Water purification system

Wild OS from Baima Snow Mountains were collected in June 2021. The bacterial membrane was first washed with distilled water, and OS was cut from the base of the stroma into two parts, the stroma and sclerotia. The stroma was labeled as OSBSz-x and the sclerotia was labeled as OSBSh-x. Three biological replicates were used for each group.
Instruments for widely targeted metabolomics include ultrahigh-performance liquid chromatograph (Sciex, Framingham, MA, USA), a high-sensitivity mass spectrometer (Sciex, Framingham, MA, USA), and a centrifuge (Thermo Scientific, Waltham, MA, USA), and a water purification system (Merck Millipore, Burlington, MA, USA). Instruments for untargeted metabolomics include ultrahigh-performance liquid chromatography (Waters, Milford, MA, USA), a high-resolution mass spectrometer (Thermo Scientific, Waltham, MA, USA), a low-temperature high-speed centrifuge (Eppendorf, Hamburg, Germany), a vortex (Qilin Bell Instrument Manufacturing Co., Ltd., Haimen, China), a water purification system (Millipore, Burlington, MA, USA), and a refrigerated vacuum concentrator (Gene company limited, Hong Kong, China).

Four commercial thin film composite (TFC) membranes, including two RO (i.e., SW30XLE and BW30 from DuPont FilmTec^TM, Wilmington, DE, USA) and two NF membranes (i.e., NF3 from RisingSun Membrane Technology (Beijing) Co., Ltd., China and Duracid from Suez (GE)) were used in this research study. Dibutyl phthalates (DBP) and benzyl butyl phthalates (BBP) with 99% and 98% purity, respectively, were purchased from Sigma-Aldrich, Saint Louis, MI, USA and were used to prepare stock solutions. Sodium sulfate (Na₂SO₄, Merck Sdn Bhd, Selangor, Malaysia) and sodium chloride (NaCl, Merck Sdn Bhd, Selangor, Malaysia) were used for the salt rejection test, while sulfuric acid (H₂SO₄, Merck Sdn Bhd, Selangor, Malaysia) and sodium hydroxide (NaOH, Merck Sdn Bhd, Selangor, Malaysia) were used for solution pH adjustment. All the solutions were prepared by using water (ASTM Type III) produced by a water purification system (Merck Millipore, Burlington, MA, USA).

Acetonitrile (ACN), methanol (MeOH), and ammonium acetate (NH₄Oac), all LC–MS grade, were purchased from Fisher Scientific (Pittsburgh, PA). Ammonium hydroxide (NH₄OH), O-methylhydroxylamine hydrochloride (MeOX), and N-Methyl-n-(tert-butyldimethylsilyl) trifluoroacetamide (MTBSTFA) were bought from Sigma-Aldrich (Saint Louis, MO). Deionized water was provided in-house by a water purification system from EMD Millipore (Billerica, MA). Phosphate buffered saline (PBS) was bought from GE Healthcare Life Sciences (Logan, UT). Standard compounds corresponding to measured aqueous metabolites/features were purchased from Sigma-Aldrich and Fisher Scientific. Lipid standards were purchased from Fisher Scientific, Sigma-Aldrich, and Avanti Polar Lipids (Alabaster, AL).