Optimization of extraction procedure and determination by high performance liquid chromatography of flavonols and phenolic acids from Hypericum Perforatum L.

Hypericum perforatum is a medicinal plant which has been known in traditional medicine as an antiinflammatory and healing agent. Nowadays the use of Hypericum extracts is concerned mainly with antidepressive applications. In the present work, HPLC – RP- C18 column chromatography with photodiode array detection was applied for the determination of the derivatives of cinnamic and benzoic acid (e.g., caffeic, chlorogenic, ferulic, sinapic, gallic acids) (Fig.1.) and flavonols - quercetine derivatives (quercetine, rhamnetine, quercitrin, mirycetine, keampferol and rutin) in Hypericum Perforatum. Phenolic compounds were extracted from the sample matrix with ethanol and ethanolwater mixture in different ratios solvent (3:7; 8:2; v/v) at 30°C and 60°C in water-bath shaker and by ultrasonic extraction and then analyzed before and after acid and basic hydrolysis. The total amount of studied flavonols and phenolic acids were compared with the total flavonoids content (TFC) and with total polyphenols content (TPC). UV-Vis spectrometry was used to investigate methods for qualitative and quantitative determination of these compounds

The influence of pH and selected cations on the spectrofluorometric determination of oxytetracycline hydrochloride

The spectrofluorometric method for the oxytetracycline hydrochloride in pure and in veterinary products Tetrox and Oxymed 50 determination is described. The influence of pH solution and presence selected cations on fluorescence intensity were studied too. It was ascertained that the highest fluorescence intensity take place at pH=9. Moreover, the quenching of fluorescence intensity was observed at presence Ca2+, Al3+ and Fe3+ in contrast to Mg2+ which caused increasing of intensity. The obtained recovery (97.23±0.12% for Tetrox and 95.21±0.10% for Oxymed 50) values meet the European Pharmacopoeia requirement. Moreover, the relative standard deviation (RSD) was below 0.23% confirmed high precise of the method. The statistical test (t-Student and F-Snedecor) used for comparison spectrofluorometric and chromatographic methods pointed that they are comparable in respect of precision but not of accuracy

Nutraceuticals: opening the debate for a regulatory framework

Currently, nutraceuticals do not have a specific definition distinct from those of other food-derived categories, such as food supplements, herbal products, pre- and probiotics, functional foods, and fortified foods. Many studies have led to an understanding of the potential mechanisms of action of pharmaceutically active components contained in food that may improve health and reduce the risk of pathological conditions while enhancing overall well-being. Nevertheless, there is a lack of clear information and, often, the claimed health benefits may not be properly substantiated by safety and efficacy information or in vitro and in vivo data, which can induce false expectations and miss the target for a product to be effective, as claimed. An officially shared and accepted definition of nutraceuticals is still missing, as nutraceuticals are mostly referred to as pharma-foods, a powerful toolbox to be used beyond the diet but before the drugs to prevent and treat pathological conditions, such as in subjects who may not yet be eligible for conventional pharmaceutical therapy. Hence, it is of utmost importance to have a proper and unequivocal definition of nutraceuticals and shared regulations. It also seems wise to assess the safety, mechanism of action and efficacy of nutraceuticals with clinical data. A growing demand exists for nutraceuticals, which seem to reside in the grey area between pharmaceuticals and food. Nonetheless, given specific legislation from different countries, nutraceuticals are experiencing challenges with safety and health claim substantiation

Determination of phenolic acids in dietary supplements containing Vitis vinifera (grape vine) by HPLC-PDA method

The procedure involving water and water-methanol extraction, RP-HPLC-C18 column chromatography with PDA detection was developed for determination of cinnamic acid and benzoic acid derivatives in grapevine’s dietary supplements (LV, RW, VIN, VIC, and DK) available on the Polish market. Phenolic acids were analysed before and after acidic and basic hydrolysis and identified against standards. Totalamount of studied phenolic acids determined by HPLC-PDA was compared with total polyphenols content (TPC) by Folin-Ciocalteu method. The average content of studied phenolic acids (70.54±0.21; 122.95±0.49; 87.67±0.10; 132.21±0.24; 266.78 ±0.39, and 18.16±0.09 mg/100 g d.m. (dry mass) for LV, RW, VIN, VIC, DK, and WW, respectively) were higher than the TPC (1489.91±0.39, 1648.19±0.14, 1574.38±0.33, 1643.64±0.12, 1984.75±0.97, and 715.55±0.36 mg/100 g d.m. for LV, RW, VIN, VIC, DK, and WW, respectively). The new developed method was validated for specifi city, repeatability, and accuracy and can be suitable for routine quality and quantity analysis of dietary supplements containing grape vine (Vitis vinifera)