5 α ‐Androst‐16‐en‐3 α ‐ol β ‐ D ‐Glucuronide, Precursor of 5 α ‐Androst‐16‐en‐3 α ‐ol in Human Sweat

5 α ‐Androst‐16‐en‐3 α ‐ol ( α ‐androstenol) is an important contributor to human axilla sweat odor. It is assumed that α ‐andostenol is excreted from the apocrine glands via a H 2 O‐soluble conjugate, and this precursor was formally characterized in this study for the first time in human sweat. The possible H 2 O‐soluble precursors, sulfate and glucuronide derivatives, were synthesized as analytical standards, i.e. , α ‐androstenol, β ‐androstenol sulfates, 5 α ‐androsta‐5,16‐dien‐3 β ‐ol ( β ‐androstadienol) sulfate, α ‐androstenol β ‐glucuronide, α ‐androstenol α ‐glucuronide, β ‐androstadienol β ‐glucuronide, and α ‐androstenol β ‐glucuronide furanose. The occurrence of α ‐androstenol β ‐glucuronide was established by ultra performance liquid chromatography (UPLC)/MS (heated electrospray ionization (HESI)) in negative‐ion mode in pooled human sweat, containing eccrine and apocrine secretions and collected from 25 female and 24 male underarms. Its concentration was of 79 ng/ml in female secretions and 241 ng/ml in male secretions. The release of α ‐androstenol was observed after incubation of the sterile human sweat or α ‐androstenol β ‐glucuronide with a commercial glucuronidase enzyme, the urine‐isolated bacteria Streptococcus agalactiae , and the skin bacteria Staphylococcus warneri DSM 20316, Staphylococcus haemolyticus DSM 20263, and Propionibacterium acnes ATCC 6919, reported to have β ‐glucuronidase activities. We demonstrated that if α ‐ and β ‐androstenols and androstadienol sulfates were present in human sweat, their concentrations would be too low to be considered as potential precursors of malodors; therefore, the H 2 O‐soluble precursor of α ‐androstenol in apocrine secretion should be a β ‐glucuronide.