Quantitative analysis of fecal bile acids by gas-liquid chromatography

Makita, Masami; Wells, William W.

doi:10.1016/0003-2697(63)90072-1

Cited by 269 publications

(30 citation statements)

References 12 publications

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“…The germinated spores then pass through the ileum and finally into the anaerobic environment of the cecum. Here, certain members of the normal microbial flora metabolize the cholate derivatives that escape enterohepatic circulation to deoxycholate, a predominant bile salt in the feces of healthy humans (4,19,36). The deoxycholate produced prevents vegetative growth of C. difficile, and the host remains uncolonized.…”

Section: Discussionmentioning

confidence: 99%

Bile Salts and Glycine as Cogerminants for Clostridium difficile Spores

2008

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Spore formation by Clostridium difficile is a significant obstacle to overcoming hospital-acquired C. difficileassociated disease. Spores are resistant to heat, radiation, chemicals, and antibiotics, making a contaminated environment difficult to clean. To cause disease, however, spores must germinate and grow out as vegetative cells. The germination of C. difficile spores has not been examined in detail. In an effort to understand the germination of C. difficile spores, we characterized the response of C. difficile spores to bile. We found that cholate derivatives and the amino acid glycine act as cogerminants. Deoxycholate, a metabolite of cholate produced by the normal intestinal flora, also induced germination of C. difficile spores but prevented the growth of vegetative C. difficile. A model of resistance to C. difficile colonization mediated by the normal bacterial flora is proposed.

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Section: Discussionmentioning

confidence: 99%

Bile Salts and Glycine as Cogerminants for Clostridium difficile Spores

2008

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“…In the case of the mixed fecal cultures where the 7a-epimerizing conversion of chenodeoxycholic acid into ursodeoxycholic acid takes place together with the 3a-epimerization of these two acids (see "RESULTS"), the methylated samples were derivatized to trimethylsilyl ethers by the method of Makita and Wells (15), and the derivatives were chromatographed on a 3% Hi Eff-8B column maintained at 230 C. This procedure gives a good separation of the four epimeric 3, 7-diols, while the chromatography of methyl esters on QF-l does not distinguish between chenodeoxycholic acid and 3p-epimerized ursodeoxycholic acid due to the closeness of retention times (see Fig. 1).…”

Section: Methodsmentioning

confidence: 99%

Transformation of Bile Acids by Mixed Microbial Cultures from Human Feces and Bile Acid Transforming Activities of Isolated Bacterial Strains

Hirano

Masuda

Oda

et al. 1981

Microbiology and Immunology

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Microbial transformation of cholic acid and chenodeoxycholic acid by anaerobic mixed cultures of human fecal microorganisms was investigated, and the results were examined in relation to the bile acid transforming activities of 75 bacterial strains isolated from the same fecal cultures. The reactions involved in the mixed cultures were dehydrogenation and dehydroxylation of the 7a-hydroxy group in both primary bile acids and epimerization of the 3a-hydroxy group in all metabolic bile acids. Extensive epimerization of the 7a-hydroxy group of chenodeoxycholic acid yielding ursodeoxycholic acid was also demonstrated by certain fecal samples. 7a-Dehydrogenase activity was widespread among the fecal isolates (88% of 16 facultative anaerobes and 51% of 59 obligate anaerobes), and 7a-dehydroxylase activity was revealed in one of the isolates, an unidentified gram-positive nonsporeforming anaerobic bacterium. 3a-Epimerization was effected by seven strains assigned to Eubacterium lentum, which were also active for 3a-and 7a-dehydrogenations. No microorganism accounting for 7a-epimerization was recovered among the isolates. Splitting of conjugated bile acid was demonstrated by the majority of obligate anaerobes but the activity was rare among facultative anaerobes.Most of the microbial transformations of bile acids in the intestinal tract are reproducible in in vitro cultures offecal microorganisms. The major reactions involved are splitting of conjugated bile acids (deconjugation), removal of the hydroxy group, mainly at C-7 (7a-dehydroxylation), oxidation of hydroxy groups at C-7, C-3, and C-12 (the respective dehydrogenations) and conversion of the hydroxy group at C-3 from a-to ,B-configuration (3a-epimerization) (19,20). With regard to the kinds of microorganisms responsible for the respective reactions, however, only a few systematic studies have been attempted. Midtvedt and Norman (17) investigated the bile acid transforming capacities of 61 laboratory strains of intestinal origin, and Dickinson et al (3) evaluated the activities of 112 isolates from the digestive tract of the rat. They found the deconjugating and dehydrogenating 271

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“…Each received a standard 2-day repeating diet (calculated from an age, height, and weight nomogram). For the gallstone patients, the diet averaged 22 kcal/kg body weight (range, [16][17][18][19][20][21][22][23][24][25][26][27][28][29][30][31][32][33][34] kcal/kg);…”

Section: Methodsmentioning

confidence: 99%

“…A second sample of bile was saponified and esterified without prior hydrolysis. One portion of the methyl ester fraction was derivatized to trimethylsilyl ethers and chromatographed on a HiEff-8BP column (18); with this derivative and column, lithocholic acid has a retention time greater than that of any of the common bile acids. A second portion was acetylated and chromatographed on a cyanosilicone column (15); with this derivative and column, lithocholic acid has a retention time less than that of any of the common bile acids.…”

Section: Methodsmentioning

confidence: 99%