Chromatographic analysis and anti-oxidative property of Naoxinqing tablet, a proprietary preparation of Diospyros kaki leaves

Magdy Kazzem, Yu-Ting Sun, Mitchell Low, Sai Wang Seto, Dennis Chang, Samiuela Lee, Harsha Suresh, Cheang S. Khoo, Alan Bensoussan, Hosen Kiat

Research output: Contribution to journalArticleResearchpeer-review

Abstract

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.

LanguageEnglish
Article number1101
Pages1-14
Number of pages14
JournalMolecules
Volume24
Issue number6
DOIs
Publication statusPublished - 20 Mar 2019

Fingerprint

Diospyros
Chromatographic analysis
tablets
Quercetin
leaves
Tablets
Chromatography
preparation
glucosides
activity (biology)
Scavenging
Liquid chromatography
Endothelial cells
acids
Metabolites
Carboxylic Acids
Cerebrovascular Disorders
Oxidants
Hydrogen Peroxide
Quality control

Bibliographical note

Copyright the Author(s) 2019. Version archived for private and non-commercial use with the permission of the author/s and according to publisher conditions. For further rights please contact the publisher.

Keywords

  • Naoxinqing
  • UPLC MS/MC analysis
  • anti-oxidative
  • endothelial cells
  • reactive oxygen species
  • Reactive oxygen species
  • Anti-oxidative
  • Endothelial cells

Cite this

Kazzem, Magdy ; Sun, Yu-Ting ; Low, Mitchell ; Seto, Sai Wang ; Chang, Dennis ; Lee, Samiuela ; Suresh, Harsha ; Khoo, Cheang S. ; Bensoussan, Alan ; Kiat, Hosen. / Chromatographic analysis and anti-oxidative property of Naoxinqing tablet, a proprietary preparation of Diospyros kaki leaves. In: Molecules. 2019 ; Vol. 24, No. 6. pp. 1-14.
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abstract = "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.",
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author = "Magdy Kazzem and Yu-Ting Sun and Mitchell Low and Seto, {Sai Wang} and Dennis Chang and Samiuela Lee and Harsha Suresh and Khoo, {Cheang S.} and Alan Bensoussan and Hosen Kiat",
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year = "2019",
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Kazzem, M, Sun, Y-T, Low, M, Seto, SW, Chang, D, Lee, S, Suresh, H, Khoo, CS, Bensoussan, A & Kiat, H 2019, 'Chromatographic analysis and anti-oxidative property of Naoxinqing tablet, a proprietary preparation of Diospyros kaki leaves', Molecules, vol. 24, no. 6, 1101, pp. 1-14. https://doi.org/10.3390/molecules24061101

Chromatographic analysis and anti-oxidative property of Naoxinqing tablet, a proprietary preparation of Diospyros kaki leaves. / Kazzem, Magdy; Sun, Yu-Ting; Low, Mitchell; Seto, Sai Wang; Chang, Dennis; Lee, Samiuela; Suresh, Harsha; Khoo, Cheang S.; Bensoussan, Alan; Kiat, Hosen.

In: Molecules, Vol. 24, No. 6, 1101, 20.03.2019, p. 1-14.

Research output: Contribution to journalArticleResearchpeer-review

TY - JOUR

T1 - Chromatographic analysis and anti-oxidative property of Naoxinqing tablet, a proprietary preparation of Diospyros kaki leaves

AU - Kazzem, Magdy

AU - Sun, Yu-Ting

AU - Low, Mitchell

AU - Seto, Sai Wang

AU - Chang, Dennis

AU - Lee, Samiuela

AU - Suresh, Harsha

AU - Khoo, Cheang S.

AU - Bensoussan, Alan

AU - Kiat, Hosen

N1 - Copyright the Author(s) 2019. Version archived for private and non-commercial use with the permission of the author/s and according to publisher conditions. For further rights please contact the publisher.

PY - 2019/3/20

Y1 - 2019/3/20

N2 - 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.

AB - 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.

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