scholarly journals Isolation and Purification of Glycoconjugates from Complex Biological Sources by Recycling High-Performance Liquid Chromatography

2013 ◽  
Vol 85 (21) ◽  
pp. 10408-10416 ◽  
Author(s):  
William R. Alley ◽  
Benjamin F. Mann ◽  
Vlastimil Hruska ◽  
Milos V. Novotny
Keyword(s):  
High Performance Liquid Chromatography ◽  
Liquid Chromatography ◽  
High Performance ◽  
Isolation And Purification ◽  
Biological Sources

High Performance Liquid Chromatography Coupled with Radioactivity Detection: A Powerful Tool for Determining Drug Metabolite Profiles in Biological Fluids

1986 ◽  
Vol 41 (1-2) ◽  
pp. 115-125 ◽  
Author(s):  
K.-O. Vollmer ◽  
W. Klemisch ◽  
A. von Hodenberg
Keyword(s):  
High Performance Liquid Chromatography ◽  
Liquid Chromatography ◽  
High Performance ◽  
Biological Fluids ◽  
Gradient Elution ◽  
Reversed Phase ◽  
Isolation And Purification ◽  
Carbon 14 ◽  
Tracer Techniques ◽  
Metabolite Profiles

Abstract High performance liquid chromatography coupled with continuous radioactivity detection rep­resents an advancement in drug metabolism research. Using radioactive substances labelled in biologically stable positions, all metabolites can be specifically detected by radioactivity measure­ment. Thus no clean-up of biological fluids is required prior to HPLC. This can prevent artefact formation from unstable metabolites, reduces recovery problems and facilitates quantitation. Separation of highly polar and unpolar metabolites is possible in a single chromatographic run using gradient elution and reversed phase materials. This technique is also well-suited for prepara­tive isolation and purification of metabolites for subsequent structure elucidation. Various metabolite profiles of drugs labelled with carbon-14 or tritium are shown. Metabolites of the following drugs are presented: norfenefrine, etozolin, thymoxamine, naloxone, and levobunolol. We review the general methodology and report our experience with this technique. In principle, this technique may be useful for all biological systems in which tracer techniques are applied.

Determination of Corticosteroids in Fish Plasma by High Performance Liquid Chromatography: Evaluation of the Method Using Striped Bass (Morone saxatilis)

1984 ◽  
Vol 41 (9) ◽  
pp. 1280-1286 ◽  
Author(s):  
R. Bruce MacFarsane
Keyword(s):  
High Performance Liquid Chromatography ◽  
Liquid Chromatography ◽  
Striped Bass ◽  
High Performance ◽  
Methylene Chloride ◽  
Morone Saxatilis ◽  
Cross Reactivity ◽  
Isolation And Purification ◽  
Bonded Phase

A sensitive, specific, and reproducible technique for the simultaneous determination of cortisol, cortisone, and corticosterone in fish plasma was developed using high performance liquid chromatography (HPLC). Corticosteroids were extracted from plasma by methylene chloride and Sep-Pak cartridges for comparative purposes. The Sep-Pak method proved more convenient and efficient. Chromatography was accomplished on a nitrile bonded phase using a methanol–water mobile phase. Steroids were identified and quantified by monitoring absorbance at 254 and 280 nm simultaneously. Sensitivity is approximately 5 ng for each steroid. An evaluation of the technique using subadult striped bass (Morone saxatilis) revealed that cortisol was the only corticosteroid detected. Plasma cortisol concentrations increased in response to brief confinement and crowding, supporting the efficacy of this measurement as an evaluator of stress in striped bass. The HPLC technique provides specific determination of each corticosteroid (thus eliminating cross-reactivity problems inherent in radioimmunoassay methods), resolves corticosteroids from androgens and estrogens, and permits the isolation and purification of individual steroids.

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