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Development and evaluation of C18 and immunosorbent solid-phase extraction methods prior immunochemical analysis of chlorophenols in human urine

AutorNichkova, Mikaela; Marco, María Pilar
Palabras claveImmunosorbent
Fecha de publicación21-mar-2005
CitaciónAnalytica Chimica Acta 533(1): 67-82 (2005)
ResumenTwo solid-phase extraction (SPE) methods, based on hydrophobic and selective (antibody–antigen) interactions, have been established and evaluated as clean-up methods prior the immunochemical analysis of 2,4,6-trichlorophenol (2,4,6-TCP) in urine samples. Without a clean-up method the extent of interferences caused by the urine matrix in the ELISA [R. Galve, M. Nichkova, F. Camps, F. Sanchez-Baeza, M.-P. Marco, Anal. Chem. 74 (2002) 468] varies depending on individual urine samples and accurate measurements are only possible when 2,4,6-TCP concentration levels are higher than 40 μg L−1. Both sample preparation methods improve detectability of the immunochemical method getting rid of the variability due to the intrinsic individual differences within the urine samples. Even though, the immunosorbent (IS)-SPE method developed has proven to be a superior sample preparation method eliminating completely matrix effects caused by both, non-hydrolyzed (NH) and hydrolyzed urine samples. The LOD reached by the C18-SPE-ELISA method (not, vert, similar4 μg L−1 for free and total chlorophenols) is sufficient for exposure assessment of the occupationally exposed population. However, the detectability (0.66 and 0.83 μg L−1 in NH and hydrolyzed urine samples, respectively) accomplished by the IS-SPE-ELISA allows also biomonitoring potential exposure of non-occupationally exposed groups. Moreover, the specificity of the IS-SPE procedure can be modulated to provide a group-specific (9 chlorophenols and 2 bromophenols are extracted with an efficacy superior to 85%) or a more selective protocol (only 2,3,4,6-TtCP, 2,4,6-TCP are extracted with a recovery superior to 80% and 2,4,6-tribromophenol with a 70% recovery). On the other hand, the IS-SPE extracts produce cleaner chromatograms allowing quantitation by GC–ECD (or GC–MS) after toluene extraction and derivatization with a LOD near 0.1 μg L−1 in NH and hydrolyzed urine samples. The IS-SPE-ELISA method has been validated with GC–ECD using spiked and real urine samples. This study also provides evidences of the general exposure of the population to organochlorinated and organobrominated substances. Measurable levels of 2,4,6-TCP, 2,4,5-TCP, 2,3,4,6-TtCP, 2,4,6-TBP and 2,4-DBP have been detected in some of the samples used in this study.
Descripción16 pages, 9 figures, 7 tables.-- Available online Dec 29, 2004.
Versión del editorhttp://dx.doi.org/10.1016/j.aca.2004.10.068
ISSN0003-2670 (Print)
1873-4324 (Online)
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