LC–MS data were acquired using an Ultimate 3000 DGP-3600RS HPLC system (Thermo Fisher Scientific, Waltham, MA, USA) coupled to an LTQ Orbitrap mass spectrometer (Thermo Fisher Scientific, Waltham, MA, USA)3 (link),12 (link). Briefly, LC separation was performed on a ZIC-pHILIC column (Merck SeQuant, Umea, Sweden; 150 mm × 2.1 mm, 5 μm particle size). Acetonitrile (A) and 10 mM ammonium carbonate buffer, pH 9.3 (B) were used as the mobile phase, with a linear gradient from 80–20% A in 30 min at a flow rate of 100 μL/min. The mass spectrometer was operated in full-scan mode with a 100–1000 m/z scan rate and automatic data-dependent MS/MS fragmentation scans. For each metabolite, we chose a singly charged, [M+H]+ or [M−H]−, peak (Supplementary Table S2 ). Peak detection and identification of metabolites were performed using MZmine 2 software (http://mzmine.github.io/ )26 (link). Detailed data analytical procedures and parameters have been described previously25 (link). Metabolite peaks were identified by comparing their m/z values and retention times with pure standards listed in previous reports3 (link),24 (link),25 (link),56 (link).
Ultimate 3000 dgp 3600rs hplc system
Manufactured by Thermo Fisher Scientific
Sourced in United States
The Ultimate 3000 DGP-3600RS HPLC system is a high-performance liquid chromatography (HPLC) instrument designed for analytical and preparative applications. It features a dual-gradient pump capable of delivering precise and accurate flow rates, ensuring reliable and reproducible separations. The system is designed to provide high-quality data and performance for a wide range of HPLC applications.
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Lab products found in correlation
4 protocols using ultimate 3000 dgp 3600rs hplc system
1
Metabolite Profiling by LC-MS/MS
2
Urinary Metabolite Analysis by HPLC-Orbitrap MS
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Urinary metabolites were analyzed using an Ultimate 3000 DGP‐3600RS HPLC system (Thermo Fisher Scientific) coupled to an LTQ Orbitrap mass spectrometer (Thermo Fisher Scientific), as described.12 , 25 Briefly, LC separation was performed on a ZIC‐pHILIC column (Merck SeQuant, Umea, Sweden; 150 mm × 2.1 mm, 5 µm particle size). Acetonitrile (A) and 10 mM ammonium carbonate buffer, pH 9.3 (B) were used as the mobile phase, with a linear gradient elution from 80% to 20% A over 30 min, at a flow rate of 100 µL/mL. The mass spectrometer was operated in full‐scan mode with a 100‐1000 m/z scan rate and automatic data‐dependent MS/MS fragmentation scans. For each metabolite, we chose a singly charged, [M + H]+ or [M‐H]−, peak (Table S2 ). Peak detection and identification of metabolites were performed using MZmine 2 software.29 Detailed data analytical procedures and parameters have been described previously.26 Normalized peak area was calculated from the following formula: Normalized peak area = (Raw peak area of each metabolite/Raw peak area of creatinine) × 1000.
3
Urinary Metabolite Profiling by HPLC-MS
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Urinary metabolites were analyzed using an Ultimate 3000 DGP-3600RS HPLC system (Thermo Fisher Scientific, Waltham, MA, USA) coupled to an LTQ Orbitrap mass spectrometer (Thermo Fisher Scientific, Waltham, MA, USA), as described (Chaleckis et al., 2016 ) (Kameda et al., 2020) . Briefly, LC separation was performed on a ZIC-pHILIC column (Merck SeQuant, Umea, Sweden; 150 mm x 2.1 mm, 5 μm particle size). Acetonitrile (A) and 10 mM ammonium carbonate buffer, pH 9.3 (B)
were used as the mobile phase, with a linear gradient elution from 80-20% A over 30 min, at a flow rate of 100 μL mL -1 . The mass spectrometer was operated in full-scan mode with a 100-1000 m/z scan rate and automatic data-dependent MS/MS fragmentation scans. For each metabolite, we chose a singly charged, [M+H]+ or [M-H]-, peak (Supplemental Table S3). Peak detection and identification of metabolites were performed using MZmine 2 software (Pluskal et al., 2010a) . Detailed data analytical procedures and parameters have been described previously (Teruya et al., 2019) .
were used as the mobile phase, with a linear gradient elution from 80-20% A over 30 min, at a flow rate of 100 μL mL -1 . The mass spectrometer was operated in full-scan mode with a 100-1000 m/z scan rate and automatic data-dependent MS/MS fragmentation scans. For each metabolite, we chose a singly charged, [M+H]+ or [M-H]-, peak (Supplemental Table S3). Peak detection and identification of metabolites were performed using MZmine 2 software (Pluskal et al., 2010a) . Detailed data analytical procedures and parameters have been described previously (Teruya et al., 2019) .
4
Non-targeted LC-MS Metabolomics Workflow
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Non-targeted LC-MS conditions were as described previously (14, 16) . Briefly, LC-MS data were obtained using an Ultimate 3000 DGP-3600RS HPLC system (Thermo Fisher Scientific, Waltham, MA, USA) coupled to an LTQ Orbitrap mass spectrometer (Thermo Fisher Scientific, Waltham, MA, USA). LC separation was performed on a ZIC-pHILIC column (Merck SeQuant, Umea, Sweden; 150 mm × 2.1 mm, 5 µm particle size). Acetonitrile (A) and 10 mM ammonium carbonate buffer, pH 9.3 (B) were used as the mobile phase, with a linear gradient from 80-20% A over 30 min, at a flow rate of 100 µL/mL. The mass spectrometer was operated in full-scan mode with a 100-1000 m/z scan rate and automatic data-dependent MS/MS fragmentation scans.
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