Feasibility study for the rapid screening of target molecules using translational diffusion coefficients: diffusion-ordered NMR spectroscopy of biological toxins

Henderson, Terry J.
February 2010
Analytical & Bioanalytical Chemistry;Feb2010, Vol. 396 Issue 4, p1465
Academic Journal
A panel of 15 biological toxins ranging between ~60–28,000 g/mol was used to evaluate the feasibility of screening aqueous samples for toxin analytes based on their translational diffusion coefficients, Dt. Toxin Dt values were measured by pulsed-field gradient 1H NMR spectroscopy using a bipolar pulse pair, longitudinal eddy current delay pulse sequence incorporating water suppression to achieve the maximum dynamic range for toxin signals. To collect data for an effective screening protocol, reference Dt values were determined from five independent measurements at both 25 and 37 °C for all toxins in the panel. In the protocol, Dt values are measured at both temperatures for a suspected toxin target in a sample, and for assignment as a potential toxin analyte, the measurements are required to fall within ±0.25 × 10−6 cm2/s of both reference Dt values for at least one toxin in the panel. Only solution viscosity was found to influence sample Dt measurements appreciably; however, the measurements are easily corrected for viscosity effects by calculating the Dt value of the suspected toxin at infinite dilution. In conclusion, the protocol provides a rapid and effective means for screening aqueous samples for all toxins in the panel, narrowing toxin identification to ≤2 possibilities in virtually all cases.


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