CN106018571A - Detection method for residual amount of chlorantraniliprole in aquatic vegetables - Google Patents

Abstract

The invention provides a detection method characterized by good stability and high sensitivity for the residual amount of chlorantraniliprole in aquatic vegetables. The method comprises the steps of: (1) preparation of a standard solution; (2) preparation of a solvent; (3) a pretreatment extraction process; and (4) determination: taking the filtrate obtained in step (1) and step (3) to conduct liquid chromatography-tandem mass spectrometry determination. The detection method for the residual amount of chlorantraniliprole in aquatic vegetables uses a mixed solvent of ethyl acetate and acetone (V/V 7:3) to extract chlorantraniliprole residue in aquatic vegetables, purification by a dispersive solid-phase extraction N-propyl ethylenediamine (PSA) adsorbent is conducted, and then the characteristics of strong selectivity and anti-interference capability of liquid chromatography-tandem mass spectrometry are utilized to determine the residual amount. By using the ethyl acetate:acetone (V/V 7:3) mixed solvent, the extraction efficiency is high, and little extraction matrix interference exists. By using the PSA filler as the adsorbent, the extracted organic acid, pigments and other interfering substances can be removed so as to achieve the purpose of purification.

Description

The method of Rynaxypyr residual quantity in detection aquatic vegetable

Technical field

The present invention relates to the detection method of a kind of aquatic vegetable residual, chlorine in especially a kind of detection aquatic vegetable The method of worm Benzoylamide residual quantity.

Background technology

Rynaxypyr is the new and effective broad spectrum pesticide of one of E.I.Du Pont Company's exploitation, to lepidopterous night Moth section, Pyralidae, Carposinidae, tortricid, miller section, diamond-back moth section, Gelechidae, Gracilariidae etc. are equal Have and well control effect, moreover it is possible to control coleoptera Culculionidae, Chrysomelidae；Diptera Agromyzidae；Bemisia tabaci Etc. multiple non-lepidoptera pest.Rynaxypyr can efficiently activate insecticide ryanodine receptor, destroys Cytoplasm Interior Ca²⁺Ambient stable, causes insecticide paralysis death, high to the larva activity of lepidoptera pest, has fast Speed stops insect feed, to be efficiently detained activity, resistance of rainwater washing against, permeability strong.

Report relatively about the method for Rynaxypyr residues detection in veterinary antibiotics the most both at home and abroad Many, main employing high performance liquid chromatography and LC/MS, its pre-treatment Extraction solvent is predominantly Acetonitrile or acid acetonitrile, and not yet have about the method for Rynaxypyr residues detection in aquatic vegetable Report.

Summary of the invention

The invention provides Rynaxypyr in a kind of good stability, highly sensitive detection aquatic vegetable residual The method of allowance.

Realize the method for Rynaxypyr residual quantity in the detection aquatic vegetable of the object of the invention, including as follows Step:

(1) preparation of standard solution:

Accurately weigh 10mg chlorine worm this Methanamide standard substance in 10mL volumetric flask, dissolve constant volume with methanol To scale, being configured to concentration is 1mg/mL Standard Stock solutions, is placed in-18 DEG C of refrigerators freezing lucifuge and protects Deposit；

Accurately a certain amount of 1mg/mL Rynaxypyr Standard Stock solutions of absorption is in volumetric flask, uses first It is 10 μ g/mL intermediate concentration standard solution that alcohol is diluted to mass concentration, is placed in 4 DEG C of refrigerators freezing lucifuge and protects Deposit；

Accurately draw a certain amount of 10 μ g/mL Rynaxypyr intermediate concentration standard solution, use first respectively Alcohol and water mixed solution (V/V 3: 7) stepwise dilution are 2ng/mL, 5ng/mL, 10ng/mL to concentration, The standard working solution of 20ng/mL, 50ng/mL and 100ng/mL, matching while using；

(2) preparation of solvent:

