AHyaluronic acid (HA) is part of the extracellular matrix of connective, epithelial and neural tissues, as well as the synovial fluid, skin, and cartilage. It is composed of repeating disaccharide units of D-glucuronic acid and N-acetyl glucosamine. Hyaluronic acid is used in abdominal surgery, ophthalmology, dermatology, rhinology; it is usable for the osteoarthritis treatment. The membranes of eggshell are a natural source of hyaluronic acid, collagen, glycosaminoglycan and collagenous proteins. In paper, we tested the possibility of extraction hyaluronic acid from the eggshell membranes by enzymatic hydrolysis. We identified optimal conditions of hydrolysis with trypsin at reaction temperature of 37 °C and pH 8; with pepsin at 40 °C and pH 3, as well as with papain at 60 °C and pH 7.5. The content of hyaluronic acid in samples was determined spectrophotometrically using the carbazole method. The experimental results showed a yield of ~ 4 -4.5 % hyaluronic acid per 1 g of dry eggshell membranes.
Sage is medicinal plant, known for its antioxidant and anti-inflammatory effects. Eight extract samples were tested in this study: extract from Salvia officinalis L. varieties from two different geographical localities (Jaslovské Bohunice and Pobedim, Slovakia), Salvia officinalis L., variety “bicolor”, Salvia officinalis L., variety “purpurescens”, Salvia apiana, Salvia divinorum, and two callus cultures of Salvia sclarea L. and Salvia aethiopis L. The highest values for composite parameters were observed for extract from Salvia apiana. It can be concluded that prepared sage extract samples are rich on polyphenolic acids (2 950±265 μg.mL−1 GAeq.) and amines (197±5.50 μg.mL−1 TRPeq.). HPLC analysis confirmed the dominant content of rosmarinic acid in the extracts; the highest content was detected in the Salvia apiana extract (1 120±15 μg.mL−1). Extract from Salvia apiana expressed too the highest antioxidant activity (1 710 – 4 669 μg.mL−1TEAC). Similarly, the highest inhibition activity was observed for this extract on thrombin (57±3.3 %) and on other proteinases (over 80 %). Spearman correlation analysis and PCA analyses revealed a coherence between antioxidant activity of samples and their content of rosmarinic acid as well as inhibitory activity towards particular proteases, and revealed the significance of thiol based secondary metabolites. Cluster analysis demonstrates the differences of Salvia apiana extract from extracts of S. officinalis L., the group of S. divinorum extract and from callus cultures.