Related Experiment Video
Updated: Sep 1, 2025

Characterization of Biological Absorption Spectra Spanning the Visible to the Short-Wave Infrared
Published on: January 10, 2025
Integrating-Sphere-Assisted Resonance Synchronous Spectroscopy for the Quantification of Material Double-Beam UV-Vis
Max Wamsley1, Pathum Wathudura1, Juan Hu2
1Department of Chemistry, Mississippi State University, Mississippi State, Mississippi 39762, United States.
Abstract:
Integrating spheres (IS) have been used extensively for the characterization of light absorption in turbid samples. However, converting the IS-based sample absorption coefficient to the UV-vis absorbance quantified with a double-beam UV-vis spectrophotometer is challenging. Herein, we report an integrating-sphere-assisted resonance synchronous (ISARS) spectroscopy method performed with conventional spectrofluorometers equipped with an integrating-sphere accessory. Mathematical models and experimental procedures for quantifying the sample, solvent, and instrument-baseline ISARS intensity spectra were provided. A three-parameter analytical model has been developed for correlating the ISARS-based UV-vis absorbance and the absorbance measured with double-beam instruments. This ISARS method enables the quantitative separation of light absorption and scattering contribution to the sample UV-vis extinction spectrum measured with double-beam UV-vis spectrophotometers. Example applications of this ISARS technique are demonstrated with a series of representative samples differing significantly in their optical complexities, from approximately pure absorbers, pure scatterers, to simultaneous light absorbers, scatterers, and emitters under resonance excitation and detection conditions.
Related Concept Videos
Ultraviolet and Visible (UV–Vis) Spectroscopy: Overview
UV–Vis Spectrometers
UV–Vis Spectroscopy: Beer–Lambert Law
Spectrophotometry: Introduction
The essential components of a spectrophotometer include a source of electromagnetic radiation, a slot for placing a material to be analyzed, and a...
UV–Vis Spectrum
The UV–Vis spectrum of a molecule is the plot of its absorbance versus wavelength. The plot is drawn by taking molar...
UV–Vis Spectroscopy of Conjugated Systems
One of the factors influencing λmax is the extent...

