Porous graphitic carbon chromatography-tandem mass spectrometry for the detection of isoprostanes in human cerebrospinal fluid
2005 (English)In: Journal of chromatography. B, ISSN 1570-0232, E-ISSN 1873-376X, Vol. 827, no 1, 39-43 p.Article in journal (Refereed) Published
F2-isoprostanes are produced by the non-enzymatic peroxidation of arachidonic acid in membrane phospholipids. This paper describes a new method for the determination of all four classes of F2-isoprostanes in human cerebrospinal fluid (CSF) involving separation on a 1 mm × 150 mm porous graphitic carbon (PGC) column and detection by triple quadrupole mass spectrometry in negative-ion electrospray mode. The sample pre-treatment consisted of an ultrafiltration step, following which 300 μl of CSF sample could be injected directly onto a 1 mm × 10 mm PGC guard column functioning as a trap for the analytes. The loading solvent was Milli-Q water at 125 μl/min. After 3 min, the sample was switched into the separation column. The F2-isoprostanes were separated in 20 min using a linear solvent gradient comprising water, methanol, acetonitrile and ammonium hydroxide at a pH of 9.5 and a flow of 50 μl/min The limit of detection (calculated as 3S/N) was approximately 40 pM (14 pg/ml). The assay was linear within the examined range (18–450 pg/ml), using CSF spiked with iPF2α-III standard (r2 > 0.995). Repeatability data were calculated for CSF spiked to 90 pg/ml and the relative standard deviation (RSD) obtained was 3% (n = 6).
Place, publisher, year, edition, pages
2005. Vol. 827, no 1, 39-43 p.
Isoprostanes; Porous graphitic carbon; Cerebrospinal fluid; Oxidative stress
IdentifiersURN: urn:nbn:se:su:diva-23522DOI: 10.1016/j.jchromb.2005.02.023OAI: oai:DiVA.org:su-23522DiVA: diva2:192468
Part of urn:nbn:se:su:diva-3222004-12-212004-12-212010-08-18Bibliographically approved