Xevo tq s mass spectrometer

Mass spectrometric analyses of ethanol washing solutions was performed on a Waters Xevo TQ-S mass spectrometer (Waters, Manchester, United Kingdom) coupled to an Acquity UPLC i-class core system (Waters, Milford, MA, United States) consisting of a binary solvent manager, sample manager, and column oven. Aliquots (2 μL) of prepared ethanolic extracts were injected into the UPLC/MS-MS system equipped with a 2.1 × 150 mm, 1.7 μm, UPLC CSH C18 column (Waters, Manchester, United Kingdom) and analyzed according to Marxen et al. (2015b) (link).

The collected plasma was deproteinized with a fourfold volume of methanol containing 2 μg/mL 4-hydroxy chalcone as an internal standard. After vortexing for 10 min, the samples were centrifuged at 20,000 × g for 15 min at 4°C. Then, the supernatants were analyzed using the LC/MS/MS technique.
The LC/MS/MS analysis was conducted using an ultra-performance LC system connected to a Xevo TQ-S mass spectrometer (Waters Corporation, Milford, MA, USA). The samples were separated using a 1.7-μm particle ACQUITY C18 column (2.1 mm × 100 mm, Waters), maintained at 50°C, under gradient mobile phase conditions with a mixture of 0.1% formic acid in water and 0.1% formic acid in acetonitrile as solvents (0–1 min 70:30 v/v, 1–3 min 70:30 to 2:98 v/v, 3–5 min 2:98 v/v, and 5–6.5 min 70:30 v/v) with a flow rate of 0.3 mL/min. The separated samples were introduced into an MS in the positive and negative electrospray ionization mode for sulfasalazine and 4-hydroxy chalcone, respectively. Each compound was quantified in the multiple reactions monitoring mode (399.24 > 119.02, Cone 20 V, Collision 46 eV for sulfasalazine; 223.15 > 117.00, Cone 50 V, and Collision 36 eV for 4-hydroxy chalcone).
The AUC of sulfasalazine was calculated as the total area of the trapezoids formed by the points of the concentration and time in the concentration-time plots.

Bile acids (BAs) were profiled by an ultra-performance liquid chromatography/multiple reaction monitoring/mass spectrometry (UPLC-MRM-MS) method as described previously [33 (link)].
50 μL of rat plasma were spiked with deuterated internal standards. Then, proteins were precipitated, and supernatants were dried and reconstituted in 50 μL methanol:water (50:50, v/v). Samples were analyzed using an Acquity UPLC system (Waters, Wilmslow, UK) equipped with an Acquity UPLC BEH C18 column (1.7 μm, 2.1 × 100 mm; Waters). The MS analysis was performed using a Waters Xevo TQ-S mass spectrometer (Waters) with an ESI source working in the negative-ion mode. This method allows the quantification of 12 unconjugated, 8 glycine-conjugated, and 11 taurine-conjugated BAs, using 5 additional deuterated BAs as internal standards in a single analytical run. These analyses were performed in the Analytical Unit, Core Facility, IIS Hospital La Fe in Valencia, Spain.

In vitro microsomal incubation and in vivo samples were analysed using a Waters Xevo TQ-S mass spectrometer by multiple reaction monitoring (Waters, Milford, MA). Conditions were 0.2% formic acid (mobile phase A) and acetonitrile (mobile phase B). Separation was achieved on a Phenomenex Kinetex C18 column (2.1 × 50 mm; 2.6 μm). The column was equilibrated at initial condition of 95% A and 5% B for 0.5 min, linear gradient over 3 min to 100% B, held over 1 min, followed by linear gradient back to 5% B over 0.1 min, at 0.6 mL/min flow rate. AO incubation samples were analysed for metabolites of 1 using an Agilent 6520 QTOF MS (Agilent, Santa Clara, CA). Using the above mobile phases, separation was achieved on a Phenomenex Kinetex C18 column (2.1 × 100 mm; 2.6 μm) (Phenomenex, Torrance, CA). The column was equilibrated at initial condition of 95% A and 5% B (0.5 min), linear gradient (15 min) to 50% B, held for 1 min, followed by linear gradient back to 5% B (0.5 min), at 0.6 mL/min flow rate.

The following chromatographic columns were assessed: Synchronis™ HILIC 100 × 2.1 mm 1.7 μm (Thermo Fisher Scientific, Waltham, MA, USA), Gemini® C6-Phenyl 100 × 2.0 mm 3 μm (Phenomenex, Utrecht, the Netherlands) and SeQuant® ZIC®-HILIC 100 × 2.1 mm 5 μm (Merck Millipore, Darmstadt, Germany).
Water with 1% formic acid and water buffered at pH 6.4 with ammonium formate were tested as mobile phase A, while acetonitrile (with and without formic acid 1%) and methanol with 1% formic acid were used as mobile phase B.
Optimization was carried out in an Acquity UPLC coupled with a Xevo TQ-S mass spectrometer (Waters, Milford, MA, USA).

Concentrations of fecal Bas were measured according to previously reported methods [11 (link)]. In short, the Waters ACQUITY ultra-performance LC system and Waters XEVO TQ-S mass spectrometer with an ESI source controlled by MassLynx 4.1 software (Waters, Milford, MA) were used for analyzing fecal extracts and BA reference standards. An ACQUITY BEH C18 column (1.7 µm, 2.1 × 100 mm internal dimensions) (Waters, Milford, MA) was used to perform chromatographic separations. UPLC-MS raw data obtained in the negative mode were analyzed using the TargetLynx applications manager version 4.1 (Waters Corp., Milford, MA) to obtain the calibration equations and the quantitative concentration of each BA in the samples. The 12αOH BAs levels were the sum of CA, DCA, taurocholic acid (TCA), glycocholic acid (GCA), taurodeoxycholic acid (TDCA), and glycodeoxycholic acid (GDCA), and the levels of non-12αOH BAs included total the concentration of CDCA, LCA, UDCA, taurochenodeoxycholic acid (TCDCA), glycochenodeoxycholic acid (GCDCA), taurolithocholic acid (TLCA), glycolithocholic acid (GLCA), tauroursodeoxycholic acid (TUDCA), and glycoursodeoxycholic acid (GUDCA).