The untargeted LC-MS metabolomics analysis was performed according to Klåvus et al. (2020) and Hanhineva et al.31 (link),67 (link). Briefly, the samples were analyzed using a UPLC–QTOF–MS system (Agilent Technologies, Santa Clara, CA, USA), which consisted of a 1290 LC system, a Jetstream electrospray ionization (ESI) source, and a 6540 UHD QTOF (quadrupole-time-of-flight) mass spectrometer. The samples were separated using both reversed-phase liquid chromatography (RPLC; Zorbax Eclipse XDB-C18, particle size 1.8 µm, 2.1 × 100 mm; Agilent Technologies) and hydrophilic interaction liquid chromatography (HILIC; Aqcuity UPLC BEH amide, 2.1 × 100 mm, 1.7 µm; Waters Corp., Milford, MA, USA), and data were acquired with both positive and negative polarity, thus yielding four runs for each sample. QC samples were injected at the beginning and at the end of the LC-MS run and after every ten injections. Automatic data-dependent MS/MS analysis was performed on samples representing each sample type and on the QC samples. The sample tray was kept at +4 °C during the analysis.
Uplc qtof ms system
Manufactured by Agilent Technologies
Sourced in United States
The UPLC–QTOF–MS system is a high-performance liquid chromatography (HPLC) instrument coupled with a quadrupole time-of-flight mass spectrometer (QTOF-MS). It is designed for accurate mass measurement and high-resolution analysis of chemical compounds.
Automatically generated - may contain errors
Lab products found in correlation
Zorbax eclipse xdb c18, by Agilent Technologies (1 mentions)
Acquity uplc beh amide column, by Waters Corporation (1 mentions)
Agilent profinder b 06, by Agilent Technologies (1 mentions)
Acquity uplc beh c18 column, by Waters Corporation (1 mentions)
Masshunter software, by Agilent Technologies (1 mentions)
Beh c18 column, by Agilent Technologies (1 mentions)
5 protocols using uplc qtof ms system
1
Untargeted Metabolomics Analysis by LC-MS
2
Plasma Metabolite Profiling Using LC-MS
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Profiling of plasma metabolites was performed using LC-MS using a UPLC-QTOF-MS System (Agilent Technologies 1290 LC, 6550 MS, Santa Clara, CA, USA) and has been described elsewhere [27 (link)]. Briefly, over-night fasted plasma samples were extracted and subsequently separated on an Acquity UPLC BEH Amide column (1.7 μm, 2.1 × 100 mm; Waters Corporation, Milford, MA, USA).
We identified metabolites by matching the measured mass-over charge ratio (m/z) and chromatographic retention times with an in-house metabolite library consisting of 111 metabolites that were measurable on all three cohorts (Additional file1 : Table S1). Out of 111 metabolites, 25 of them, mostly consisting of acylcarnitines had putative identities based on their fragmentation spectra and the rest had confirmed identities (Additional file 1 : Table S1). Metabolite peak areas were integrated using Agilent Profinder B.06.00 (Agilent Technologies, Santa Clara, CA, USA). The normalisation process of metabolite levels is described in the supplementary method (Additional file 1 : Supplementary method) [28 (link)].
We identified metabolites by matching the measured mass-over charge ratio (m/z) and chromatographic retention times with an in-house metabolite library consisting of 111 metabolites that were measurable on all three cohorts (Additional file
3
Metabolomics Serum Analysis by UPLC-Q-TOF/MS
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Metabolomics serum samples were analyzed by an UPLC-Q-TOF/MS system (Agilent, United States). An Acquity UPLC BEH C18 column (2.1 mm × 100 mm, 1.7 μm, Waters, United States) was used for separation. The gradient mobile phase was a mixture of 0.1% formic acid in water (A) and acetonitrile (B) with a flow rate of 0.35 ml·min−1 at 35°C. The linear gradient was as follows: 90–70% A (0–3 min) and 70–5% A (3–25 min).
The following MS parameters were employed: full scan range, 50–1,200 m/z; drying gas flow, 6 L·min−1; source drying gas temperature, 300°C; sheath gas temperature, 320°C, sheath gas flow, 12 L·min−1; nebulizer pressure, 1.0 bar; capillary voltage, 3.5 kV.
During the whole analytical process, quality control (QC) samples were analyzed every 10 samples. The stability of the analytical method was evaluated according to the relative standard deviation (RSD) of the retention time and intensity of 10 randomly selected characteristic ion peaks of QC samples.
The following MS parameters were employed: full scan range, 50–1,200 m/z; drying gas flow, 6 L·min−1; source drying gas temperature, 300°C; sheath gas temperature, 320°C, sheath gas flow, 12 L·min−1; nebulizer pressure, 1.0 bar; capillary voltage, 3.5 kV.
During the whole analytical process, quality control (QC) samples were analyzed every 10 samples. The stability of the analytical method was evaluated according to the relative standard deviation (RSD) of the retention time and intensity of 10 randomly selected characteristic ion peaks of QC samples.
4
UPLC-Q-TOF-MS Analysis of Plant Metabolites
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
UPLC-Q-TOF-MS system (Agilent Co.) was used to identify the components in RS samples with the ESI-MS spectra obtained in both the negative and positive ion modes. The conditions of the capillary voltage (CV) of the positive ion mode and negative ion mode were +3.5 kV and -3.5 kV, respectively. The nebulization and auxiliary gas was high-purity nitrogen (>0.9999) with pressure of nebulizer gas of 35 psig and the flow rate of drying gas of 8.0 L min-1 at temperature of 350°C. Fragmentor voltage (FV) and skimmer voltage (SV) were set at 175 and 65 V, respectively. The collision energy (CE) selected ranged from 15 V to 40 V. Data was collected for MS at MS scan ranging 100–1700 Da and for MS/MS at MS/MS scan ranging 50–1500 Da. The Mass Hunter software (Agilent Technologies) was employed to acquire data including the exact mass and to analyze the elemental compositions. The mobile phase used in UPLC-Q-TOF-MS analysis comprised formic acid aqueous solution (0.1%, v/v) (A phase) and acetonitrile (B phase). The separation condition was identical to that of the UPLC-UV-FC system.
5
UPLC-Q-TOF-MS Analysis of Metabolites
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
An UPLC‐Q‐TOF‐MS system (Agilent, USA) was used for analysis with a BEH C18 column (2.1 mm × 100 mm, 1.7 µm). 0.1% formic acid in water (A) and acetonitrile (B) was used as a mobile phase with the following elution program: 0–3 min, 10%–30% B, 3–25 min, and 30%–95% B. Column temperature, 35℃; flow rate, 0.35 ml/min. The full scan range was 50 to 1,200 m/z; sheath gas temperature, 320℃; sheath gas flow, 12 L/min; drying gas temperature, 300℃; drying gas flow, 6 L/min; capillary voltage, 3.5 kV; and nebulizer pressure, 1.0 bar.
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!