Set of secondary emission standards for calibration of the spectral responsivity in emission spectroscopy

Technical spectra are reported for a series of six secondary emission standards based on the commercially available fluorophors: tryptophan, alpha-NPO, tetraphenylbutadiene, coumarin 153, DCM, and LDS 751, These spectra cover the wavelength range between 300 and 800 nm with significant spectral overlap and conform to the requirements of secondary emission standards. Standard emission spectra are determined by averaging the technical spectra obtained from three independently calibrated fluorescence spectrometers. A method for generating the wavelength-dependent correction file by using these standards is outlined. Comparison to the National Bureau of Standards (NBS)-reported quinine sulfate spectrum illustrates the accuracy of both the emission standards and the method of generating a complete emission correction file. With the use of the standards and method reported here, the accuracy of the emission correction is estimated to be better than 10% for the entire wavelength range and significantly better over the wavelength region of a typical spectrum.