Quantitative analysis of bile acids in serum and bile, using gas -liquid chromatography

Henegouwen, G. P. van Berge; Ruben, A. Th.; Brandt, K H

doi:10.1016/0009-8981(74)90243-5

Cited by 110 publications

(18 citation statements)

References 14 publications

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“…However, these authors expressed concern at over-estimation by their procedure when compared with a GLC procedure. Cholate concentrations in normal sera also showed good agreement with reported concentrations determined by GLC (Makino et al, 1969;Laatikainen and Hesso, 1975;Ross et al, 1977) but were lower than the range reported by van Berge Henegouwen et al (1974). The radioimmunoassays of Simmonds et al (1973) and Matern et al (1976) gave similar ranges for normal subjects.…”

Section: Discussionsupporting

confidence: 78%

“…Chenodeoxycholate concentrations in normal sera are similar to those reported by GLC (van Berge Henegouwen et al, 1974;Laatikainen and Hesso, 1975;Ross et al, 1977) but are lower than those reported by Schalm et al (1977) using a similar radioimmunoassay procedure. However, these authors expressed concern at over-estimation by their procedure when compared with a GLC procedure.…”

Section: Discussionsupporting

confidence: 78%

“…Sensitive assays for individual serum bile salt conjugates may provide further useful diagnostic information , but until recently only gas liquid chromatography (GLC) was capable of assaying individual bile salts in serum (van Berge Henegouwen et al, 1974;Ross et al, 1977). This methodology is technically complex and time-consuming, and consequently there is the need for a rapid assay procedure that demonstrates sensitivity, precision, and accuracy similar to gas liquid chromatography.…”

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confidence: 99%

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Radioimmunoassay of primary bile salts in serum.

Baqir¹,

Murison²,

Ross³

et al. 1979

J Clin Pathol

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SUMMARY Rapid, sensitive radioimmunoassays have been developed for the conjugated primary bile salts, cholate and chenodeoxycholate, using immunogens prepared by the mixed anhydride procedure. Antibodies produced showed equal specificity for glycine and taurine conjugates. Crossreactivities were comparable with those from other published radioimmunoassays. The assays were routinely performed on unextracted sera and the concentrations correlated well with concentrations determined by gas-liquid chromatography.Accuracy, determined by the addition of bile salt to charcoal-extracted serum, and precision, determined by replicate analysis of a normal sample, were both less than + 10 %. These figures are comparable with those obtained by both gas-liquid chromatography and other radioimmunoassays for bile salts.Normal sera were found to contain 0-49-1-32 ,umol/l of cholate and 0-55-2 02 ,mol/l of chenodeoxycholate. Serum concentrations in patients with liver disease were higher than this normal range. Three patients with mild liver disturbance were found to have one bile salt in the upper limit of normal, but in each case the other primary bile salt was outwith the normal range.Elevation of serum bile salt concentration in hepatobiliary disease (Sherlock and Walshe, 1948) has been shown to be a sensitive test for liver disease (Korman et al., 1974). Sensitive assays for individual serum bile salt conjugates may provide further useful diagnostic information , but until recently only gas liquid chromatography (GLC) was capable of assaying individual bile salts in serum (van Berge Henegouwen et al., 1974;Ross et al., 1977). This methodology is technically complex and time-consuming, and consequently there is the need for a rapid assay procedure that demonstrates sensitivity, precision, and accuracy similar to gas liquid chromatography. Radioimmunoassay provides such a potential, and several methods have now been reported for bile salt conjugates (Simmonds et al., 1973;Murphy et al., 1974 acid, 5-0 Ci/mmol specific activity, were supplied by New England Nuclear Ltd. Non-radioactive bile salts were analysed by GLC and thin-layer chromatography (TLC) and radioactive bile salts by TLC alone. No impurities exceeding 1 % were found, and purification was not attempted. Scintillation liquid NE260 was obtained from Nuclear Enterprises Ltd, Freund's complete adjuvant from Difco Laboratories Ltd, bovine serum albumin from Hoechst Pharmaceuticals Ltd, and porcine y-globulin from Koch-Light Ltd. Activated charcoaluntreated powder (230-250 mesh size) was supplied by Sigma Chemical Co Ltd. All other chemicals were analytical grade. REAGENTSAn albumin buffer was prepared which comprised 0-1 % w/v albumin and 0-025 % w/v porcine yglobulin in saline buffered at pH 74 by 0-01 M phosphate.Ammonium sulphate was prepared as a saturated solution, and the pH was determined to ensure that it fell within the range 4 5-5 0.Bile salt free serum was obtained by charcoal extraction of 50 ml serum by 2-5 g charcoal. After incubation at room temperature ...

