Development and validation of an UPLC-ESI-MS/MS method for determination of dehydroevodiamine, limonin, evodiamine, and rutaecarpine in Evodiae Fructus

Abstract

Objective: Evodiae Fructus (EF), one of the most widely used traditional Chinese medicines, mainly consists of alkaloids, is widely used for the treatments of headache and gastrointestinal disorders. In this study, a sensitive and reliable UPLC-ESI-MS/MS method was developed for qualitative determination of dehydroevodiamine, limonin, evodiamine, and rutaecarpine.

Materials and methods: Chromatographic separations were accomplished on a Phenomenex Kinetex XB-C18 column (2.1 × 150 mm, 1.7 μm) by using a gradient elution profile with a mobile phase consisting of 0.5% formic acid in water (A) and acetonitrile (B). Detection was performed using multiple reactions monitoring mode under ESI in the positive ion mode.

Results: The results showed good linearity over the investigated concentration ranges (R (2)>0.9900) for the analytes. The limit of quantitations (LOQs) were 6.88 ng/mL for dehydroevodiamine, 18.6 ng/mL for limonin, 6.24 ng/mL for evodiamine, and 2.56 ng/mL for rutaecarpine, respectively. Intraday and interday precisions (relative standard deviations, %) were <5% and accuracies ranged from 92% to 106%.

Conclusion: The validated method was successfully applied to assay the contents of the four compounds in EF samples from different regions, with which just 10 min was needed to analyze each sample.

Keywords: Evodiae Fructus; Quantitative determination; UPLC-ESI-MS/MS.

Figure 1

Representative MRM chromatograms of (a) 80% ethanol blank and (b) mixed standards, and (c) sample solution. (a) Dehydroevodiamine; (b) limonin; (c) evodiamine; and (d) rutaecarpine

Figure 2

Spectra of product ions of (a) dehydroevodiamine, (b) limonin, (c) evodiamine, and (d) rutaecarpine

Figure 3

The cumulative variance of the four generated principal components. The first two components account for 94.98% of the total variance

Figure 4

PCA scores plot of the 19 EF samples using the contents of the four analytes as input data

Figure 5

PC1 (a) and PC2 (b) loadings plot of the four variables