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Title: Application of capillary zone electrophoresis to the separation of metallothionein isoforms

Abstract

Metallothioneins (MTs) from various eukaryotic species were subjected to capillary zone electrophoresis (CZE) performed under the following conditions: Capillary: untreated polyimide-clad fused silica 75 [mu]m I.D. [times] 94 cm; Loading and Running Voltage: 30 kV; Electrode Buffer: 5s0 mM TrisHCI pH 9.1; Sample Buffer: 10 mM Tris-HCI pH 9.1; Sample Volume: 250 [mu]I; Detection: UV absorbance at 214 nm; Loading Method: electrokinetic migration; Loading Times: 1-6 seconds. Complete separation of MT-1 and MT-2 isoforms was achieved in less than 6 min. Using a rabbit liver Cd,Zn-MT standard (0.5 mg/ml), a linear relationship was found to exist between the voltage used to load the MT sample and the integrated peak area of the individual MT isoforms. Similarly, the integrated peak area of the separated MT isoforms was a linear function of the time used to load the sample. These findings suggest that with appropriate standardization CZE is capable of both qualitative and quantitative determinations of MT isoforms. In CZE, the concentration of MT in solution determines the limit of sensitivity. MT could be accurately detected at a concentration as low as 10 [mu]g/ml. CZE was applied to assess thermal and proteolytic degradation of rabbit liver MT. CZE represents a rapid analyticalmore » technique for the characterization of MT isoforms that requires very little sample yet provides excellent resolution of individual MT isoforms.« less

Authors:
 [1]; ;  [2]
  1. Dept. of Agriculture, Beltsville, MD (United States)
  2. Rowett Research Inst., Aberdeen (United Kingdom)
Publication Date:
OSTI Identifier:
6428460
Resource Type:
Journal Article
Journal Name:
Journal of Liquid Chromatography; (United States)
Additional Journal Information:
Journal Volume: 16:9-10; Journal ID: ISSN 0148-3919
Country of Publication:
United States
Language:
English
Subject:
63 RADIATION, THERMAL, AND OTHER ENVIRON. POLLUTANT EFFECTS ON LIVING ORGS. AND BIOL. MAT.; METALLOTHIONEIN; ELECTROPHORESIS; METALS; METABOLISM; LIVER; RATS; ANIMALS; BODY; DIGESTIVE SYSTEM; ELEMENTS; GLANDS; MAMMALS; METALLOPROTEINS; ORGANIC COMPOUNDS; ORGANS; PROTEINS; RODENTS; VERTEBRATES; 560300* - Chemicals Metabolism & Toxicology

Citation Formats

Richards, M P, Beattie, J H, and Self, R. Application of capillary zone electrophoresis to the separation of metallothionein isoforms. United States: N. p., 1993. Web. doi:10.1080/10826079308019919.
Richards, M P, Beattie, J H, & Self, R. Application of capillary zone electrophoresis to the separation of metallothionein isoforms. United States. https://doi.org/10.1080/10826079308019919
Richards, M P, Beattie, J H, and Self, R. Fri . "Application of capillary zone electrophoresis to the separation of metallothionein isoforms". United States. https://doi.org/10.1080/10826079308019919.
@article{osti_6428460,
title = {Application of capillary zone electrophoresis to the separation of metallothionein isoforms},
author = {Richards, M P and Beattie, J H and Self, R},
abstractNote = {Metallothioneins (MTs) from various eukaryotic species were subjected to capillary zone electrophoresis (CZE) performed under the following conditions: Capillary: untreated polyimide-clad fused silica 75 [mu]m I.D. [times] 94 cm; Loading and Running Voltage: 30 kV; Electrode Buffer: 5s0 mM TrisHCI pH 9.1; Sample Buffer: 10 mM Tris-HCI pH 9.1; Sample Volume: 250 [mu]I; Detection: UV absorbance at 214 nm; Loading Method: electrokinetic migration; Loading Times: 1-6 seconds. Complete separation of MT-1 and MT-2 isoforms was achieved in less than 6 min. Using a rabbit liver Cd,Zn-MT standard (0.5 mg/ml), a linear relationship was found to exist between the voltage used to load the MT sample and the integrated peak area of the individual MT isoforms. Similarly, the integrated peak area of the separated MT isoforms was a linear function of the time used to load the sample. These findings suggest that with appropriate standardization CZE is capable of both qualitative and quantitative determinations of MT isoforms. In CZE, the concentration of MT in solution determines the limit of sensitivity. MT could be accurately detected at a concentration as low as 10 [mu]g/ml. CZE was applied to assess thermal and proteolytic degradation of rabbit liver MT. CZE represents a rapid analytical technique for the characterization of MT isoforms that requires very little sample yet provides excellent resolution of individual MT isoforms.},
doi = {10.1080/10826079308019919},
url = {https://www.osti.gov/biblio/6428460}, journal = {Journal of Liquid Chromatography; (United States)},
issn = {0148-3919},
number = ,
volume = 16:9-10,
place = {United States},
year = {1993},
month = {1}
}