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

Pentafkluorobenzyloxime-trimethylsilyl derivatives of androgens, progestoge ns and corticosteroids were prepared and used for the analysis of these ste roids in bovine aqueous humour and vitreous humour by GC-MS method. Appropr iate deuteriated isotopomers of the parent steroids were labelled with deut erium via simple synthetic procedure and used as internal standards. The co ncentration (ng ml(-1), +/- S.E.M.) of these steroids in bovine aqueous hum our 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 = 3; 0.14 +/- 0.03); and (2) bovine vitreous humour (n = 19). hydrocortiso ne (n = 19; 1.78 +/- 0.25), progesterone (n = 18; 0.09 +/- 0.01). 4-androst ene-3,17-dione (n = 19, 0.11 +/- 0.02), 11-deoxycorticosterone (n = 12; 29. 27 +/- 6.42), 17 alpha -hydroxyprogesterone (12 = 6; 5.55 +/- 3.12). The co ncentration of corticosterone, 11-deoxycorticosterone and 17 alpha -hydroxy progesterone and testosterone one and corticosterone were below the limit o f detection in aqueous humour and vitreous humour, respectively. (C) 2001 E lsevier Science B.V. All rights reserved.