquid Chromatography-Mass Spectrometry further include：Ion source：EFI
Mist ionizes (ESI), cation scan pattern；Monitoring pattern：Multiple-reaction monitoring (MRM)；Ion source temperature：280℃；Gas stream
Amount：9L/min；Atomizer：40psi；Sheath temperature degree：350℃；Sheath throughput:11L/min；Capillary separation voltage：4000v.
Preferably in scheme, the Mass Spectrometry Conditions of the gas chromatography-mass spectrometry include：Ionization source uses electron bombardment
Ionization, ionizing energy 70eV, ion source temperature are 200 DEG C, and interface temperature is 250 DEG C.
Preferably in scheme, the chromatographic column of the gas chromatography-mass spectrometry is DB-17MS, 30m × 0.25mm,
0.25um。
Preferably in scheme, the chromatographic condition of the gas chromatography-mass spectrometry further includes：Carrier gas is helium, described
The purity of helium is more than 99.999%.
In further scheme, a kind of above-mentioned method for detecting persticide residue in Radix Ophiopogonis further includes following step
Suddenly：
Step (1), it is appropriate that precision weighs Radix Ophiopogonis to be measured, using QuEChERS pretreatment technologies to the Radix Ophiopogonis at
Reason, is made the first test solution；
Step (2), precision weighs ribose acid lactone and appropriate sorbierite, adds acetonitrile and water dissolution, as analysis protectant；
Step (3), precision measurement first test solution is appropriate, is blown to nitrogen and is closely done, and adds acetone solution, and add
It is appropriate to enter the analysis protectant, mixing, as the second test solution；
Step (4), it is appropriate that precision weighs 107 kinds of agricultural chemical reference substances, and respectively plus the first reference substance solution is made in acetonitrile；It is accurate
It is appropriate to measure the first reference substance solution, respectively plus the second reference substance solution is made in acetone.
Preferably, the QuEChERS pretreatment technologies include extraction process and purification process, wherein
The extraction process is：Radix Ophiopogonis powder to be measured is taken, No. three sieves are crossed, precision weighs in right amount, sets in tool plug centrifuge tube, adds
It is appropriate to enter glacial acetic acid solution, vortex makes the Radix Ophiopogonis powder complete wetting, and stands a period of time；Then it is appropriate that acetonitrile is added,
It sets and is vibrated on oscillator；The mixed of a certain proportion of anhydrous magnesium sulfate and anhydrous sodium acetate is added in cooling a period of time in ice bath
Powder is closed, shakes immediately scattered, then sets and is vibrated on oscillator, it is finally cooling in ice bath, and a period of time is centrifuged, take supernatant conduct
Extracting solution；
The purification process is：The precision measurement extracting solution is appropriate, sets the dispersed solid phase extraction for being pre-loaded with scavenging material
It takes in purification pipe, vortex makes extracting solution mix well, then sets and vibrated on oscillator, finally centrifugation a period of time, supernatant is taken to use
Miillpore filter filters, and subsequent filtrate is taken, as the first test solution.
Preferably, the frequency of the oscillator is 500 times/min；The rotating speed of the centrifugation is 5000 turns/min.
Preferably, the scavenging material includes anhydrous magnesium sulfate, N- propyl ethylenediamines, octadecylsilane chemically bonded silica,
Silica gel and Graphon.
In a kind of preferably scheme, the extraction process can be：Radix Ophiopogonis powder to be measured is taken, No. three sieves are crossed, precision claims
3.0g is taken, sets in 50ml polystyrene tool plug centrifuge tube, 1% glacial acetic acid solution 15ml of people, vortex is added to keep the Radix Ophiopogonis powder complete
Full infiltration, and stand 30 minutes；Then accurate that 15ml acetonitriles are added, it sets and is vibrated 5 minutes on oscillator, it is 30 points cooling in ice bath
Clock, it is 4 that 7.5g mass ratioes, which are added,:1 anhydrous magnesium sulfate and the mixed-powder of anhydrous sodium acetate, shake scattered immediately, then set oscillator
Upper oscillation 3 minutes, it is finally 10 minutes cooling in ice bath, and centrifuge 5 minutes, take supernatant as extracting solution.
