868 JOURNAL OF THE SOCIETY OF COSMETIC CHEMISTS Although the pressed cake contained only 4% moisture (or less), it was not regarded as an anhydrous product. It was easily wetted because of its water-soluble binder. The products were obtained from stock. Results are shown in Table IV. Fourteen subjects used the cake eyeliner. Thirteen cakes showed no plate count after use. One sample had 10 fungus/g. Thirteen brushes were collected. Twelve had a count of 8 or less and one had a count of 420. Pressed Powder Blusher (Dry Application) Pressed powder blusher contained 95% pigment-talc mixture and 5% of an anhydrous binder. It was applied dry with a nylon bristle brush. The material used was prepared in the laboratory without special handling. The plate count prior to use was 50/g. This un- preserved product was used by 6 subjects and both the product and brush were collected after 3 weeks of use. Results are shown in Table V. All used products had plate counts of less than 70/g. The enrichment and streaking procedures showed the presence of staphylococci on two units, gram-positive bacilli on a third, and no recovery from three. Actually, the profile of the product was practically the same after use as before use. The bristles of the brushes were washed with Tween- saline solution and the washings were plated. Counts ranged from 6 to 1000 per brush. Brushes before use had counts of 4 to 5. Table V Microbiological Profile of a Dry-Applied Pressed Powder Blusher without Preservatives after Use No. of Isolations From Product Subjects Plate Count Enrichment Blusher cake 6 70/g Staph, gram q- bacilli Brush 6 6-1000 Not done Liquid Eyeliner The liquid eyeliner was an oil-in-water, anionic emulsion of pH 8.2. All test samples were prepared in the laboratory with extreme care and aseptically filled into sterile containers. The pigments used were ethylene oxide-treated. At the time of consumer use, all samples showed a negative plate count. Controls were kept to assure that all unused products exhibited a negative plate count for 3 months. Three groups

MICROBIOLOGICAL PROFILE OF COSMETICS Table VI Microbiological Profile of a Liquid Eyeliner, with and without Preservatives, after Use 869 Plate Count s Isolation From Enrichment b No. of Upon After 3 Upon Preservative a Subjects collection months collection After 3 months None 10 8 = 10/g 9 = 100/g Staph 2 = TNTC 1 = TNTC Gram bacilli 0.075% MeP 8 N.R. N.R. 5 = N.R. 0.04% PP 3 = staph 0.2% MeP 10 N.R. N.R. N.R. 0.1% PP Staph Gram q- bacilli, mold 7 = N.R. 1 = mold N.R. a MeP = methyl paraben PP = propyl paraben. b N.R. = no recovery TNTC = too numerous to count. of eyeliners were prepared as follows: unpreserved with 0.075% methyl and 0.04% propyl paraben with 0.2% methyl and 0.1% propyl paraben. The second group represented a marginal or possible subopti- mum preservative level. Table VI shows that after use, the unpreserved liner had a plate count of from 10/g to TNTC. The product containing the low preservative level and the higher preservative level had negative plate counts immediately after use and 3 months after use. After enrichment streaking on differential media, the unpreserved samples showed the presence of gram-positive bacilli and Staphylococcus. Enrichment of the product containing the low preservative level showed Staphylococcus in two units and coagulase q- staphylocci from one unit. All other units were negative. After 3 months, the unpreserved units which had less than 10/g immediately after use were less than 100/g. Another unit went from TNTC to 130/g. One of the units which was TNTC upon collection remained TNTC after 3 months. The enrichment procedure showed recovery of gram-positive bacilli and mold in all unpreserved units. For the low preservative product, the enrichment procedure showed that one unit contained only fungi 3 months after use. All units containing the higher preservative level showed no recovery after 3 months. Emulsion Products The facial cleanser cream was a relatively high oil (45%), nonionic, oil-in-water emulsion supplemented with 5% of a natural ingredient (avocado oil). The pH was 5.9. The preserved product contained