The diluted urine samples (30 μL urine and 470 μL MilliQ) were subjected to solid‐phase extraction (SPE) using Oasis Prime HLB (Waters). After loading of the samples, the column was washed with 500 μL methanol (5%). Bile acids were recovered from the cartridge by elution with 500 μL acetonitrile/methanol (90%). A volume of 2 μL was injected and analysed by flow injection analysis (flow rate = 0.15 mL/min) using 90% acetonitrile. ESI‐MS measurements in negative mode were carried out using an Acquity UPLC system, Xevo TQ‐S micro MS (Waters Co). In each run, a urine sample from a CTX patient (before treatment) was measured as a positive control. Subsequently, semiquantitative analysis of m/z 627 (corresponding to the major bile alcohol glucuronide excreted by CTX patients or isomers thereof)7 was performed in a single run using a calibration curve with taurocholic acid (Sigma‐Aldrich Co.) and taurocholic acid‐D4 (Cambridge Isotope Laboratories) as internal standard.
Xevo tq s micro ms
Manufactured by Waters Corporation
The Xevo TQ-S micro MS is a triple quadrupole mass spectrometer designed for analytical applications. It provides high-sensitivity quantitative analysis with enhanced selectivity and speed. The core function of the Xevo TQ-S micro MS is to accurately measure and analyze the mass-to-charge ratio of ionized molecules in a sample.
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Lab products found in correlation
4 protocols using xevo tq s micro ms
1
Bile Acid Profiling in Urine Samples
2
Lipidomic Analysis of Biological Samples
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Concentrations of lipids (ceramides, hexosylceramides and lysophosphatidylcholines-LPCs) were determined by liquid chromatography coupled to tandem mass spectrometry (LC–MS/MS), as previously described113 (link) with slight modifications. Briefly, 50 µL of sample were spiked with 200 µL of methanolic ice-cold and 50 µl of internal standard solution. After vortexing and centrifugation (5 min, 2917 × g, 4 °C), the supernatant was transferred to a HPLC vial and 5 µl were injected into LC–MS/MS system.
The chromatographic separation of the lipid species was performed using an Acquity UPLC instrument (Waters Associates, Milford, Massachusetts, USA) equipped with an Acquity UPLC® (BEH C18, 1.7 µm, 2.1 × 100 mm) column (Waters Associates). The flow rate was 0.3 ml/min and the temperature of the column was set at 55 °C. An isocratic method was used, with a solution of 1 mM ammonium formate and 0.01% formic acid in methanol as the mobile phase solvent. The detection of the lipid species was performed with a triple quadrupole (Xevo TQS-Micro MS, Waters) mass spectrometer equipped with an electrospray ionization source operated in the positive ion mode. The monitoring and quantification of the lipids was performed in the multiple reaction monitoring mode.
The chromatographic separation of the lipid species was performed using an Acquity UPLC instrument (Waters Associates, Milford, Massachusetts, USA) equipped with an Acquity UPLC® (BEH C18, 1.7 µm, 2.1 × 100 mm) column (Waters Associates). The flow rate was 0.3 ml/min and the temperature of the column was set at 55 °C. An isocratic method was used, with a solution of 1 mM ammonium formate and 0.01% formic acid in methanol as the mobile phase solvent. The detection of the lipid species was performed with a triple quadrupole (Xevo TQS-Micro MS, Waters) mass spectrometer equipped with an electrospray ionization source operated in the positive ion mode. The monitoring and quantification of the lipids was performed in the multiple reaction monitoring mode.
3
Bile Acid Profiling of Cerebrotendinous Xanthomatosis
4
Targeted LC-MS Metabolomics Workflow
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For LC-MS analysis, in analytical triplicate, 5 μL of the sample was injected on an XBridge BEH amide column through an Acquity H-class UPLC system (Waters Corporation). MS was done using a Waters Xevo-TQS-micro MS with polarity-switching (positive mode 3 kV, negative mode 2 kV), and multiple reaction monitoring modes were used to acquire data with a randomized injection order. Before, during, and after the run, quality control (QC) samples were injected. Data were processed through TargetLynx (v4.1) to identify peaks from Total Intensity Chromatograms. Peaks were then integrated, and ion counts were obtained and exported for further processing in R. Metabolites found in < 50% QC samples or those with a coefficient of variation > 30% were dropped. Besides, QC samples were used to fit a cross-validated locally estimated scatterplot smoothing (LOESS) function to each metabolite. This accounted for instrumental drift and was used for ion count normalization. The raw data of the LC-MS analysis was uploaded in Metabolights (https://www.ebi.ac.uk/metabolights/index ) and the study number is MTBLS2806.
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