Comparative chromatographic and spectral study of the alkaloid composition of the biomass of tissue culture and the herb of Glaucium Flavum

Introduction. The culture of plant cells and tissues is an effective technology for obtaining various secondary metabolites, including for use in the pharmaceutical industry. Compounds synthesized in callus and suspension cultures may be identical to plant ones, and the yield of secondary metabolites, in some cases, exceeds plant sources. In this regard, yellow hornpoppy (Glaucium flavum Crantz.) is of great interest, containing glaucin, which has a unique antitussive effect and is a source of a number of medications.

The purpose of this study is a comparative phytochemical study of the alkaloid composition of the tissue culture and the herb of Glaucium flavum.

Material and methods. Callus tissue cultures of yellow hornpoppy yellow hornpoppy (Glaucium flavum Crantz.) were obtained in the laboratory of the Institute of Pharmacy of Kazan State Medical University. The herb of the Glaucium flavum cultivated on the territory of the Botanical Garden of Samara University was collected during the flowering period of plants in 2021. A comparative study of the alkaloid composition of the callus culture and the herb of Glaucium flavum was carried out by methods of luminescent microscopy on an Altami LUM-2 microscope (Russia), thin-layer chromatography (TLC) on Sorbfil-PTSH-AF-A-UV plates, as well as spectrophotometry. A mixture of solvents n-butanol-acetic acid-water 4:1:2 was used as a solvent system. The detection of separated substances was carried out by scanning chromatographic plates in UV light at a wavelength of 366 and 254 nm, followed by the manifestation of Dragendorf reagent. The identification of the isolated glaucin hydrochloride was carried out on the basis of UV, ¹H-NMR, ¹³C-NMR spectroscopy data. ¹H NMR spectra were obtained on the "JNM-ECX 400" device (399.78MHz), ¹³C NMR spectra were obtained on the "JNM–ECX 400" device (100.52 MHz). The spectra were recorded using a Specord 40 spectrophotometer (Analytik Jena AG, Germany) in the wavelength range of 190-500 nm.

Results. During the luminescent analysis of the tissue culture of the Glaucium flavum Crantz., as well as intact plants, the type of luminescence characteristic of isoquinoline alkaloids was noted. Using TLC analysis, isoquinoline alkaloids, including glaucin, were found in samples of callus cultures and the aerial parts of Glaucium flavum, however, a lower glaucin content was noted in the sample of the callus culture compared to that of the intact plant. The spectrophometric analysis of the studied samples showed that the nature of the absorption curve in intact plants and crops of the so-called Glaucium flavum has both common and distinctive features. The UV spectrum of water-alcohol extraction from a sample of Glaucium flavum tissue culture has a similar spectrophotometric profile, however, it contains only one absorption maximum (288 nm), coinciding with that of the UV spectrum of water-alcohol extraction from the herb of this plant.

Conclusions. Therefore, the final conclusion regarding the content of glaucin in the studied samples can be made only during the preparative isolation of substances from the biomass of Glaucium flavum using column chromatography.