A rapid, sensitive method for accurate analysis of individual bile acids in biological fluids by high performance thin-layer chromatography and densitometry.
1. A rapid new micromethod for quantitative analysis of individual bile acids in duodenal juice by high performance thin-layer chromatography (HPTLC) and densitometry is described and evaluated by comparison with standard TLC and spectrophotometry. 2. Advantages of HPTLC over TLC include more rapid separation, better resolution and more sensitive detection (5 - 10 fold), without the need for prior extraction. Densitometry provides simple, direct and rapid quantitation. 3. The method is accurate and reliable over a range of bile acid concentrations. In the 0.5 mM range, recovery was greater than 89%, and coefficients of variation for within-day analysis were 2 - 12% and for between-day analysis were 6 - 18% for the individual bile acids. Twenty analyses can be performed by one worker in a single day. 4. We conclude that the method offers several advantages over most currently described techniques, is suitable for routine use and is deserving of wider application.
D001647:Bile Acids and Salts / Q000032:analysis*; D002635:Chenodeoxycholic Acid / Q000032:analysis; D002793:Cholic Acids / Q000032:analysis; D002855:Chromatography, Thin Layer / Q000379:methods*; D003720:Densitometry / Q000379:methods; D003840:Deoxycholic Acid / Q000032:analysis; D004386:Duodenum; D005069:Evaluation Studies as Topic; D005999:Glycochenodeoxycholic Acid / Q000032:analysis; D006000:Glycocholic Acid / Q000032:analysis; D006801:Humans; D007223:Infant; D007419:Intestinal Secretions / Q000032:analysis*; D008095:Lithocholic Acid / Q000032:analysis; D013056:Spectrophotometry, Ultraviolet; D013655:Taurochenodeoxycholic Acid / Q000032:analysis; D013656:Taurocholic Acid / Q000032:analysis
D001647:Bile Acids and Salts; D002793:Cholic Acids; D003840:Deoxycholic Acid; D002635:Chenodeoxycholic Acid; D013655:Taurochenodeoxycholic Acid; D013656:Taurocholic Acid; D008095:Lithocholic Acid; D005999:Glycochenodeoxycholic Acid; D006000:Glycocholic Acid