Title: Optimization of LC separation of anthraquinones in Sennae fructus and Sennae folium supported by an in-silico tool
Authors : Peter, Samuel
Meier, Nadja
Josic, Gojko
Meier, Beat
Wolfram, Evelyn
Published in : Planta Medica : Journal of Medicinal Plant and Natural Product Research
Volume(Issue) : 81
Issue : S 01
Publisher / Ed. Institution : Georg Thieme Verlag
Issue Date: 2016
License (according to publishing contract) : Licence according to publishing contract
Type of review: Peer review (Publication)
Language : English
Subjects : Cassia senna L.; Ph Eur; Anthraquinone; Sennoside; Aloe emodin; Rhein
Subject (DDC) : 615: Pharmacology and therapeutics
Abstract: The European Pharmacopeia (Ph. Eur.) contains herbal monographs for Sennae folium and fructus. The current spectrophotometric assay is time consuming and unspecific. An HPLC method for the official Sennae monographs is under discussion. The aim of this study is to optimize an HPLC method for anthraquinone aglyca to be robust and valid with optimal separation and analysis time. The currently proposed Ph. Eur. draft method is compared to a method developed by [2]: Senna pods and leaves from different suppliers were extracted by ACN/0.1% NaHCO3. Solid-Phase-Extraction is applied for method [2], only. Nucleodur 100 – 3 C18 column and H2O/ACN/MeOH in a gradient was used as stationary and mobile phase with detection at 435nm. It was found that the Ph Eur draft method delivers in the range of 5 – 20% w/w higher results for anthraquinone content in the herbal drug material calculated as Sennoside B. Overestimation due to coeluting peaks or analyte losses due to SPE could explain the differences in content. Therefore, a rational approach for optimization was attempted using the software Drylab®. Based on 12 initial runs with varying conditions in the entire variable parameter space (column temperature, mobile phase and gradient), the in-silico tool suggests further experiments. The tracking of the changes in retention time of the target analyte peaks is a crucial prerequisite and requires both DAD and MS data of each peak. The result of the in-silico supported optimization is the identification of a suitable operation region of the method parameters shown in Fig. 1a. The transfer of the method from UHPLC to HPLC resulted in an acceptable separation at 43 °C and a gradient of A. water and B. MeOH/ACN (98/2 v/v) both containing 0.1% formic acid of 0/30, 8.5/30, 28/48, 30/99 [min/%B] (Fig. 1b). From our study we conclude that Drylab® is a useful LC method optimization tool. However, the peak assignment of all peaks in the chromatograms is a challenge for plant extracts.
Departement: Life Sciences und Facility Management
Organisational Unit: Institute of Chemistry and Biotechnology (ICBT)
Publication type: Article in scientific Journal
DOI : 10.1055/s-0036-1597006
ISSN: 0032-0943
URI: https://digitalcollection.zhaw.ch/handle/11475/2952
Appears in Collections:Publikationen Life Sciences und Facility Management

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