Phytochemical screening and effect of Viscum album L. on monoamine oxidase A and B activity and serotonin, dopamine and serotonin receptor 5-HTR1A levels in Galleria mellonealla (Lepidoptera).

ETHNOPHARMACOLOGICAL RELEVANCE: Viscum album L. (European mistletoe), a member of the Santalaceae, is a hemiparasitic, evergreen shrub growing on deciduous and coniferous trees. In traditional and folk medicine, mistletoe was used for the treatment of central nervous system disorders such as epilepsy, hysteria, insomnia, nervous excitability, neuralgia, headache, dizziness and fatigue. However, relatively little is known of its neuropharmacological activity. AIM OF THE STUDY: The aim of the present study was to evaluate the effect of treatment with aqueous and hydroethanolic extracts from Viscum album L. parasitizing birch, linden and pine, on MAO-A and MAO-B activity as well as serotonin, dopamine and serotonin receptor 5-HTR1A levels in Galleria mellonella (Lepidoptera) larvae. MATERIALS AND METHODS: The phytochemical composition of the extracts was characterised using UPLC-DAD-ESI-MS/MS. To investigate the neuropharmacological activity of Viscum album L. extracts, Galleria mellonella (Lepidoptera) larvae were used as a model organism. The inhibitory potential of the extracts against MAO-A and MAO-B was determined by fluorometry. The serotonin, dopamine and serotonin receptor 5-HTR1A levels in larvae hemolymph after treatment were quantified by ELISA. RESULTS: UPLC-DAD-ESI-MS/MS analysis allowed the identification of 88 compounds, either full or in part. Most of the characterised phytochemicals were flavonoids, hydroxycinnamic acids and lignans. Screening found that aqueous and hydroethanolic mistletoe extracts inhibited the enzymatic activity of either MAO-A or MAO-B or both. Additionally, mistletoe extract administration increased the levels of serotonin and serotonin receptor 5-HTR1A. None of the tested extracts had any significant effect on dopamine level. CONCLUSIONS: A key novel finding was that the aqueous and hydroethanolic extracts from Viscum album L. inhibited monoamine oxidase activity and increased the levels of serotonin and serotonin receptor 5-HTR1A in Galleria mellonella (Lepidoptera) larvae. These properties may be due to the presence of phenolic constituents, particularly flavonoids. Further research based on bioassay-guided fractionation of mistletoe is needed to identify CNS-active molecules.

TESTING PHARMACEUTICAL RELEASE OF ACTIVE SUBSTANCES FROM MEDICINAL PRODUCTS CONTAINING ST. JOHN'S WORT.

The aim of this study was to determine the content of hypericins and flavonoids in tablets and capsules containing the extract or powdered herb of St. John's wort, in herbs for infusion and herbal infusions and to release of these compounds from tablets and capsules. HPLC method was used to determine the assay of hypericins and flavonoids in all tested products. The hypericins content was between 0.35 mg and 1.44 mg per tablet or capsule. The release of hypericins from these products in the phosphate buffer of pH 6.8 is between 30 and 60% of the determined content. The degree of hypericins release from herbs into infusions was 15% on average, which corresponds to 0.64 mg of hypericins per infusion of 4 g of herbs. The flavonoids content was between 8.79 and 36.3 mg per tablet or capsule. The release of flavonoids in the phosphate buffer of pH 6.8 is between 63 and 85% of the determined content. The degree of flavonoids release was 76% on average, which corresponds to 77.0 mg per infusion of 4 g of herbs. The test results confirmed that infusions from the St. John's wort constitute are a rich source of flavonoids. At the same time, the universally accepted opinion that aqueous infusions contain only trace amounts of hypericins was not confirmed. Infusions from Herba hyperici may also be a source of hypericins in amounts comparable with the minimum dose recommended for the treatment of mild to moderate depressive episodes.

Determination of total hypericins in St. John's wort and herbal medicinal products.

The work aimed to determine the levels of hypericins expressed as hypericin in the herbal substance of St. John's wort, in capsules and tablets containing the extract of St. John's wort, tablets containing powdered herb and in tincture and juice from fresh St. John's wort, by HPLC method with spectrophotometric detection. In addition, the amount of hypericins in the infusion prepared from St. John's wort was determined by HPLC and spectrophotometry methods. According to traditional indications aqueous infusions from St. John's wort containing mainly hydrophilic components are used in gastrointestinal diseases. On the other hand, ethanolic extracts containing hypericin and hyperforin affect the CNS and are indicated for the treatment of episodes of mild depressive disorders. The results obtained in the work indicate that the daily dose of hypericins taken by a patient as infusions is 0.328 mg on average for herbs in sachets and in bulk form. It can be compared to the daily dose of hypericins contained in tablets and capsules based on the alkoholic extract of St. John's wort and tablets containing powdered St. John's wort herb. For solid dosage forms, this dose ranges from 0.288 mg to 0.636 mg. The assays were performed using consistent analytical methods for all tested pharmaceutical products and consequently it was possible to compare doses taken by patients and their strength of action.