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SELECTIVE TRACE ENRICHMENT BY IMMUNOAFFINITY CAPILLARY ELECTROCHROMATOGRAPHY ON-LINE WITH CAPILLARY ZONE ELECTROPHORESIS - LASER-INDUCED FLUORESCENCE
Citation:
Thomas, D. H., D. J. Rakestraw, J. S. Schoeniger, V. LopezAvila, AND J M. Van Emon. SELECTIVE TRACE ENRICHMENT BY IMMUNOAFFINITY CAPILLARY ELECTROCHROMATOGRAPHY ON-LINE WITH CAPILLARY ZONE ELECTROPHORESIS - LASER-INDUCED FLUORESCENCE. ELECTROPHORESIS 20:57-66, (1999).
Impact/Purpose:
More cost-effective field screening and monitoring methods will be provided to increase the amount of information available concerning the location, source, and concentration of pollutants. Rapid and sensitive immunoassays such as enzyme-linked immunosorbent assays (ELISAs) to monitor remediation and cleanup activities at Superfund sites will be developed. Each new method will be tested on real-world samples from monitoring studies. Field studies will be conducted when time and resources permit. The feasibility and application of immunosensors to provide field analytical methods for the dynamic monitoring of hazardous substances of interest to the EPA will also be investigated. Concern has been expressed for the potential exposure of individuals to toxic compounds who live near hazardous waste sites or who may become exposed through other means. Thus, the development of methods for measuring biomarkers for human exposure assessment studies is also addressed.
During the remainder of the Task several projects will be undertaken including:
- Complete the development and evaluation of bioanalytical methods for dioxin and related compounds
- Perform dioxin immunoassay analysis on samples from a dioxin SITE demonstration
- Comparison of an ELISA with gas chromatography (GC) for monitoring polychlorinated biphenyls (PCBs) in soils/sediments collected from a Superfund field demonstration
- Survey of bioanalytical methods and sensor technologies for environmental monitoring
- Development of immunoaffinity chromatography sample preparations for PCBs
- Preparation of standard operating procedures (SOPs) for each bioanalytical method developed
- Conduct yearly research meeting
- Conduct survey of high priority chemicals at National Priorities List (NPL) sites for bioanalysis suitability
- Develop new bioanalytical methods for hazardous compounds of public concern
- Perform PCP immunoassay analysis on soil and sediment samples from a Superfund site and compare with GC data
- Preparation of fact sheets and journal articles
Description:
Limited by the lack of a sensitive, universal detector, many capillary-based liquid-phase separation techniques might benefit from techniques that overcome modest concentration sensitivity by preconcentrating large injection volumes. The work presented employs selective solid-phase extraction by immunoaffinity capillary electrochromatography (ACEC) to enhance detection limits. A model analyte, fluorescein isothiocyanate (FITC) biotin, is electrokinetically applied to a capillary column packed with an immobilized anti-biotin-IgG support. After selective extraction by the immunoaffinity capillary, the bound analyte is eluted, migrates by capillary zone electrophoresis (CZE), and is detected by laser-induced fluorescence. The column is regenerated and reused many times. We evaluate the performance of IACEC for selective trace enrichment of analytes prior to CZE. The calibration curve for FITC-biotin bound versus application time is linear from 10 to 300 seconds. Recovery of FITC-biotin spiked into a diluted urinary metabolites solution was 89.4% versus spiked buffer, with a precision of 1.8% relative standard deviation (RSD).