Spectrophotometric Determination Of Total Content Research Articles

BIOCOLLECTION OF PHARMACEUTICAL PARASITIC STRAINS OF CLAVICEPS PURPUREA – BASE FOR SELECTION OF NEW LINES PRODUCING ERGOALKALOIDS IN VITRO

Relevance. Ergot Claviceps purpurea is a very important source of pharmaceutical raw materials (ergoalkaloids). Aim of the study – for biocollection maintenance two parasitic strains of Claviceps purpurea (Fries) Tulasne were reproduced by in planta cultivation on winter rye: ergotoxin A-6, ergotoxin VKM-F-2450-D. Material and methods. Spectrophotometric determination of total content of indole alkaloids (with the van Urk reagent) and qualitative-quantitative selective determination of the alkaloid composition by thin-layer chromatography (TLC) with densitometric scanning of chromatograms were used. Results. Maternal sclerotia contained from 0.6 to 1 g of alkaloids per 100 g of sample for VKM-F-2450-D, from 0.5 to 0.7 g per 100 g of sample for A-6. VKM-F-2450-D and A-6 contained from 30% to 40% of the sum of α-ergocryptine and ergocornine and approximately 20% of β-ergocryptine. New gen-eration sclerotia contained from 0.31 to 0.76 g of alkaloids per 100 g of sample for VKM-F-2450-D, from 0.21 to 0.62 g per 100 g of sample for A-6. VKM-F-2450-D and A-6 contained from 40% to 62% and from 21% to 66% of the sum of α-ergocryptine and ergocornine, from 13% to 22% and from 13% to 36% of β-ergocryptine respectively. Conclusion. Two stable lines (ergotoxin A-6-S, ergotoxin ВKM-F-2450-D-S) with in vitro biosynthesis of ergoalkaloids with purple pigmentations features were detected, isolated and subcultivated by morphological selection in axenic culture. Accumulation and retention of Claviceps genetic resources is very actual: biocollection of parasitic strains is a good reserve for selection of new saprophytic lines.

Phenolic compounds of the liquid extract from cleavers herb (galium aparine l.)

Aim. To study the qualitative composition and the quantitative content of phenolic compounds of the liquid extract obtained from cleavers (Galium aparine L.) herb.Materials and methods. The extract of cleavers herb was obtained by triple extraction of the raw material with 60% ethanol when heating, followed by the concentration of the combined extracts to the ratio of the raw material: finished product of 1:1. The content of extractives was determined by the gravimetric method. Phenolic compounds were studied by the methods of high-performance liquid chromatography (HPLC) and UV-spectrophotometry.Results. In the chromatographic study of the extract from cleavers herb 11 compounds of phenolic nature were reliably identified, including 5 hydroxycinnamic acids – caffeic, сhlorogenic, neoсhlorogenic, 3,5- and 4,5-dicaffeoylquinic acids; and 6 flavonoids – catechin, quercetin, quercitrin, isoquercitrin, rutin and hyperoside. Rutin (1.89 mg/ml) and сhlorogenic acid (0.78 mg/ml) were predominant compounds. In the spectrophotometric determination of total content of phenolic compounds it was found that the amount for hydroxycinnamic acids calculated with reference to сhlorogenic acid was 1.9 %, for flavonoids calculated with reference to rutin – 0.25 %, for polyphenolic compounds calculated with reference to gallic acid – 1.35 %. The dry residue was 24.63 %.Conclusions. The significant content of extractive substances, flavonoids and у hydroxycinnamic acids with predominance of rutin and сhlorogenic acid in alcohol-water extract from cleavers herb indicate the feasibility of its further pharmacological study.