Formic acid (200 μL) and acetonitrile (2.5 mL) were added to 500 μL of human serum or 15 mL of urine for protein precipitation. Supernatants were purified by solid-phase extraction using an OASIS WAX cartridge (Waters). Final samples were evaporated to 150 μL under a gentle stream of nitrogen. For quantitation, standards were spiked in blank calf serum or human urine previously tested as PFAS-free. Details on extraction procedures and quantitation methods are available in the Supporting Information . Limits of quantitation and recoveries for human serum and urine are available on Table S.4 .
Oasis wax cartridge
Manufactured by Waters Corporation
Sourced in United States
Oasis WAX cartridges are solid-phase extraction (SPE) devices used for the selective isolation and purification of analytes from complex sample matrices. These cartridges contain a hydrophilic-lipophilic balanced (HLB) sorbent material that can retain a wide range of polar and non-polar compounds. The cartridges are designed to provide efficient and reproducible sample preparation for various analytical applications.
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Lab products found in correlation
Nexera liquid chromatograph, by Shimadzu (1 mentions)
Qtrap 6500 tandem mass spectrometer, by AB Sciex (1 mentions)
Vacuum centrifuge, by Labconco (1 mentions)
Sequant zic chilic column, by Merck Group (1 mentions)
Trizol reagent, by Thermo Fisher Scientific (1 mentions)
Srebp2, by Santa Cruz Biotechnology (1 mentions)
Thp 1, by American Type Culture Collection (1 mentions)
Hepg2, by American Type Culture Collection (1 mentions)
Adenosine 3 phosphate 5 phosphosulfate, by Merck Group (1 mentions)
Tphp d 15, by Merck Group (1 mentions)
Uridine 5 diphosphoglucuronic acid, by Merck Group (1 mentions)
Reduced β nicotinamide adenine dinucleotide 2 phosphate, by Merck Group (1 mentions)
12 protocols using oasis wax cartridge
1
Extraction and Quantitation of PFAS
2
Quantification of Bile Acids by LC-MS/MS
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The total BA levels in the PV and IVC were examined using a total BA test kit (Wako, Osaka, Japan). Each BA concentration was analyzed using liquid chromatography-mass spectrometry (LC–MS/MS). Plasma samples were diluted with methanol to prepare a sample solution, whereas intestinal contents were diluted after homogenization and used as a sample solution. Subsequently, 50 μL of the internal standard solution and 900 μL of water were added to 50 μL of the sample solution, followed by pretreatment using an OASIS WAX cartridge (Waters Co, Milford, MA, USA). The Nexera liquid chromatograph (Shimadzu Co., Ltd., Kyoto, Japan) and InertSustain C18 column (GL Science Co., Tokyo, Japan) were connected to a QTRAP 6500 tandem mass spectrometer (SCIEX, Framingham, MA, USA). Purified sample solutions were then analyzed using LC–MS/MS.
3
HILIC-based Metabolite Extraction
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Frozen cell extracts were defrosted by 1 to 2 min of incubation in a 37°C water bath and sonicated for 10 min in a water-ice slurry. After 10 min of centrifugation at 20,000 × g, samples were loaded on a 1 ml/30 mg Oasis Wax cartridge (Waters) preconditioned with 1 ml of MeOH and 1 ml 50 mM ammonium acetate buffer (pH 4.5). After washing with 1 ml ammonium acetate buffer was performed, analytes were eluted with 200 μl of 2.8% ammonium hydroxide–MeOH/ACN/H2O 50:30:20 (vol/vol/vol). After addition of 10 μl of 5% trehalose and brief vortex mixing, the samples were dried in a vacuum centrifuge (Labconco). Dried trehalose-stabilized extracts were redissolved in 20 μl of MeOH/ACN/H2O (50:30:20 [vol/vol/vol]) and moved to an autosampler vial for analysis. Separation was performed on an iHilic column (Hilicon) (2.1 mm by 100 mm, 3.5-μm pore size) or a SeQuant Zic-cHILIC column (Merck) (2.1 mm by 100 mm, 3-μm pore size) at 0.3 ml/min using the following binary gradient: 100% B, ramp to 85% B in 1.5 min followed by 10 min of isocratic hold at 85% B and a linear decrease to 30% B in 3 min with a 2-min hold at 30% B and 8 min reequilibration under the initial conditions (A, 3.75 mM ammonium acetate–1.25 mM acetic acid–2 mM acetylacetone–Milli-Q water, B, 11.25 mM ammonium acetate–3.75 mM acetic acid–2 mM acetylacetone–80% ACN). The injection volume was 2 μl.
4
PFOS Sorption in Paddy Soil
5
Solid-Phase Extraction Cartridge Comparison
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Two types of SPE cartridge were used in this study: the ISOLUTE cartridge (ENV+, 50 mg, 1ml) from Biotage (Uppsala, Sweden), and the OASIS WAX cartridge (30 mg, 30 μm, 1 ml) from Waters (Milford, Massachusetts,USA) .
