We present the construction and performance of a 20-μL active volume probe that utilizes zerosusceptibility wire for the detection transceiver coil and a 3.5 mm outer diameter thin-wall bubble flow cell to contain the sample. The probe shows good rf homogeneity, resolution, line shape and sensitivity. The sensitivity and resolution of the 20-μL probe was compared to those for several other coil configurations, including smaller detection volumes, a thin wire copper coil immersed in susceptibility matching perfluorocarbon FC-43 (fluorinert) fluid, and a standard 5 mm probe. In particular, the 1 H mass sensitivity, S m (SNR per micromole), was 3-4 fold higher than that for the standard 5 mm probe. Finally, the use of the zero-susceptibility wire in smaller volume probes is discussed along with potential future improvements and applications.

Improved NMR detection of mass limited samples can be obtained by taking advantage of the mass sensitivity of microcoil NMR, while throughput issues can be addressed using multiple, parallel sample detection coils. We present the design and construction of a double resonance 300-MHz dual volume microcoil NMR probe with thermally-etched 440-nL detection volumes and fused silica transfer lines for high-throughput stopped-flow or flow-through sample analysis. Two orthogonal solenoidal detection coils and the novel use of shielded inductors allowed the construction of a probe with negligible radio-frequency cross talk. The probe was resonated at 1 H-2 D (upper coil) and 1 H-13 C (lower coil) frequencies such that it could perform 1D and 2D experiments with active locking frequency. The coils exhibited line widths of 0.8 to 1.1 Hz with good mass sensitivity for both 1 H and 13 C NMR detection. 13 C directly detected 2D HETCOR spectra of 5% v/v 13 C labeled acetic acid were obtained in less than 5 min. Demonstration of the probe characteristics as well as applications of the versatile two-coil double resonance probe are discussed.

A new HPLC-NMR system is described that performs analytical separation, pre-concentration, and NMR spectroscopy in rapid succession. The central component of our method is the online preconcentration sequence that improves the match between post-column analyte peak volume and the micro-coil NMR detection volume. Separated samples are collected on to a C18 guard column with a mobile phase composed of 90% D 2 O/10% acetonitrile-D3, and back-flashed to the NMR microcoil probe with 90% acetonitrile-D3/10% D 2 O. In order to assess the performance of our unit, we separated a standard mixture of 1 mM ibuprofen, naproxen, and phenylbutazone using a commercially available C18 analytical column. The S/N measurements from the NMR acquisitions indicated that we achieved signal enhancement factors up to 10.4 (±1.2)-fold. Furthermore, we observed that pre-concentration factors increased as the injected amount of analyte decreased. The highest concentration enrichment of 14.7 (±2.2)-fold was attained injecting 100 μL solution of 0.2 mM (~4 μg) ibuprofen.

Recent advances in microcoil NMR have provided commercially available, robust methodologies for analyzing mass and volume limited samples in the low microliter regime, and the technology has been applied in a number of areas. Unfortunately, due to constraints on sample size and the limited solubility of some compounds of interest, the application of this approach to certain areas of development, such as the structural analysis of chromatography eluates, is restricted. A current challenge is to provide an option within a previously unexplored sample size regime (tens of microliters) while still taking advantage of the increase in mass sensitivity afforded by solenoidal microcoil NMR. In this article, we present the design and construction of a microcoil NMR probe with a custom detection cell for the routine analysis of 20-lL samples. The detection cell is comprised of a CO 2 -laser-heated HF-etched borosilicate active volume with fused silica transfer lines added to provide sample input and output. This setup produces an enlarged sample bubble within the detection coil and provides easy connection with 1/16 in. standard LC connections, lending itself to applications with HPLC-NMR, online SPE and similar separation techniques, as well as higher-throughput robotic automation. NMR performance characteristics determined using standard compounds showed the probe exhibited reasonable resolution (<0.01 ppm), although sensitivity was less than optimal due to tuning constraints. Future improvements and opportunities are also discussed.

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