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This work reports on methods to detect and estimate anionic detergents, which are important ingredients of synthetic milk formulations, based on the extraction of a detergent-Methylene Blue complex in chloroform which can be subsequently quantified by measuring the optical density at 653 nm. A base value of 0.52 for optical density was found to represent pure cows' milk. Any result above this value suggested the presence of an anionic detergent in milk.