Quantitative surface-enhanced Raman spectroscopy of dipicolinic acid—towards rapid anthrax endospore detection

Abstract

Dipicolinic acid (DPA) is an excellent marker compound for bacterial spores, including those of Bacillus anthracis (anthrax). Surface-enhanced Raman spectroscopy (SERS) potentially has the sensitivity and discrimination needed for trace DPA analysis, but mixing DPA solutions with citrate-reduced silver colloid only yielded measurable SERS spectra at much higher (>80 ppm) concentrations than would be desirable for anthrax detection. Aggregation of the colloid with halide salts eliminated even these small DPA bands but aggregation with Na₂SO₄(aq) resulted in a remarkable increase in the DPA signals. With sulfate aggregation even 1 ppm solutions gave detectable signals with 10 s accumulation times, which is in the sensitivity range required. Addition of CNS⁻ as an internal standard allowed quantitative DPA analysis, plotting the intensity of the strong DPA 1010 cm⁻¹ band (normalised to the ca. 2120 cm⁻¹ CNS⁻ band) against DPA concentration gave a linear calibration (R² = 0.986) over the range 0–50 ppm DPA. The inclusion of thiocyanate also allows false negatives due to accidental deactivation of the enhancing medium to be detected.