Abstract

Daegunjoong-tang (DGJT) is an oriental medicine consisting of four medicinal herbs (Zingiber officinale Rosc., Panax ginseng C.A.Mey., Oryza sativa L., and Zanthoxylum schinifolium Sieb. et Zucc.) that is used to treat intestinal- and cancer-related diseases. In this study, a protocol for quality control of DGJT based on reverse-phase high-performance liquid chromatography (HPLC) and liquid chromatography tandem mass spectrometry (LC–MS/MS) analysis were developed. In HPLC analysis, the marker analytes (hyperoside, quercitrin, ginsenoside Rg1, and 6-gingerol) were separated, verified, and quantified using a mobile phase of 0.1% (v/v) aqueous formic acid–0.1% (v/v) formic acid in acetonitrile system, and a C18 reverse-phase column (4.6 mm × 250 mm, particle size; 5 m) maintained at 40 °C. In LC–MS/MS analysis, all analytes were separated using a Waters Acquity UPLC BEH C18 column (2.1 mm × 100 mm, particle size; 1.7 μm). Using the developed HPLC and LC–MS/MS methods, the four marker analytes were found in the samples at 0.95–13.86 mg/g (HPLC) and 0.27–2.42 mg/g (LC–MS/MS). The assay will be useful for evaluating the quality of DGJT.

Highlights

  • Daegunjoong-tang (DGJT; Daikenchuto in Japanese; Da-Jian-Zhong-Tang in Chinese) is a herbal medicine prescription first recorded in Gumgoeyorak (金匱要略, Jin Gui YaoLue in Chinese), a famous traditional oriental medicine book written by Zhongjing Zhang in the Han Dynasty, and consists of four herbal medicines, Zingiber officinale Rosc., Panax ginseng C.A.Mey., Oryza sativa L., and Zanthoxylum schinifolium Sieb. et Zucc. [1]

  • Each herbal medicine contains a range of compound types including phenolic compounds, gingerols, and shogaols in Z. officinale [3]; triterpenoid saponins and ginsenosides in P. ginseng [4]; polysaccharides and maltose in O. sativa [5]; and essential oils oleic acid, palmitic acid, and estragole, flavonoid glycosides hyperoside and quercitrin, and the alkaloids schinifolin in Z. schinifolium [6,7,8]

  • A simultaneous reverse-phase high-performance liquid chromatography (RP–HPLC) and liquid chromatography tandem mass spectrometry (LC–MS/MS) multiple reaction monitoring (MRM) methods were developed for the quality control of DGJT using four marker components: hyperoside, quercitrin, ginsenoside Rg1, and 6-gingerol

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Summary

Spectrometry Methods for Quality

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Introduction
Plant Materials
Chemicals and Reagents
DGJT Sample Extraction
HPLC Quantification of the Four Marker Analytes
Method Validation of Developed Assay
Identification of the Main Component of Each Herbal Medicine
Selection of Marker Analytes for Quality Standardization
Establishment of Optimal Analysis Conditions
Conclusions
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