Chromatographic separation, determination and identification of ecdysteroids: Focus on Maral root (Rhaponticum carthamoides, Leuzea carthamoides)

Abstract

The review presents general principles for choosing optimal conditions for ecdysteroid separation, identification, and isolation using HPLC/TLC techniques in RP, NP‐HILIC or NP modes. Analytics of ecdyteroids pose a still insufficiently resolved problem. Plant‐derived ecdysteroids are a point of interest of pharmaceutical industry and sport medicine due to their postulated adaptogenic and anabolic properties. In insects, ecdysteroids regulate larval transformation. Maral root (Rhaponticum carthamoides, Leuzea carthamoides), traditional Siberian folk‐medicine plant used as stimulant to boost overall health and fitness, is a particularly rich source of a wide variety of phytoecdysteroids. The similarity of molecular structures of ecdysteroids present in its extracts together with high content of unrelated compounds of similar chromatographic characteristics makes optimization of separation, identification and isolation of ecdysteroids a difficult analytical task. In that respect, two‐dimensional separations, two‐dimensional separations, 2D HPLC or 2D TLC, could be of use. For identification, the hyphenated techniques are particularly important. Thus, comprehensive overview of MS spectral parameters of ecdysteroids is provided. Described principles could easily be applied for separation of ecdysteroids in extracts from other sources. They are also useful for development of separation procedures for isolation of ecysteroids in preparative‐scale applications.

Various in vitro systems of Ragged Robin (Lychnis flos-cuculi L.): a new potential source of phytoecdysteroids?

“…In most plants the content varies from 0.001 to 0.1% of dry weight, with only several taxa reported surpassing 1% (equal to 10 mg g −1 ),-for example, Rhaponticum carthamoides herb-ca. 1% (Głazowska et al 2018), Serratula inermis inflorescence-2%, Cyanotis arachnoidea roots-2.9%, and Diploclisia glaucescens stems-3.2% (Baltaev 2000). Genus Silene, closely interrelated to Lychnis, includes numerous species also exceptionally rich in diverse complex of ecdysteroids, with 20HE as dominant constituent (Báthori et al 2000;Mamadalieva et al 2014).…”

“…To further confirm the presence of ecdysteroids, the plate was sprayed with an anisaldehyde reagent [p-anisaldehyde 0.5 ml (Merck, Darmstadt, Germany); glacial acetic acid 10 ml; concentrated sulfuric acid 4.5 ml; methanol 85 ml (POCh, Gliwice, Poland)]. After heating at 103 °C for about 1 min, the spots of ecdysteroids (under visible light) turned bluishpurple, and after 24 h they changed the color to yellowishgreen, which is typical for ecdysteroids (Nowak et al 2013;Głazowska et al 2018).…”

“…The autosampler injected 15 μl of each sample in triplicate. The UV absorption was measured at λ max = 242 nm in methanol, characteristic for ecdysteroid cholest-7-ene-6-one chromophore (Głazowska et al 2018). The mobile phase contained water and methanol; the chromatography step was conducted using gradient elution.…”

“…Ecdysteroids are primarily obtained from the plant material harvested from the natural site or from cultivated plants (Lafont and Dinan 2003;Głazowska et al 2018). They are, however, also produced employing plant in vitro cultures, a source that is often richer in those compounds and where numerous methods increasing ecdysteroid content can be applied under controlled conditions (Thiem et al 2017).…”

Effect of plant adaptogens on piglets and sows

“…Исходя из результатов публикаций зарубежных ученых [8], фокус исследований следует сосредотачивать на использовании R. carthamoides, как главного компонента, придающего уникальную активность экдистероид содержащим составам. S. coronata следует рассматривать в качестве дополняющего фитокомплекс иными компонентами (отлич-ными от R. carthamoides).…”