Accelerated 13C detection by concentrating the NMR sample in a biphasic solvent system

Abstract

CDCl₃ is the most frequently used solvent for the NMR investigation of organic compounds. Busy chemistry labs need to investigate hundreds of compounds daily. While ¹H NMR investigation takes a couple of minutes, recording ¹³C NMR spectra necessitates hours of signal averaging due to the low abundance and low sensitivity of ¹³C nuclei. The longer acquisition time for ¹³C NMR results in a loss of precious spectrometer time in a shared multi-user environment. A regular 5 mm o.d. NMR tube is the most commonly used tube for NMR in organic chemistry labs and is also the cheapest option. We show that for analytes soluble in the CDCl₃ solvent using a regular 5 mm o.d. NMR tube, the speed of ¹³C observation can be enhanced by a factor of two by resorting to a sample preparation method that employs a biphasic system made of H₂O or D₂O at the top of another layer of CDCl₃. By using the biphasic system of two immiscible solvents, the analyte can be concentrated in the CDCl₃ layer (within the more sensitive volume of the NMR coil), resulting in the improvement of the signal to noise ratio (SNR) by a factor of up to 1.8 for ¹³C and 2D ¹H–¹³C HSQC spectra, which results in more than two-fold reduction in the experimental time. ¹H NMR and other 2D NMR also get a sensitivity boost. The amount of CDCl₃ required for sample preparation can also be reduced by 40% using this biphasic system (CDCl₃/H₂O). Sample preparation in such an immiscible biphasic system is effortless and straightforward. The performance of such biphasic samples is closer to that of Shigemi tubes and better than that of 3 mm o.d. tubes.