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Section: Discussionsupporting

confidence: 78%

Section: Discussionsupporting

confidence: 78%

mentioning

confidence: 99%

See 1 more Smart Citation

Radioimmunoassay of primary bile salts in serum.

Baqir¹,

Murison²,

Ross³

et al. 1979

J Clin Pathol

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“…As these studies indicated that the proportion of bile acids present as sulphates in serum and bile was quite low, the solvolysis step was not included routinely in the analysis of bile acids in serum or bile. Recovery for conjugated reference bile acids was satisfactory (84 5 to 96-3 %), and good separation between the individual bile acids-cholic acid, chenodeoxycholic acid, deoxycholic acid, and lithocholic acid-and the internal standard 7ketodeoxycholic acid was achieved by gas/liquid chromatography (GLC) (van Berge Henegouwen et al, 1974). Determinations of conjugated serum cholic and chenodeoxycholic acid were also performed by radioimmunoassay (Simmonds et al, 1973;and Schalm et al, 1975), and the values correlated well with our GLC method.…”

Section: Patientsmentioning

confidence: 71%

“…After refluxing for two hours with 2 % methanolic KOH, the solution was neutralised using Dowex 50W-X4, filtered with methanol and evaporated. The residue was dissolved in water; enzymatic hydrolysis and preparation of trifluoroacetate methyl ester derivatives of bile acids for GLC were carried out as described (van Berge Henegouwen et al, 1974). Recoveries for conjugated bile acids in urine carried through the entire procedure with solvolysis were 80-0 to 84-5 % and 84-0 to 90 0% for the entire procedure without solvolysis (van Berge Henegouwen, 1974).…”

Section: Patientsmentioning

confidence: 99%

Sulphated and unsulphated bile acids in serum, bile, and urine of patients with cholestasis.

Henegouwen¹,

Brandt²,

Eyssen³

et al. 1976

Gut

172

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SUMMARY Samples of serum, bile, and urine were collected simultaneously from patients with cholestasis of varying aetiology and from patients with cirrhosis; their bile acid composition was determined by gas/liquid chromatography and mass spectrometry In cholestasis, the patterns in all three body fluids differed consistently and strikingly. In serum, cholic acid was the major bile acid and most bile acids (> 93 %) were unsulphated, whereas, in urine, chenodeoxycholic was the major bile acid, and the majority of bile acids (> 60 %) were sulphated. Secondary bile acids were virtually absent in bile, serum, and urine. The total amount of bile acids excreted for 24 hours correlated highly with the concentration of serum bile acids; in patients with complete obstruction, urinary excretion averaged 71 6 mg/24 h. In cirrhotic patients, serum bile acids were less raised, and chenodeoxycholic acid was the predominant acid. In healthy controls, serum bile acids were consistently richer in chenodeoxycholic acid than biliary bile acids, and no bile acids were present in urine. No unusual monohydroxy bile acids were present in patients with primary biliary cirrhosis, but, in several patients, there was a considerable amount of hyocholic acid present in the urinary bile acids. The analyses of individual bile acids in serum and urine did not appear to provide helpful information in the differential diagnosis of cholestasis. Thus, in cholestasis, conjugation of chenodeoxycholic acid with sulphate becomes a major biochemical pathway, urine becomes a major route of bile acid excretion, and abnormal bile acids are formed.

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Autosomal dominant benign recurrent intrahepatic cholestasis (BRIC) unlinked to 18q21 and 2q24

et al. 2000

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Benign recurrent intrahepatic cholestasis (BRIC) is an autosomal recessive liver disease characterized by multiple episodes of cholestasis without progression to chronic liver disease. On the basis of recent evidence of locus heterogeneity, we studied 19 subjects (7 affected members) of a BRIC family. Male-to-male transmission and the presence of affected females suggested autosomal dominant inheritance. Blood samples were collected after informed consent. Subjects were genotyped by using markers mapping to 18q and 2q24 region, respectively, where the genes FIC1 and FIC2 have been mapped. Segregation of haplotypes excluded the two regions in our family. These findings suggest further genetic heterogeneity of the origin of BRIC.

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Quantitative analysis of bile acids in serum and bile, using gas -liquid chromatography

Cited by 110 publications

References 14 publications

Radioimmunoassay of primary bile salts in serum.

Radioimmunoassay of primary bile salts in serum.

Sulphated and unsulphated bile acids in serum, bile, and urine of patients with cholestasis.

Autosomal dominant benign recurrent intrahepatic cholestasis (BRIC) unlinked to 18q21 and 2q24

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