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Porous graphitic carbon chromatography-tandem mass spectrometry for the detection of isoprostanes in human cerebrospinal fluid
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- Claeson Bohnstedt, Kristina (author)
- Stockholms universitet,Institutionen för analytisk kemi
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- Karlberg, Bo (author)
- Stockholms universitet,Institutionen för analytisk kemi
- Basun, Hans (author)
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- Schmidt, Staffan (author)
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- (creator_code:org_t)
- Elsevier BV, 2005
- 2005
- English.
- In: Journal of chromatography. B. - : Elsevier BV. - 1570-0232 .- 1873-376X. ; 827:1, s. 39-43
- Journal article (peer-reviewed)
Abstract
Subject headings
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- 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).
Subject headings
- NATURVETENSKAP -- Kemi -- Analytisk kemi (hsv//swe)
- NATURAL SCIENCES -- Chemical Sciences -- Analytical Chemistry (hsv//eng)
Keyword
- Isoprostanes; Porous graphitic carbon; Cerebrospinal fluid; Oxidative stress
- Analytical chemistry
- Analytisk kemi
Publication and Content Type
- ref (subject category)
- art (subject category)
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