Pharmaceutical Sciences Faculty Publications

The Depletion of Protein Signals in Metabonomics Analysis with the WET-CPMG Pulse Sequence

Document Type

Article

Publication Date

2-21-2003

Journal Title

Biochemical and Biophysical Research Communications

ISSN

0006-291X

Volume

301

Issue

4

First Page

952

Last Page

959

DOI

10.1016/s0006-291x(03)00079-2

PubMed ID

12589805

Abstract

Nuclear magnetic resonance (NMR) spectroscopy is a powerful analytical tool capable of providing a comprehensive metabolic profile of biofluids such as urine, plasma, and serum. Unfortunately, when measuring serum and plasma, the high protein concentration can obscure the signals originating from low molecular weight metabolites. We evaluated the use of different parameters within the Carr-Purcell-Meiboom-Gill (CPMG) pulse train of fast spin-echoes to remove the macromolecular signal contribution in one-dimensional proton (1H) NMR spectra. Experimental parameters such as the refocusing delay in the CPMG pulse train, pulse miscalibration, and recycle time were examined to assess the ability to remove the protein signals from the spectrum without causing a deleterious effect on the signals originating from free, low molecular weight metabolites. The 1H-NMR spectra of a variety of serum samples spiked with 2'-deoxyadenosine were acquired using various acquisition parameters. Our results show that the delay used in the CPMG spin-echo and the combination of the acquisition pulse flip angle and recycle time are the two major factors affecting the observed metabolite signal amplitudes in the resulting 1H-NMR spectrum.

Keywords

Body fluids, deoxyadenosines, humans, nuclear magnetic resonance, biomolecular, proteins, protons, serum albumin

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