Detection of bio-constituents in complex biological tissue using Raman microscopy. Application to human nail clippings

Raman spectra of human nail clippings from various sources were collected and then deconvoluted to obtain the pure component spectra of the underlying constituents present. This blind-deconvolution was performed using a self-modeling curve resolution technique, namely band-target entropy minimization (BTEM). The aim was to simplify the complexity of the Raman spectra and hence to identify the underlying biological molecules in more detail. BTEM analysis could recover 13 pure component Raman spectral estimates from the collected 438 spectra measured from 113 nail samples. Six recovered pure component spectral estimates correspond to proteins or polypeptides that contain various amino acids such as phenylalanine, tyrosine, tryptophan, and cysteine. Two are associated with the secondary structures of proteins, and five are associated with two carotenoid species, lipid, ferulic acid, and calcium phosphate. Subsequently, the relative concentrations of these bio-constituents were calculated from the measured mixture spectra and the pure component BTEM estimates. These profiles indicated that the concentrations of some bio-constituents are correlated while others are not. A further analysis using target transformation factor analysis (TTFA) revealed the possible presence of curcumin in the human nails. Since the present approach and analysis is rather general, it might be extended to many other biological tissues in a rather straightforward and similar manner, thus revealing more detailed underlying biochemical information such as biomarkers that may be useful for diagnostic purposes.