The digested patient samples were reconstituted in 2% ACN/0.1% FA and spiked with 5 fmol/μL heavy peptides for a final concentration of 0.25 μg/μL, and 2 μL of the resulting samples were analyzed by LC-SRM using a Waters nanoACQUITY UPLC system (Waters Inc.) coupled to a Thermo Scientific TSQ Altis triple quadrupole mass spectrometer (ThermoFisher Scientific Inc.). A 100 μm i.d. × 10 cm, BEH 1.7-μm C18 capillary column (Waters Inc.) was operated at a temperature of 44 ºC. The mobile phases were (A) 0.1% FA in water and (B) 0.1% FA in ACN. The peptide samples were separated at a flow rate of 400 nL/min using a 110-min gradient profile as follows (min:%B): 7:1, 9:6, 40:13, 70:22, 80:40, 85:95, 93:50, 94:95 and 95:1. The parameters of the triple quadruple instrument were set with 0.7 fwhm Q1 and Q3 resolution, and 1.2 s cycle time. Data were acquired in time-scheduled SRM mode (retention time window: 15 min).
Tsq altis triple
Manufactured by Thermo Fisher Scientific
The TSQ Altis triple quadrupole mass spectrometer is a high-performance instrument designed for a wide range of analytical applications. It provides accurate, reliable, and sensitive detection and quantification of target compounds in complex sample matrices.
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2 protocols using tsq altis triple
1
Quantitative Peptide Analysis of Patient Samples
2
Quantification of Sphingolipid Biomarkers
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Ceramides, sphingosine and S1P were analyzed as described in our previous work (49 (link)). In brief, these sphingolipids were quantified on a Thermo Fisher TSQ Altis triple quadrupole mass spectrometer, operated in positive ion mode, coupled to a Vanquish UHPLC system (Thermo Fisher). Lipids were separated on an Agilent Eclipse Plus C8 column. Peaks were integrated using Xcalibur software (Thermo Fischer). Individual sphingolipid species were normalized to class-specific internal standards (Avanti Polar Lipids).
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