Metabolomic Analysis of Cell Samples

Cell samples were resuspended in high-performance liquid chromatography (HPLC) grade water for mass spectrometry as described previously [14 (link)]. Briefly, the solutions were injected and analyzed using a hybrid 5500 QTRAP triple quadrupole mass spectrometer (AB/SCIEX) coupled to a Prominence UFLC HPLC system (Shimadzu) via selected reaction monitoring (SRM) of a total of 274 unique endogenous water-soluble metabolites for steady-state analyses of samples. Some metabolites were targeted in both positive and negative ion mode for a total of 306 SRM transitions using positive/negative ion polarity switching. Peak areas from the total ion current for each metabolite SRM transition were integrated using MultiQuant v2.1 software (AB/SCIEX). Comprehensive metabolomic data analysis was performed using MetaboAnalyst 4.0 [104 (link)].

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Zhang W.C., Skiados N., Aftab F., Moreno C., Silva L., Corbilla P.J., Asara J.M., Hata A.N, & Slack F.J. (2022). MicroRNA-21 guide and passenger strand regulation of adenylosuccinate lyase-mediated purine metabolism promotes transition to an EGFR-TKI-tolerant persister state. Cancer Gene Therapy, 29(12), 1878-1894.

Publication 2022

5500 QTRAP triple quadrupole mass spectrometer Prominence UFLC HPLC system MultiQuant v2.1 software

Cell High performance liquid chromatography Hybrid Ion current Mass spectrometry

Corresponding Organization :

Other organizations : Beth Israel Deaconess Medical Center, Harvard University, University of Central Florida, Massachusetts General Hospital

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Variable analysis

independent variables

Not explicitly mentioned

dependent variables

Steady-state levels of 274 unique endogenous water-soluble metabolites

control variables

Cell samples were resuspended in high-performance liquid chromatography (HPLC) grade water
Metabolites were analyzed using a hybrid 5500 QTRAP triple quadrupole mass spectrometer (AB/SCIEX) coupled to a Prominence UFLC HPLC system (Shimadzu) via selected reaction monitoring (SRM)
Peak areas from the total ion current for each metabolite SRM transition were integrated using MultiQuant v2.1 software (AB/SCIEX)
Comprehensive metabolomic data analysis was performed using MetaboAnalyst 4.0

controls

No positive or negative controls were explicitly mentioned in the provided information.

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