Lcms 8030

The analysis of PFASs (i.e., PFOA, PFOS, and PFBS) in plant and river water samples was conducted using a liquid chromatograph (LC) coupled with triple quadrupole linear ion trap tandem mass spectrometer (Shimadzu LCMS-8030, Canby, OR, USA) equipped with an electrospray ionization (ESI) source, which was in a negative ion mode. The targeted PFASs were quantified using multiple reaction monitoring (MRM) mode of analysis. The chromatographic separation of analytes was achieved with a Luna^® Omega Polar C18 column (2.1 × 100 mm, 3.0 µm, Phenomenex, Aschaffenburg, Germany). The column temperature was set at 40 °C. A gradient elution program was applied and was made of 20 mM ammonium acetate (solvent A) and 100% MeOH (solvent B), at a flow rate of 0.3 mL/min and an injection volume of 10 µL used for individual samples. The linear gradient elution program started at 20% B and increased to 80% B after 5 min, then increased to 95% B for 15 min; it was kept to 100% B for 17–27 min, before being 20% B for 30–40 min. The total run time for each injection was 40 min. Argon gas was used as the collision gas. The LC system was a LCMS-8030 Shimadzu system with a DGU-20A_3R degassing unit, coupled with an LC-20AD liquid chromatograph, a CTO-20AC column oven, a SIL-20AC autosampler and a NM32LA nitrogen gas generator.

A 10 µL aliquot of the sPAH stock solution (10 mM in DMSO) was diluted in 990 µL of a mixture of acetonitrile and water. This solution was then diluted 100-fold in phosphate buffer containing mice liver microsomes (0.5 mg/mL), 1 mM NADPH, and 3 mM MgCl₂, and incubated at 37 °C. After 2, 10, 20, 40, and 60 min, 70 µL aliquots were collected and mixed with 70 µL acetonitrile at 0 °C. Equivalent experiments were performed without NADPH in order to identify chemical instability or enzymatic process not depending on NADPH, and with testosterone as a positive control. The enzymatic reaction was stopped by addition of acetonitrile. Samples were analyzed by LC–MS/MS on an UHPLC LC–MS 8030 (Shimazu, Kyoto, Japan).
For metabolite identification, 50 µM of sPAH were incubated with mice liver microsomes and NADPH. Two samples were prepared: one in which acetonitrile was added immediately (t0) and one in which acetonitrile was added after 30 min. After centrifugation for 5 min at 15,000× g, 2 samples of each supernatant were analyzed by LC–MS/MS on a LC–MS 8030 (Shimazu, Kyoto, Japan) and detected by selected ion monitoring (SIM).

We used LCMS-8030 (Shimazu, Japan). A Sim-pack XR-ODSIII 2.2 μm C18 150 mm × 2.0 mm HPLC column (Shimazu, Japan) was used for HPLC separation. The column was kept at 40 °C. Mobile phase A was 0.1% formic acid in water, and mobile phase B was acetonitrile. LC gradient was as follows: 45%B → 75%B (15 min) → 75%B (15 min) → 45%B (15 min) → 45%B (10 min). The flow rate was 0.2 mL/min. A typical injection volume was 18 μL using partial loop injection mode.