Despite consistent evidence that greater consumption of fruits and vegetables (FV) is associated with significant reductions in chronic disease morbidity and mortality, the majority of adults in the United States consume less than the amounts recommended by public health agencies. As such, there is a critical need to design and implement effective programs and policies to facilitate increases in FV consumption for the prevention of these diseases. To accomplish this, an accurate, inexpensive, and convenient method for estimating the dietary FV intake is required. A promising method for quantifying the FV intake via proxy that has gained interest in recent years is the measurement of skin carotenoid levels via spectroscopy-based devices. However, there exist certain dietary and non-dietary factors that may affect the skin carotenoid levels independently of the dietary intake of carotenoids. In order to validate the ability of this method to accurately estimate the FV intake among diverse demographics, these factors must be identified and taken into consideration. Therefore, this narrative review seeks to summarize the available research on factors that may affect the skin carotenoid levels, determine current gaps in knowledge, and provide guidance for future research efforts seeking to validate spectroscopy-measured skin carotenoid levels as a means of accurately estimating the FV intake among various populations.