Primary secondary amine

Certified reference material (CRM) was procured from Dr. Ehrenstorfer (Augsburg, Germany). LC-MS grade acetonitrile and methanol (≥99.9%) were procured from J. T. Baker (New Jersey, USA), Ammonium formate and formic acid (≥90.00%) were bought from Empart (Hyderabad, India). Ethyl acetate (≥99.9%) was procured from Merck Mumbai, India. Ultrapure water of 18.2 MΩ was obtained using a Milli-Q water purification system (Merck Millipore, Mumbai, India). Anhydrous magnesium sulphate (≥99.90%), anhydrous sodium sulphate, graphitized carbon black (GCB), anhydrous sodium chloride (≥99.90%) was obtained from Himedia (Bangalore, India). The primary secondary amine (PSA, 40 μm particle size) was obtained from Agilent Technologies, (California, CA, USA).

LC-MS-grade acetonitrile (MeCN) and H₂O were purchased from Fluka (Steinheim, Germany). A Milli-Q purification device (Millipore Direct-QR 3, Millipore International, Merck KGaA, Darmstadt, Germany) was used for obtaining the water used in the analyses. Acetone (>99%) and hexane (HPLC grade) were obtained from Centralchem (Bratislava, Slovakia) and VWR Chemicals (Radnor, PA, USA), respectively. Magnesium sulphate (MgSO₄), graphitized carbon black (GCB), C18 sorbents, and primary secondary amine (PSA) were supplied by Agilent Technologies (Santa Clara, CA, USA). The aniline standards, namely 3,4-dichloroaniline (3,4-DCA) and 3,5-dichloroaniline (3,5-DCA), were obtained from Riedel de Haën (Seelze, Germany), with purity higher than 99.9%. Phthalate analytical standards, such as dimethyl phthalate (DMP), di-n-butyl phthalate (DnBP), mono-butyl phthalate (mBP), benzyl butyl phthalate (BBP), di-n-octyl phthalate (DnOP), and di-(2-ethyl hexyl) phthalate (DEHP)), were supplied by Sigma Aldrich (Steinheim, Germany).

Analytical grade FIP (≥ 95%) and THIO (≥ 98%) were procured from Sigma–Aldrich (Supplementary Table S1). All solvents used in this research, such as methanol (MeOH), acetone (Ace), and acetonitrile (MeCN), were of analytical grade quality (99.9%) and obtained from Merck, Germany. Anhydrous magnesium sulfate (MgSO₄) and sodium chloride (NaCl) of reagent grade purity at 99% were supplied by BioShop, Burlington, Canada. Primary–secondary amine (PSA) was supplied by Agilent Technologies, United States. Stock solutions of FIP and THIO (1 g/L) were prepared in Ace and MeOH for soil/culture media spikings and chromatographic analyses, respectively. These solutions were stored in amber glass vials at −20°C to ensure stability.
A mineral salt medium (MSM) (Cycoń et al., 2009 (link)) composed of 2 g (NH₄)₂SO₄, 0.2 g MgSO₄.7H₂O, 0.01 g CaCl₂.2H₂O, 0.001 g FeSO₄.7H₂O, 1.5 g Na₂HPO₄.12H₂O, 1.5 g KH₂PO₄, and 0.5 g K₂HPO₄ per liter of deionized water was employed. The pH of the medium was adjusted to 7.2. Before usage, the medium was sterilized at 121°C for 20 min.

The leaves from each replicate were lyophilized (Lyophilizer SL-404/B) for 24 h until the complete removal of water. The leaves were ground in a knife mill (Model SKU: SP227-01) for 1 min, until a fine powder was obtained. For extraction, the QuEChERS original method with modifications was used, in which 0.3g of sample was weighed on a precision scale and placed in a Falcon tube (CRAL—Articles for Laboratory Ltda.). In each Falcon tube containing a leaf sample, we added 1 mL of acetonitrile and a package of the original QuEChERS Kit (Bond Elut—Agilent Technologies) [22 ]. The sample was shaken for 1 min in vortex and then centrifuged (Eppendorf 5810 R centrifuge) for 3 min at 3400 rpm. After this, we collected 1ml of the supernatant solution, which was shaken in vortex for 30.0 seconds along with 0.3g of MgSO₄ (Synth), 0.05g of primary secondary amine (PSA) (Agilent Technologies), and 0.004g of graphite carbon (Sigma-Aldrich). The samples were centrifuged again for 3 min at 3400 rpm and then 1 mL of the supernatant was removed and filtered in Aura MT Syringe Filter, PTFE, Hydrophilic, 0.45 Pore Size, 25 mm Diameter and taken for chromatographic analysis.

The purities of the ten pesticide standards were from 97% to 99%, which were obtained from the Institute of Control of Agrochemicals, Ministry of Agriculture People’s Republic of China. Standard stock solutions (500 mg/L) for mixture of the ten pesticides were prepared in Acetonitrile and stored at −20 °C. The work solution was prepared daily. Acetonitrile was of chromatography grade and obtained from Fisher Scientific (Fair Lawn, NJ, USA). Sodium chloride (NaCl) and anhydrous magnesium sulfate (MgSO₄) were of analytical grade and purchased from Sinopharm Chemical Reagent Co., Ltd. (Beijing, China). Multi-walled carbon nanotubes (MWCNTs) with average external diameters of 10–20 nm and primary secondary amine (PSA) (40 µm) were acquired from Agilent Technology Co., Ltd. (Beijing, China). Micron calcium was provided by Bai Jia An Bioengineering Co., Ltd. (Liaoning, China). Active oxygen was provided by Guangzhou Zao Gu Biotechnology Co., Ltd. (Guangzhou, China). Alkaline electrolyzed water and ozonated water were prepared by the Specialized preparation machine.
Centrifugation was performed in an Anke TDL–40 B centrifuge equipped with a bucket rotor (8 × 100 mL) (Shanghai, China). An ATARGIN VX–III multitube vortexer was used in sample preparation (Beijing, China).