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There are many aspects of UV detection for HPLC which are poorly understood and could be improved or optimized in order to improve your analyses. Key information on the instrumentation and the use and optimization of UV detectors for modern HPLC applications will be discussed. Fundamental UV theory will be revised with suggestions on how to choose optimum acquisition parameters based on analyte spectra and UV instrument design. Key practical concepts will be explored and the optimization of each acquisition variable will be explained in detail in terms of both the instrumentation and chromatographic data sensitivity and specificity. Instrument limitations and dependencies will be discussed so that optimum method robustness might be obtained in everyday use. Aspects of modern HPLC instrument design will be explored and related to improve chromatographic performance.


Topics include: 

  • Review of fundamental UV absorbance theory
  • Understanding the optical arrangements of common UV detectors
  • Data acquisition parameter optimization – including bandwidth, slit width, and time constant
  • Detector linearity – limitations imposed by analyte spectra, bandwidth, and slit width settings
  • Choosing an appropriate reference wavelength to reduce gradient baseline drift and noise
  • Sensitivity – lamp effects, optical and electronic noise, solvent effects etc.
  • Flow cell design – modern light-pipe designs, illuminated volume, internal reflection principles and dispersion
  • Detection robustness - optimizing settings based on chromatographic data and improving your analytical robustness/sensitivity