EFFECTS OF LIPID EXTRACTION AND SOAKING 19 amino acids other than those from fi laggrin proteolysis are likely, we believe that the present method measures NMF, since citrulline is not generated in SC other than from fi laggrin proteolysis, which is the source of the bulk of the free amino acids found in SC (23–25). In addition, the other amino acids, as well as the sum, follow the same patterns of signifi cance as citrulline in these studies. TREATMENT EFFECTS Signifi cant differences between the treatments were also observed (Figure 2). For the amino acids of greater prevalence in NMF and the SC, the fi ndings were as follows: (a) for citrulline at tape 1, all treatments were signifi cantly different (b) for the summed AAs at tape1, the control was signifi cantly different from the extracted and soaked (ES) site and (c) the extracted site was different from all others. For tape 3, (a) signifi cant differences between ES and the other three treatments as well as between the extracted (E) and soaked (S) sites were seen for the summed AAs, Cit, His, and Phe (b) for leu- cine, ES was signifi cantly different from C, S, and E, and for serine, ES was signifi cantly different from E. At strip 5, the summed AAs, His, Leu, and Phe all showed a signifi - cant difference between ES and the other three sites for citrulline, all comparisons were signifi cant. Because all treatments converged by tape 10, tapes 1, 3, and 5 were summed to examine treatment effects over the upper portion of the SC. For the summed AAs, the sum of tapes 1, 3, and 5 confi rms the trends shown in the individual tape strips, with site ES differing from all others and E differing from S, but not from C. E and ES are different for serine. ES is different from all for histidine. For citrulline, E is different from all. For both leu- cine and phenylalanine, E is different from all and S and ES are different. Table III pro- vides a summary of the signifi cant differences in NMF. Figure 2. Signifi cant reduction in NMF levels was caused by extraction and soaking, with the combination producing especially low levels of NMF. *Indicates signifi cant difference from control, p 0.001.

Table III Signifi cant NMF Results All AAs Ser Gly His Arg Cit Phe Strip 1 E vs S, ES, C ES vs C E vs ES E vs ES, S E vs S, ES, C ES vs C NS E vs ES, C, S C vs S S vs ES E vs S, ES, C ES vs C Strip 3 ES vs C, E, S E vs S E vs ES E vs ES ES vs S, E, C E vs S ES vs E, C E vs C, S, ES E vs S ES vs C, E, S E vs S Strip 5 ES vs S, E, C None ES vs ES ES vs E, C, S ES vs E, S E vs ES ES vs E, C, S Sum 1, 3, 5 ES vs E, C, S E vs S E vs ES ES vs E, C ES vs E, C, S E vs ES E vs ES, C, S E vs ES, C, S ES vs S Sum 1, 3, 5, 10 ES vs E, C, S NS E vs ES ES vs E, C, S ES vs E, C, S ES vs E, C ES vs E, C, S Control 1 vs 3, 5, 10 3 vs 10 1 vs 3, 5, 10 1 vs 3, 5, 10 1 vs 3, 5, 10 3 vs 10 1 vs 3, 5, 10 1 vs 3, 5, 10 3 vs 10 1 vs 3, 5, 10 3 vs 10 Extracted 1 vs 3, 5, 10 1 vs 3, 5, 10 1 vs 3, 5, 10 1 vs 3, 5, 10 1 vs 3, 5, 10 1 vs 3, 5, 10 1 vs 3, 5, 10 Soaked 1 vs 3, 5, 10 3 vs 5, 10 1 vs 3, 5, 10 3 vs 5, 10 1 vs 3, 5, 10 1 vs 3, 5, 10 3 vs 5, 10 1 vs 3, 5, 10 3 vs 10 1 vs 3, 5, 10 3 vs 10 1 vs 3, 5, 10 3 vs 10 Extracted and soaked 1 vs 5, 10 3 vs 5, 10 1 vs 5, 10, 3 vs 10, 5 vs 10 1 vs 3, 5, 10 3 vs 5, 10 5 vs 10 1 vs 3, 5, 10 3 vs 5, 10 5 vs 10 1 vs 3, 5, 10 3 vs 5, 10 1 vs 3, 5, 10 3 vs 5, 10 5 vs 10 1 vs 3, 5, 10 3 vs 5, 10 C = control E = extracted only S = soaked only ES = extracted and soaked site. Tape strip numbers are listed as numbers onlye.g., , 1 for tape strip 1. All comparisons listed are signifi cant at p 0.05. JOURNAL OF COSMETIC SCIENCE 20