Feasibility of discrimination of gall bladder (GB) stone and GB polyp using voltage-applied SERS measurement of bile juice samples in conjunction with two-trace two-dimensional (2T2D) correlation analysis.

Voltage-applied SERS measurement of bile juice in conjunction with two-trace two-dimensional (2T2D) correlation analysis was demonstrated as a potential tool to enhance discrimination of gall bladder (GB) stone and GB polyp. When SERS spectra of the aqueous phases extracted from raw bile juice samples were measured with applying external voltage from -300 to +300 mV (100 mV intervals), subsequent spectral variations of the adsorbed components (bilirubin-containing compounds) on the SERS substrate were minute, and discrimination of the two GB diseases in a principal component score domain was difficult. Therefore, 2T2D correlation analysis, effectively identifying asynchronous (dissimilar) spectral behaviors in the voltage-induced SERS spectra, was used to improve the discrimination. When two spectra of a sample collected with application of +100 and +300 mV were adopted, the features of subsequent 2T2D slice spectra were characteristic, and discrimination of GB stone and GB polyp substantially improved. External voltage application and recognition of the voltage-induced spectral features by 2T2D correlation analysis were key factors for the improvement. Since the demonstrated method relied on only a few SERS-active compounds, infrared (IR) spectroscopy featuring all the present components in the samples was also evaluated for comparison. However, the IR-based discrimination was inferior because the metabolite compositions in the samples between the GB diseases were not noticeably different.