Bile acid analysis in biological fluids: a novel approach.
In contrast to current methods of bile acid analysis that require the separation of bile acids into different groups prior to their analysis, the HPLC method using a reverse phase column and gradient elution that we developed permits the separation and detection of nonconjugated, glycine-conjugated, and esterified bile acids as their fluorescent dimethoxycoumarin esters. The mild conditions for ester formation make possible the identification of allylic bile acids characteristic of metabolic errors in bile acid synthesis. Quantification is obtained using 7 alpha,12 alpha-dihydroxy-5 beta-cholanoic acid as an internal standard. In addition to identification based on retention time, peak-shift strategy is used by treatment of aliquots with cholyglycine hydrolase and/or solvolysis. Loss of the parent peak and appearance of the derivative provide further assurance of the identity of each bile acid in biologic fluids that contain other organic acids.
Budai
K
K
Division of Hepatic Diseases, New York University Medical Center, NY 10016, USA.
Javitt
N B
NB
D000328:Adult; D001647:Bile Acids and Salts / Q000032:analysis* / Q000737:chemistry / Q000592:standards; D001826:Body Fluids / Q000737:chemistry; D002648:Child; D002793:Cholic Acids / Q000032:analysis; D002851:Chromatography, High Pressure Liquid / Q000379:methods* / Q000592:standards; D005069:Evaluation Studies as Topic; D005260:Female; D006801:Humans; D007202:Indicators and Reagents; D007231:Infant, Newborn; D012015:Reference Standards; D014556:Urine / Q000737:chemistry
D001647:Bile Acids and Salts; D002793:Cholic Acids; D007202:Indicators and Reagents