Detection of Separated Analytes in Subnanoliter Volumes Using Coaxial Thermal Lensing

Abstract

A collinear-beam thermal lens detector has been constructed and its properties were characterized. Its application to the high-performance liquid chromatography (HPLC) separation of a mixture of five anthraquinone dyes dissolved in water shows a linear response over 3.5 orders of magnitude and a detection limit that is subnanomolar in the dye concentrations. These results are compared with those obtained previously using cavity ring-down spectroscopy (CRDS) in a Brewster's angle flow cell (Bechtel, K. L.; Zare, R. N.; Kachanov, A. A.; Sanders, S. S.; Paldus, B. A. Anal. Chem. 2005, 77, 1177-1182). The peak-to-peak baseline noise of the thermal lensing detection is 3.5 x 10(-8) absorbance units (AU) with a path length of 200 microm, whereas the peak-to-peak baseline noise of CRDS detection is approximately 2 x 10(-7) AU with a path length of 300 microm. Both of these figures of merit should be compared to the peak-to-peak baseline noise of one of the best commercial UV-vis HPLC detection systems, which is approximately 5 x 10(-6) AU with a path length of 10 mm (1-s integration time). Therefore, the thermal lensing technique has a demonstrated sensitivity of subnanomolar detection that is approximately 140 times better than that of the best commercial UV-vis detector and approximately 5 times better than that of CRDS.