Abstract
We report problems encountered during preparation of tritium-labeled unconjugated
bilirubin (3H-UCB) from precursor 3H-5-aminolevulinic acid (3H-ALA) in 2 dogs with external biliary drainage installed into the animals under general
anesthesia. Under prolonged sedation, 12.9 or 14.0 mCi of 3H-ALA was administered intravenously in two divided doses, and bile was collected
for 9 hours. In one animal, taurocholate (TC) infusion was needed to maintain bile
flow. 3H-UCB was isolated from the bile and recrystallized with the improved method of Webster
et al (Webster CC, Tiribelli C, Ostrow JD. J Lab Clin Med 2001;137:370-3). Based on
radioactivity and pigment content, hourly bile collections were pooled to optimize
specific activities. Surprisingly, in the first dog, only 2.9% of injected radioactivity
was recovered in bile and only 14.1% in urine, and the specific activities of the
crystalline 3H-UCB from the two pools were only 39.5 and 30.0 × 103 dpm/μg. High-performance liquid chromatography analysis revealed that only 4% of
ALA degraded during 5 minutes in injection solution at pH 6.8. The low incorporation
of 3H-ALA and low specific activity of 3H-UCB was apparently caused mainly by prior degradation and exchange of labile tritium
of the 3H-ALA and probably by enhanced endogenous ALA synthesis caused by the anesthetic/sedative
agents. Revised procedures in the second dog improved the incorporation of 3H-ALA to 11.9% excreted in bile and the specific activity of the crystalline 3H-UCB to 122.0 and 50.8 × 103 dpm/μg, while urinary excretion of tritium increased to 28.5%. These experiences
emphasize possible pitfalls in preparing 3H-UCB by biosynthetic labeling from 3H-ALA administered to dogs. (J Lab Clin Med 2001;138:313-21)
Abbreviations:
ALA (5-aminolevulinic acid), 3H-ALA (3H-labeled 5-aminolevulinic acid), AUC (area under the curve), HPLC (high-performance liquid chromatography), PBG (porphobilinogen), TC (sodium taurocholate), UCB (unconjugated bilirubin)To read this article in full you will need to make a payment
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Article info
Publication history
Accepted:
July 20,
2001
Received in revised form:
June 8,
2001
Received:
June 28,
2000
Footnotes
☆Supported by special grants from the University of Trieste; by a grant from the Junta de Castilla y León, Spain; and by the Seattle Institute for Biomedical and Clinical Research. Dr Tiribelli was supported by grants from the Italian Ministry of University and Research (MURST-Cofin 98) and Ministry of Health (ICS060.1/RF98.67). Dr Stelzner was supported by a Merit Review Grant and Dr Ostrow and Ms Webster by a Medical Investigator Award from the US Department of Veterans Affairs. Dr Ostrow was supported also by the Gastroenterology Foundation at the Academic Medical Center of the University of Amsterdam, the Netherlands.
☆☆*Current address: J. Enriqué Bayón, MD, Escuela Universitaria de Enfermeria, Campus Universitario de Ponferrada, Avenida Astorgasn, E-24400 León, Spain.
★*Current address: J. Donald Ostrow (151L),GI/Hepatology Lab, Research Service, Seattle VA Medical Center, 1660 South Columbian Way, Seattle, WA 98108-1597.
★★Reprint requests: Javier González-Gallego, MD, Department of Physiology, University of León, Spain. E-mail: [email protected]
Identification
Copyright
© 2001 Mosby, Inc. Published by Elsevier Inc. All rights reserved.
