Bile acids thin layer chromatography

Kazumo, T., Thin-layer chromatography of bile acids, Kagaku No Ryoiki Zokan, 64, 19, 1964. 

Hara, S. and Takeuchi, M., Systematic analysis of bile acids and their derivatives by thin layer chromatography, J. Chromatogr., 11, 565, 1963 Chem. Abs., 60, 838f, 1964. 

Anthony, W.L. and Beher, W.T., Color detection of bile acids using thin layer chromatography,. /. Chromatogr., 13, 570, 1964 Chem. Abs., 60, 13546c, 1964. 

Bll. Batta, A. K., Salen, C., and Shefer, S., Thin-layer chromatography of conjugated bile acids. J. Chromatogr. 168, 557-561 (1979). 

Iwata, T., and Yamasaki, K., Errzymatic deterrrtination and thin-layer chromatography of bile acids in blood. J. Biodtem. (Tokyo) 56, 424-431 (1964). 

O Moore, R. R. L., and Percy-Robb, I. W., Analysis of bile acids and their conjugates in jejunal juice by thin-layer chromatography and direct densitometry, Clin. Chim. Acta 43, 39-47 (1973). 

P4. Panveliwalla, D., Lewis, B., Wootton, I. D. P., and Tabaqchali, S., Determination of individual bile acids in biological fluids by thin-layer chromatography and fluorimetry. /. Clin. Pathol. 23, 309-314 (1970). 

Shepherd, R. W., Bunting, P. S., Khan, M., Hill, J. G., Soldin, S. J., and Gall, D. G., A rapid, sensitive method for accurate analysis of individual bile acids in biological fluids by high performance thin-layer chromatography and densitometry. CUn. Biochem. (Ottawa) 11, 106-111 (1978). 

Touchstone, J. C., Levitt, R. E., Levin, S. S., and Soloway, R. D., Separation cf conjugated bile acids by reverse-phase thin-layer chromatography. Lipids 15, 386-387... 

Van den Ende, A., Radecker, C. E., and Mairuhu, W. M., Microanalysis of free and conjugated bile acids by thin-layer chromatography and in situ spectrofluorimetry. Anal. Biochem. 134, 153-162 (1983). 

Figure 8 Influence of linker structure in HMG-CoA reductase inhibitor-bile acid prodrugs on the biliary secretion profile. The bile acid conjugates of the HMG-CoA reductase inhibitor HR 780 (1 mM) dissolved in 10 mM Tris/Hepes buffer (pH 7.4)/ 300mM mannitol/5% ethanol were injected as bolus into a mesenteric vein of anesthetized rats. After cannulation of the common bile duct, bile was fractionated and analyzed for the prodrugs or metabolites by thin-layer chromatography. (Adapted from Ref. 27.)...

Separation of Bile Acids by Normal-Phase Thin Layer Chromatography (NP-TLC) on Unmodified Silica Gel and Mixtures of Silica Gel and Kieselguhr at Room Temperature "" ... 

Fig. 2 The 1-D (A) and 2-D (B) chromatograms of the mixture of the seven bile acids investigated on thin-layer chromatography (TLC) silica gel 60 (E. Merck, 1.05553). I. Eluent n-hexane-ethyl acetate-acetic acid (25 20 5, v/v). II. Eluent chloroform-n-butanol-acetic acid-water (2 32 2 2, v/v) M represents bile acids mixture where 1 indicates chenodeoxycholic acid, 2 glyco-deoxycholic acid, 3 deoxycholic acid, 4 lithocholic acid, 5 cholic acid, 6 glycocholic acid, and 7 glycolithochoUc acid.

SEPARATION OF BILE ACIDS BY NORMAL PHASE HIGH-PERFORMANCE THIN-LAYER CHROMATOGRAPHY (NP-HPTLC) ON CYANO-AND DIOL-MODIFIED SILICA GEL AT ROOM... 

Zarzycki, P.K. Wierzbowska, M. Lamparczyk, H. Retention and separation studies of cholesterol and bile acids using thermostated thin-layer chromatography. J. Chromatogr. A, 1999, 857, 255-262. 

Pyka, A. Dolowy, M. Separation of selected bile acids by TLC. VII. Separation by reversed partition high performance thin layer chromatography, J. Liq. Chromatogr. Relat. Technol. 2005, 28 (10), 1573-1581. 

To recover bile acids or their conjugates from biological fluids, such as bile or intestinal contents, several different procedures are available. When the bile acid concentration is sufficiently high (above 1 meq/liter), as, e.g., in bile, 5-100 p. of the fluid may be directly applied to the starting line for paper or thin-layer chromatography. When low concentrations of bile acids are present, bile may be added drop by drop into 10-20 volumes of ethanol... 

TABLE IV. Thin-Layer Chromatography of Bile Acids on Silica Gel G ... 

TABLE VII. Thin-Layer Chromatography of Disubstitiited Bile Acids... 

In a recent study Hofmann et al. (116) studied the bile acid composition of bile from germ-free rabbits. Deproteinized bile was first analyzed by thin-layer chromatography for a tentative identification of the conjugated bile acids. After alkaline hydrolysis and methylation, bile acid methyl esters were analyzed by thin-layer and gas chromatography. Trifluoroacetate esters and trimethylsilyl ethers of methyl cholanoates were run on QF-1 or Hi-Eff-8B columns, respectively. Gas chromatography-mass spectrometry was used for the final identifications. 

In quantitative studies, Stiehl et al. (91) extracted bile acids from duodenal bile with methanol-acetone and the conjugates were hydrolyzed according to the method of Nair et al. (118). The free bile acids were then separated with thin-layer chromatography and determined spectrophoto-metrically after reaction with a sulfuric acid reagent. 

After deproteinization of bile, Turnberg and Anthony-Mote (94) were able to quantitate bile acids directly with a NAD-dependent steroid oxido-reductase from a Pseudomonas strain. The suitable concentration range was 10-80 mg of bile acids per 100 ml. The amounts of individual bile acids could be determined after preparative thin-layer chromatography, either before or after hydrolysis of bile acid conjugates. 

Quantitative recoveries of endogenously labeled bile acids in homogenized human feces can be obtained by continuous extraction for 48 hr with hot chloroform-methanol, 1 1 (18). After saponification, acidification, and continuous diethyl ether extraction, the bile acids are purified on silicic acid (Section IIIB 4 and Ref. 18) to give one mono- and disubstituted, and one trisubstituted bile acid fraction. For identification purposes further subfractionation can be made [see Table V (69, 77, 126, 127)]. The subfrac-ticns are subsequently subjected to small-scale preparative thin-layer chromatography of methylated bile acids. The fractions eluted from the thin-layer plates are next subjected to peak-shift analyses followed by final identification by gas chromatography-mass spectrometry. When the fecal bile acid composition has been elucidated in this way the mono-, di- and trisubstituted bile acids from the first silicic acid column may be quantitated after methyla-tion and by analysis on QF-1. These results are then compared with those obtained after trifluoroacetylation of the bile acid methyl esters. (18). 

Ali et al. (128 and 129) suspended freeze-dried human feces in alkaline-50 % ethanol and extracted with petroleum ether-diethyl ether, 1 1, to remove neutral lipids. The bile acids in the aqueous phase were next subjected to conditions for hydrolysis of conjugates. After acidification, the free bile acids were extracted with diethyl ether, methylated, and purified by preparative thin-layer chromatography. Mono-, di-, and trisubstituted methyl cholanoates are eluted together from the thin-layer plate and then quantitated on QF-1 columns before and after conversion into trifluoroacetates. 

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