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An accurate and sensitive method by high-performance liquid chromatography connected with 3 alpha-hydroxysteroid dehydrogenase immobilized column for the measurement of urinary bile acids of noncholestatic and cholestatic infants was performed in this study. The purification and group separation of urinary bile acids were done through a piperidinohydroxypropyl Sephadex LH-20 column. By ultraviolet spectromonitor, non 3 alpha-hydroxy bile acids such as 3 beta-hydroxy-5-cholenate were determined, and 3 alpha-hydroxy bile acids by fluorescence spectromonitor. Urinary bile acids in the specimens obtained from noncholestatic and cholestatic infants were determined by this procedure. Major bile acids were cholate, chenodeoxycholate conjugates and cholate unconjugate in healthy infants. In cholestatic infants, total and individual bile acids were extremely increased, and 3 beta-hydroxy-5-cholenate conjugates appeared in the sulfated fraction.

Direct measurement of urinary bile acids of infants by high-performance liquid chromatography connected with an enzyme immobilized column.