Quantitative determination of cucurbitacin E and cucurbitacin I in homoeopathic mother tincture of Gratiola officinalis L. by HPLC.

"Hedgehyssop" Gratiola officinalis L. (Scrophulariaceae) is found as an ingredient in homeopathic remedies. Among the active compounds found in G. officinalis, the cucurbitacines constitute a group of triterpenoid substances which are well-known for their bitterness and toxicity. Due to the toxicity of the cucurbitacin's aglycones it becomes necessary to determine the content of the aglycones and glycosides that may be responsible for the pharmacological activity and toxicity in homeopathic tinctures according to the European Pharmacopeia guidelines. In this context a HPLC method was developed for the identification and determination of cucurbitacin E and I in homeopathic mother tinctures. To evaluate the total concentration of the aglycones, cucurbitacin E and I formed after hydrolysis we determined the concentration of both compounds after enzymatic hydrolysis with beta-glucosidase in vitro. Reversed-phase HPLC with a Eurospher C18 column with precolumn and acetonitrile-water gradient system as the mobile phase proved to be suitable for direct determination of both aglycons, cucurbitacin E and I in Gratiola-mother tinctures. The contents of cucurbitacin E, cucurbitacin I, cucurbitacin E glycoside and cucurbitacin I glycoside were found as 0.0065%, 0.0031%, 0.0011% and 0.0006%, respectively in Gratiola-mother tincture prepared according to method 2a HAB.

Quantitative and cytotoxic activity determinations on Galanthus nivalis subsp. cilicicus.

Aerial and underground parts of Galanthus nivalis subsp. cilicicus, a wild-growing species in Turkey, were collected during two different vegetation periods in flowering and fruiting seasons. Herba and bulbus Galanthi were prepared from each specimen. With the aim of collecting data for prospective monographs on this drug, contents of humidity, ash, sulphated ash and total alkaloids were determined according to DAB 10. The specimens were also analyzed quantitatively for two of the principal alkaloids of the genus, galanthamine and lycorine, by using a method based on spectrophotometry complemented with TLC. LC50 values were determined for the ethanolic and alkaloidal extracts of each of the specimens using brine shrimp lethality bioassay.