Targeted Metabolomics Using LC-MS/MS

Samples were re-suspended in 20 μL HPLC-graded water for targeted tandem mass spectrometry (LC-MS/MS) and 5 μL were injected and analyzed using a hybrid 5500 QTRAP triple quadrupole mass spectrometer (AB/SCIEX) coupled to a Prominence UFLC system (Shimadzu) using an Amide HILIC column (Waters) and analyzed with selected reaction monitoring (SRM) with positive/negative polarity switching. Peak areas from the total ion current for each of 297 metabolite SRM transition were integrated using MultiQuant v2.1 software (AB/SCIEX)23 (link). The resulting raw data from the MultiQuant software was uploaded to MetaboAnalyst (http://www.metaboanalyst.ca/MetaboAnalyst) for subsequent data processing and analyses17 (link)24 (link)

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McKay T.B., Lyon D., Sarker-Nag A., Priyadarsini S., Asara J.M, & Karamichos D. (2015). Quercetin Attenuates Lactate Production and Extracellular Matrix Secretion in Keratoconus. Scientific Reports, 5, 9003.

Publication 2015

hybrid 5500 QTRAP triple quadrupole mass spectrometer Prominence UFLC system Amide HILIC column MultiQuant v2.1 software

Amide Hplc Hybrid Ion current Ms ms

Corresponding Organization :

Other organizations : University of Oklahoma Health Sciences Center, Dean McGee Eye Institute, Beth Israel Deaconess Medical Center, Harvard University

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Variable analysis

independent variables

Not explicitly mentioned

dependent variables

Peak areas from the total ion current for each of 297 metabolite SRM transition

control variables

Samples were re-suspended in 20 μL HPLC-graded water
5 μL were injected and analyzed using a hybrid 5500 QTRAP triple quadrupole mass spectrometer (AB/SCIEX) coupled to a Prominence UFLC system (Shimadzu) using an Amide HILIC column (Waters)
Analyzed with selected reaction monitoring (SRM) with positive/negative polarity switching

controls

Not explicitly mentioned
Not explicitly mentioned

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