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In order not to invalidate the test for bacterial sterility of a biological product, the growth inhibitory effect of added bactericides or bacteriostatics should be either neutralized by some way or other or diluted out in a large volume of medium. Organo-mercuric antiseptics are used in various kinds of biological products as the preservative, and several procedures to make the antiseptic ineffective in sterility test media have been proposed (Brewer, 1944; Pittman, 1946; Bonnel and Raby, 1957; Didenko, 1957; Romanov et al., 1959). The medium established by Brewer (1944 and 1944) and Pittman (1946) is excellent and considered sufficient to inactivate the mercuric antiseptic (National Institutes of Health, 1954). This medium has been adopted in many countries (Public Health Service Regulations, 1962; Scheibel and Bentzon, 1957; Requirements, 1960), and our minimum Requirements for sterility test (Minimum Requirements of Biologic Products, 1962) was originally described on the basis of their studies. Our experiences, however, suggested that the method was not always effective to some species of microorganisms sensitive to the mercuric antiseptics. On the other hand, though fungi are known to be, in general, sensitive to mercuric preservatives, no device has been proposed for use in the test for mycotic sterility of products preserved with mercuric antiseptics, except that of Scheibel and Bentzon (1957). In this situation, a study has been performed to directly follow up the inactivation of the antiseptic in a sterility test medium. For this study a method of measuring a minute amount of the antiseptic with regard to its antiseptic activity is indispensable. The colorimetric method using dithizone solution in CC14 is inadequate for the purpose, not only because its sensitivity is insufficient but also because it cannot detect the antiseptic activity. Engley (1950) employed a paper disc method to compare mercuric antiseptics, but it was only qualitative. More recently, Hara, et al. (1959) established a microbiological assay method for organo-mercuric antiseptics, and proposed an analytical method for determination of organo-mercuric antiseptics in biological products using the chemical and microbiological assays collaterally. Their method overcame one of the weak points of the chemical method, and has opened a way to directly pursue the antiseptic activity

THE INACTIVATION OF SODIUM ETHYLMERCURITHIOSALICYLATE IN STERILITY TEST MEDIA WITH SPECIAL REFERENCE TO MYCOTIC STERILITY TEST