In a kind of preferably scheme, the purification process can be：Precision measures the extracting solution 8ml, sets advance dress
Have in the dispersive solid-phase extraction purification pipe of scavenging material, wherein scavenging material includes anhydrous magnesium sulfate 900mg, N- propyl second two
Amine 300mg, octadecylsilane chemically bonded silica 300mg, silica gel 300mg and Graphon 90mg；Vortex makes extracting solution fill
Divide mixing, then set and vibrated 5 minutes on oscillator, so that extracting solution is fully cleaned up, finally centrifuges 5 minutes, take supernatant micropore
Filter membrane filters, and subsequent filtrate is taken, as the first test solution.
Preferably, in the step (2), in the analysis protectant, the mass ratio of ribose acid lactone and sorbierite is 2:
1。
In a kind of preferably scheme, the preparation process of the analysis protectant can be：Precision weighs ribose acid lactone
0.5g adds acetonitrile 25ml to make dissolving, and the solution of a concentration of 20mg/ml of ribose acid lactone is made；Another precision weighs sorbierite
0.25g adds water 12.5ml to dissolve, and adds acetonitrile to 25ml, the solution that sorbitol concentration is 10mg/ml is made, finally by above-mentioned two
Kind solution mixing, shakes up, as analysis protectant.
Preferably, in the step (3), the preparation process of second test solution is：Described in the accurate measurement
First test solution 3ml is blown to nitrogen and is closely done, added acetone solution and be settled to 2ml, shake up；Precision measures 1ml, is added
The analysis protectant 0.3ml, mixing, as the second test solution.
Preferably, in the step (4), the concentration of first reference substance solution can be 100ppm；Second reference substance
The concentration of solution can be 500ppb.
Compared with prior art, using a kind of method for detecting persticide residue in Radix Ophiopogonis provided by the invention, tool
There is following advantageous effect：
1, method provided by the present invention, using Liquid Chromatography-Mass Spectrometry and gas chromatography-mass spectrometry to wheat
Winter carries out chromatographic isolation and detection, and equipment popularity rate is high, and use cost is low.
2, in method provided by the present invention, the pre-treating method of Radix Ophiopogonis to be measured is simple, and sample usage amount is small, Ke Yifang
Just, quickly it is prepared into test solution.
3, in method provided by the present invention, gas-chromatography separation preferentially uses DB-17MS, 30m × 0.25mm, 0.25 μm
Chromatographic column, liquid chromatogram separation preferentially using octadecylsilane chemically bonded silica column (CORTECSTM UPLC C18,2.1mm ×
150mm, 1.6 μm) so that the separating degree between each impurity and ingredient to be measured (remains of pesticide) is good, and this method also has
The features such as system stability is good, repeated good, and accuracy is high, and appearance time is fast, and analysis time is short, saving experimental cost.
It 4, can be quick, effective, easy to 107 pesticide index (portions in Radix Ophiopogonis using detection method provided by the present invention
Point index further includes isomer) it is measured, provide reliable data monitoring for Radix Ophiopogonis quality safety.
Description of the drawings
In order to illustrate the technical solution of the embodiments of the present invention more clearly, below will be to needed in the embodiment attached
Figure is briefly described, it should be understood that the following drawings illustrates only certain embodiments of the present invention, therefore is not construed as pair
The restriction of range for those of ordinary skill in the art without creative efforts, can also be according to this
A little attached drawings obtain other relevant attached drawings.
Fig. 1 is a kind of for detecting in Radix Ophiopogonis in the method for persticide residue, using gas phase provided in 1 using implementing
The chromatogram that chromatograph-mass spectrometer coupling method is determined.
