Abstract

Rhizoma Alismatis (RA), widely known as “Ze-Xie” in China, is the tuber of Alisma orientale (Sam.) Juzep (Alismaceae), a Chinese herbal medicine that has been used to treat hyperlipidemia, diabetes, hypertension, dysuria, and inflammation. In this study, a sensitive and reliable method based on an ultra-performance liquid chromatography (UPLC) couple with two ionisation modes, including electrospray ionisation (ESI) and atmospheric pressure chemical ionisation (APCI) tandem mass spectrometry (MS), namely, UPLC-ESI/APCI-MS/MS was developed and validated to simultaneously determine 8 triterpenoids (ESI mode) and 2 sesquiterpenoids (APCI mode) in RA. Ten marker compounds were analysed with a Waters' CORTECS UPLC C18 column (200 mm × 2.1 m, 1.6 μm) and gradient elution with water (contained 0.1% formic) and acetonitrile within 7 min. The established method was validated for linearity, intra- and interday precisions, accuracy, recovery, and stability. The calibration curve for 10 marker compounds showed good linear regression (r > 0.9971). The limits of detection and quantification for analytes were 0.14–1.67 ng/mL and 0.44–5.65 ng/mL, respectively. The relative standard deviations (RSD, %) and accuracy (RE, %) of intra- and interday precisions were less than 3.83% and 1.21% and 3.22% and 1.46%, repeatability and stability for real samples were less than 2.78% and 3.19%, respectively. All recoveries of the 10 marker compounds ranged from 97.24% to 102.49% with RSDs less than 4.05%. The developed method efficiently determined the 10 marker compounds in RA and was subsequently applied to optimise harvest time and crude processing temperature. The result indicated the 90% wilted phase and 70°C (or lower) may be the best harvest time and the processing temperature of RA.

Highlights

  • Rhizoma Alismatis (RA), widely known as “Ze-Xie” in China, is the tuber of Alisma orientale (Sam.) Juzep (Alismaceae) [1]

  • Some articles investigated the harvesting or processing of RA [31,32,33,34,35], they focused only on triterpenoids such as alisol B 23-acetate or/and alisol A 24-acetate. us, the content accumulation process of the two categories in the growth process and the processing of RA was not fully characterised. Us, both sesquiterpenoids and triterpenoids are considered necessary for the quantitative analysis in RA. is work aims to develop an ultra-performance liquid chromatography (UPLC)-electrospray ionisation (ESI)/atmospheric pressure chemical ionisation (APCI)-mass spectrometry (MS)/MS for simultaneous determination triterpenoids and sesquiterpenoids in RA and subsequently apply it to optimise the harvest time and crude processing temperature of RA

  • Alismol was purchased from Yunnan Xi Li Biotechnology Co., Ltd. (Yunnan, China). e purity of each standard was higher than 98% by using high-performance liquid chromatography-ultraviolet detection (HPLC-UV) and their structures were confirmed by NMR

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Summary

Introduction

Rhizoma Alismatis (RA), widely known as “Ze-Xie” in China, is the tuber of Alisma orientale (Sam.) Juzep (Alismaceae) [1]. E main active ingredients of RA are classified into triterpenoids and sesquiterpenoids [9, 10] Triterpenoids, such as alisol B 23-acetate, exhibit hypolipidemic, hypoglycaemic, and diuretic effects [2, 6, 7]. Gas chromatography-mass spectrometry (GCMS) was used in the qualitative analysis of RA for the volatile oil, containing sesquiterpenoids [25] These methods are only used for the qualitative or quantitative analysis of one type terpenoid (triterpenoid or sesquiterpenoid). Is work aims to develop an UPLC-ESI/APCI-MS/MS for simultaneous determination triterpenoids and sesquiterpenoids in RA and subsequently apply it to optimise the harvest time and crude processing temperature of RA Us, both sesquiterpenoids and triterpenoids are considered necessary for the quantitative analysis in RA. is work aims to develop an UPLC-ESI/APCI-MS/MS for simultaneous determination triterpenoids and sesquiterpenoids in RA and subsequently apply it to optimise the harvest time and crude processing temperature of RA

Materials and Methods
Validation of Quantitative Method
Results and Discussion
II III IV V VI Stage
Conclusions
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