No abstract

A simple, precise and sensitive method for separation and determination of total bile acid sulfates in human urine is described. The sulfate fraction of urinary bile acids was separated with lipophilic anion exchange gel, piperidinohydroxypropyl Sephadex LH-20 after sample clean-up with Sep-Pak C18 cartridge. The obtained sulfate fraction was submitted to solvolysis with a small volume of dimethoxypropane-HC1 solution and subjected to enzymatic-fluorimetrical assay using 3a-hydroxysteroid dehydrogenase and resazurin. In this method, no influence of existing salts in the reaction mixture on fluorescence intensity was observed and solvolysis reaction was almost complete. Overall recoveries of glycine-and taurine-conjugated bile acid 3-sulfates from normal urine ranged from 90.5 to 93.7% and those of unconjugates from 48.7 to 78.0%. The sensitivity of the described method enabled to estimate total bile acid sulfates with 0.5 ml of normal urine and precision tests showed the satisfactory accurasy. The level of total urinary bile acid sulfates was estimated on some patients with hepatobiliary diseases and healthy subjects. bile acid sulfates ; 3a-hydroxysteroid dehydrogenase ; solvolysis ; piperidinohydroxypropyl Sephadex urine Since the sulfate ester of bile acid in man was initially described (Palmer 1967), the metabolic significance of bile acid sulfates in hepatobiliary diseases has been discussed by many investigators (Stiehl 1974;Stiehl et al. 1980Stiehl et al. , 1982Makino et al. 1975;Van Berge Henegouwen et al. 1976 ;Summerfield et al. 1977 ;Bremmelgaard and Sjovall 1979). The sulfate group enhances the water solubility of bile acids rendering them easy to excrete in the urine. From these facts, the role of sulfation is considered to be one of protection against the toxity of bile acids and measurement of bile acid sulfates in biological materials gives important information regarding bile acid metabolism in pathological situations. Although the determination of bile acid sulfates has been performed by gas-liquid chromatography or gas-liquid chromatography mass-spectrometry, these methods are not

To elucidate the role of conjugated biliary bile acids in gallstone dissolution, the acids in bile were determined by high-performance liquid chromatography before and after the treatment with ursodeoxycholic acid for 3-26 months in patients with gallstone. The stone-dissolving effect of ursodeoxycholic acid was confirmed in 7 of 10 patients and the lithogenic index lowed significantly after the treatment.The compositions of cholate, chenodeoxycholate and ursodeoxycholate were about 33, 45 and 4%, respectively, in the control and pre-treatment groups. In the post-treatment group, a markedly low value was observed in primary bile acids both glycine-conjugates and taurine-conjugates, especially in cholate, with a significantly high value of ursodeoxycholate (p<0.001) of both conjugates. On the other hand, no difference was observed in the composition of deoxycholate with significantly low percentage of taurine-conjugates compared with that in the pre-treatment group. The ratio of glycine-to taurine-conjugated bile acids showed a significantly higher value in the post-treatment group than in the pretreatment group (p <0.001) and the control group (p<0.005). The bile specimens were measured concomitantly by gas-liquid chromatography and the results were compared with those of high-performance liquid chromatography. The mean value of total bile acids, the ratio of cholate to chenodeoxycholate and the ratio of glycine-to taurine-conjugated bile acids obtained by the former analysis procedure represented about 57, 80 and 115% of those of the latter. It is concluded that the high G/T value seems to have a role in the dissolution mechanism. bile acid; ursodeoxycholic acid; cholesterol gallstone; high-performance liquid chromatography; ratio of glycine-to taurine-conjugated bile acids (G/ T ratio)Since the dissolution of cholesterol gallstones by chenodeoxycholic acid (CDCA) (Bell et al. 1972;Danzinger et al. 1972) and ursodeoxycholic acid (UDCA) (Sugata and Shimizu 1974;Makino et al. 1975) was demonstrated, an attention has been

Total sulfated bile acids of forty two serum samples and eleven urine samples from eight patients with acute intrahepatic cholestasis were analized with a newly established enzymatic fluorimetrical assay. As the degree of cholestasis advanced, the level of total serum sulfated bile acids elevated, but increasing rate of them gradually decreased. At the advanced stage of cholestasis, the proportion of total sulfated bile acids in total serum bile acids was significantly lower than that of healthy subjects. The level of total serum sulfated bile acids highly correlated with urinary excretion of sulfated bile acids. The renal clearance of sulfated bile acids was not altered during the course of cholestasis and