Abstract

A method for high-performance liquid chromatography coupled with linear ion trap quadrupole Orbitrap high-resolution mass spectrometry (HPLC-LTQ-Orbitrap MS) was developed and validated for the qualitative and quantitative assessment of Shejin-liyan Granule. According to the fragmentation mechanism and high-resolution MS data, 54 compounds, including fourteen isoflavones, eleven ligands, eight flavonoids, six physalins, six organic acids, four triterpenoid saponins, two xanthones, two alkaloids, and one licorice coumarin, were identified or tentatively characterized. In addition, ten of the representative compounds (matrine, galuteolin, tectoridin, iridin, arctiin, tectorigenin, glycyrrhizic acid, irigenin, arctigenin, and irisflorentin) were quantified using the validated HPLC-LTQ-Orbitrap MS method. The method validation showed a good linearity with coefficients of determination (r2) above 0.9914 for all analytes. The accuracy of the intra- and inter-day variation of the investigated compounds was 95.0–105.0%, and the precision values were less than 4.89%. The mean recoveries and reproducibilities of each analyte were 95.1–104.8%, with relative standard deviations below 4.91%. The method successfully quantified the ten compounds in Shejin-liyan Granule, and the results show that the method is accurate, sensitive, and reliable.

Highlights

  • Pharyngitis is an inflammation of the pharyngeal mucous membrane and submucous lymphoid tissues that affects people of all ages around the world

  • The illness is not caused by an infection; in such a setting, antibiotics may be the wrong choice for treatment or they may only offer a modest improvement in symptoms [1,2]

  • This is the first time that a technique using high-performance liquid chromatography coupled with linear ion trap quadrupole Orbitrap high-resolution mass spectrometry (HPLC-LTQ-Orbitrap MS) has been applied to identify and quantify the components in SJLYKL

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Summary

Introduction

Pharyngitis is an inflammation of the pharyngeal mucous membrane and submucous lymphoid tissues that affects people of all ages around the world. Shejin-liyan Granule (SJLYKL) is a traditional Chinese herbal medicine that has been used to treat pharyngitis in clinical practice. It consists of six herbs: Rhizoma Belamcandae, Arctium lappa, radix. We developed a simple and rapid method for the comprehensive qualitative and quantitative analyses of the major constituents of SJLYKL. This is the first time that a technique using high-performance liquid chromatography coupled with linear ion trap quadrupole Orbitrap high-resolution mass spectrometry (HPLC-LTQ-Orbitrap MS) has been applied to identify and quantify the components in SJLYKL

Reagents and Materials
Sample Preparation
Chromatographic and Mass Conditions
Method Validation for Quantitative Analysis
Development of the Extraction Method
Profiles of Ingredients in the SJLYKL Extract
Quantitative Analysis
Conclusions
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