Fig. 2 is a kind of for detecting in Radix Ophiopogonis in the method for persticide residue, using liquid phase provided in 1 using implementing
The chromatogram that chromatograph-mass spectrometer coupling method is determined.
Specific implementation mode
Below in conjunction with attached drawing in the embodiment of the present invention, technical solution in the embodiment of the present invention carries out clear, complete
Ground describes, it is clear that described embodiments are only a part of the embodiments of the present invention, instead of all the embodiments.Usually exist
The component of the embodiment of the present invention described and illustrated in attached drawing can be arranged and be designed with a variety of different configurations herein.Cause
This, the detailed description of the embodiment of the present invention to providing in the accompanying drawings is not intended to limit claimed invention below
Range, but it is merely representative of the selected embodiment of the present invention.Based on the embodiment of the present invention, those skilled in the art are not doing
The every other embodiment obtained under the premise of going out creative work, shall fall within the protection scope of the present invention.
Embodiment 1
A kind of method for detecting persticide residue in Radix Ophiopogonis is provided in the present embodiment, is included the following steps：
Step (1), the preparation of the first test solution；
It is appropriate that precision weighs Radix Ophiopogonis to be measured, is handled the Radix Ophiopogonis using QuEChERS pretreatment technologies, is made
One test solution；
Preferably, the QuEChERS pretreatment technologies include extraction process and purification process.
Further, the extraction process can be：Radix Ophiopogonis powder to be measured is taken, No. three sieves are crossed, precision weighs in right amount, sets tool
It fills in centrifuge tube, addition glacial acetic acid solution is appropriate, and vortex makes the Radix Ophiopogonis powder complete wetting, and stands a period of time；Then
It is appropriate that acetonitrile is added, sets and is vibrated on oscillator；Cooling a period of time, is added a certain proportion of anhydrous magnesium sulfate and nothing in ice bath
The mixed-powder of water sodium acetate, shakes scattered immediately, then sets and is vibrated on oscillator, finally cooling in ice bath, and centrifuges a period of time,
Take supernatant as extracting solution.
In a kind of preferably scheme, the extraction process can be：Radix Ophiopogonis powder to be measured is taken, No. three sieves are crossed, precision claims
3.0g is taken, sets in 50ml polystyrene tool plug centrifuge tube, 1% glacial acetic acid solution 15ml of people, vortex is added to keep the Radix Ophiopogonis powder complete
Full infiltration, and stand 30 minutes；Then accurate that 15ml acetonitriles are added, it sets and is vibrated 5 minutes on oscillator, it is 30 points cooling in ice bath
Clock, it is 4 that 7.5g mass ratioes, which are added,:1 anhydrous magnesium sulfate and the mixed-powder of anhydrous sodium acetate, shake scattered immediately, then set oscillator
Upper oscillation 3 minutes, it is finally 10 minutes cooling in ice bath, and centrifuge 5 minutes, take supernatant as extracting solution.
Preferably, the frequency of the oscillator is 500 times/min.
Preferably, the rotating speed of the centrifugation is 5000 turns/min.
Further, the purification process can be：The precision measurement extracting solution is appropriate, sets and is pre-loaded with scavenging material
Dispersive solid-phase extraction purification pipe in, vortex makes extracting solution mix well, then sets and is vibrated on oscillator, when finally centrifuging one section
Between, it takes supernatant to be filtered with miillpore filter, takes subsequent filtrate, as the first test solution；
Preferably, the scavenging material includes anhydrous magnesium sulfate, N- propyl ethylenediamines, octadecylsilane chemically bonded silica,
Silica gel and Graphon.
In a kind of preferably scheme, the purification process can be：Precision measures the extracting solution 8ml, sets advance dress
Have in the dispersive solid-phase extraction purification pipe of scavenging material, wherein scavenging material includes anhydrous magnesium sulfate 900mg, N- propyl second two
Amine 300mg, octadecylsilane chemically bonded silica 300mg, silica gel 300mg and Graphon 90mg；Vortex makes extracting solution fill
Divide mixing, then set and vibrated 5 minutes on oscillator, so that extracting solution is fully cleaned up, finally centrifuges 5 minutes, take supernatant micropore
Filter membrane filters, and subsequent filtrate is taken, as the first test solution.
