Rapid quantification of 11 prostanoids by combined capillary column gas chromatography and negative ion chemical ionization mass spectrometry: Application to prostanoids released from normal human embryonic lung fibroblasts WI38 in a culture medium

The rapid and simultaneous quantification of 11 prostanoids has been carried out with a short‐capillary gas chromatograph and negative ion chemical ionization (ammonia) mass spectrometer. The methoxime‐trimethylsilyl ether‐pentafluorobenzyl esters (MO‐TMS‐PFB) of nine prostanoids, PGA 1 , PGA 2 , PGB 1 , PGB 2 , PGD 2 , PGE 1 , PGE 2 , 6‐oxo‐PGF 1α and TXB 2 and the TMS‐PFB of two prostanoids, PGF 1α and PGF 2α , were separated in less than 5.5 min on a bonded OV‐1 capillary column 0.25 mm i.d. × 6 m (0.15 μm thickness) using hydrogen as a carrier gas. PGD 2 , PGE 2 , PGF 2α , 6‐oxo‐PGF 1α and TXB 2 were quantified up to 2.5 fmol injected (0.1 pmol derivatized) and both PGA 2 and PGB 2 up to 25 fmol injected (1 pmol derivatized). In order to maintain the stability of the prostanoids containing a carbonyl group, such as TXB 2 during the purification and derivatization steps of biological materials, methyl acetate was used in place of methyl formate as an eluant for Sep‐Pak C 18 purification. Normal human embryonic lung fibroblasts W138 (5.63 × 10 5 cells in a log phase) produced: PGA 2 15.28, PGB 2 13.48, PGD 2 7.95, PGE 1 2.62, PGE 2 177.76, PGF 2α 25.14, 6‐oxo‐PGF 1α 27.33 and TXB 2 61.00 pmol in 10 ml of Eagle minimal essential medium.