Ethyl acetate: acetone (V/V 7: 3) solvent: measure 700mL ethyl acetate and 300mL acetone, Mix homogeneously；

Methanol and water mixed solution (V/V 3: 7): measure 300mL methanol and 700mL water, mix homogeneously；

(3) pre-treatment extraction process:

Weigh 10.00g sample (being accurate to 0.05g) to be placed in tool plug plastic centrifuge tube, addition ethyl acetate: Acetone (v/v 7: 3) 20mL, ultrasonic 10min, add the mixing of 3g sodium chloride, and 4000r/min is centrifuged 10min, Taking the supernatant and add 0.1g PSA powder vortex, it is clear that standing takes 2mL upper strata after PSA powder precipitation Liquid, is blown to do in 45 DEG C of water-bath rotary evaporations or nitrogen, accurately adds 1.00mL methanol and water mixed solution After (V/V 3: 7) dissolves, in organic filter membrane of mistake 0.22 μm to sample injection bottle, for Instrumental Analysis；

(4) measure: take step (1) and filtrate that step (3) obtains carries out liquid chromatography-tandem mass spectrometry Instrument measures.

The having the beneficial effect that of method of Rynaxypyr residual quantity in the detection aquatic vegetable of the present invention:

The method of Rynaxypyr residual quantity, use ethyl acetate: third in the detection aquatic vegetable of the present invention Ketone (V/V 7: 3) mixed solvent extracts Rynaxypyr residual in aquatic vegetable, is dispersed through type Solid-Phase Extraction After N-propyl group ethylenediamine (PSA) adsorbent purifies, utilize the strong selectivity of liquid chromatography-tandem mass spectrometry and resist dry Disturb the characteristic of ability, measure its residual quantity.Use ethyl acetate: acetone (V/V 7: 3) mixed solvent extracts Efficiency is high, and the matrix interference of extraction is few；Use PSA filler as adsorbent, having of extraction can be removed The interfering materials such as machine is sour, pigment, reach to purify purpose.Present invention application linear optimizing control substitutes solid phase Extraction, it is possible to reduce time for sample pretreatment, reduces the consumption of organic solvent, cost-effective, reduces environment Pollute；The inventive method good stability, highly sensitive, measure for Rynaxypyr residual in aquatic vegetable Surely the most quickly analysis means is provided.

Accompanying drawing explanation

Fig. 1 is the chromatogram adding 5.0 μ g/kg in blank Rhizoma Nelumbinis.

Fig. 2 is the chromatogram adding 5.0 μ g/kg in blank Typha latifolia L..

Detailed description of the invention

In the detection aquatic vegetable of the present invention, the method for Rynaxypyr residual quantity, comprises the steps:

(1) preparation of standard solution:

Accurately weigh 10mg chlorine worm this Methanamide standard substance in 10mL volumetric flask, dissolve constant volume with methanol To scale, being configured to concentration is 1mg/mL Standard Stock solutions, is placed in-18 DEG C of refrigerators freezing lucifuge and protects Deposit；

Accurately a certain amount of 1mg/mL Rynaxypyr Standard Stock solutions of absorption is in volumetric flask, uses first It is 10 μ g/mL intermediate concentration standard solution that alcohol is diluted to mass concentration, is placed in 4 DEG C of refrigerators freezing lucifuge and protects Deposit；

Accurately draw a certain amount of 10 μ g/mL Rynaxypyr intermediate concentration standard solution, use first respectively Alcohol and water mixed solution (V/V 3: 7) stepwise dilution are 2ng/mL, 5ng/mL, 10ng/mL to concentration, The standard working solution of 20ng/mL, 50ng/mL and 100ng/mL, matching while using；

(2) preparation of solvent:

Ethyl acetate: acetone (V/V 7: 3) solvent: measure 700mL ethyl acetate and 300mL acetone, Mix homogeneously；

Methanol and water mixed solution (V/V 3: 7): measure 300mL methanol and 700mL water, mix homogeneously；

