Method for Determining Residues of Benzovindiflupyr in Animal-Derived Foods Using Anyeep TQ9120
This study established a method for detecting flusilazole residues in milk using the Anyeep TQ9120 ultra-high-performance liquid chromatography-tandem mass spectrometry system from Anyeep. The method demonstrated excellent linearity, with detection and quantification limits meeting relevant regulatory standards. This approach enables reliable detection and quantification of flusilazole in animal-derived food products.
benzovindiflupyr（Benzovindiflupyr）YesOne typeA novel pyrazole amide fungicide with high efficacy, broad-spectrum activity, and long residual effect. It is primarily used to control plant diseases caused by Phoma species, including leaf blight, powdery mildew, rust, take-all, basal rot, black spot, small spot, and gray mold.Benzoyl trifluoromethyl azole exhibits high acute oral toxicity and is significantly toxic to aquatic organisms.。
This article follows theSN/T 5648-2024 Benzenesulfonylfluorine in Animal-Derived Foodrequirements of the "Determination of Residues by Liquid Chromatography-Tandem Mass Spectrometry" have been met, and development has been initiated.Fluxapyroxad in milkLiquid chromatography-tandem mass spectrometry method for residue determinationThis method exhibits high sensitivity, good linearity, and precision.Recovery RateAll method parameters meet the required standards.
Instruments and Reagents
Anyeep TQ9120 Ultra-High Performance Liquid Chromatography-Tandem Mass SpectrometrySystem
Acetonitrile, Methanol(MS-grade, from Thermo Fisher Scientific, USA)
Formic acid, acetic acid, ammonium formate (mass spectrometry grade, ANPEL)
Calcium Hydroxide (Analytical Grade, Shanghai Anpel)
Benzene Fluopyram Standard (Shanghai Anpel)
Desalination Salt Pack (SBEQ-CA8010-BZ, Anhydrous Magnesium Sulfate 4 g, Sodium Chloride 1 g, Sodium Citrate 1 g, Disodium Citrate 0.5 g, ANPEL Shanghai)
Depleted PPR Mini-column (SBEQ-CA6485, ANPEL, Shanghai)
Liquid Chromatography Conditions
Mobile PhaseA: 5 mM ammonium formate, 0.1% formic acid, water
Mobile Phase B: Methanol
Chromatography column: CNW Athena UHPLC C18, 2.1 mm × 100 mm, 1.8μm
Column Temperature：40 ℃
Flow rate: 0.3 mL/min
Injection volume：2 μL
Mass Spectrometry Conditions
Ionization Source： Electrospray Ionization Source (ESI)+）
Source Temperature： 550 ℃
Spray Voltage： 4500 V
air curtain： 35 psi
Mist gas： 50 psi
Auxiliary Gas： 70 psi
Crash： 8
Scan Mode：MRM
Gradient elution program
Timemin | A% | B% | Flow RatemL/min |
0.0 | 80 | 20 | 0.3 |
2.5 | 5 | 95 | 0.3 |
4.0 | 5 | 95 | 0.3 |
3.1 | 80 | 20 | 0.3 |
7.0 | 80 | 20 | 0.3 |
Analyte MRM Parameters
Test Item | Q1 Ion | Q3 Ion | Residency Time (ms) | CE (V) | DP (V) |
| 397.9 | 341.9* | 100 | 12 | 100 |
benzovindiflupyr | 397.9 | 321.9 | 100 | 20 | 100 |
| 397.9 | 377.9 | 100 | 8 | 100 |
Note: * indicates the quantification ion pair.
Sample Processing
Sample Processing:Weigh 5.0 g of the sample to a precision of 0.01 g into a 50 mL centrifuge tube. Add 10 mL of 1% acetonitrile, vortex for 1 min, and add the cleanup salt pack (containingMagnesium sulfate anhydrous 4 g, sodium chloride 1 g, sodium citrate 1 g, disodium citrate 0.5 g), vortex thoroughly and oscillate-extract for 10 min, centrifuge at 4000 rpm for 5 min; collect the supernatant for later use.
Transfer 2 mL of the supernatant to phospholipid PPR.Pipette the eluate into a small column, mix well, and pass through 0.22.μm filter, ready for injection.
Matrix Standard Solution:Process a batch of blank matrix samples using the "Sample Preparation" procedure. Prepare a series of matrix-matched standard solutions with benaloxazole concentrations of 0.2, 0.5, 1.0, 2.0, 5.0, 10, 20, 50, and 100 ng/mL by mixing the blank matrix and standard solution at a ratio of 9:1 (v/v).
Recovery Rate Sample:Add a certain amount of benzovindiflupyr standard to the test sample, following "Sample ProcessingPerform the operation to configure the final content as 5, 10, and 100.μg/kgBenzo-ene-fluorine-fungazole recovery samples. Prepare 6 replicates for each concentration.
Linear Range
Test Item | Retention Time min | Limit of Detection μg/kg | Limit of Quantitation μg/kg | Linear Range n mL | Linear Equation {x} weighted | correlation coefficient r2 |
benzovindiflupyr | 3.599 | 0.1 | 0.4 | 0.2-100 | Y = 44013.6 * X + 1883.35 | 0.9988 |
Precision and Recovery
Test Item | background μg/kg | Add label μg/kg | 1 | 2 | 3 | 4 | 5 | 6 | Precision | Recovery Rate |
benzovindiflupyr | 0.00 | 5 | 4.807 | 4.891 | 4.927 | 4.743 | 4.648 | 4.571 | 2.90% | 95.29% |
0.00 | 10 | 9.398 | 9.140 | 9.056 | 8.888 | 8.793 | 9.051 | 2.32% | 90.55% | |
0.00 | 100 | 89.632 | 90.725 | 90.494 | 87.798 | 88.374 | 87.597 | 1.53% | 89.10% |
Summary
This article establishes Anyeep TQ9120 Ultra-High Performance Liquid Chromatography-Tandem Mass Spectrometry SystemMethod for Determining Benomyl Residues in MilkAs the results show，The method exhibits excellent sensitivity, stability, and a wide linear range.All relevant metrics meet the required standards.Suitable forResidue levels of benzovindiflupyr in animal-derived foodsofStandard analysis and testing.