This review summarizes the possibility to diagnosing the chronic diseases and application of determination of metabolites of the noninvasive human nails. A target derivatization UPLC-ESI-MS method is a powerful tool for the analysis of bioactive compounds with high sensitivity and selectivity. We developed target derivatization UPLC-ESI-MS methods to analyze the biological trace components in human fingernail. Moreover, we have used these methods to detect the metabolites in human fingernails, such as DL-amino acids, polyamines and Advanced Glycation End products (AGEs). Through detecting the biological trace components in the real samples from human, we tested the possibility to use these new methods to diagnosing the chronic diseases, i.e., diabetes, lung cancer. Some applications utilizing these target derivatization methods for the analyses determination of metabolites in the human fingernails are also described in this paper.