Hipersolv

UHPLC-HR-MS data for dereplication was recorded using an Acquity I-class UPLC (Waters,Milford, MA, USA) coupled to a PDA detector and a Vion IMS QToF (Waters). The chromatographic separation was performed using an Acquity C-18 UPLC column (1.7 μm, 2.1 mm × 100 mm; Waters). Mobile phases consisted of acetonitrile (HiPerSolv, VWR) for mobile phase A and pH₂O produced by the in-house Milli-Q system as mobile phase B, both containing 0.1% formic acid (v/v; 33015, Sigma). The gradient was run from 10 to 90% B over 12 min at a flow rate of 0.45 ml/min. Samples were run in ESI+ and ESI-ionization mode. The data was processed and analyzed using UNIFI 1.9.4 (Waters). Exact masses were calculated using ChemCalc (Patiny and Borel, 2013 (link)). For dereplication extracts of the respective cultivation media were prepared to exclude media components from consideration.

Acetonitrile, methanol, and water (HiPerSolv)
were purchased from VWR International GmbH (Darmstadt, Germany). Chloroform
(Rotipuran) was purchased from Carl Roth GmbH + Co. KG (Karlsruhe,
Germany). 2,5-Dihydroxybenzoic acid (DHB), ethanol, glacial acetic
acid, and trifluoroacetic acid were purchased from Merck (Darmstadt,
Germany). Hematoxylin, eosin Y, Eukitt, and α-cyano-4-hydroxycinnamic
acid (CHCA) were purchased from Sigma-Aldrich (Darmstadt, Germany).
1,5-Diaminonapthalene (DAN) was purchased from Acros Organics (Geel,
Belgium). 2,5-Dihydroxyacetophenone (DHAP) and ammonium acetate were
purchased from Alfa Aesar (Kandel, Germany). 9-Aminoacridine (9-AA)
was purchased from TCI (Eschborn, Germany).

The standards of pesticides were purchased from Dr. Ehrenstorfer GmbH (Augsburg, Germany). For acetamiprid, chlorantraniliprole, and iprovalicarb, the mix DRE-Q60 004725 of 10 µg/mL each was used, while for the other three pesticides, custom individual 10 µg/mL standard solutions in acetonitrile were used, also from Dr. Ehrenstorfer. Dilutions were conducted in acetonitrile to the desired concentrations, and then a final dilution was made by transferring 0.25 mL of the standard solution, adding 10 µL of the internal standard (TPP 5 µg/mL), and then 0.74 mL of ultrapure water. The Quick QuEChERS-Medium Cartridge was purchased from United Chemical Technologies (Bristol, PA, USA); the extraction salts including magnesium sulphate (MgSO₄), sodium chloride (NaCl), acetonitrile, 2-propanol, acetone were from Merck KGaA (Darmstadt, Germany); formic acid (HCOOH; purity ≥ 95%) was from Fisher Chemical; and ammonium formate (HCO₂NH₄) from Sigma Aldrich (St. Louis, MO, USA). Methanol was supplied by HiPerSolv, VWR Chemicals (Radnor, PA, USA). A Kinetex biphenyl chromatography column (2.6 μm, 4.6 × 150 mm) was purchased from Phenomenex (Torrance, CA, USA) and ultrapure water was obtained by a Milli-Q water purification system from Millipore (Bedford, MA, USA).

Acetonitrile and water (HiPerSolv) were purchased
from VWR International GmbH (Darmstadt, Germany). 2,5-Dihydroxybenzoic
acid (DHB) was purchased from Merck (Darmstadt, Germany). Trifluoroacetic
acid (TFA) was purchased from AppliChem GmbH (Darmstadt, Germany).
Girard’s reagent T (GirT) was purchased from Merck (Darmstadt,
Germany). Formic acid (FA) was purchased from Fisher Scientific (Schwerte,
Germany).

UPLC-HR-MS/MS data for dereplication and structure elucidation was recorded. The chromatographic separation was performed using an Acquity C18 UPLC column (1.7 μm, 2.1 mm × 100 mm) (Waters). Mobile phases consisted out of ddH₂O produced by the in-house Milli-Q system for mobile phase A and acetonitrile (HiPerSolv, VWR) as mobile phase B, both containing 0.1% formic acid (v/v) (33,015, Sigma). The gradient was run from 10 to 90% B over 12 min at a flow rate of 0.45 mL/min. Samples were run in ESI+ and ESI- ionization modes. The data was processed and analyzed using UNIFI 1.9.4 (Waters). Calculation of exact ion masses was done by using ChemCalc (Patiny and Borel, 2013 (link)).

The following compounds were obtained as
bromide salts from Sigma-Aldrich (Buchs, Switzerland): tetraethylammonium
(T2), tetrabutylammonium (T4), tetrahexylammonium (T6), tetraoctylammonium
(T8), tetradecylammonium (T10), and tetradocecylammonium (T12). Benzyldimethyldodecylammonium
(BAC-C12), benzyldimethyltetradecylammonium (BAC-C14), and benzyldimethylhexadecylammonium
(BAC-C16) chloride salts were also purchased from Sigma-Aldrich. Standards
with a concentration of 5 μM were prepared in 50% v/v methanol
(HPLC grade, HiPerSolv, VWR Chemicals, Schlieren, Switzerland) and
water with a resistivity of 18.2 MΩ·cm (Milli-Q, Merck
Millipore, Schaffhausen, Switzerland) from 1 mM stock solutions.

The samples were analyzed with ESI+ and ESI- ionization mode on a UPLC-QToF-MS for dereplication. The system (all from Waters) consisted of an Acquity UPLC I-class coupled to a PDA detector and a Vion IMS QToF. An Acquity C18 UPLC column (1.7 µm, 2.1 mm × 100 mm) was used for the separation. Milli-Q water was used for mobile phase A and acetonitrile (HiPerSolv, VWR) for mobile phase B, both containing 0.1% formic acid (v/v) (33015, Sigma). A 12-min gradient increasing from 10% to 90% acetonitrile with flow rate 0.45 mL/min was used. UNIFI 1.9 (Waters) was used to process the data.