6
Extraction and Purification of PFASs from Soil
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All samples were dried by a freeze dryer prior to extraction and extracted in triplicate. The soil extraction method was similar to the approach used by Liu et al. (2018) (link). Briefly, a 2000 mg subsample of a homogenized soil sample was placed in a 50 mL polypropylene (PP) centrifuge tube containing 15 mL of methanol. Tubes were vortexed for 30 s, sonicated for 20 min at 30–35°C. The extract was separated from the soil by centrifugation at 8000 rpm for 10 min. The supernatant was transferred to a 500 mL PP beaker with a 10 mL pipette. The extraction process was repeated two more times to ensure complete removal of all PFASs. The combined extract was diluted to 300 mL with deionized water and then was purified by a solid-phase extraction column. Waters Oasis WAX cartridges (150 mg, 6 mL) were sequentially pre-conditioned with 6 mL of 0.5% NH4OH in methanol, 6 mL of methanol and 6 mL of deionized water. The extract was loaded into the cartridges with a flow 3–5 mL min−1. The cartridges were sequentially washed with 5 mL of deionized water and 5 mL of 25 mM ammonium acetate buffer solution at pH four and vacuumed at 65 kPa to remove any residual water. The analytes were eluted 4 mL of 0.5% NH4OH in methanol. The eluent was evaporated to dryness under a gentle stream of high-purity N2 and resolved into 1000 μL 50:50 methanol: water for PFASs analysis.
7
Brain Penetration of 20 kDa PSCMA in Mice
8
Lipid Metabolism Regulation Pathway
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Cell culture reagents and supplies were purchased from GIBCO BRL (Grand Island, NY); 25-hydroxycholesterol from New England Nuclear (Boston, MA). THP-1 and HepG2 cells were obtained from American Type Culture Collection (Rockville, MD). The reagents for real time RT-PCR were from AB Applied Biosystems (Warrington WA1 4 SR, UK). The chemicals used in this research were obtained from Sigma Chemical Co. (St. Louis, MO) or Bio-Rad Laboratories (Hercules, CA). Polyclonal rabbit antibodies against SREBP-1, SREBP-2, and HMG-CoA reductase were purchased from Santa Cruz Biotechnology (Santa Cruz, CA). All solvents were obtained from Fisher (Fair Lawn, NJ) unless otherwise indicated. The enhanced chemiluminescence (ECL) reagents were from Amersham Biosciences (Piscataway, NJ). Oasis®Wax Cartridges were purchased from Waters Corporation (Milford, MA). The testosterone and 27-hydroxycholesterol were obtained from Research Plus Inc. (Bayonne, NJ). LK6 20×20 cm normal phase thin layer chromatography (TLC) plates were purchased from Whatman Inc. (Clifton, NJ).
9
Salicylic Acid Quantification in Leaves
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SA content was determined according to a previously described method with specified modifications (41 (link)). Briefly, fresh leaves were ground in liquid nitrogen, and 0.1 g of sample was suspended in 4 ml of extraction buffer [1% (v/v) acetic acid in acetonitrile/water (4:1)] with stable isotope-labeled internal standards. Suspended samples were extracted, centrifuged, and concentrated as described previously (41 (link)). Samples were purified by solid-phase extraction using Oasis WAX cartridges (Waters Corp., Milford, MA, USA) from which SA was eluted with 3% (v/v) formic acid in acetonitrile. Following evaporation of each fraction, samples were analyzed on an Agilent 1260-6410 Triple Quad LC/MS system (Agilent Technologies Inc., Santa Clara, CA, USA) equipped with a Capcell Pak ADME-HR S2 column (Osaka Soda Co. Ltd., Osaka, Japan).
10
Sampling and Analyzing PFAS in Seawater
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SML and SSL samples were taken simultaneously to the samples used for bacterial analysis. Seawater was analyzed as reported previously (Casal et al., 2017 (link)). Briefly, samples were filtered through pre-combusted glass fiber filters (47 mm, GF/F Whatman). The analytes were extracted using solid phase extraction (SPE) with OASIS WAX cartridges (Waters). After loading 500 mL of SML water and 2 L of SSL water through the cartridges, these were stored at −20°C in sealed bags until their elution with methanol, followed by methanol containing 0.1% ammonia in an ultraclean laboratory. PFAS analysis was performed by Ultra Performance Liquid Chromatography tandem triple quadrupole mass spectrometry (UPLC-MS/MS) based on an established method with minor modifications (González-Gaya et al., 2014 (link)). Details of methods and results of PFAS in the SML and SSL are reported and discussed elsewhere (Casas et al., 2020 (link)) and used here when needed.
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