Abstract
The concentration and fluctuation of BA in serum may be the most sensitive index of hepatic dysfunction. Of existing methods, gas-liquid chromatography (GC) is complex and radioimmunoassay (RIA) is limited by availability of specific antibodies. We have modified the DBSF method (Siskos, et.al.,J Lipid Res 18:666,′77) to measure NS+S BA by differences in fluorescence (f) between sample and reference cuvettes. Paired extraction with 1) isopro-panol-reduces f due to protein, and with 2) ethanol/acetone-causes solvolysis, allowing enzymatic oxidation at 3α-position. Total solvolysis of standards was confirmed by TLC. Recovery of 14C-taurocholate was >92%. BA concentration by DBSF correlated with GC (r=0.97) and with RIA for cholylglycine (r=0.91). Normal fasting total BA (8.6±2.5 SD μmol/1) was followed by a postprandial (90′) two-fold increase due to influx via the enterohepatic circulation. No overlap with normals was found in acute (27±10.5) or chronic (79±43) hepatitis. There were no significant differences in values obtained at comparable ages in 9 patients with neonatal hepatitis (105±51) and 16 patients with biliary atresia (132±61). S comprised a varying percentage of total BA, being virtually absent (2-5%) in normals, increasing to 15-25% with severe cholestasis.
Conclusion: A valid modified DBSF assay for S+NS, which is reproducible, rapid and easily performed on 0.1ml of serum may be a sensitive screen for liver disease.
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Balistreri, W., Shapiro, M. & Soloway, R. 394 SERUM SULFATED (S) AND NONSULFATED (NS) BILE ACID (BA) CONCENTRATION VIA DUAL-BEAM SPECTROPHOTOFLUORI-IMETRY (DBSF). Pediatr Res 12 (Suppl 4), 429 (1978). https://doi.org/10.1203/00006450-197804001-00399
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DOI: https://doi.org/10.1203/00006450-197804001-00399