(3) pre-treatment extraction process:

Weigh 10.00g sample (being accurate to 0.05g) to be placed in tool plug plastic centrifuge tube, addition ethyl acetate: Acetone (v/v 7: 3) 20mL, ultrasonic 10min, add the mixing of 3g sodium chloride, and 4000r/min is centrifuged 10min, Taking the supernatant and add 0.1g PSA powder vortex, it is clear that standing takes 2mL upper strata after PSA powder precipitation Liquid, is blown to do in 45 DEG C of water-bath rotary evaporations or nitrogen, accurately adds 1.00mL methanol and water mixed solution After (V/V 3: 7) dissolves, in organic filter membrane of mistake 0.22 μm to sample injection bottle, for Instrumental Analysis；

(4) measure: take step (1) and filtrate that step (3) obtains carries out liquid chromatography-tandem mass spectrometry Instrument measures.

Compound concentration is 2ng/mL, 5ng/mL, 10ng/mL, 20ng/mL, 50ng/mL and 100ng/mL Standard working solution, be analyzed under the following instrument parameter optimized:

Liquid phase chromatogram condition

Chromatographic column: Agilent ZORBAX SB-C18 post 150mm × 2.1mm (i.d.)；

Flowing phase: A 0.1% aqueous formic acid, B methanol；

The gradient elution program optimized；

Flow velocity: 0.25mL/min；

Sample size: 25 μ L；

Column temperature: 30 DEG C；

Mass Spectrometry Conditions

Ion source: ESI source

Scan mode: cation scans

Monitoring mode: multiple-reaction monitoring

The mass spectrometry parameters optimized under multiple-reaction monitoring pattern

* quota ion is represented

Ion transfer tube temperature: 350 DEG C

Sheath gas (high pure nitrogen) 40Arb

Auxiliary gas (high pure nitrogen) 10Arb

Collision gas (high-purity argon gas) 1.5m Torr

Spray voltage: 3Kv.

The present invention is according to using ethyl acetate on the basis of the physicochemical property of Rynaxypyr and consulting literatures: Acetone (v/v 7: 3) mixed solvent is as Extraction solvent, by experimental verification ethyl acetate: acetone (v/v 7: 3) mixes Bonding solvent is higher than acetonitrile and acid acetonitrile, the matrix interference simultaneously extracted to the extraction efficiency of Rynaxypyr The most less.The advantage of the method be easy and simple to handle, time-consumingly the shortest, highly sensitive, detection limit is low.

Embodiment 1: the detection of Rynaxypyr residual quantity in Rhizoma Nelumbinis

Sample pre-treatments

Weigh 10.00g Rhizoma Nelumbinis sample (being accurate to 0.05g) to be placed in tool plug plastic centrifuge tube, add acetic acid second Ester: acetone (v/v 7: 3) 20mL, ultrasonic 10min, adds the mixing of 3g sodium chloride, and 4000r/min is centrifuged 10min, takes the supernatant and adds 0.1g PSA powder vortex, stand and take 2mL after PSA powder precipitation The supernatant, is blown to do in 45 DEG C of water-bath rotary evaporations or nitrogen, accurately adds 1.00mL methanol: water mixes After solution (V/V 3: 7) dissolves, in organic filter membrane of mistake 0.22 μm to sample injection bottle, for Instrumental Analysis.

Chromatogram is as shown in Figure 1.

Liquid phase chromatogram condition

Chromatographic column: Agilent ZORBAX SB-C18 post 150mm × 2.1mm (i.d.)；Flowing phase: A 0.1% aqueous formic acid, B methanol, gradient elution program is shown in Table 3；Flow velocity: 0.25mL/min；Sample introduction Amount: 25 μ L；Column temperature: 30 DEG C.

