Source: Laboratory of Dr. Paul Bower - Purdue University
The method of standard additions is a quantitative analysis method, which is often used when the sample of interest has multiple components that result in matrix effects, where the additional components may either reduce or enhance the analyte absorbance signal. That results in significant errors in the analysis results.
Standard additions are commonly used to eliminate matrix effects from a measurement, since it is assumed that the matrix affects all of the solutions equally. Additionally, it is used to correct for the chemical phase separations performed in the extraction process.
The method is performed by reading the experimental (in this case fluorescent) intensity of the unknown solution and then by measuring the intensity of the unknown with varying amounts of known standard added. The data are plotted as fluorescence intensity vs. the amount of the standard added (the unknown itself, with no standard added, is plotted ON the y-axis). The least squares line intersects the x-axis at the negative of the concentration of the unknown, as shown in Figure 1.
Figure 1. Graphic representation of method of standard addition.
1. Preparing the Reagents
A scan of the excitation wavelength from 335–435 showed the highest absorption at 399 nm, so the excitation monochromator was set for that value. Then the emission scan was performed from 450–550 nm, and the strongest signal was found to be at 520 nm. These are the wavelengths that are used for all of the samples.
Sample | Application and Summary The method of standard additions is often the technique utilized when accurate quantitative results are desired, used in analytical analysis such as atomic absorption, fluorescence spectroscopy, ICP-OES, and gas chromatography. This is often used when there are other components in the sample of interest that causes either a reduction or enhancement of the absorbance desired for quantitative results. When this is the case, one cannot simply compare the analytes signal to standards using the traditional calibration curve a Tags Method Of Standard AdditionQuantitative AnalysisMatrix EffectsAnalyte Measurement SignalsUnknown Component ConcentrationsAnalytical TechniquesLight SpectroscopyMass SpectrometryElectrochemistryMatrixSignal Reduction enhancementErrors In AnalysisKnown Analyte SolutionBasics Of Standard Addition MethodFluorescence MeasurementComplex SamplesMolecule InteractionsFluoresce Ability ChangeIonic Strength ChangeMitigate Effects 건너뛰기... JoVE 소개 Copyright © 2024 MyJoVE Corporation. 판권 소유 |