The tremendous growth of fluorescence applications in the life and material sciences has proceeded largely without sufficient concern for the reliability and uncertainties related to the characterization and performance validation of fluorescence instruments and measurements. To assess the state-of-the-art comparability and trueness of corrected emission spectra accomplishable with the aid of commercially available spectral fluorescence standards, in part II of this study, twelve field laboratories were randomly selected from academia and industry based upon their fluorescence measuring equipment. These laboratories performed a reference material (RM)-based fluorometer calibration following a standard operation procedure provided with the fluorescence standards and used this emission correction curve for the determination of the corrected emission spectra of three test dyes X, QY, and Y. This revealed an average accuracy of the correctedemission spectra of the test dyes of minimum 6.8 % for common commercial spectrofluorometers, routine measurement conditions, and the data evaluation software LINKCORR developed and provided by BAM. These results only slightly exceed the relative standard uncertainties of 4.2 % for physical standard (PTS)-based spectral corrections and 2.4 % for RM-based spectral corrections demonstrated in part I of this study by an international interlaboratory comparison of four expert laboratories.
Citation: Analytical Chemistry
Pub Type: Journals
biosystems and health, calibration, fluorescence, instrument qualification, method validation, reference materials, spectral correction