This article describes a unique noninvasive capability to determine the concentration (in mg/cm3) and total amount of topically applied materials in the skin (in μg/cm2 of skin surface). It is based on in vivo confocal Raman spectroscopy. A theoretical derivation is given of a general method to calculate a concentration ratio from a Raman spectrum of a material in a medium, which can be a solvent or other matrix, such as the skin. A practical implementation of the method is then presented along with a clarification of the assumptions used and applied to a quantitative analysis of the in vivo skin penetration of trans-retinol and propylene glycol (PG). A comparison was made between the concentrations profiles of retinol and PG found in the skin and the concentrations of retinol and PG that had been applied to the skin. Determination of the amount of these materials in the skin at different timepoints after topical application also enabled a straightforward calculation of the flux of materials into the skin (in μg cm−2 h).
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