Visible Light Spectroscopy of Liquid Solutes from Femto- to Attoliter Volumes Inside a Single Nanofluidic Channel.

UV-vis spectroscopy is a workhorse in analytical chemistry that finds application in life science, organic synthesis, and energy technologies like photocatalysis. In its traditional implementation with cuvettes, it requires sample volumes in the milliliter range. Here, we show how nanofluidic scattering spectroscopy (NSS), which measures visible light scattered from a single nanochannel in a spectrally resolved way, can reduce this sample volume to the attoliter range for solute concentrations in the mM regime, which corresponds to as few as 10 probed molecules. The connection of the nanochannel to a microfluidic in-and-outlet system enables such measurements in continuous flow conditions, and the integrated online optical reference system ensures their long-term stability. On the examples of the nonabsorbing solutes NaCl and HO and the dyes Brilliant Blue, Allura Red, and Fluorescein, we demonstrate that spectral fingerprints can be obtained with good accuracy and that solute concentrations inside the nanochannel can be determined based on NSS-spectra. Furthermore, by applying a reverse Kramers-Kronig transformation to NSS-spectra, we show that the molar extinction coefficient of the dye solutes can be extracted in good agreement with the literature values. These results thus advertise NSS as a versatile tool for the spectroscopic analysis of solutes in situations where nanoscopic sample volumes, as well as continuous flow measurements are critical, e.g., in single particle catalysis or nanoscale flow cytometry.