Step (2), the preparation of analysis protectant；
Precision weighs ribose acid lactone and appropriate sorbierite, adds acetonitrile and water dissolution, and ribose acid lactone and sorbierite is made
Mass ratio be 2:1 solution, as analysis protectant.
In a kind of preferably scheme, the preparation process of the analysis protectant can be：Precision weighs ribose acid lactone
0.5g adds acetonitrile 25ml to make dissolving, and the solution of a concentration of 20mg/ml of ribose acid lactone is made；Another precision weighs sorbierite
0.25g adds water 12.5ml to dissolve, and adds acetonitrile to 25ml, the solution that sorbitol concentration is 10mg/ml is made, finally by above-mentioned two
Kind solution mixing, shakes up, as analysis protectant.
Step (3), the preparation of the second test solution；
Precision measurement first test solution is appropriate, is blown to nitrogen and is closely done, and adds acetone solution, and described divide is added
It is appropriate to analyse protective agent, mixing, as the second test solution.
In a kind of preferably scheme, the preparation process of second test solution is：It is described accurate to measure described the
One test solution 3ml is blown to nitrogen and is closely done, added acetone solution and be settled to 2ml, shake up；Precision measures 1ml, and institute is added
State analysis protectant 0.3ml, mixing, as the second test solution.
Step (4), the preparation of reference substance solution；
It is appropriate that precision weighs 107 kinds of agricultural chemical reference substances, and respectively plus the first reference substance solution is made in acetonitrile；Precision measures first
Appropriate reference substance solution, respectively plus the second reference substance solution is made in acetone.
Preferably, a concentration of 100ppm of first reference substance solution；A concentration of 500ppb of second reference substance solution.
Step (5), measuring method；
Precision measures first test solution and the first reference substance solution is appropriate, respectively sample introduction, using liquid chromatogram-
Mass Spectrometry is measured, and records chromatogram；
Precision measures second test solution and the second reference substance solution is appropriate, respectively sample introduction, using gas-chromatography-
Mass Spectrometry is measured, and records chromatogram；
Step (6) calculates；
The content of pesticide in first test solution and second test solution is calculated by external standard method.
Preferably, in the step (5), the chromatographic condition of the Liquid Chromatography-Mass Spectrometry includes：Using octadecane
Base silane bonded silica gel column, using 0.1% formic acid solution containing 5mmol/L ammonium formates as mobile phase A, to contain 5mmol/L first
The methanol solution of sour ammonium and 0.1% formic acid is Mobile phase B, and according to the form below carries out gradient elution；
It will be appreciated by those skilled in the art that the meaning of upper table, such as in table, at 5~15 minutes, mobile phase A is 95 →
5, the meaning that Mobile phase B is 5 → 95 is：In 5~15 minutes, the content of mobile phase A is gradually lowered to 5% from 95%,
At the same time, the content of Mobile phase B progressively increases to 95% from 5%, any time, the sum of the content of mobile phase A and Mobile phase B
It is 100%；It is appreciated that in 17~17.5 minutes, the content of mobile phase A progressively increases to 95% from 5%, at the same time,
The content of Mobile phase B is gradually lowered to 5% from 95%, any time, and the sum of the content of mobile phase A and Mobile phase B is 100%.
Further, the chromatographic condition of the Liquid Chromatography-Mass Spectrometry further includes：The column temperature of chromatographic column is 45 DEG C,
Sample size is 2 μ l, and the flow velocity of the mobile phase A and B are 0.2ml/min.
Preferably, the chromatographic column of the Liquid Chromatography-Mass Spectrometry be CORTECSTM UPLC C18,2.1mm ×
150mm, 1.6 μm.
