The quantification of endogenous steroids in bovine aqueous humour and vitreous humour using isotope dilution GC-NCI-MS

Abstract

Pentafluorobenzyloxime-trimethylsilyl derivatives of androgens, progestogens and corticosteroids were prepared and used for the analysis of these steroids in bovine aqueous humour and vitreous humour by GC-MS method. Appropriate deuteriated isotopomers of the parent steroids were labelled with deuterium via simple synthetic procedure and used as internal standards. The concentration (ng ml(-1), +/- S.E.M.) of these steroids in bovine aqueous humour and vitreous humour were found to be as follow: (1) aqueous humour (n = 17); hydrocortisone (n = 17; 2.40 +/- 0.54), progesterone (n = 15; 0.06 +/- 0.01), 4-androstene-3,17-dione (n = 8; 0.15 +/- 0.07) and testosterone (n = 4; 0.14 +/- 0.04); and (2) bovine vitreous humour (n = 19); hydrocortisone (n = 19; 1.78 +/- 0.25), progesterone (n = 18; 0.09 +/- 0.01), 4-androstene-3,17-dione (n = 19; 0.11 +/- 0.02), 11-deoxycorticosterone (n = 12; 29.27 +/- 6.42), 17alpha-hydroxyprogesterone (n = 6; 5.55 +/- 3.12). The concentration of corticosterone, 11-deoxycorticosterone and 17alpha-hydroxyprogesterone and testosterone and corticosterone were below the limit of detection in aqueous humour and vitreous humour, respectively.