Abstract
Determination of prostaglandins by UV detection after chromatographic separation requires derivatization if detection at low wavelengths (190–195 nm) is to be avoided. The present study includes optimization of a previously reported one-step UV derivatization procedure in which prostaglandins are oxidized to their corresponding 15-oxo derivatives with pyridinium dichromate in acetonitrile. A dection limit for PGE 1, and PGE 2 of 0.14 pmol was obtained by using a photometric detector equipped with a cadmium UV emission source. As the emission energy maximum of cadmium coincides with the UV absorption maxima of the prostaglandin enone chromophores, signal-to-noise ratios were improved relative to conventional deuterium lamp spectrophotometric detectors. Further reduction of detection limits was obtained by using C 18 narrow- and micro-bore columns. 15-Oxo-PGF 2α, -PGF 1α, -PGE 2 and -PGE 1 were separated by isocratic elution on C 18 columns with acetonitrile—aqueous 10 m M phosphoric acid. Signal-to-noise ratios resulting from columns of different internal diameter as well as flow-cells of different volumes were compared.
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