Table 3

Mass Spectrometry Conditions

Ion source: ESI source；Scan mode: cation scans；Monitoring mode: multiple-reaction monitoring, the most instead The mass spectrometry parameters optimized under monitoring pattern is answered to be shown in Table 4；Ion transfer tube temperature: 350 DEG C；Sheath gas (High Purity Nitrogen Gas) 40Arb；Auxiliary gas (high pure nitrogen) 10Arb；Collision gas (high-purity argon gas) 1.5m Torr；Spray voltage: 3Kv。

Table 4

* quota ion is represented

Embodiment 2: the detection of Rynaxypyr residual quantity in Typha latifolia L.

Sample pre-treatments

Weigh 10.00g Typha latifolia L. sample (being accurate to 0.05g) to be placed in tool plug plastic centrifuge tube, add acetic acid second Ester: acetone (v/v 7: 3) 20mL, ultrasonic 10min, adds the mixing of 3g sodium chloride, and 4000r/min is centrifuged 10min, takes the supernatant and adds 0.1g PSA powder vortex, stand and take 2mL after PSA powder precipitation The supernatant, is blown to do in 45 DEG C of water-bath rotary evaporations or nitrogen, accurately adds 1.00mL methanol: water mixes After solution (V/V 3: 7) dissolves, in organic filter membrane of mistake 0.22 μm to sample injection bottle, for Instrumental Analysis.

Chromatogram as shown in Figure 2, liquid phase chromatogram condition is with embodiment 1, and Mass Spectrometry Conditions is with embodiment 1.

Embodiment described above is only to be described the preferred embodiment of the present invention, not to this Bright scope is defined, under design spirit premise without departing from the present invention, and this area ordinary skill technology people Member's various deformation of making technical solution of the present invention and improvement, all should fall into claims of the present invention true In fixed protection domain.

Claims (1)

1. detect a method for Rynaxypyr residual quantity in aquatic vegetable, comprise the steps:

(1) preparation of standard solution:

Accurately weigh 10mg chlorine worm this Methanamide standard substance in 10mL volumetric flask, dissolve constant volume with methanol To scale, being configured to concentration is 1mg/mL Standard Stock solutions, is placed in-18 DEG C of refrigerators freezing lucifuge and protects Deposit；

Accurately a certain amount of 1mg/mL Rynaxypyr Standard Stock solutions of absorption is in volumetric flask, uses first It is 10 μ g/mL intermediate concentration standard solution that alcohol is diluted to mass concentration, is placed in 4 DEG C of refrigerators freezing lucifuge and protects Deposit；

Accurately draw a certain amount of 10 μ g/mL Rynaxypyr intermediate concentration standard solution, use first respectively Alcohol and water mixed solution (V/V 3: 7) stepwise dilution are 2ng/mL, 5ng/mL, 10ng/mL to concentration, The standard working solution of 20ng/mL, 50ng/mL and 100ng/mL, matching while using；

(2) preparation of solvent:

Ethyl acetate: acetone (V/V 7: 3) solvent: measure 700mL ethyl acetate and 300mL acetone, Mix homogeneously；

Methanol and water mixed solution (V/V 3: 7): measure 300mL methanol and 700mL water, mix homogeneously；

(3) pre-treatment extraction process:

Weigh 10.00g sample (being accurate to 0.05g) to be placed in tool plug plastic centrifuge tube, addition ethyl acetate: Acetone (v/v 7: 3) 20mL, ultrasonic 10min, add the mixing of 3g sodium chloride, and 4000r/min is centrifuged 10min, Taking the supernatant and add 0.1g PSA powder vortex, it is clear that standing takes 2mL upper strata after PSA powder precipitation Liquid, is blown to do in 45 DEG C of water-bath rotary evaporations or nitrogen, accurately adds 1.00mL methanol and water mixed solution After (V/V 3: 7) dissolves, in organic filter membrane of mistake 0.22 μm to sample injection bottle, for Instrumental Analysis；

(4) measure: take step (1) and filtrate that step (3) obtains carries out liquid chromatography-tandem mass spectrometry Instrument measures.