Reiko Kiyonami1, Michael G. Harrington2, Alfred N. Fonteh2, Roger G. Biringer3 and Andreas F. Huhmer1
ASMS 2019 Poster
Purpose: Developing an unbiased discovery to targeted verification approach to detect low abundant endogenous cardiac steroids from biological fluids with high confidence on a Thermo Scientific™ Orbitrap ID-X™ Tribrid™ mass spectrometer.
Methods: Cerebrospinal fluid (CSF) samples from healthy participants were used as the model of biological fluids. High resolution MS and MS2/MS3 data were collected with an Orbitrap Tribrid mass spectrometer. The comprehensive structure relative fragment ion information from MS2 and MS3 was processed using Thermo Scientific™ Mass Frontier™ 8.0 software and Thermo Scientific™ Compound Discoverer™ 3.0 software for cardiac steroid annotation. A targeted PRM (parallel reaction monitoring) assay was developed based on the discovery phase results and used for confirming that the cardiac steroids annotated from the discovery phase could reproducibly detected across multiple CSF samples.
Results: Two endogenous cardiac steroids (Digoxigenin & Marinobufagenin) were detected with the discovery experiments and their existences across multiple CSF samples were confirmed with targeted verification experiments.
1. Thermo Fisher Scientific, San Jose, CA
2. Huntington Medical Research Institutes, Pasadena, CA
3. LECOM Bradenton, Bradenton, FL
United States
