Calibration of an integrating sphere for determining the absorption coefficient of scattering suspensions

Measuring the absolute absorption of suspensions of absorbing particles with unknown scattering characteristics is not possible in conventional spectrophotometers or in integrating spheres that have the sample located outside the sphere. A method for the calibration and use of an integrating sphere with a centrally located sample to measure absolute absorption coefficients of scattering suspensions is presented. Under the tested conditions the integrating sphere used in this study was insensitive to changes in the scattering coefficient of the sample but had a nonlinear response to increasing absorption of the sample, which could be corrected with an empirically derived function. This response was analyzed by using a Monte Carlo simulation, and results indicated that amplification of the absorption signal was primarily due to photons reflected from the sphere surface and the baffle reentering the cuvette. The calibration procedure described here may be generally applicable to spheres of different configurati n. An example of the use of the sphere for determining the absorption and scattering coefficients of marine phytoplankton samples is presented.