GLC analysis of hydrocortisone, triamcinolone acetonide, and desonide in culture media of mouse and human dermal fibroblasts using flame-ionization detection.
A quantitative GLC assay with flame-ionization detection capable of detecting nanogram quantities of hydrocortisone, triamcinolone acetonide, and desonide in biological fluids was developed. This assay consisted of two extractions of the glucocorticoids from 1 N sodium chloride-treated cell culture media into ethyl acetate and subsequent double derivatization with methoxyamine and N-trimethylsilylimidazole. The chemical structures of methoxime-trimethylsilyl derivatives were confirmed by GLC-mass spectrometry. The methoxime-trimethylsilyl derivatives were stable for 24 hr. The applicability of this assay was demonstrated by studies of the glucocorticoid levels in L-929 and human dermal fibroblasts cell culture media over prolonged incubation (0--96 hr).
D000328:Adult; D000818:Animals; D002849:Chromatography, Gas; D003470:Culture Media; D003898:Desonide / Q000032:analysis*; D005347:Fibroblasts / Q000378:metabolism; D005410:Flame Ionization; D008401:Gas Chromatography-Mass Spectrometry; D005938:Glucocorticoids / Q000378:metabolism; D006801:Humans; D006854:Hydrocortisone / Q000032:analysis*; D008297:Male; D051379:Mice; D011246:Pregnadienetriols / Q000032:analysis*; D012867:Skin / Q000378:metabolism*; D014222:Triamcinolone Acetonide / Q000032:analysis*
D003470:Culture Media; D005938:Glucocorticoids; D011246:Pregnadienetriols; D014222:Triamcinolone Acetonide; D003898:Desonide; D006854:Hydrocortisone