EPR method for the measurement of cellular sulfhydryl groups

An EPR method that can measure the concentration of sulfhydryl groups in intact cells has been developed using a specially designed stable nitroxyl biradical. The biradical, RS-SR, contains a disulfide bond and readily undergoes thiol-disulfide exchange reactions with thiols resulting in a characteristic EPR spectrum which can be analyzed to provide a quantitative measure of sulfhydryl groups. The data obtained from the EPR method are in good agreement with those obtained from the conventional optical method using Ellman's reagent. The advantages of the EPR method are that the measurement can be carried out on intact cells or any other highly colored, absorbing and/or scattering solutions and the sensitivity is such that only a few cells (approximately 100) are needed for each quantitative measurement.