Is n-pentane really an index of lipid peroxidation in humans and animals? A methodological reevaluation.
Volatile hydrocarbons such as ethane and n-pentane are known to originate from peroxidation of polyunsaturated fatty acids in membrane lipids and they are accepted as a sensitive and direct index of lipid peroxidation both in vitro and in vivo. Until now, an appropriate and commonly accepted method for the analysis of volatile hydrocarbons in exhalation air has not been described. We therefore developed a methodology for routine application in humans that is based on cryofocusing in combination with gas chromatography and is adaptable to mass spectrometry. The samples may be stored in stainless steel bombs up to 3 weeks, and sample volumes necessary to analysis are variable and can be adapted to analytical requirements. The interference by water and carbon dioxide, always present in excess, is strongly reduced. Mass spectroscopic analysis of exhalation air in human control subjects demonstrates, however, the presence of isoprene as the major constituent hitherto identified as n-pentane. The commonly used columns fail to separate n-pentane and isoprene. Based upon studies of the diverse methodologies reported in literature, it must be assumed that the reported responses of the gas chromatographic "n-pentane" peak in exhalation air of humans and animals, hitherto identified exclusively by authentic reference gases, are actually responses to isoprene or, at least, a mixture of both n-pentane and isoprene.
University of Heidelberg, Faculty of Medicine, Children's Hospital, Central Laboratory of Clinical Chemistry, Germany.
D
D
Kohlmüller
W
W
Kochen
D000328:Adult; D000393:Air Pollutants / Q000032:analysis*; D000818:Animals; D001944:Breath Tests / Q000379:methods; D002070:Butadienes / Q000032:analysis; D002849:Chromatography, Gas / Q000379:methods; D005069:Evaluation Studies as Topic; D045782:Hemiterpenes; D006801:Humans; D015227:Lipid Peroxidation / Q000502:physiology*; D013058:Mass Spectrometry / Q000379:methods; D010420:Pentanes / Q000032:analysis*
D000393:Air Pollutants; D002070:Butadienes; D045782:Hemiterpenes; D010420:Pentanes; C005059:isoprene; C033353:pentane