Ultrapure milliq water

The diatom Phaeodactylum tricornutum (CIB-41 from CIBNOR (https://www.cibnor.gob.mx/, accessed on 22 November 2021) La Paz, Mexico) was obtained from the Mexican Company “Microalgas Oleas de México S.A.” (Guadalajara, Mexico). The diatom was cultured in F/2 culture medium as reported previously by Guillard [69 ] and harvested at the end of the exponential phase. The microalga (diatom) was provided freeze-dried (freeze-dry system Labconco Freezone 18 L Benchtop Freeze Dry System, Kansas, MO, USA), packed in vacuum-sealed plastic bags and then was stored at 4 ± 2 °C in the dark until use. All solutions used in this study were of analytical grade except for the standard solutions and chromatographic solvents. For supercritical fluid extraction, carbon dioxide and ethanol were purchased from Indura Air Products Group (Santiago, Chile) and Sigma-Aldrich (Santiago, Chile), respectively. Bovine Serum Albumin (BSA) (as the standard in protein analysis) and glucose (as the standard in carbohydrate determination) were acquired from Sigma-Aldrich (Santiago, Chile). For fucoxanthin identification and quantification, a standard of >99% purity (Sigma-Aldrich, Santiago, Chile) was used. Ethyl acetate, acetonitrile, and ultrapure (MilliQ) water were of chromatographic grade (Sigma-Aldrich, Santiago, Chile) for high-performance liquid chromatography (HPLC).

Hydrogen tetrachloroaurate (HAuCl₄), penicillin, streptomycin, and L-glutamine were bought from Pro Sci Tech, Kirwan, Australia. Cold pressed cacao powder was obtained from Forest Super Foods, Melbourne, Australia, and stored in an air-tight container (Goodguys, Adelaide, Australia). NaOH pellets, phosphate buffer saline (PBS) tablets, foetal bovine serum (FBS), nitric acid (70%), and Dulbecco’s Modified Eagle Medium (DMEM) were purchased from Sigma-Aldrich, Sydney, Australia. Hydrochloric acid (36%) was procured from Ajax Finechem Pty. Ltd., Sydney, Australia. All of the reagents were used as received. Ultra-pure MilliQ water (resistivity 18.2 Ω, Sigma-Aldrich, Sydney, Australia) was used for all of the experimental and cleaning procedures.