Universität zu Köln

Kohler, Maxie, Thomas, Andreas ORCID: 0000-0003-1199-0743, Walpurgis, Katja, Schaenzer, Wilhelm and Thevis, Mario (2010). Mass spectrometric detection of siRNA in plasma samples for doping control purposes. Anal. Bioanal. Chem., 398 (3). S. 1305 - 1313. HEIDELBERG: SPRINGER HEIDELBERG. ISSN 1618-2642

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Abstract

Small interfering ribonucleic acid (siRNA) molecules can effect the expression of any gene by inducing the degradation of mRNA. Therefore, these molecules can be of interest for illicit performance enhancement in sports by affecting different metabolic pathways. An example of an efficient performance-enhancing gene knockdown is the myostatin gene that regulates muscle growth. This study was carried out to provide a tool for the mass spectrometric detection of modified and unmodified siRNA from plasma samples. The oligonucleotides are purified by centrifugal filtration and the use of an miRNA purification kit, followed by flow-injection analysis using an Exactive mass spectrometer to yield the accurate masses of the sense and antisense strands. Although chromatography and sensitive mass spectrometric analysis of oligonucleotides are still challenging, a method was developed and validated that has adequate sensitivity (limit of detection 0.25-1 nmol mL(-1)) and performance (precision 11-21%, recovery 23-67%) for typical antisense oligonucleotides currently used in clinical studies.

Item Type:	Journal Article
Creators:	Creators Email ORCID ORCID Put Code Kohler, Maxie UNSPECIFIED UNSPECIFIED UNSPECIFIED Thomas, Andreas UNSPECIFIED orcid.org/0000-0003-1199-0743 UNSPECIFIED Walpurgis, Katja UNSPECIFIED UNSPECIFIED UNSPECIFIED Schaenzer, Wilhelm UNSPECIFIED UNSPECIFIED UNSPECIFIED Thevis, Mario UNSPECIFIED UNSPECIFIED UNSPECIFIED
URN:	urn:nbn:de:hbz:38-494327
DOI:	10.1007/s00216-010-4013-0
Journal or Publication Title:	Anal. Bioanal. Chem.
Volume:	398
Number:	3
Page Range:	S. 1305 - 1313
Date:	2010
Publisher:	SPRINGER HEIDELBERG
Place of Publication:	HEIDELBERG
ISSN:	1618-2642
Language:	English
Faculty:	Unspecified
Divisions:	Unspecified
Subjects:	no entry
Uncontrolled Keywords:	Keywords Language SMALL INTERFERING RNA; CHEMICAL-MODIFICATION; CIRCULATING MICRORNAS; MYOSTATIN; STABILITY; ANTISENSE; OLIGONUCLEOTIDES; QUANTIFICATION; THERAPEUTICS; BARRIERS Multiple languages Biochemical Research Methods; Chemistry, Analytical Multiple languages
URI:	http://kups.ub.uni-koeln.de/id/eprint/49432