Ethyl acetate

Ethanol and mEthanol were purchased from Merck (Darmstadt, Germany). GC–FID standards: ethyl acetate (CAS N° 141-78-6; purity ≥ 99.8%) and acetic acid (CAS 64-19-7; purity 99.8%) were purchased from Riedel-de-Haen (Seelze, Germany), mEthanol (CAS N° 67-56-1; purity ≥ 99.9%) was purchased from Merck (Darmstadt, Germany). 2-methylbutan-1-ol (CAS N° 137-32-6; purity ≥ 98%), 3-methylbutan-1-ol (CAS N° 123-51-3; purity ≥ 98.5%), butan-1-ol (CAS N° 71-36-3; purity ≥ 99.5%), 2-methylpropan-1-ol (CAS N° 78-83-1; purity ≥ 99.5%), propan-1-ol (CAS N° 71-23-8; purity ≥ 99.5%), 2-propen-1-ol (CAS N° 107-18-6; purity ≥ 98%), butan-2-ol (CAS N° 78-92-2; purity ≥ 99.5%), 4-methylpentan-2-ol (CAS N° 108-11-2; purity ≥ 98%), and acetaldehyde (CAS N° 75-07-0; purity ≥ 99.5%) were purchased from Fluka (Buchs, Switzerland).

Certified pesticide standards (purity 94–99%) were obtained from Dr. Ehrenstorfer LGC Standards (Augsburg, Germany), Toronto Research Chemicals (Toronto, ON, Canada) and Sigma Aldrich (Seelze, Germany). Individual stock solutions of pesticides at concentrations of 1000 µg/mL were prepared in acetone or Dimethylformamide and stored in amber screw-capped glass vials at below −18 °C. Mixed standard solutions for validation and calibration were prepared by appropriate dilutions of stock standard solutions with acetonitrile. LC-MS purity-grade acetonitrile, acetone, cyclohexane, and ethyl acetate were supplied by Honeywell (Charlotte, North Carolina, USA). Dimethylformamide, sodium sulphate (anhydrous), and sodium chloride were obtained from Sigma Aldrich (Bellefonte, Pennsylvania, USA). Tributyl phosphate (Sigma-Aldrich, Seelze, Germany) was used as an internal standard and prepared as a spiking solution at concentrations of 10 µg/mL in acetonitrile. Ultrapure water (18.2 MΩ/cm) was produced by the Direct-Q^® 5 UV System (Millipore Corporation Merck, Darmstadt, Germany).

Alpha-amylase from human salivary, alpha-glucosidase (1 unit/mL, Sigma), the ACE solution from rabbit lung, 2,2′-Azino-bis (3-ethylbenzthiazoline-6-sulfonic acid sulfonic acid) (ABTS^·+), dibutylphthalate polystyrene xylene (DPX), 4-dimethylamino cinnamaldehyde (DMACA), 3,5-dinitrosalicylic acid (DNS), 2,2-diphenyl-1-picrylhydrazyl (DPPH), ferric chloride, hippuroyl-His-Leu hydrate, 6-hydroxy-2,5,7,8-tetramethylchroman-2-carboxylic acid (Trolox), Melanin from Sepiaofficinalis, p-nitrophenyl alpha-d-glucopyranoside (pNPG), phosphate buffer, potassium ferricyanide, potassium permanganate, potassium persulfate, potassium phosphate buffer, sodium borate buffer, sodium carbonate, sodium hydroxide, sodium potassium tartrate, starch, and xylene were from Sigma Chemical Company (St. Louis, Missouri, USA). Analytical-grade acetic acid, acetone, chloroform, ethanol, ethyl acetate, hydrochloric acid, hydrogen peroxide, and methanol were from Honeywell (Seelze, Hanover, Germany).

Two commercial biopesticides, Cinna (Hortalan; El Ejido, Spain) and Prevam® (ORO AGRI; Palmela, Portugal), based on cinnamon extracts and orange essential oil respectively, were obtained.
Ethyl acetate (EtOAc, ≥99.7%) and methanol (MeOH, ≥99.9%) were provided from Honeywell (Charlotte, NC, USA), whereas formic acid (99.0%) and water (H₂O, LiChrosolv®) were from Merck (Darmstadt, Germany). All solvents were HPLC grade.
Analytical standards used were thymol provided by Tokyo Chemical Industry (Tokyo, Japan), (R)-(+)-limonene and m-cymene by Sigma Aldrich (Saint Louis, MO, USA), and trans-cinnamaldehyde by Dr. Ehrenstorfer (Augsburg, Germany). Internal standards (IS) were triphenyl phosphate provided by Supelco (Darmstadt, Germany) for UHPLC and biphenyl (Dr. Ehrenstorfer) for GC.
For each compound, individual stock solutions were prepared at 1000 mg/L in EtOAc. From the stock solutions, individual intermediate solutions at 10 and 1 mg/L in EtOAc were made. These solutions were kept at −18 °C.
Extracts were filtered with an Econofltr nylon filter 0.2 µm, 13 mm (Agilent Technologies; Santa Clara, CA, USA).

Standards of gallic acid, quercetin, cyanidin chloride, caffeic acid, quercetin-3-O-glucoside and tryptophan were purchased from Sigma-Aldrich (Saint-Quentin Fallavier, France). Folin-Ciocalteu’s reagent (10%), aluminum chloride (AlCl₃) (10%), potassium acetate (1 M), DPPH, Trolox (6-hydroxy-2,5,7,8-tetramethylchroman-2-carboxylic acid), ABTS (7 mM), potassium persulfate (2.45 mM), ferric chloride (FeCl₃·6H₂O), 2,4,6-tri(2-pyridyl)-s-triazine (TPTZ), methyl-tert-butyl-ether (MTBE),2,2’-Azobis(2-amidinopropane) dihydrochloride (AAPH), fluorescein, sodium nitroprusside (5 mM), sulphanilamide, n-(1-naphthyl) ethylenediamine dihydrochloride phosphate buffered saline (PBS) acetate buffer, phosphate buffer, and mEthanol were supplied by Sigma-Aldrich (Saint-Quentin Fallavier, France). Ethanol, hexane, acetone, formic acid, and sodium carbonate (75 g/L), were purchased from Carlo Erba Reagents (Val de Reuil, France); hydrochloric acid, butanol, acetonitrile, ethyl acetate, phosphoric acid, and ferric ammonium sulfate (2%) were from HoneyWell Riedel-de Haën (Seelze, Germany). All solvents were of the highest analytical grade.