TESTING DEODORANTS WITH CHLOROPHYLL AND DERIVATIVES 37 spiration was the fact that the sample used in the experiments of Table 2 contained greater amounts of non-protein nitrogenous com- pounds than did the sample upon which the experiments of Table 1 had been carried out. These differ- ences are shown in the tabulation on page 36. It is evident from the data pre- sented above that the second pooled sample of perspiration provided more nutriment for growth and reproduction of bacteria than did the first. Also, it furnished a richer substrate, in the form of nitroge- nous compounds for the produc- tion of odorous end products of the putrefactive reactions of the cu- taneous micro6rganisms. Results of both series of experi- ments indicate that anaerobic bac- teria survived exposure of the samples of perspiration to air during forty-eight hours at 37øC. and, vice versa, the capacities for growth and reproduction of aerobic organisms were not inhibited by storage in contact with vapor phases contain- ing less than 0.1 per cent of oxygen. These findings are in agreement with the observation of Pillsbury (15) that, in the extensive studies of cutaneous microiSrganisms, he and his collaborators had not found any species of bacteria which was either an obligative aerobe or an obliga- tire anaerobe. Maintenance of reserves of acid over base during incubation in- hibited, to a substantial degree, rates of reproduction of micro- organisms and, also, the develop- ments of offensive odors. Refrigerated aliquots of both pooled samples of perspiration yielded quantities of volatile acids in excess of the amounts of volatile bases. In this respect, the findings for the refrigerated aliquots are representative of results obtained in comparable analyses of fresh samples of perspiration (5). These analytical data suggest that in the refrigerated aliquots, as well as in fresh samples of perspiration, fatty acids, having relatively low molec- ular weights, are present as free acids or they are bound by non- volatile bases. TABULATION B--COMPARATIVE INCREASES IN VOLATILE BASES AND VOLATILE ACIDS OF PER- SPIRATION DURING INCUBATION FOR 48 HouRs A•r 37øC. UNDER ErrHER AEROBIC OR ANAErO- BIC CONDITIONS Range of Reac- Pooled Sam- ticns of Aliquots ple of Perspiration as pH Average Ratios of Concentrations in Incubated Aliquots to Concen- trations in Refrigerated Aliquots Volatile Bases Volatile Acids Of Table 1 Of Table 2 4.32-4.76 40,0 1 ß 80 7,16-7.21 102.5 2.46 8.45 80.0 2.76 4.19-4.98 13.5 1.27 7.34-7.37 32.5 2.97 7.97-7.99 38.5 2.14

38 JOURNAL OF THE SOCIETY OF COSMETIC CHEMISTS During storage either in contact with air or under anaerobic condi- tions, increases in both volatile bases and volatile acids were ob- served. However, the more signifi- cant result was the fact that the increments in volatile bases exceed the increases in volatile acids as shown in the tabulation on page 37. In view of the fact that aliquots of the two pooled samples of per- spiration, at any one of the three levels of acid-base balances listed in the preceding tabulation, exhibited approximately equivalent increases in both base and acids, volatile with steam, during storage either in contact with air or in the absence of oxygen, data for the two aliquots were combined in the calculations of the average ratios reported above. It will be seen that aliquots of both pooled samples of perspiration which were buffered at levels of pH = 7.00 or above yielded greater increments in both volatile bases and acids than did the aliquots buffered at levels of pH = 4.00. Also, the former aliquots were malodorous at the ends of periods of storage whereas the weakly acid aliquots developed no demonstrable odors during periods of incubation. In this respect, results of the two series of experiments reported in Tables 1 and 2 corroborate the find- ings of earlier investigations (5). Subsequently, several consecutive experimental studies were under- taken for purposes of identification of the volatile bases and volatile acids produced during incubation of perspiration either exposed to air or exposed to similar volumes of gases from which oxygen had been eliminated. Only a brief summary of the results of these later investiga- tions will be presented in this intro- duction. In samples of perspiration which were analyzed immediately after collection, the total nitrogen varied from about 70 to 150 mg. per cent and the non-protein nitrogen made up about 80 per cent of the total nitrogen. The principal constitu- ent of the non-protein nitrogen was urea. Ammonia accounted for rela- tively small fractions, i.e., from 3 to 4 per cent of the total nitrogen. Approximately 10 per cent of the total nitrogen of fresh perspiration was volatile with steam and about 70 per cent of this volatile fraction was found to be ammonia nitrogen. Primary amines or amides ac- counted for the remaining 30 per cent of the nitrogen which was volatilized with steam. In contrast with freshly collected perspiration, samples of stale, malo- dorous perspiration yielded quanti- ties of volatile nitrogen which were equivalent to from 50 to 70 per cent of the total nitrogen and to about 90 per cent of the non-protein fraction. Nitrogenous compounds made up the total volatile base of which about 90 per cent was found to be ammonia. Amines and amides were identified as constituents of the remaining 10 per cent. Components of the fraction of acids which have been identified to date in steam distillates from stale perspiration but were not found in