The Mass Spectrometry Conditions of the Liquid Chromatography-Mass Spectrometry include：Ion source：Electron spray ionisation (ESI), cation is swept
Retouch pattern；Monitoring pattern：Multiple-reaction monitoring (MRM)；Ion source temperature：280℃；Gas flow：9L/min；Atomizer：
40psi；Sheath temperature degree：350℃；Sheath throughput:11L/min；Capillary separation voltage：4000v.This Mass Spectrometry Conditions can make mesh
It marks object and obtains preferably response.
The chromatographic condition of the gas chromatography-mass spectrometry includes：Using quartz capillary column, column flow rate
For wire velocity control pattern, initial flow rate 1.3ml/min；Using Splitless injecting samples, injector temperature is 240 DEG C；Heat up journey
Sequence is：60 DEG C of initial temperature is kept for 1 minute, first rises to 120 DEG C with 30 DEG C/min, then rise to 160 DEG C with 10 DEG C/min, then
230 DEG C are risen to 2 DEG C/min, then 300 DEG C are risen to 15 DEG C/min, 6min is kept, finally rises to 320 DEG C with 20 DEG C/min, protect
Hold 3min.
The Mass Spectrometry Conditions of the gas chromatography-mass spectrometry include：Ionization source uses electron impact ionization, ionizing energy
For 70eV, ion source temperature is 200 DEG C, and interface temperature is 250 DEG C.This Mass Spectrometry Conditions can make object obtain preferably sound
It answers.
Preferably, the chromatographic column of the gas chromatography-mass spectrometry is DB-17MS, 30m × 0.25mm, 0.25um.
Further, the chromatographic condition of the gas chromatography-mass spectrometry further includes：Carrier gas is helium, the helium
Purity be more than 99.999%.
Embodiment 2
The present embodiment is mainly investigated, and the first test solution is prepared according to the method implemented described in step (1) described in 1
Afterwards, whether there is also the ingredients for interfering component (i.e. remaining farm chemical ingredients in Radix Ophiopogonis) to be measured in the first test solution.
By a large amount of investigations and experiment of early period, to screening, screening pesticide product have been carried out in Radix Ophiopogonis the case where contained pesticide
Kind include limiting with pesticide, high poison persistent pesticide and common pesticides etc., is found by screening, may usually be contained in Radix Ophiopogonis
It is several in 107 kinds of farm chemical ingredients (part farm chemical ingredients further include one or more isomers)；In detecting Radix Ophiopogonis
Persticide residue when, this 107 kinds of farm chemical ingredients can be predominantly detected.
Therefore in this example, accurate to measure 107 kinds of (including a variety of isomers) appropriate agricultural chemical reference substances, respectively with base
Matter is configured to reference substance solution；Precision measures appropriate reference substance solution, and mixing contrast solution is made, using in embodiment 1 in mixing
The measuring method of step (5) is measured, wherein when being measured using gas chromatography-mass spectrometry, gas chromatography-mass spectrum
The parameter of combined instrument is as shown in table 1, and it is as shown in Figure 1 to measure obtained chromatogram.
Location parameter when table 1 is measured using gas chromatography-mass spectrometry
In table, CE1, CE2, CE3 respectively represent collision energy；The identical ingredient of quota ion pair isomer each other,
Such as the quota ion pair of propiconazole -1 and propiconazole -2 is all：259.0>173.0, show propiconazole -1 and propiconazole -2 each other
Isomer can be regarded as a kind of farm chemical ingredients in the present embodiment；Remaining is repeated no more.
When being measured using Liquid Chromatography-Mass Spectrometry, the parameter of liquid chromatograph-mass spectrometer is as shown in table 2,
It is as shown in Figure 2 to measure obtained chromatogram.
