Surface-enhanced Raman detection of RNA and DNA bases following flow-injection analysis or HPLC separation

Therese M. Cotton; Rong-Sheng Sheng; Fan Ni

doi:10.1117/12.22922

Abstract

The goal of this study is to develop Surface-enhanced Raman scattering (SERS) detection methods for flow injection analysis (FIA) and high performance liquid chromatography (HPLC). Nucleic acid bases have been chosen for analysis because of their importance in life processes. The advantages to the use of SERS-based detection include its sensitivity, specificity and versatility. With the development of improved methodology, the detection limits should be comparable to UV spectroscopy. However, the specificity is considerably superior to that obtained with electronic spectroscopy in that the Raman spectrum provides a molecular fingerprint of the individual analytes. Raman spectroscopy is very versatile: aqueous samples, gases and solids can be analyzed with equal facility. The results presented here demonstrate that SERS can be used as a detection method for both FIA and HPLC detection. In the following experiments Ag sols have been used as the active substrate. The effect of various parameters such as temperature, pH, flow rate, and the nature of the interface between the HPLC system and the Raman spectrometer have been examined. One of the most significant findings is that the temperature of the Ag sol/HPLC effluent mixture has a dramatic effect on the SERS intensities. This effect is a result of increased colloid aggregation at higher temperatures. Aggregation is known to produce greater enhancement in SERS and proceeds much more rapidly at elevated temperatures. An increase in the temperature of the Ag sol enables SERS detection under flowing conditions and in real time. This is a substantial improvement over many of the previous attempts to interface SERS detection to FIA or HPLC. In most of the previous studies, it was necessary to stop the flow as the analyte eluted from the chromatogram and measure the SERS spectra under static conditions.

Citation Download Citation

Therese M. Cotton, Therese M. Cotton, Rong-Sheng Sheng, Rong-Sheng Sheng, Fan Ni, Fan Ni, "Surface-enhanced Raman detection of RNA and DNA bases following flow-injection analysis or HPLC separation", Proc. SPIE 1336, Raman and Luminescence Spectroscopies in Technology II, (1 November 1990); doi: 10.1117/12.22922; https://doi.org/10.1117/12.22922

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