Photoelectric Light-Scattering Photometer for Determining High Molecular Weights*

TLDR

The authors present a photoelectric photometer that measures absolute turbidity, dissymmetry, and depolarization of dilute high‑molecular‑weight solutions to determine their molecular weights. The device uses a monochromatic parallel beam, a six‑sided scattering cell at 0°, 45°, 90°, and 135°, a multiplier phototube and galvanometer, an opal glass diffusor, and removable polarizer/analyzer, with turbidity calculated from the ratio of 90° scattering deflection to the primary beam deflection, and includes corrections for refraction, reflection, and imperfect diffusion. The photometer’s multiplier tubes show polarization‑dependent response, and its performance is validated by comparing molecular weights of polystyrene fractions, proteins, sugars, and other analytes with values obtained by other methods.

Abstract

A photoelectric photometer designed for the measurement of absolute turbidity, dissymmetry, and depolarization of dilute solutions of high molecular weight materials, and hence determination of their molecular weights, is described. The photometer comprises essentially a monochromatic parallel primary beam of radiation, a six-sided scattering cell for measurements at 0°, 45°, 90°, and 135°, a multiplier phototube and galvanometer, a standard opal glass diffusor, and removable polarizer and analyzer. Turbidity is determined in terms of a ratio of deflections for the 90° scattering and for the primary beam reduced in intensity by neutral filters and diffused by an opal glass plate. Working relationships leading to determination of absolute turbidity are developed. These relationships include corrections for refraction and reflection effects, and for imperfect diffusion by the opal glass. The latter is evaluated by comparison of the opal glass with reflecting diffusors corrected for specular component of reflectance. The response of some multiplier photo-tubes is shown to be dependent on the plane of polarization of the incident radiation. Data illustrating the performance of the photometer include comparison of molecular weights of polystyrene fractions, beta-lactoglobulin, bovine serum albumin, lysozyme, sucrose octaacetate, Raleigh’s ratio and depolarization for benzene, turbidity of a “standard” polystyrene, and particle size of a GR-S latex, with data obtained by other methods or other investigators.