Plasma globotriaosylsphingosine (lyso-GB3) was measured by liquid chromatography tandem mass spectrometry (LC-MS/MS) using a published method with minor modifications [20 (link)]. Dimethyl psychosine was used as internal standard. Acquity UPLC BEH C18 column (2.1 mm X 50 mm with 1.7 μm particle size) was used for chromatography. Plasma glucosylsphingosine (lyso-GL1) was measured by a published LC-MS/MS method with minor modifications [21 (link)]. D5-glucosylsphingosine was used as internal standard. Lyso-GL1 is separated from its isomer galactosylsphingosine (psychosine) using Acquity UPLC BEH amide column (2.1 mm X 100 mm with 1.7 μm particle size). Agilent 6490 triple quadruple mass spectrometer coupled with Agilent 1290 liquid chromatography system was used for both analyses. Measurements were performed in cell pellets and culture supernatants as indicated in each figure.
Agilent 1290 liquid chromatography system
Manufactured by Agilent Technologies
Sourced in United States
The Agilent 1290 liquid chromatography system is a high-performance liquid chromatography (HPLC) instrument designed for analytical and preparative separations. It features advanced technology to deliver fast and precise chromatographic separations. The system includes a solvent delivery module, an autosampler, a column compartment, and a detector.
Automatically generated - may contain errors
Lab products found in correlation
5 protocols using agilent 1290 liquid chromatography system
1
Quantification of Plasma Glycosphingolipids
2
HPLC-MS/MS Analysis of Metabolites
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The supernatant was analyzed by HPLC-MS/MS on TripleTOF™ 6600plus mass spectrometer (AB SCIEX, USA), coupled to an Agilent 1290 liquid chromatography system (Agilent, USA). For LC separation, the ACQUITY UPLC, BEH C18 column was used. 5 μL sample was injected and separated with a 12 min gradient. The electrospray ionization mass spectra were acquired in positive and negative ion mode, respectively. The ion spray voltage was set to 5000 V for positive mode and 4000 V for negative mode.
3
Quantification of Intracellular Rifabutin in A. baumannii
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Extractions and measurements were done as previously described 33 (link),34 (link). Log-phase A. baumannii culture was incubated 0.79 or 0.38 μg/mL RBT in the presence or absence of amino acid mixture at 37 °C. Bacteria were harvested 0, 1, 8, and 24 hours and CFUs were determined by plating serial dilutions on agar media. The cell free supernatant was collected by filtration through a 0.22 μm filter. RBT were extracted by adding LC-MS grade acetonitrile:methanol:water (40:40:20) solution that was precooled to − 40 °C. Liquid chromatography mass spectrometry (LC-MS) differentiation and detection of RBT was performed using a Cogent Diamond Hydride Type C column (Microsolve Technologies) with an Agilent Accurate Mass 6230 TOF coupled with an Agilent 1290 Liquid Chromatography system as previously published.33 (link),35 (link) An isocratic pump was used for continuous infusion of a reference mass solution to allow mass axis calibration. Detected RBT ion was validated based on unique accurate mass-retention time identifiers for masses. RBT level was analyzed using Agilent Qualitative Analysis B.08.00 (Agilent Technologies) with a mass tolerance of <0.005 Da. The intracellular RBT was calculated as the [RBT]drug only control – [RBT]filtrate. 3 biological replicates were tested per group.
4
Quantification of Intracellular Rifabutin in A. baumannii
5
Untargeted Plasma Lipidomic Analysis
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The plasma lipidome of patients was determined using untargeted lipidomic analysis. The lipids were extracted based on a previously published and validated method [22 (link)]. Lipid extracts were analysed via ultrahigh-performance liquid chromatography (UHPLC) coupled with electrospray ionization quadrupole time of flight (ESI-Q-TOF) tandem mass spectrometry (MS/MS) according to a previously published method [23 (link), 24 (link)] using an Agilent 1290 liquid chromatography system (Agilent Technologies, Santa Clara, CA, USA) coupled with a 6520 ESI-Q-TOF mass spectrometer (Agilent Technologies, Santa Clara, CA, USA) was used. Data were acquired in both positive and negative ionization modes.
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!