Sodium sulphate na2so4

Ethyl Acetate, hexane and acetone were HPLC gradient solvents from Fisher Scientific UK.
Sodium sulphate (Na2SO4) was obtained from Sigma Aldrich and alumina was obtained from Merck in Germany. The recovery standard labeled d-10 chlorpyrifos diethyl, toclofos, diazinon, pirimicarb, cyprodinil, azinphos; deltamethrine was obtained from Cambridge Isotope Laboratories Inc. The stock solution of the recovery standard was 100 ug/ml in nonane and was diluted to 1 ug/ml in hexane. Analytical grade triphenyl phosphate (TPP-d15) was purchased from QMX Laboratories Ltd, Thaxted, UK. TPP was used as an internal standard. A 100 ug/ml TPP was prepared using acetone as the solvent. A stock solution of 1000 ug/ml chlorpyrifos was prepared in acetone using analytical grade chlorpyrifos-ethyl obtained from Sigma-Aldrich UK.
From mix stock solution seven calibration curves were prepared and solvent exchanged to hexane. A blank containing hexane was included.

Phycocyanin was supplied as the commercial product Linablue G1 from DIC Spirulina (Chuo-ku, Tokyo 103-8233, Japan). This formulation contained 40% Spirulina colour, including phycocyanin and other proteins, 5% tri-sodium citrate, and 55% D-trehalose.
Citric acid (Sigma-Aldrich) in demineralised water was used to prepare standard aqueous solutions with pH 6, 5, 4, 3, 2.5, 2, and 1.9, all ± 0.1. The final pH was measured using a Metrohm 827 pH lab. SDS, and sodium sulphate (Na2SO4) were from Sigma-Aldrich.

Individual standards of siloxanes (L2: hexamethyldisiloxane, L3: octamethyltrisiloxane, L4: decamethyltetrasiloxane, L5: polydimethylsiloxane, D3: hexamethylcyclotrisiloxane, D4: o c t a m e t h y l c y c l o t e t r a s i l o x a n e , D 5 : d e c a m e t h y l c y c l o p e n t a s i l o x a n e , D 6 : dodecamethylcyclohexasiloxane) and of the internal standard (M4Q: tetrakis(trimethylsiloxy)silane) were purchased from Sigma-Aldrich (St. Louis, MI, USA). All standards, stock solutions and mixes prepared subsequently were stored in the dark in amber glass vials at -20 °C until use. Analytical grade Prolabo solvents dichloromethane (DCM) and n-hexane (Hex) were supplied by VWR (Fontenay-sous-Bois, France). Sodium sulphate (Na2SO4) was acquired from Merck (Darmstadt, Germany), anhydrous magnesium sulphate (MgSO4) and sodium acetate (NaCH3COO) from Sigma-Aldrich and primary secondary amine (PSA)-bonded silica and C18 from Supelco (Bellefonte, PA, USA). Na2SO4 and MgSO4 were baked at 450 °C overnight before use. Commercial Qu EChERS (Q1, 6 g Mg SO 4 +1 . 5 g NaCH3COO; Q2, 900 mg MgSO4+300 mg PSA+150 mg C18) were obtained from United Chemical Technologies (Bristol, PA, USA). Helium (99.9999 %) used as carrier gas in the GC-MS system and nitrogen (99.995 %) for solvent evaporation were supplied by Air Liquide (Maia, Portugal).

Annatto seeds were obtained from Java, with a tropical monsoon climate, between January and June 2020. Its altitude is between 500-600 meters, and barometric pressure is about 95KPa (Figure S1).
Botanical identification was performed at the Department of Pharmacy, STIFAR Yayasan Pharmacy Semarang by identifying fresh plant using guidance from Flora of Java {Spermatophytes only). Vol 1.
N.V.P. Noordhoff, Netherlands (specimen identification No: 1777/IS-LBF/Stifar/S.Ket-Det/II/2021).
The seeds were portioned (500 g), vacuum packed and stored away from direct light conditions at room temperature until analysis. The acetone and methanol (analytical grade) used on the extraction procedure, 0,22 µm PTFE filter membrane were purchased from Merck (Darmstadt, Germany).
Calcium carbonate (CaCO3), L-ascorbic acid, sodium sulphate (Na2SO4), and Thin Layer Chromatography Silica Gel F254 (Catalog No. 1.51022) were purchased from Merck (Darmstadt, Germany). Chromatographic grade ethanol and acetonitrile were obtained from Sigma-Aldrich (Phillipsburg, USA) and ultrapure water was obtained from the Millipore system (Billerica, USA).
Formic acid was purchased from Merck (Darmstadt, Germany). Pure bixin was obtained from Sigma-Aldrich Co. (St. Louis, USA) and served as standard compound.