Serum amino acid calibration standards were prepared with AccQ•Tag™ Ultra Amino Acid Analysis Solution (AA) kit from Waters according to instructions with slight modifications for detection on a mass spectrometer [21 (link)]. A 5-point standard concentration curve was made from the calibration standard solution to calculate amino acid concentrations in serum samples. Serum samples of 10 μl were spiked with an internal standard, norvaline then derivatized according to AccQ•Tag instructions. High-resolution separation was done using an ACQUITY UPLC system and injecting 1 μl, with an Amino Acid Analysis column from Waters. Mass detection was completed on a XEVO TQ-S Mass Spectrometry, Waters in ESI positive mode.
Xevo tq s mass spectrometry
Manufactured by Waters Corporation
Sourced in United States
The XEVO TQ-S is a high-performance triple quadrupole mass spectrometry system designed for sensitive and precise quantitative analysis. It features advanced ion optics and a robust, reliable design to deliver accurate and reproducible results.
Automatically generated - may contain errors
3 protocols using xevo tq s mass spectrometry
1
Serum Amino Acid Quantification
2
Liver Metabolomics and Lipid Profiling
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The liver samples were harvested and underwent targeted metabolomics profiling analysis. Firstly, Q300 Kit (Metabo-Profile, Shanghai, China) was used to generate all raw metabolites data by UPLC-MS/ MS. Then data was processed at iMAP platform. For another, lipid profiling of liver was also generated on a Waters ACQUITY Ultra-Performance LC (UPLC) system. A Waters XEVO TQ-S mass spectrometry and MassLynx 4.1 software (Waters, Milford, MA) were also used to generate and process data. At first, distribution characteristics of metabolite profile between groups was identified by Principal component analysis (PCA) and orthogonal partial least squares discriminant analysis. According to the OPLS-DA model, VIP (variable importance in projection) was obtained and was used for following process. Metabolites with p-value < 0.05 and VIP ≥ 1 were screened as statistically significant metabolites. Following analysis of enriched pathway was performed with selected metabolites.
3
UHPLC-MS/MS Targeted Metabolomics
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LC-MS analysis were carried out on an ultra-high performance liquid chromatography system equipped with a Xevo TQ-S mass spectrometry (Waters, USA). The mobile phase consisted of 0.1% formic acid in ultrapure water (A) and acetonitrile (B) was in the following gradients: 0–4 min, 10% B; 4–15 min, 50% B; 15–23 min, 60% B; 23–30 min, 80% B; 30–35 min, 95% B; 35–50 min, 10% B. The flow rate was 0.2 mL/min and the injection volume of the sample was 20 μL. The positive electrospray ionization mode (ESI+) with multiple reaction monitoring (MRM) was carried out in MS analysis. Qualitative and quantitative analysis based on internal standards and standard curves.
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