关键词: 液相色谱-四极杆-飞行时间质谱, 筛查与确证, 欧盟能力验证, 农药残留, 紫甘蓝

Abstract:

An analytical method for the simultaneous rapid screening and accurate confirmation of 415 pesticide residues in red cabbages was established using liquid chromatography-quadrupole-time of flight-mass spectrometry (LC-QTOF/MS) with single acquisition. In the established method, the pesticides in red cabbage were extracted using acetonitrile-acetic acid (99∶1, v/v) and salted-out using anhydrous magnesium sulfate and sodium chloride. The resultant solution was then cleaned-up by automatic solid phase extraction using a Carbon/NH₂ cartridge. The SPE cartridge was activated with 4 mL acetonitrile-toluene (3∶1, v/v) and the effluents were discarded. The resultant solution was transferred to the Carbon/NH₂ cartridge, using 3×2 mL acetonitrile-toluene (3∶1, v/v) to wash the test sample concentrate bottle, and waited until the surface of the test sample concentrate liquid reached the top layer of anhydrous Na₂SO₄ before transferring the washing liquid to the cartridge. A 30-mL reservoir was attached to the upper part of the SPE cartridge and 25 mL acetonitrile-toluene (3∶1, v/v) was used to wash the SPE cartridge again. The eluent was evaporated in the glass tube in a water bath at 37 ℃ and shaking speed 150 r/min to reduce the volume to 0.5 mL. Nitrogen was used to dry the concentrates, and the residues were dissolved in 1.0 mL acetonitrile-water (3∶2, v/v), homogenized by ultrasonication, and passed through 0.22-μm filtering membrane before determination. The dissolved sample solution was loaded onto a ZORBAX SB-C18 column (100 mm×2.1 mm, 3.5 μm) and separated under gradient elution using 0.1% (v/v) formic acid aqueous solution containing 5 mmol/L ammonium acetate and acetonitrile as the binary mobile phase. The eluent from the column was further detected by QTOF/MS under electrospray positive ionization in the MS/MS scanning mode. A matrix-matched external calibration method was used for quantitation. By optimizing the different parameters under Auto MS/MS and All Ions MS/MS acquisition modes, the optimal conditions for All Ions MS/MS under each acquisition mode were obtained, which were then compared for selection of a better mode. The results demonstrated that the developed method can be used to accurately screen and quantify all 415 pesticides in red cabbage. The linear regression correlation coefficients (r²) for the 415 pesticides were all greater than 0.990 in the corresponding linear concentration range. In addition, the screening detection limits (SDL) of 411 pesticides were no more than 5 μg/kg, and the limits of quantification (LOQs) of 413 pesticides were no more than 10 μg/kg. At the spiked levels of LOQ, two-fold LOQ, and 10-fold LOQ, the recoveries were in the ranges of 65.7%-118.4%, 72.0%-118.8% and 70.2%-111.2%, with relative standard deviations (RSDs) in the ranges of 0.9%-19.7%, 0.2%-19.9% and 0.6%-19.9%, respectively. The method was applied to detect pesticide residues in the red cabbage samples provided by the 2019 European proficiency test project for unknown pesticide screening (EUPT-SM-11) and accurate quantitation (EUPT-FV-21). For EUPT-SM-11, all the spiked and incurred pesticides in red cabbage were qualified accurately, without false positives or false negatives. This is completely consistent with the final results published by the EU official. For EUPT-FV-21, there were 19 non-volatile pesticides that can be detected by LC-MS, which were then accurately quantitated with the corresponding pesticide standard. The results demonstrate that the proposed method is accurate and reliable. It is also rapid and time-saving, and can be used for high-throughput screening and quantitative determination of pesticide residues in cabbage. It can also be extended to other fruits and vegetable matrices.

Key words: liquid chromatography-quadrupole-time of flight-mass spectrometry (LC-QTOF/MS), screening and confirmation, European proficiency test, pesticide residues, red cabbage