Xevo tq s triple quadrupole tandem mass spectrometer

The UPLC-MS/MS system consisted of an ACQUITY I-Class UPLC system and an XEVO TQ-S triple-quadrupole tandem mass spectrometer (Waters Corp., Milford, MA, USA) equipped with an ESI source. Chromatographic separation was performed using a UPLC BEN C₁₈ column (2.1 × 50 mm, 1.7 m; Waters Corp., Milford, MA, USA) at 40 °C. The mobile phase consisted of water containing 0.2% formic acid (B) and acetonitrile containing 0.2% formic acid (A) delivered at 0.3 mL/min as a gradient as follows: 0–0.5 min, 10% A; 0.5–3 min, 90% A; 3–4 min, 90% A; 4–4.5 min, 10% A; 4.5–5 min, 10% A. Waters XEVO TQ-S triple-quadrupole mass spectrometer equipped with an ESI source was used for the mass analysis and detection. All analytes were quantified in the MRM mode, and the optimised MS parameters are listed in Table 8. The main working parameters were set as follows: source temperature, 120 °C; desolvation temperature, 350 °C; and nebuliser gas (N₂), 650 L/h. The scan time for all analytes was set at 0.2 s and acquisition and processing were conducted on a Micromass Masslynx4.1.

Measurement of carnitine in the fat body were performed by using ultra‐high‐performance liquid chromatography with tandem mass spectrometry (UPLC‐MS/MS). The fat bodies from two larvae at the L3 stage were used per sample and samples were processed according to previous description (Nishida et al, 2021 (link)). The detection was carried out on a XEVO TQ‐S triple quadrupole tandem mass spectrometer coupled with electrospray ionization source (Waters). Precursor ion was scanned at m/z (MH⁺: 162.073 > 102.825 for Carnitine) by multiple reaction monitoring and established methods using individual authentic compounds and biological samples. The peak area of a target metabolite was analyzed using MassLynx 4.1 software (Waters). The insoluble pellets were heat‐denatured with 0.2 N NaOH and used to quantify total protein using a BCA protein assay kit (Thermo Fisher Scientific).

A Waters ACQUITY UPLC instrument was used for the chromatographic analysis. Chromatographic separation was performed on a Waters ACQUITY UPLC BEH C18 column (50 mm × 2.1 mm, 1.7 μm) and a pre-column. Then, The temperature of the column chamber was maintained at 40°C, the autosampler (FTN) was set at 10°C, the temperature of the sample chamber was 4°C, the injection volume was 2 μL, and the flow rate was constant at 0.3 mL/min. Gradient elution was performed using 0.1% formic acid and acetonitrile as the mobile phases; the gradient elution procedure is presented in Table 1.
Finally, a positive ion scan was performed in multiple reaction-monitoring (MRM) acquisition modes in combination with a Waters Xevo TQ-S triple quadrupole tandem mass spectrometer (Milford, MA, USA) and an electrospray ionization source (ESI). The parameters and data of the MS/MS system acquired using Masslynx 4.1 software (Milford, Massachusetts, USA) are presented in Table 2.

A Waters Xevo TQ-S triple quadrupole tandem mass spectrometer (Milford, MA, USA) and a Waters ACQUITY UPLC I-Class system (Milford, MA, USA) constituted this UPLC-MS/MS system, which used electrospray ionization (ESI) source. In the experiment, 0.1% formic acid solution and acetonitrile were used as the mobile phase in the instrument, and the model of the column was ACQUITY UPLC BEH C18 column (2.1 mm × 50 mm, 1.7 μm). A gradient elution was used to separate derazantinib and IS, and the linear gradient elution procedure was as follows: 0–0.5 min (10% acetonitrile was eluted at a flow rate of 0.30 mL/min); 0.5–1.0 min (acetonitrile, 10–90%); 1.0–1.4 min (acetonitrile, 90%) and 1.4–1.5 min (acetonitrile, 90–10%). The single injection volume was 2 μL, and the single analysis time was 2.0 min. In addition, the injection temperature was 10 °C, and the column temperature was 40 °C.
A Xevo TQ-S triple quadrupole tandem mass spectrometer was used for mass spectrometry monitoring, combined with an ACQUITY UPLC system for analysis. In positive ion mode, the collision energy of derazantinib and IS were 25 eV and 15 eV, respectively, and the cone voltage were 20 V and 30 V, respectively. In addition, the selective reaction monitoring (SRM) modes of ion transitions of derazantinib and IS were m/z 468.96 → 382.00 and m/z 488.01 → 400.98, respectively.

Targeted and semi-quantitative metabolomics analyses were performed on a Waters Xevo TQ-S triple quadrupole tandem mass spectrometer coupled to the ultra-pressure liquid chromatography (UPLC-MS) platform using the previously published protocol in FIMM (Institute for Molecular Medicine Finland) [25 ]. Briefly, metabolites were extracted from 100 μL serum, 20 mg muscle, and 20 mg WAT samples respectively using protein precipitation by adding acetonitrile +1% formic acid. The collected extracts were dispensed in Ostro 96-well plates (Waters Corporation, Milford, USA) and filtered by applying a vacuum at a delta pressure of 300–400 mbar for 2.5 min on robot’s vacuum station. Filtered sample extract (5 μL) was injected in an Acquity UPLC system coupled to a Xevo TQ-S triple quadrupole mass spectrometer (Waters Corporation, Milford, MA, USA) which was operated in both positive and negative polarities with switching time of 20 ms. Multiple Reaction Monitoring (MRM) acquisition mode was selected for the quantification of metabolites. MassLynx 4.1 software was used for data acquisition, data handling, and instrument control. Data processing was done using TargetLynx 4.1 software. Eighty-nine metabolites in serum, 71 in muscle, and 71 in WAT were identified in whole samples.