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ABSTRACT Pharmacological activities as anti-inflammatory, anti-hyperuricemic, anti-gouty arthritis, antitumor and trypanocidal activities of the aerial parts from Lychnophora trichocarpha (Spreng.) Spreng. ex Sch.Bip., Asteraceae (Brazilian arnica) have already been proved. Eremantholide C is a sesquiterpene lactone and one of the active chemical constituents responsible for these activities presented by L. trichocarpha. Therefore, the aim of this work was to develop and validate a stability indicating HPLC method for eremantholide C. Eremantholide C stability was evaluated in L. trichocarpha ethanolic extract and in its isolated form. Analytical conditions employed C18 column, acetonitrile/water in gradient elution, flow of 0.8 ml/min at 30 ºC. To correct for the loss of analyte during sample preparation the use of coumarin as an internal standard was necessary. The developed method provides good separation and resolution of the peaks, allowing quantification of eremantholide C, isolated or directly in the ethanolic extract, in internal standard presence. Validation results showed that this method is linear in the concentration range 2-180 µg/ml, precise, accurate and specific. Stability studies showed that L. trichocarpha ethanolic extract and eremantholide C remain stable for 6 months when stored at room temperature and impermeable glass bottle, therefore they can be used safely and effectively within this period. While at 40 ºC there was stability loss, at 8 ºC a stability increase was observed for the extract and the isolated eremantholide C. Forced degradation studies showed that eremantholide C degraded under acidic and alkaline conditions and was stable for three days under neutral and oxidative conditions, and when exposed to high temperature. Thus, with the development of a stability indicative method and the application of it in eremantholide C stability studies, the results can guide the development of new products that adequately preserve the original features of the biologically active substance with quality, safety and efficacy.
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Introduction: Calendula officinalis L. is aromatic herbaceous yearling of the family of Asteraceae. Ethanol extract, decoction and ointment of the plant is used to treat or relieve injury, trauma, erosion, purulent trauma or slow healing abrasions, furuncle, carbuncle, congelation, burn, bed sore, herpes and lichen as cream and spray. Goal :To define biologically active substances in cultivated calendula officinalis Materials and Methods: Calendula officinalis has been harvested from Monos pharmacological institute, garden of medical plants and prepared according to the appropriate standards. β –carotene and flavonoids were quantified by spectrophotometer, Alkaloid, tannin and ascorbic acids were quantified by tetrameter, Extractive substances, ash and humidity were quantified by weight analysis Results: Quantitative analysis of the flower of calendula officinalis has been carried out following first Mongolian national pharmacopeia and Russian National pharmacopeia XI and defined that β -carotene 1.4313%, alkaloids 0.1229%, flavonoids 2.8817%, tannin 1.2376%, ascorbic acid 0.0702%, extractive substances 40.18%, ash content 11.75% and humidity 5.95%. Flower of calendula officinalis has been extracted by water, 30%, 50% and 80% ethanol, then made comparative analysis on content of β–carotiene. When extracted by 80% ethanol, content of β – carotene was the highest or 150 mg. Therefore optimum extraction solvent quantity has been defined to be 80% ethanol. Microbiological analysis has no revealed any organisms and bacteria in solid extract of the plant. Conclusions: 1. Quality and countable analysis of biologically active substance in the flower of calendula officinalis has been completed. 2. β –carotene the main active substance in cultivated calendula officinalis, is found to be 1.4 gr which that meets Mongolian National Standards of medicine. 3. The 80% ethanol extract of calendula officinalis contained 150mg β –carotene, the maximum content of β –carotene. Hence optimum extraction solvent was found to be 80%ethanol and it will be and used for future research. 4. Microbiological parameters of 80% solid extract of the plant has met quality requirements. Key words: β –carotene, Biologically active substance, Calendula officinalis,