Journal
JOURNAL OF PHARMACEUTICAL AND BIOMEDICAL ANALYSIS
Volume 25, Issue 5-6, Pages 803-809Publisher
PERGAMON-ELSEVIER SCIENCE LTD
DOI: 10.1016/S0731-7085(01)00342-9
Keywords
Beer's Law deviation; polychromatic radiation; systematic errors in spectrophotometric assays; nonexponential power attenuation
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It is well established that the use of polychromatic radiation in spectrophotometric assays leads to excursions from the Beer-Lambert limit, This Note models the resulting systematic error as a function of assay spectral width, slope of molecular extinction coefficient, and analyte concentration. The theoretical calculations are compared with recent experimental results: a parameter is introduced which can be used to estimate the magnitude of the systematic error in both chromatographic and nonchromatographic spectrophotometric assays. It is important to realize that the polychromatic radiation employed in common laboratory equipment can yield assay errors up to similar to4% even at absorption levels generally considered 'safe' (i.e. absorption < 1). Thus careful consideration of instrumental spectral width, analyte concentration, and slope of molecular extinction coefficient is required to ensure robust analytical methods. (C) 2001 Elsevier Science B.V. All rights reserved.
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