Deacylated products (lyso-Gb3 and its eight analogs) were detected using a Xevo TQD MS with an Acquity UPLC system (Waters, Milford, MA), as described previously (27 (link)). All MS parameters were optimized to achieve the highest sensitivity for lyso-Gb3 analogs. Liquid chromatography and MS parameters are summarized in Table S1 . The peak areas corresponding to lyso-Gb3 and its analogs in the multiple-reaction-monitoring (MRM) chromatogram were calculated using MassLynx software (Waters).
Xevo tqd ms
Manufactured by Waters Corporation
Sourced in United States
The Xevo TQD MS is a triple quadrupole mass spectrometer produced by Waters Corporation. It is designed for high-sensitivity, high-throughput quantitative analysis of small molecules in complex matrices.
Automatically generated - may contain errors
Lab products found in correlation
Acquity uplc system, by Waters Corporation (2 mentions)
I class uplc, by Waters Corporation (2 mentions)
Masslynx software, by Waters Corporation (1 mentions)
Uplc beh c18, by Waters Corporation (1 mentions)
Acquity uplc beh c18 column, by Waters Corporation (1 mentions)
Acquity system, by Waters Corporation (1 mentions)
Oasis prime hlb 1 cc cartridges, by Waters Corporation (1 mentions)
5 protocols using xevo tqd ms
1
Quantification of Lyso-Gb3 Analogs
2
UPLC-MS/MS Analysis of Vonoprazan and Metabolite
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ACQUITY I-Class UPLC and Waters XEVO TQD MS (Milford, MA, USA) were used for the detection of vonoprazan and M-I with an ACQUITY UPLC BEH C18 chromatographic column (50 × 2.1 mm, 1.7 μm) at 40 °C. The mobile phase consisted of acetonitrile and 0.1% formic acid in water in gradient proportions, as shown in Table 1 , with a flow rate of 0.4 mL/min. The injection volume of the sample was 2 μL. The scanning method was multiple reaction monitoring (MRM) with detection in positive ion mode and an ESI + ion source. Other mass spectrometry parameters were as follows: capillary voltage 2.0 kV, ion source temperature 150 °C, dissolvent temperature 500 °C, argon flow rate 0.15 mL/min, cone hole gas flow 50 L/h, solvent gas flow 1000 L/h. The precursor ion and product ion were m/z 346.04→314.97 for vonoprazan, m/z 347.50→205.00 for M-I, and m/z 285.10→193.10 for internal standard (IS), diazepam.
3
UPLC-MS/MS Quantification of Vonoprazan
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ACQUITY I-Class UPLC and Waters XEVO TQD MS (Milford, MA, USA) were used for the detection of vonoprazan and M-I with ACQUITY UPLC BEH C18 column (50 mm×2.1 mm, 1.7 μm) (Milford, MA, USA) at temperature of 40°C. The mobile phase consisted of acetonitrile and 0.1% formic acid in gradient proportion with running time of 3 minutes at flow rate of 0.4 mL/min. The injection volume of sample was 2 μL. The scanning method was multiple reaction monitoring (MRM) with detection in positive ion mode and an ESI+ ion source. Other mass spectrometry parameters were listed as following: capillary voltage 2.0 kV; ion source temperature 150 °C; argon flow rate: 0.15 mL/min. The other conditions of mass spectrometry were referred to Shen.10 (link)
4
UHPLC-MS Analysis of Compounds
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UHPLC-MS experiments were conducted on an Acquity system from Waters (Milford, MA, USA) comprising a binary pump, autosampler and column oven, which was coupled to a Xevo TQD MS (Waters). The separation conditions were the same as for the Agilent instrument. ESI parameters were set as follows: capillary voltage 3.50 kV, cone voltage 50 V, desolvation gas flow 650 L/h and desolvation gas and source temperature 350 °C and 150 °C, respectively. The MS was operated in positive ionization mode.
5
Quantifying Fecal Bile Acid Levels
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BA levels in feces were determined following a previously described protocol [25 ,26 (link)]. Lyophilized feces (approximately 50 mg) were finely ground and mixed with 0.2 M NaOH (1 mL). The mixtures were then purified from lipids by extraction with hexane (1 mL). The extraction step was repeated three times. The samples were centrifuged (20,000×g, 10 min, 4 °C) and supernatants were further cleaned using Oasis PRiME HLB 1 cc cartridges (Waters, Milford, MA, USA) that were conditioned with methanol (1 mL) followed by ultrapure water (3 mL). The loaded cartridges were washed with ultrapure water (500 μL), and the analytes were eluted with methanol-acetonitrile (1:1, v/v, 1 mL) for liquid chromatograph-mass spectrometry (LC-MS/MS) analysis. BA was analyzed on an Acquity UPLC system and a Waters Xevo TQD MS (Waters). The analytes were quantified using external standards, and calibrators were prepared in methanol–acetonitrile (1:1, v/v) within a range of 0.001–1.0 μg/mL, with quality controls at 0.1 and 1.0 μg/mL.
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