EFFECTS OF RESOLUTION, PEAK RATIO AND SAMPLING FREQUENCY IN DIODE-ARRAY FLUORESCENCE DETECTION IN LIQUID-CHROMATOGRAPHY

In this work, different methods of quantification that use both fluore scence spectral and retention time data were evaluated for accuracy an d precision in diode-array fluorescence detection in liquid chromatogr aphy. The methods evaluated include the peak height and peak area meth ods of quantification, an adaptive Kalman filter and the generalized r ank annihilation method. All of these methods were used to estimate th e concentration of a model led component in the presence of other part ially resolved unmodelled components. The peak area and peak height me thods of quantification gave the largest errors as the chromatographic resolution was decreased. The generalized rank annihilation method ga ve the most reliable results when the chromatographic resolution was g reater than 0.3. However, this method was affected by retention time s hifts between the calibration and unknown samples and the rate at whic h the spectra were acquired.