1290 infinity binary uplc

The liquid chromatography–mass spectrometry was performed on a 1290 Infinity Binary UPLC coupled with a 6540 UHD Accurate-Mass Q-TOF (Agilent Technologies, Santa Clara, CA, USA) as described previously by Hanhineva et al.^{43 (link)} Briefly, a Zorbax Eclipse XDB-C18 column was used for the reversed-phase separation and an Aqcuity UPLC BEH amide column (Waters, Milford, MA, USA) for the HILIC separation. After each chromatographic separation, the ionization was carried out using jet stream electrospray ionization (ESI) in the positive and negative mode, yielding four data files per sample. The collision energies for the MS/MS analysis were chosen as 10, 20 and 40 V, for compatibility with the spectral databases.

The samples were analyzed by liquid chromatography-mass spectrometry consisting of a 1290 Infinity Binary UPLC coupled with a 6540 UHD Accurate-Mass Q-TOF (Agilent Technologies), as described previously [49 (link)]. In brief, a Zorbax Eclipse XDB-C18 column (2.1 × 100 mm, 1.8 μm; Agilent Technologies) was used for the reversed-phase (RP) separation and an Acquity UPLC BEH amide column (Waters) for the HILIC separation. After each chromatographic run, the ionization was carried out using jet stream electrospray ionization (ESI) in the positive and negative mode, yielding four data files per sample. The collision energies for the MS/MS analysis were selected as 10, 20, and 40 V, for compatibility with spectral databases.

The liquid chromatography–mass spectrometry (LC–MS) analysis was performed according to Klåvus et al. [25 (link)]. Briefly, liquid chromatography was performed on a 1290 Infinity Binary UPLC (Agilent Technologies). Reversed-phase separation was employed using a Zorbax Eclipse XDB-C18 column (dimensions 2.1 × 100 mm, particle size 1.8 µm) as the stationary phase. The mobile phase consisted of ultra-pure water (solution A) and methanol (solution B), both containing 0.1% v/v formic acid. The flowrate of the eluent was 0.6 mL/min. The elution gradient profile was as follows (t [min], %B): (0, 2), (10, 100), (14.5, 100), (14.51, 2), (16.5, 2). A measure of 1 µL of each standard or sample was injected for analysis.
Mass spectrometry was performed on an Agilent 6540 Q-TOF with a Jet Stream ESI ion source. The fragmentor voltage used was 100 eV and scan range 20–1600 m/z. From every precursor scan cycle (400 milliseconds), four most abundant precursor ions were automatically selected for MS/MS fragmentation, excluded after two acquired MS/MS spectra, and released again from the exclusion list after 0.25 min. The collision energies used for the MS/MS analysis were (±)10, 20, and 40 eV, for compatibility with online databases such as METLIN and HMDB database [46 (link),49 (link)]. Both negative and positive ionization modes were used in the study.