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Multivariate analysis to separate the signal given by cross-reactants in immunoassay with sample matrix dilution
Catalin Nistor1, Jakob Christensen, Natalia Ocio
1Department of Analytical Chemistry, Lund University, P.O. Box 124, 221 00 Lund, Sweden.
Analytical and Bioanalytical Chemistry
|November 30, 2004
Summary
This study introduces a novel immunoassay method to accurately quantify two cross-reactive compounds, 2,4-dinitrophenol (2,4-DNP) and 4-nitrophenol (4-NP), in complex samples using sequential dilution and advanced data modeling.
Area of Science:
- Analytical Chemistry
- Biochemical Assays
- Chemometrics
Background:
- Immunoassays often face challenges with cross-reactivity, limiting accurate quantification of multiple analytes simultaneously.
- Complex sample matrices further complicate the selective detection of similar compounds.
Purpose of the Study:
- To develop a novel immunoassay approach for simultaneous and selective quantification of two cross-reactive analytes.
- To overcome signal interference between 2,4-dinitrophenol (2,4-DNP) and 4-nitrophenol (4-NP) in complex samples.
Main Methods:
- A sequential dilution strategy combined with spiking with a reference compound.
- Detection using enzyme-linked immunosorbent assay (ELISA).
- Multivariate data analysis employing Principal Component Analysis (PCA) and Partial Least Squares Regression (PLSR) for signal separation and calibration.
Main Results:
- Successful separation of signals from 2,4-DNP and 4-NP was achieved.
- Accurate calibration models were developed for individual analyte quantification.
- Prediction concentration errors were 1.4 µM for 2,4-DNP and 72 µM for 4-NP.
Conclusions:
- The proposed method enables selective and simultaneous quantification of cross-reactive analytes in complex matrices.
- This approach enhances the utility of immunoassays for analyzing mixtures of similar compounds.