Ultrasensitive detection of thyrotropin-releasing hormone based on azo coupling and surface-enhanced resonance Raman spectroscopy.

Surface-enhanced resonance Raman scattering (SERRS) has been used to establish a rapid and quantitative assay based on the diazotization coupling reaction for thyrotropin-releasing hormone (TRH). Ultrahigh sensitivity of this approach originates from two factors: changing TRH to an azo compound and the SERRS effect with the addition of silver nanoparticles (AgNPs) at 532 nm excitation wavelength. The lowest detectable concentration of TRH was found to be as low as 1 pg mL(-1), which is 10-fold lower than the lowest normal reference value in human serum reported in previous literature. The quantitative measurements in human serum based on this method were conducted, and the results showed its feasibility for detection in complex biological samples. In comparison with conventional TRH identification and quantification methodologies, radioimmunoassay (RIA) and subsequent various hyphenated techniques, the main advantages of this study are simplicity, rapidness (2 minutes), time effectiveness, no additional steps required to further characterize the immunogenic material, highest sensitivity (57.1 fg), high selectivity, practicality and reliability. Thus, this work puts forward a research tool that may be applied to the determination of TRH in practical assays.