Plasma amino acids were measured by flow injection tandem mass spectrometry using stable isotope dilution techniques as described previously (Haqq et al., 2005 ; Laferrère et al., 2011 ; Lien et al., 2009 ; Newgard et al., 2009 ). Data were acquired using a Waters triple quadrupole detector equipped with Acquity UPLC system and controlled by the MassLynx 4.1 MS software platform (Waters, Milford, MA).
Triple quadrupole detector
Manufactured by Waters Corporation
Sourced in Spain
The Triple Quadrupole Detector is a highly sensitive mass spectrometry instrument designed for quantitative and qualitative analysis. It consists of three quadrupole mass filters arranged in series, allowing for the detection and identification of target analytes with high specificity and accuracy.
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3 protocols using triple quadrupole detector
1
Plasma Amino Acid Profiling by Mass Spectrometry
2
Quantifying Acylcarnitine Profiles in Mice
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Hearts were rapidly harvested from WT and KO mice fed ad libitum or fasted for two days. Tissue homogenates (50 mg/mL) were prepared using a polytron in 50% acetonitrile and 0.3% formic acid. 45 acylcarnitine species (including short, medium, and long-chain) were analyzed by tandem mass spectrometry (MS/MS) using sample preparation methods described previously [13 (link), 14 (link)]. The data were acquired using a Waters triple quadrupole detector equipped with AcquityTM UPLC system and controlled by MassLynx 4.1 software platform (Waters, Milford, MA).
3
Ultra-Sensitive UHPLC-MS/MS Quantification
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An ultra-high performance liquid chromatography system coupled to a triple quadrupole detector (UHPLC-MS/MS) has been used. It consists of an ACQUITY Quaternary Solvent Manager used to load samples as well as to wash and recondition the analytical column, an autosampler capable of injecting volumes up to 25 μL per injection for up to 21 vials, a column manager and a triple quadrupole detector, which were all from Waters (Barcelona, Spain). The detection parameters for each compound are shown in Table 2.
The analytical column was a 50 mm × 2.1 mm, ACQUITY UHPLC BEH Waters C 18 column with a particle size of 1.7 μm (Waters, Barcelona, Spain) operating at a temperature of 30°C.
The sample volume injected was 10 µL, and the analyte separation was carried out using water with 0.1% (v/v) of ammonia and methanol without additives at a flow rate of 0.3 mL•min -1 in gradient mode. The gradient started at an 80:20 (v/v) mixture of water:MeOH, which changed to 40:60 (v/v) in 1.5 minutes and to 25:75 (v/v) in 1.25 minutes more. Then, the gradient changed to 0:100 (v/v) in 1 minute. Finally, it returned to 80:20 in 2.25 minute and stayed at that mixture for calibration for an additional 0.5 minutes. Thus, the chromatographic separation was completed in 6.5 minutes.
The analytical column was a 50 mm × 2.1 mm, ACQUITY UHPLC BEH Waters C 18 column with a particle size of 1.7 μm (Waters, Barcelona, Spain) operating at a temperature of 30°C.
The sample volume injected was 10 µL, and the analyte separation was carried out using water with 0.1% (v/v) of ammonia and methanol without additives at a flow rate of 0.3 mL•min -1 in gradient mode. The gradient started at an 80:20 (v/v) mixture of water:MeOH, which changed to 40:60 (v/v) in 1.5 minutes and to 25:75 (v/v) in 1.25 minutes more. Then, the gradient changed to 0:100 (v/v) in 1 minute. Finally, it returned to 80:20 in 2.25 minute and stayed at that mixture for calibration for an additional 0.5 minutes. Thus, the chromatographic separation was completed in 6.5 minutes.
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