Location parameter when table 2 is measured using Liquid Chromatography-Mass Spectrometry
Wherein, Fragmentor indicates cracking voltage, can be seen that from table 1, table 2 and Fig. 1, Fig. 2 according to embodiment 1
The measuring method (including chromatographic condition and Mass Spectrometry Conditions) of middle step (5) is measured, mix contrast solution in 107 kinds (including
Isomer) ingredient in agricultural chemical reference substance can reach and efficiently separate, separating degree is good, and agricultural chemical reference substance to it is to be measured at
Divide noiseless；In summary, a kind of specificity for detecting the method for persticide residue in Radix Ophiopogonis provided by the present invention is strong.
Embodiment 3
The methodological study of the present invention：Linear and range
Linearly the basis of quantitative determination, such as impurity quantitatively detect, content and dissolving-out method be required to verification linearly and
The range of linearity, so that in the measurement range of design, sample concentration is in a linear relationship with response.
In this example, 107 kinds of agricultural chemical reference substance solution are prepared (including with point different according to the method described in embodiment 1
Structure body, including the first reference substance solution and the second reference substance solution), and a variety of concentration levels are diluted to, difference sample introduction, using reality
The detection method applied in example 1 is measured, to determine the linear equation of 126 kinds of farm chemical ingredients, linear phase respectively
Shown in relationship number (r) and the range of linearity, table 3 and table 4, wherein r is linearly dependent coefficient, r for characterize studied variable it
Between linearly related degree；Linear equation in the range of linearity is standard curve.
The ingredient that table 3 is gone out using gas chromatography/mass spectrometry
As shown in Table 3, when being measured using gas chromatography-mass spectrometry, 54 kinds can be effectively detected out (at this
In embodiment, isomer can be regarded as a kind of farm chemical ingredients) farm chemical ingredients, in the range of linearity of each farm chemical ingredients, each pesticide
The linearity of ingredient is good, and linearly dependent coefficient r is all higher than 0.999, and the range of linearity is wider.
The ingredient that table 4 is determined using Liquid Chromatography-Mass Spectrometry
As shown in Table 4, when being measured using Liquid Chromatography-Mass Spectrometry, can be effectively detected out 54 kinds (including
1 kind of isomer ingredient) farm chemical ingredients, in the range of linearity of each farm chemical ingredients, the linearity of each farm chemical ingredients is good, line
Property correlation coefficient r is all higher than 0.999, and the range of linearity is wider.
Embodiment 5
The methodological study of the present invention：Accuracy
In the present embodiment, using sample-adding recovery test method, to calculate the rate of recovery result of each ingredient, wherein be loaded back
The process of acceptance test method is：Precision measure embodiment 2 in the mixing contrast solution it is appropriate, be separately added into 107 kinds of pesticides at
Point (in the present embodiment, there are the farm chemical ingredients of isomer, isomer to distinguish sample introduction and measure, therefore in total including 126 kinds
Ingredient) in right amount, the measuring method in embodiment 1 described in step (5) is then used, continuous sample introduction 6 times, is measured respectively respectively,
And the area of each chromatographic peak in corresponding chromatogram is calculated, to calculate the rate of recovery, meanwhile, calculating relative standard deviation (RSD,
Relative standard deviation), result of calculation is as shown in table 5 and table 6.
Table 5 uses the rate of recovery of each ingredient of gas chromatography/mass spectrometry
As shown in Table 5, using gas chromatography/mass spectrometry go out 72 kinds of (including a variety of isomers) pesticides at
The rate of recovery of timesharing, each ingredient is higher, and (in 72 ingredients, the RSD of only 4 ingredients is slightly larger than relative standard deviation RSD
15%, remaining is respectively less than or much smaller than 15%) relatively low.
Table 6 measures the rate of recovery of each ingredient using Liquid Chromatography-Mass Spectrometry
As shown in Table 6,54 kinds of (including a kind of isomer) farm chemical ingredients are measured using Liquid Chromatography-Mass Spectrometry
When, the rate of recovery of each ingredient is higher, and relative standard deviation RSD (RSD of 54 ingredients be respectively less than or be much smaller than 15%) also compared with
It is low.
