Ion‐pair high‐performance liquid chromatography of bile salt conjugates: Application to pig bile

Legrand-Defretin, Véronique; Juste, Catherine; Henry, Robert J.; Corring, T.

doi:10.1007/bf02536421

Cited by 36 publications

(11 citation statements)

References 40 publications

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“…After deconjugation, cholate becomes the main component. The three main conjugated bile salts of porcine bile were glycodeoxycholate, glycohyodeoxycholate (dihydroxyconjugated bile salts) and glycohyocholate (trihydroxyconjugated bile salt) ( Legrand‐Defretin et al . 1991 ).…”

Section: Discussionmentioning

confidence: 99%

Isolation and characterization of a Lactobacillus amylovorus mutant depleted in conjugated bile salt hydrolase activity: relation between activity and bile salt resistance

Grill¹,

Cayuela²,

Antoine³

et al. 2000

J Appl Microbiol

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. 2000. Growth experiments were conducted on Lactobacillus amylovorus DN-112 053 in batch culture, with or without pH regulation. Conjugated bile salt hydrolase (CBSH) activity was examined as a function of culture growth. The CBSH activity increased during growth but its course depended on bile salts type and culture conditions. A Lact. amylovorus mutant was isolated from the wild-type strain of Lact. amylovorus DN-112 053 after mutagenesis with N-methyl-N H -nitro-N-nitrosoguanidine. An agar plate assay was used to detect mutants without CBSH activity. In resting cell experiments, the strain showed reduced activity. Differences between growth parameters determined for wild-type and mutant strains were not detected. Comparative native gel electrophoresis followed by CBSH activity staining demonstrated the loss of proteins harbouring this activity in the mutant. Four protein bands corresponding to CBSH were observed in the wild-type strain but only one was detected in the mutant. The speci®c growth rate of the mutant strain was affected more by bile salts than the wild-type strain. Nevertheless, bile was more toxic for the wild-type strain. In viability studies in the presence of nutrients, it was demonstrated that glycodeoxycholic acid exerted a higher toxicity than taurodeoxycholic acid in a pH-dependent manner. No difference was apparent between the two strains. In the absence of nutrients, the wild-type strain died after 2 h whereas no effect was observed for the mutant. The de-energization experiments performed using the ionophores nigericin and valinomycin suggested that the chemical potential of protons (ZDpH) was involved in Lactobacillus bile salt resistance.

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Section: Discussionmentioning

confidence: 99%

Isolation and characterization of a Lactobacillus amylovorus mutant depleted in conjugated bile salt hydrolase activity: relation between activity and bile salt resistance

Grill¹,

Cayuela²,

Antoine³

et al. 2000

J Appl Microbiol

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“…Then, turning to a biomedical purpose, the pig was used as a model of gall-stone formation in humans and the group tried to develop cumbersome methods for purification and identification of the various constituents of bile possibly involved in the crystallisation of biliary cholesterol [82,118]. A model was developed to induce cholesterolic lithiasis in the pig fed a diet enriched with cholesterol and cyclodextrine, while the beneficial effect of soy proteins was shown to result from a change in the bile acid pool leading to reduced hydrophobicity [22,83].…”

Section: Gastric Emptyingmentioning

confidence: 99%

Forty years of achievement in french research on digestive physiology in the pig

Laplace¹,

Aumaı̂tre²,

Rérat³

2001

Reprod. Nutr. Dev.

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-The contribution of the research group set up by Rérat in the early 1960s in Jouy-en-Josas, and then expanded to Rennes, is summarised. As digestive processes are a major key to the nutritional status of monogastric animals, original methodologies based on advanced experimental surgery were developed in the pig to quantify the digestion and absorption yield, to know their factors of variation, and to understand their physiological and nutritional consequences. The group also extended its expertise to the control of food intake, the role of the nervous system and regulatory peptides, and several biomedical gut-related topics. pig / digestion / absorption / neurohumoral regulation / adaptationRésumé -Réalisations et succès de la recherche française : quarante ans de physiologie digestive chez le porc. Dès le début des années 60, un groupe de recherches a été constitué à l'initiative de A. Rérat à Jouy-en-Josas pour étudier divers aspects de la physiologie digestive chez le porc. Leur exploration a fourni des clés essentielles pour la maîtrise du statut nutritionnel chez cette espèce monogastrique, et un instrument d'amélioration de la production porcine. Ce groupe s'est élargi au fil des années et a même essaimé dans les années 80, notamment à Rennes. Il a apporté une contribution essentielle par le développement de méthodologies nouvelles généralement basées sur une chirurgie expérimentale très élaborée. Ces approches ont fourni de nombreux travaux originaux qui ont permis de décrire les processus de digestion (motricité, transit, sécrétions, hydrolyses enzymatiques

