Abstract:

An ultra-high performance liquid chromatography-triple quadrupole mass spectrometry (UPLC-QQQMS) method
was developed and validated to simultaneously determine seven constituents including chebumeinin A, chebumeinin B,
chebulic acid, pentagalloyl glucose, corilagin, chebulagic acid and gallic acid in dried whole pomegranate (containing
peels and seeds, a traditional Mongolian medicinal material), dried pomegranate peels (a traditional Chinese medicinal
material), and fresh pomegranate peels and seeds (Punica granatum L.). 4β-(L-acetylcysteinyl)-epicatechin was used
as an internal standard for the quantification. The internal standard was added to methanol before being used to extract
samples. The separation was performed on an Agilent ZORBAX Eclipse XDB-C18 column (2.1 mm × 50 mm, 1.8 μm) with
methanol-0.1% formic acid as the mobile phase at a flow rate of 0.4 mL/min. As a result, the linear ranges of chebumeinin
A, chebumeinin B, chebulic acid, pentagalloyl glucose, corilagin, chebulagic acid and gallic acid were 0.003 4–13.875 0,
0.013 5–13.875 0, 0.003 4–13.875 0, 0.216 8–55.500 0, 0.867 2–55.500 0, 0.216 8–55.500 0, and 0.003 4–13.875 0 μg/mL,
respectively, with correlation coefficients higher than 0.99. The average recovery rate of the developed method ranged
from 99.44% to 105.54% and the intra-day and inter-day relative standard deviations (RSDs) were 0.11%–0.95% and
0.14%–1.21%, respectively. This method is sensitive, simple, fast, and accurate, and can be used for the quality control of
pomegranate for medicinal use and related food products or pharmaceuticals.

Key words: Punica granatum, whole pomegranate, pomegranate peel, ultra-high-performance liquid chromatography triple quadrupole mass spectrometry (UPLC-QQQMS), simultaneous determination