Volatile organic compounds (VOCs) emanating from human skin contribute to an individual’s body odour. Understanding the modulation of human odour by a fragrance is of significant importance to the cosmetic sector in the design, development and evaluation of new products. The present research describes an in vivo approach for passive headspace sampling of skin volatile emissions in human participants. A wearable headspace solid-phase microextraction (HS-SPME) method has been employed to investigate baseline endogenous skin volatiles and the subsequent modulation of skin volatile profiles after application of a fragrance to skin. Coupled with gas chromatography-mass spectrometry (GC-MS) this method enables characterisation of scent profiles and fragrance longevity in vivo. A total of 51 compounds were identified in participants’ skin, including 19 endogenous and 32 fragrance-derived compounds. The temporal variation in volatile profiles at different times after fragrance application was investigated. Fragrance diffusion from skin varied between participants resulting in diversified scent profiles over time. This non-invasive approach could be employed during cosmetic product development for in vivo evaluation of fragrance profiles and for assessment of the retention of fragrance components in skin to reduce reliance on expert panels during product development.