Validated Quantitative 1H NMR Method for Simultaneous Quantification of Indole Alkaloids in Uncaria rhynchophylla.

Abstract

Uncariae Ramulus Cum Uncis, known as “Gou-Teng” in Chinese, is derived mainly from the dried hook-bearing stems of Uncaria rhynchophylla. Quantitative determination of monoterpenoid indole alkaloids is critical for controlling its quality. In the present study, a rapid, accurate, and precise method was developed for the simultaneous quantitation of four characteristic components, namely, rhynchophylline (1), isorhynchophylline (2), corynoxeine (3), and isocorynoxeine (4), through 1H NMR spectrometry techniques. This method was performed on a 600 MHz NMR spectrometer with optimized acquisition parameters for performing quantitative experiments within 14 min. The highly deshielded signal of NH was at δH 10–11 in the aprotic solvent DMSO-d6, which enables satisfactory separation of the signals to be integrated. Validation of the quantitative method was also performed in terms of specificity, linearity, sensitivity, accuracy, and precision. The method is linear in the concentration range of 25–400 μg/mL. The lower limit of quantification is 25 μg/mL. The intra- and interday relative standard deviation across three validation runs over the entire concentration range is less than 2.51%. The accuracy determined at three concentrations was within ±4.4% in terms of relative error. The proposed qNMR method was demonstrated to be a powerful tool for quantifying the alkaloids in traditional Chinese medicines (TCMs) due to its unique advantages of high precision, rapid analysis, and nonrequirement of standard compounds for calibration curve preparation. Moreover, qNMR represents a feasible alternative to high-performance liquid chromatography-based methods for the quality control of TCMs.