1 290 infinity series uhplc system

The supernatant in the sampler bottle was analyzed using a 1,290 Infinity series UHPLC System equipped with an UPLC BEH Amide column (2.1 × 100 mm, 1.7 μm, Waters) (Agilent Technologies) with a mobile phase comprising 25 mmol L⁻¹ ammonium acetate and 25 mmol L⁻¹ ammonia hydroxide in water (pH = 9.75) (A) and acetonitrile (B). The analysis was carried out with an elution gradient as follows: 0 ∼ 0.5 min, 95% B; 0.5 ∼ 7.0 min, 95 ∼ 65% B; 7.0 ∼ 8.0 min, 65 ∼ 40% B; 8.0 ∼ 9.0 min, 40% B; 9.0 ∼ 1 min, 40 ∼ 95% B; and 9.1 ∼ 12.0 min, 95% B. The column temperature was 25°C, and the injection volume was 2 μL (positive) or 2 μL (negative).
A triple TOF 6600 mass spectrometer (AB Sciex) was used to acquire MS/MS spectra on an information-dependent basis (IDA). The acquisition software (Analyst TF1.7, AB Sciex) continuously evaluates the full-scan survey MS data as it collects and triggers the acquisition of MS/MS spectra depending on preselected criteria. The ESI source conditions were as follows: Gas 1 was 60 psi, Gas 2 was 60 psi, curtain gas was 35 psi, source temperature was 600°C, declustering potential was 60 V, and ion spray voltage floating (ISVF) was 5000 V or -4000 V in positive or negative modes, respectively.

The untargeted metabolomic analysis was performed by utilizing a 1,290 Infinity series UHPLC System (Waters Corporation, Milford, MA, USA). The mobile phase was composed of 25 mmol/L ammonium acetate in water was applied as phase A, and 25 mmol/L ammonia in acetonitrile was used as phase B. The analysis procedure was processed as previously described (Zhao et al., 2019 (link)). The Triple TOF 6600 mass spectrometry (AB Sciex, Boston, MA, USA) was used to obtain spectra data, and the acquisition software (Analyst TF 1.7, AB Sciex, Framingham, MA, USA) continuously evaluated the full-scan survey MS data. In each cycle, the most intensive 12 precursor ions (intensity > 100) were chosen for MS/MS at collision energy (CE) of 30 eV. The cycle time was 0.56 s. Electrospray ionization (ESI) source conditions were set as previous study described (Liu et al., 2016 (link); Zhao et al., 2019 (link)).

Samples were analyzed on a 1290 Infinity series UHPLC System (Waters Corporation, Milford, MA, USA) as previous study described [17 (link)]. Briefly, 10 μL of reconstituted sample was injected on a UPLC BEH Amide column (2.1 mm × 100 mm, 1.7 μm). The mobile phase consisted of 25 mmol/L ammonium acetate and 25 mmol/L ammonia hydroxide in water (pH = 9.75) (A) and acetonitrile (B). Each sample was analyzed in the positive ion mode and negative ion mode. The Triple TOF 6600 mass spectrometry (AB Sciex, Boston, MA, USA) was used for its ability to acquire MS/MS spectra on an information-dependent basis (IDA) during an LC/MS experiment. In this mode, the acquisition software (Analyst TF 1.7, AB Sciex, Framingham, MA, USA) continuously evaluates the full scan survey MS data as it collects and triggers the acquisition of MS/MS spectra depending on the preselected criteria. In each cycle, the most intensive 12 precursor ions with intensity above 100 were chosen for MS/MS at a collision energy (CE) of 30 eV. The cycle time was 0.56 second. Electrospray ionization (ESI) source conditions were set as follows: gas 1 as 60 psi, gas 2 as 60 psi, curtain gas as 35 psi, source temperature as 600°C, declustering potential as 60 V, and ion spray voltage floating (ISVF) as 5000 V or -4000 V in the positive or negative modes, respectively.