Smart2pure water purification system

The process of preparing the extract involved the use of ethanol (96%) sourced from Kalsnavas Elevators Ltd. (Jaunkalsnava, Latvia). LC-MS grade acetonitrile, methanol, and formic acid were procured from Fisher Scientific (Loughborough, UK), while water used for LC-qTOF-MS analysis was purified via a Smart2Pure water purification system (Thermo Scientific, Dreieich, Germany). Gallic acid, cyclohexane, xylene, ascorbic acid, and chlorogenic acid were obtained from Acros Organics (Geel, Belgium), and Na₂CO₃ was sourced from Honeywell (Charlotte, NC, USA). In addition, 2,2-diphenyl-1-picrylhydrazyl (DPPH) and phenol were purchased from Alfa Aesar (Kandel, Germany). We used Folin–Ciocalteu reagent, H₂SO₄, HNO₃, HCl, and D-glucose, all of which were purchased from Fisher Scientific (Loughborough, UK). Standards of rosmarinic acid and leucine were obtained from Sigma-Aldrich (St. Louis, MO, USA).

Chloroform (for HPLC, >99.8%, amylene-stabilized, Sigma-Aldrich, Saint-Quentin-Fallavier, France), dichlorometane (for HPLC, isocratic grade, stabilized with ethanol, Carlo Erba, France), acetonitrile (LiChrosolv^®, hypergrade for LC-MS, Merck, Darmstadt, Germany), and deionized water (18.2 MΩcm⁻¹, Barnstead™ Smart2Pure™ Water Purification System, Thermo Scientific™, Waltham, MA, USA) were used for sample extraction, dissolution, and preparation of the mobile phases for the LC-ESI QTOF MS analysis. Ammonium formate (puriss. p.a., eluent additive for LC-MS, Fluka, Honeywell International, Inc., Charlotte, NC, USA) and formic acid (eluent additive for LC-MS, Fluka Analytical) were used for the preparation of eluent additives for LC-ESI QTOF MS.

Horseradish peroxidase (HRP)-labeled anti-PfHRP2 antibody was obtained from Immunology Consultants Laboratory, Inc (Lake Oswego, OR) and diluted to 10 μg mL^–1 in phosphate buffered saline (PBS) (Fisher Scientific, Pittsburgh, PA). 3′,3′,5′,5′ tetramethylbenzidine substrate (TMB/H₂O₂) was obtained from Neogen (Lexington, KY), and deionized (DI) water (18.3 MΩ cm) was generated using a Thermo Scientific Smart2Pure water purification system.