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Chromatographic Analysis and Anti-Oxidative Property of Naoxinqing Tablet, a Proprietary Preparation of Diospyros Kaki Leaves

NICM Health Research Institute, Western Sydney University, Penrith, NSW 2751, Australia
National Measurement Institute, North Ryde, NSW 2113, Australia
School of Medicine, Western Sydney University, Penrith, NSW 2571, Australia
Wentworth Institute, Surry Hills, NSW 2010, Australia
Faculty of Medicine, University of New South Wales, Kensington, NSW 2052, Australia
Faculty of Medicine and Health Sciences, Macquarie University, Macquarie Park, NSW 2113 Australia
Authors to whom correspondence should be addressed.
Academic Editors: Susana M. Cardoso and Alessia Fazio
Molecules 2019, 24(6), 1101;
Received: 31 October 2018 / Revised: 8 March 2019 / Accepted: 11 March 2019 / Published: 20 March 2019
(This article belongs to the Special Issue The Antioxidant Capacities of Natural Products 2019)
PDF [1145 KB, uploaded 20 March 2019]


The Naoxinqing (NXQ) tablet is a standardised proprietary herbal product containing an extract of persimmon leaves (Diospyros kaki) for the management of cardio- and cerebrovascular diseases. Although previous reports suggested that the efficacy of NXQ is at least partly mediated by its anti-oxidative property, the anti-oxidative effect of the major components of NXQ has not been studied systematically. For quality control purposes, only analytical methods limited to 3 marker analytes have been reported, the extent to which the other components affect efficacy has not been explored. In this study, we developed an ultra-performance liquid chromatography-tandem mass spectrometry (UPLC MS/MS) method for the identification of seven analytes (kaempferol-3-O-glucoside (astragalin), quercetin-3-O-galactoside (hypericin), quercetin-3-O-glucoside (isoquercitin), kaempferol, 3,4-dihydroxybenzoic acid (protocatechuic acid), and furan-2-carboxylic acid (pyromucic acid) and quercetin) in the NXQ. This is the first method reported and validated for the quantification of the seven major secondary metabolites in NXQ. The results for the quantified analytes were then compared in 15 different batches of NXQ. The variation observed in the seven components highlights the need to quantify key bioactive components to ensure product consistency. Radical scavenging activity and abundance was used to rank the analytes. The anti-oxidative effects of NXQ were examined using cultured human vascular endothelial cells (EA.hy926). Corrected 2,2-di(4-tert-octylphenyl)-1-picrylhydrazyl (DPPH) activity results revealed that quercetin and kaempferol have the strongest anti-oxidant capacity in the extract. Both quercetin and kaempferol significantly inhibited the hydrogen peroxide (H2O2)-induced EA.hy926 cell injury and intracellular reactive oxygen species (ROS) generation. In conclusion, we established and validated an UPLC-MS/MC method for the analysis of major bioactive components in the NXQ and demonstrated that its anti-oxidative property may play a critical role in cerebrovascular protection. View Full-Text
Keywords: Naoxinqing; UPLC MS/MC analysis; anti-oxidative; endothelial cells; reactive oxygen species Naoxinqing; UPLC MS/MC analysis; anti-oxidative; endothelial cells; reactive oxygen species

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Kazzem, M.; Sun, Y.-T.; Low, M.; Seto, S.W.; Chang, D.; Lee, S.; Suresh, H.; Khoo, C.S.; Bensoussan, A.; Kiat, H. Chromatographic Analysis and Anti-Oxidative Property of Naoxinqing Tablet, a Proprietary Preparation of Diospyros Kaki Leaves. Molecules 2019, 24, 1101.

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