To sum up, through this embodiment the result shows that：It is provided by the present invention a kind of for detecting pesticide residue in Radix Ophiopogonis
The method of amount, the rate of recovery is good, and accuracy is high.
Embodiment 6
The methodological study of the present invention：Method precision
In the present embodiment, using by the mixing contrast solution of substrate preparation, utilizing step in embodiment (1) in embodiment 2
Suddenly the measuring method of (5), continuous sample introduction 6 times record peak area, as a result show：The RSD values of each Component peak area are respectively less than
15.0%；
Further, same lot number Radix Ophiopogonis sample is taken, according to 1 the method for embodiment, prepares the first test solution respectively
With 6 parts of the second test solution, and sample introduction is surveyed in accordance with the law using measuring method (including chromatographic condition and Mass Spectrometry Conditions) described in embodiment 1
It is fixed, as a result show：The relative standard deviation (RSD) of each pesticide detected level is respectively less than 15.0%.
To sum up show a kind of method for detecting persticide residue in Radix Ophiopogonis provided by the present invention, method precision
It is good.
The above description is merely a specific embodiment, but scope of protection of the present invention is not limited thereto, any
Those familiar with the art in the technical scope disclosed by the present invention, can easily think of the change or the replacement, and should all contain
Lid is within protection scope of the present invention.
Claims (10)
1. a kind of method for detecting persticide residue in Radix Ophiopogonis, which is characterized in that including step (5) and step (6),
In,
Step (5), precision measures the first test solution and the first reference substance solution is appropriate, respectively sample introduction, using liquid chromatogram-
Mass Spectrometry is measured, and records chromatogram；
Precision measures the second test solution and the second reference substance solution is appropriate, respectively sample introduction, using gas chromatography-mass spectrography
Method is measured, and records chromatogram；
Wherein, the chromatographic condition of the Liquid Chromatography-Mass Spectrometry includes：Using octadecylsilane chemically bonded silica column, column
Temperature be 45 DEG C, using 0.1% formic acid solution containing 5mmol/L ammonium formates as mobile phase A, with contain 5mmol/L ammonium formates and
The methanol solution of 0.1% formic acid is Mobile phase B, flow velocity 0.2ml/min, according to the form below progress gradient elution；
Step (6) is calculated the content of pesticide in first test solution and second test solution by external standard method.
2. the method according to claim 1 for detecting persticide residue in Radix Ophiopogonis, which is characterized in that the liquid phase color
The chromatographic column of spectrum-Mass Spectrometry is CORTECSTM UPLC C18,2.1mm × 150mm, 1.6 μm.
3. the method according to claim 2 for detecting persticide residue in Radix Ophiopogonis, which is characterized in that the gas phase color
The chromatographic condition of spectrum-Mass Spectrometry includes：Using quartz capillary column, column flow rate is wire velocity control pattern,
Initial flow rate is 1.3ml/min；Using Splitless injecting samples, injector temperature is 240 DEG C；Temperature program is：60 DEG C of initial temperature,
It is kept for 1 minute, first rises to 120 DEG C with 30 DEG C/min, then 160 DEG C are risen to 10 DEG C/min, then rise to 230 DEG C with 2 DEG C/min,
300 DEG C are risen to 15 DEG C/min again, keeps 6min, finally rises to 320 DEG C with 20 DEG C/min, keeps 3min.
4. according to any methods for detecting persticide residue in Radix Ophiopogonis of claim 1-3, which is characterized in that described
The Mass Spectrometry Conditions of Liquid Chromatography-Mass Spectrometry further include：Ion source：Electron spray ionisation (ESI), cation scan pattern；Prison
Survey pattern：Multiple-reaction monitoring (MRM)；Ion source temperature：280℃；Gas flow：9L/min；Atomizer：40psi；Sheath temperature
Degree：350℃；Sheath throughput:11L/min；Capillary separation voltage：4000v.
