Purity determination of pyributicarb by internal standard correction-high-performance liquid chromatography-quantitative nuclear magnetic resonance.

Abstract

An internal standard correction-high-performance liquid chromatography-quantitative nuclear magnetic resonance (ISC-HPLC-qNMR) procedure was established as a reliable quantitative method for complex organic compounds with low purity in order to solve the risk of qNMR inaccuracy because of insufficient resolution of impurity peaks from the selected quantitative peak. This method collects a small quantity of target analyte from low-purity organics by LC. After drying and re-dissolving in deuterated solvent containing internal standard, the solution was analyzed by 1H NMR and HPLC. Another solution prepared by accurately weighing unpurified low-purity substance and internal standard was analyzed by HPLC. Based on the theoretical derivation derived from the Beer-Lambert law, using the ratio of the HPLC peak areas of two solutions as correction, the purity was then calculated without the same reference as target analyte. Compared to previous methods with similar selectivity and accuracy, it has advantages such as a less purified sample is required, time for lyophilization is reduced by half, and sample preparation is more controllable. The proposed method was verified by analysis of a suite of six commercially available, high-purity compounds, and the difference of results between it and direct qNMR was within 0.1%. The result of pyributicarb using ISC-HPLC-qNMR was 97.6% (U = 0.5%; k = 2), and the reference value was 97.61% (U = 0.22%; k = 2). The results demonstrate that the proposed method provides a new way for reference material producers to calibrate lower-purity organics and has the potential advantage of accurate quantification of lower-purity organics.