Determination of Triterpenic Acids and Screening for Valuable Secondary Metabolites in Salvia sp. Suspension Cultures

Abstract

Plant in vitro cultures are a prospective alternative for biochemicals production, for example the triterpenes oleanolic and ursolic acid present in plants and cell cultures of Salvia sp. Our objective was to develop a suitable analysis protocol for evaluation of triterpenic acid yield in plant raw material and in vitro cultures supporting selection processes. Moreover, valuable bioactive compounds had to be revealed. Thus, different strategies enhancing the separation for a sensitive and effective HPLC-UV method were investigated and the developed method was validated for linearity, precision, accuracy, limits of detection and quantification. A baseline separation of these isomers enabled detection limits of below 0.4 µg/mL and quantification limits of about 1.2 µg/mL. Over the tested concentration range a good linearity was observed (R² > 0.9999). The variations in the method were below 6% for intra-and inter-day assays of concentration. Recoveries were between 85-98% for both compounds using ethanol as extraction solvent. Additionally, metabolite profiling of cell suspension culture extracts by GC-MS has shown the production variability of different plant metabolites and especially the presence of plant phenols and sterols. These studies provide a method suitable for screening plant and cell culture productivity of triterpenic acids and highlighted interesting co-products of plant cell cultures.

Production of bioactive plant secondary metabolites through in vitro technologies—status and outlook

“…g −1 DW oleanolic acid and ursolic acid, respectively, in suspension cultures of Salvia fruticosa. This was about 1/2 and 1/3 of the content found in the intact plants (Kümmritz et al 2014). Lantana camara L. (Verbenaceae) is a popular ornamental plant which, however, is also known as a notorious weed.…”

Establishment and triterpenoid production of Ocimum basilicum cambial meristematic cells

“…Basil and its essential oil are often used in traditional medicine (Irondi et al 2016;Ezeani et al 2017;Bae et al 2020), cosmetics (Vivas Castaño et al 2016;Volpe et al 2018;Yeşil et al 2020), and pharmacology (Sestili et al 2018;Zhan et al 2020). Among the pharmacologically interesting compounds found in basil are linalool (Medeiros Venancio et al 2016), rosmarinic (Kwon et al 2019), oleanolic (OA; Qamar et al 2020) and ursolic acid (UA; Arshad Qamar et al 2010;Kümmritz et al 2014). The latter are triterpenoids, a class of isoprenoid-based secondary metabolites.…”

The Studies on Seed Germination and in Vitro Cultures of Salvia L.Species from Turkish Flora

“…The production variability of different metabolites (phenols and sterols) from cell suspension culture (Kümmritz et al, 2014) S Cell cultures obtained leaf…”