Second derivative-high-performance liquid chromatographic-fluorometric detection of porphyrins in chick embryo liver cell culture medium.
A high-performance liquid chromatographic system is described which is suitable for the separation and quantitative determination of a mixture of porphyrin esters in nanogram quantities from the culture medium used for maintenance of a monolayer culture of chick embryo liver cells. The desired sensitivity was obtained by coupling a high-performance liquid chromatograph with a second derivative-fluorometric detection system. The most effective method for preparation of the porphyrin methyl esters prior to chromatography was found to be lyophilization of the culture medium prior to esterification with 5% sulphuric acid in methanol.
D000818:Animals; D002478:Cells, Cultured; D002642:Chick Embryo; D002645:Chickens; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D003470:Culture Media / Q000032:analysis; D008099:Liver / Q000032:analysis / Q000196:embryology*; D011166:Porphyrins / Q000032:analysis*
D003470:Culture Media; D011166:Porphyrins