Determination of oestrone, $17{\alpha}$-and $17{\beta}$-oestradiol in bovine aqueous humor using gas chromatography-negative ion chemical ionization mass spectrometry

Perfluorotolyl (PFT)-ether and perfluorotoly-trimethylsilyl (PFT-TMS) ether derivatives of oestrone, $17{\alpha}$- and $17{\beta}$oestradiol were prepared under phase transfer conditons. The former derivatives under negative ion chemical ionization conditions gave significant ions in the mass spectrometer but $17{\alpha}$- and $17{\beta}$ -oestradiol gave poor resolution. However, the PFT-TMS derivatives of 17.${\alpha}$- and$17{\beta}$-oestradiol showed good resolution. These derivatives were used for the analysis of oestrogens in bovine aqueous humour, vitreous humour and retina. The mean $({\pm}SEM)$ concentrations of oestrone in bovine aqueous humour (n=18), vitreous humour (n=18) and bovine retina (n=4) were $0.47{\pm}0.11$, $0.46{\pm}0.14$ and $1.10{\pm}0.24 ng.ml^{-1}$, respectively. $17{\alpha}$-Oestradiol was detected in 16 out of 18 samples of bovine aqueous humour and vitreous humour and the mean $({\pm}SEM)$ concentrations were $0.30{\pm}0.10$ and $0.08{\pm}0.02 ng.ml^{-1}$, respectively. The mean $({\pm}SEM)$ concentration of 17.betha.-oestradiol in aqueous humour (n=7) and vitreous homour (n=11) $0.83{\pm}0.26 ng ml^{-1}$ and $0.39{\pm}0.09 ng ml^-1$, respectively. In retina the concentrations of both steroids were below the detection limit.