For LC–MS analyses, extracts were dissolved in MeOH to a final concentration of 1 mg/mL, and 1 μL was injected into Waters Acquity UPLC system coupled to Agilent 6530 QTOF MS. Samples were analyzed according to the protocol that was previously published29 (link). LC–MS/MS acquisition of the pure compounds was performed using Shimadzu Nexera X2 UHPLC system coupled to Shimadzu 9030 QTOF mass spectrometer as previously described61 (link). LC–MS/MS acquisition for molecular networking was performed using Thermo Instruments MS system (LTQ Orbitrap XL, Bremen, Germany) equipped with an electrospray ionization source (ESI) as described29 (link).
Acquity uplc system
Manufactured by Agilent Technologies
Sourced in United States
The ACQUITY UPLC system is a high-performance liquid chromatography (HPLC) instrument designed for ultra-high performance liquid chromatography (UPLC) applications. It is capable of generating high-pressure liquid flows and delivering samples to a separation column for the analysis of complex mixtures.
Automatically generated - may contain errors
Lab products found in correlation
Hss t3 column, by Agilent Technologies (2 mentions)
6530 accurate mass q tof, by Agilent Technologies (2 mentions)
Nexera x2 uhplc system, by Shimadzu (1 mentions)
9030 qtof mass spectrometer, by Shimadzu (1 mentions)
6530 qtof ms, by Agilent Technologies (1 mentions)
Masshunter workstation software, by Agilent Technologies (1 mentions)
4 protocols using acquity uplc system
1
LC-MS Analysis of Compounds
2
Bile Acid Profiling by UPLC-QToF
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Bile acid extraction was performed by adding methanol containing internal standards to 50 μL plasma. Samples were analyzed using an ACQUITY UPLC system with an HSS T3 column coupled to a 6530 Accurate Mass QToF (Agilent Technologies).
Following liquid chromatography‐mass spectrometry analysis, data was preprocessed by performing peak integration, background correction, and determination of the relative ratios between metabolites and their corresponding internal standards. The metabolite units were expressed as the peak area ratios of the target analyte to the respective internal standard. An in‐house written tool was applied using the QC samples to compensate for shifts in the sensitivity of the mass spectrometer throughout the batches. Both internal standard correction and QC correction were applied to the dataset before reporting results. Quality assurance of metabolite measurements was performed using the QC relative standard deviation (RSDqc). For amines, reported compounds had an RSDqc <15%, for positive and negative lipids RSDqc <20%, and for bile acids RSDqc <30%.
Following liquid chromatography‐mass spectrometry analysis, data was preprocessed by performing peak integration, background correction, and determination of the relative ratios between metabolites and their corresponding internal standards. The metabolite units were expressed as the peak area ratios of the target analyte to the respective internal standard. An in‐house written tool was applied using the QC samples to compensate for shifts in the sensitivity of the mass spectrometer throughout the batches. Both internal standard correction and QC correction were applied to the dataset before reporting results. Quality assurance of metabolite measurements was performed using the QC relative standard deviation (RSDqc). For amines, reported compounds had an RSDqc <15%, for positive and negative lipids RSDqc <20%, and for bile acids RSDqc <30%.
3
Lipid Extraction and UPLC-QToF Analysis
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
We took 10‐μL and 20‐μL plasma samples that were spiked with calibration and internal standards and extracted using isopropyl alcohol (for the positive lipid platforms) or methanol (for the negative lipid platforms). Samples were analyzed using an ACQUITY UPLC system with an HSS T3 column coupled to a 6530 Accurate Mass QToF (Agilent Technologies, Santa Clara, CA).32
4
UPLC-QTOF-MS Metabolite Profiling
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Each sample was analyzed on a Waters ACQUITY UPLC system (Milford, MA, USA) equipped with an Agilent UPLC column (ZORBAX Extent-C18 50 mm × 2.1 mm, 1.8 μm). The mobile phase system composed of 0.1% formic acid in water (A) and acetonitrile (B). The flow rate was controlled at 0.3 mL/min with a gradient program of 0–8 min, 5–30% B; 8–16 min, 30–32% B; 16–20 min, 32–40% B; 20–25 min; 40–90% B; 25–28 min, 90-5% B.
The electrospray ionization (ESI) was used as the ionization source by Agilent 6385UHD Accurate-Mass Q-TOF/MS G2 High Definition Mass Spectrometer. The separation was performed by an Agilent orbax Extend-C18 Column (2.1 mm × 50 mm, 1.8 μm) at 35 °C. In ESI-MS analysis, the parameters were set as source temperature at 120 °C, and desolvation gas temperature at 350 °C. In positive ion mode, the cone voltage was set as 40 V, the capillary voltage was set as 4.0 kV and the extraction cone voltage was set as 4.0 V. In negative ion mode, the cone voltage was set as 30 V, the capillary voltage was set as 3.5 kV and the extraction cone voltage was set as 5.0 V. To obtain the fragment information of the metabolites, argon was used as the collision gas in MS/MS mode and the collision energy was operated at alternative voltages of 10–70 eV. The mass data were analyzed by Agilent Mass Hunter Workstation software (version B.03.01).
The electrospray ionization (ESI) was used as the ionization source by Agilent 6385UHD Accurate-Mass Q-TOF/MS G2 High Definition Mass Spectrometer. The separation was performed by an Agilent orbax Extend-C18 Column (2.1 mm × 50 mm, 1.8 μm) at 35 °C. In ESI-MS analysis, the parameters were set as source temperature at 120 °C, and desolvation gas temperature at 350 °C. In positive ion mode, the cone voltage was set as 40 V, the capillary voltage was set as 4.0 kV and the extraction cone voltage was set as 4.0 V. In negative ion mode, the cone voltage was set as 30 V, the capillary voltage was set as 3.5 kV and the extraction cone voltage was set as 5.0 V. To obtain the fragment information of the metabolites, argon was used as the collision gas in MS/MS mode and the collision energy was operated at alternative voltages of 10–70 eV. The mass data were analyzed by Agilent Mass Hunter Workstation software (version B.03.01).
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!