Known pheromones are not present in the bioactive NP-SPE fraction from three different strains.

<p>(<b>A</b>) Male urine of BALB/cJ, C57BL/6J, and CD-1 each diluted 1∶3 in PBS promotes uterine weight increase in female mice (Dunnett's). (End body masses are not significantly different, p = 0.839). Averages of body mass (g): PBS, 17.29; BALB/cJ, 17.47; C57, 17.40; CD-1, 17.26. n = 27 per group. (<b>B</b>) Females exposed to the NP-SPE amino-column elutions from three different mouse strains display uterine weight gain (Dunnett's). (End body masses are not significantly different p = 0.662.) Averages of body mass (g): PBS, 17.41; BALB/cJ, 17.40, C57, 17.39; CD-1, 17.11.n = 14–15 per group. (<b>C</b>) GCMS chromotograph of pheromone standards HMH (1), SBT (3), and β-farnesene (4) as indicated. Fragmentation patterns of standards were used for analysis. (<b>D</b>) GCMS total ion chromatographs for known pheromones from total urine (black trace) and bioactive NP-SPE fractions (red trace) from BALB/c, C57BL/6J, and CD-1 male donors. Detection of HMH (1), DHB (2), and SBT (3) are indicated. No extractable compounds, including these known pheromones, were detectable in NP-SPE samples. The NP-SPE red trace is offset below the urine trace to visualize the lack of detectable peaks, the y-axis is otherwise identical. (<b>E</b>) LCMS/MS analysis of bioactive NP-SPE samples from BALB/c, C57BL/6J, and CD-1 male donors does not detect the presence of IAA and IBA using fragmentation patterns obtained from standards (top) as indicated. Arrow indicates the exact mass of the parent ion. (<b>F</b>) Analysis of EEARSM peptide using the mass of the transition state during dissociation of the standard (top). EEARSM was not detectable in any of the NP-SPE samples from any strain. In panels E-F the y-axis are identical between standards and NP-SPE samples. *p<0.05, **p<0.01.