5. the method according to claim 4 for detecting persticide residue in Radix Ophiopogonis, which is characterized in that the gas phase color
The Mass Spectrometry Conditions of spectrum-Mass Spectrometry include：Ionization source uses electron impact ionization, ionizing energy 70eV, ion source temperature
It it is 200 DEG C, interface temperature is 250 DEG C.
6. the method according to claim 5 for detecting persticide residue in Radix Ophiopogonis, which is characterized in that the gas phase color
The chromatographic column of spectrum-Mass Spectrometry is DB-17MS, 30m × 0.25mm, 0.25um.
7. the method according to claim 6 for detecting persticide residue in Radix Ophiopogonis, which is characterized in that further include following
Step：
Step (1), it is appropriate that precision weighs Radix Ophiopogonis to be measured, is handled the Radix Ophiopogonis using QuEChERS pretreatment technologies, makes
At the first test solution；
Step (2), precision weighs ribose acid lactone and appropriate sorbierite, adds acetonitrile and water dissolution, as analysis protectant；
Step (3), precision measurement first test solution is appropriate, is blown to nitrogen and is closely done, adds acetone solution, and institute is added
It is appropriate to state analysis protectant, mixing, as the second test solution；
Step (4), it is appropriate that precision weighs 107 kinds of agricultural chemical reference substances, and respectively plus the first reference substance solution is made in acetonitrile；Precision measures
First reference substance solution is appropriate, and respectively plus the second reference substance solution is made in acetone.
8. the method according to claim 7 for detecting persticide residue in Radix Ophiopogonis, which is characterized in that described
QuEChERS pretreatment technologies include extraction process and purification process, wherein
The extraction process is：Radix Ophiopogonis powder to be measured is taken, No. three sieves are crossed, precision weighs in right amount, sets in tool plug centrifuge tube, ice is added
Appropriate acetum, vortex make the Radix Ophiopogonis powder complete wetting, and stand a period of time；Then it is appropriate that acetonitrile is added, sets and shakes
It swings and is vibrated on device；Cooling a period of time, is added the mixed powder of a certain proportion of anhydrous magnesium sulfate and anhydrous sodium acetate in ice bath
End, shakes scattered immediately, then sets and is vibrated on oscillator, finally cooling in ice bath, and centrifuges a period of time, takes supernatant as extraction
Liquid；
The purification process is：Precision measures that the extracting solution is appropriate, set be pre-loaded with scavenging material dispersive solid-phase extraction it is net
Change in pipe, vortex makes extracting solution mix well, then sets and vibrated on oscillator, finally centrifugation a period of time, takes supernatant micropore
Filter membrane filters, and subsequent filtrate is taken, as the first test solution.
9. the method according to claim 8 for detecting persticide residue in Radix Ophiopogonis, which is characterized in that the oscillator
Frequency be 500 times/min；The rotating speed of the centrifugation is 5000 turns/min.
10. the method according to claim 9 for detecting persticide residue in Radix Ophiopogonis, which is characterized in that the purification
Material includes anhydrous magnesium sulfate, N- propyl ethylenediamines, octadecylsilane chemically bonded silica, silica gel and Graphon.
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| CN113203823B (en) * | 2021-04-21 | 2024-05-31 | 中国烟草总公司郑州烟草研究院 | Application of hydroxyl-containing compound, matrix improver and analysis method of flavor components in tobacco |
| CN113945656A (en) * | 2021-09-29 | 2022-01-18 | 中国农业科学院农产品加工研究所 | Method for detecting pesticide and metabolite residues thereof in traditional Chinese medicinal materials |
| CN113933432A (en) * | 2021-10-22 | 2022-01-14 | 中国烟草总公司郑州烟草研究院 | High-sensitivity and high-precision analysis method suitable for detecting alkaloid in tobacco leaves |
| CN113933432B (en) * | 2021-10-22 | 2023-08-25 | 中国烟草总公司郑州烟草研究院 | High-sensitivity and high-precision analysis method suitable for detecting alkaloid in tobacco leaves |
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