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“…These different mobilities can be interpreted rationally -addition of Me-[3-CD to the mobile phase results in the formation of inclusion complexes between the bile acids and the Me-[3-CD, and the presence of a 12Gt-hydroxyl group in the substrates might contribute to a weakening of complex formation [6,12]. This be- Table I.…”

Section: Resultsmentioning

confidence: 95%

“…This generalization suggests that conversion of polar and ionic compounds into less polar and nonionic derivatives results in increased hydrophobicity and thus greater solubility in the C18-bonded silica gel. Addition of a cationic ion-pair reagent such as TBAP, as a counter ion, to an aqueous mobile phase is expected to result in the formation of the corresponding nonionic hydrophobic complexes with anionic bile acid conjugates [12]. In fact, when 5 % 0.5 mol L -1 TBAP solution was added to the mobile phase the hydrophobicity of the bile acid conjugates increased significantly and the amount of water in the mobile phase could be much reduced, particularly for analysis of the double conjugates with taurine and sulfuric acid.…”

Section: Resultsmentioning

confidence: 99%

Separation of double conjugates of bile acids by two-dimensional high-performance thin-layer chromatography with tetra-n-butylammonium phosphate and methyl β-cyclodextrin

et al. 1999

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SummaryThis paper describes a method for the separation, by high-performance thin-layer chromatography (HPTLC), of a series of polar, ionic, and hydrophilic double conjugates of bile acids amidated at the C-24 carboxyl group with glycine or taurine and sulfonated or glucosylated at hydroxyl groups in the 513-steroid nucleus. The method involves two-dimensional (2D) reversed-phase (RP) HPTLC with the combined use of tetra-n-butylammonium phosphate (TBAP) and methyl as mobile phase additives. Complete separation of the hydrophilic bile acid conjugates, particularly of the recalcitrant pairs of chenodeoxycholic acid and deoxycholic acid conjugates in each group, was achieved by 2D inclusion RPHPTLC by developing with methanol-water-0.5 tool L -1 TBAP, 90:10:5-75:25:5 (v/v) in the first dimension and the same mobile phase containing 5 mM Meq3-CD in the second dimension. The method could be usefully applied to biosynthetic and metabolic studies of bile acids in biological materials.necessary. TLC is, therefore, widely used for the preliminary and/or routine analysis of a wide variety of compounds. Several studies have reported the TLC separation of biologically important bile acids in connection with hepatobiliary diseases [1][2][3]. The compounds separated include unconjugated cholic acid (CA), chenodeoxycholic acid (CDCA), ursodeoxycholic acid (UDCA), deoxycholic acid (DCA), and lithocholic acid (LCA), and their conjugates with glycine, taurine or sulfuric acid. As far as we are aware, however, there have been few reports of the TLC analysis of more polar and ionic double conjugates of bile acids, amidated at the C-24 carboxyl group with glycine (or taurine) and sulfonated or glycosylated at the hydroxyl groups in the 513-steroid nucleus, on normal phase (NP) TLC plates with alcohol-acetic acid-water mixtures as the mobile phase [4,5]. According to these papers, separation of all of the double-conjugated pairs of CDCA and DCA was unsuccessful. We have previously reported an efficient method for the separation of the five major bile acids in both the unconjugated and conjugated forms [6]. The method involved the use of two-dimensional (2D) reversed-phase (RP) high-performance thin-layer chromatography (HPTLC) [7,8] with aqueous methanol (10-30 %) in the first direction and the same mobile phase containing 5 mM methyl 13-cyclodextrin (Me-J3-CD) in the second direction. This paper described the application of this 2D inclusion RPHPTLC method to the complete separation of various types of double-conjugated bile acids.

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Ion‐pair high‐performance liquid chromatography of bile salt conjugates: Application to pig bile

Cited by 36 publications

References 40 publications

Isolation and characterization of a Lactobacillus amylovorus mutant depleted in conjugated bile salt hydrolase activity: relation between activity and bile salt resistance

Isolation and characterization of a Lactobacillus amylovorus mutant depleted in conjugated bile salt hydrolase activity: relation between activity and bile salt resistance

Forty years of achievement in french research on digestive physiology in the pig

Separation of double conjugates of bile acids by two-dimensional high-performance thin-layer chromatography with tetra-n-butylammonium phosphate and methyl β-cyclodextrin

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