SIMULTANEOUS DETERMINATION OF FOUR FLAVONOID AGLYCONE ANALOGS IN THE INFLORESCENCE OF JUGLANS REGIA L. BY HIGH-PERFORMANCE LIQUID CHROMATOGRAPHY-DIODE ARRAY DETECTION

Abstract

A reversed-phase high-performance liquid chromatography coupled with a diode array detection method and a high-performance liquid chromatography-electrospray ionization tandem mass spectrometry method were first developed and validated for the simultaneous determination of four flavonoid aglycone analogs in the inflorescence of Juglans regia L. grown in different areas of Gansu province, China, surmounting the challenges rising from the complexity of the matrices owing to different origins of the plant. The separation of the four compounds was accomplished through a ternary mobile phase, with good specificity and satisfactory resolution. The chromatographic peaks of the four investigated compounds were identified by comparing their retention time and mass spectrometric profile with the corresponding reference compounds. The method showed a good linearity (r² > 0.9998), high precision (RSD <5%), and recoveries (ranged from 96.6–102.7%) were acceptable. The limits of detection (LOD) and the limits of quantification (LOQ) are ranged from 0.26–0.58 µg/ml and from 0.86–1.90 µg/ml, respectively. The results demonstrated that this analytical approach is ideal for the determination of such bioactive compounds as flavonoid aglycones, and that it constructed a basis for the comprehensive evaluation of the quality of Juglans regia L.

Keywords:

• flavonoid aglycones analogs
• high-performance liquid chromatography-diode array detection
• high-performance liquid chromatography– electrospray tandem mass spectrometry
• Juglans regia L
• simultaneous determination

ACKNOWLEDGMENT

The authors gratefully acknowledge the Fund of Scientific and Technical research of Traditional Chinese Medicine of Gansu province (GZK-2010-57) and the Fund of the Scientific and Technical Innovation Project of Gansu province (1104FKCA122) for financially supporting this research. The authors thank Prof. Chong Li for providing reference standards.

Notes

UV is ultraviolet.

^a x, amount of substance (µg); y, peak area.

^b LOD, limit of detection (S/N = 3).

^c LOQ, limit of quantification (S/N = 10).

^a (SD/mean) × 100. RT is retention time.

^a [(Mean of measured concentration-spiked concentration)/spiked concentration] × 100.

^b (S.D./mean) × 100.

^a RD, relative deviation (%) = (|measured value mean|/mean) × 100.

^b Inflorescences of Julans regia L. from Gansu province.

^c The data were presented as average of three replicates.

^d The sum of four compounds including Quercetin-3-O-β-D-glucoside, Quercetin-3-O-α-L-arabinopyranoside, Kaempferol-3-O-β-D-glucoside, and Quercetin-3-O-α-L-arabinofuranoside.