Comparative Fingerprint of Aromatic Herbs and Yeast Alcoholic Extracts used as Ingredients for Promen, a Prostate Preventive Nutraceutical

The aim of this study was to characterize and identify different bioactive compounds in plant sources and yeast powders  to obtain an original nutraceutical (Promen) which has beneficial effects in prostate disease prevention. Seven plant and fruit sources, namely nettle (Urtica dioica), green tea (Camellia sinensis), fluff with small flowers (Epilobium parviplorum), tomato (Solanum licopersicum),  sea buckthorn (Hippophae rhamnoides), pumpkin (Cucurbita maxima), sunflower (Helianthus annus) and lyophilized beer yeast (Saccharomyces cerevisiae) were investigated. Methanolic extracts were prepared using 15% plant concentration and the purified fractions were analyzed using high throughput techniques like UV-VIS spectroscopy, high performance liquid chromatography coupled with photodiode array detection (HPLC-DAD) and mass spectrometry LC-QTOF -MS. The majority of the investigated plants were rich in phenolic derivatives, polyphenols (flavonoid glucosides), while yeast was rich in aminoacids, peptides and vitamins B. The major compounds identified were: Juglone, Resveratrol, Quercetin, Epigallocatechin, Gallocatechin, Biochanin A, Isorhamnetin 3-O-glucoside 7-O-rhamnoside, Quercetin 3-O-galactoside 7-O-rhamnoside, Kaempferol 3,7-O-diglucoside and p-Coumaroylquinic acid. The specific biomarkers were identified for both plant extracts used as ingredients to obtain an nutraceutical  Promen. Combined UV-Vis spectroscopy, HPLC-PDA chromatography and LC-MS spectrometry are recommended as accurate, sensible and reliable tools to investigate the plants and nutraceutical fingerprints and to predict the relation between ingredients composition and their health effects

Preparation and Comparative Characterization of Alginate-Made Microcapsules and Microspheres Containing Tomato, Seabuckthorn Juices and Pumpkin Oil

Recent studies have shown the benefits of tomatoes, seabuckthorn juices and pumpkin oil, rich in bioactives with antioxidant capacity, in the prevention of prostate diseases. To stabilize their antioxidant activity, microencapsulation represent a good technological alternative, improving the stability and bioavailability of bioactive molecules ( phenolic derivatives, carotenoids, phytosterols, vitamins).   The aim of the study was to prepare and characterize microspheres and microcapsules based on emulsions made of natural polymers like Natrium alginate mixed with tomato and/or seabuckthorn juices, with or without pumpkin oil.  The viscosity of emulsions, the morphology of microcapsules and microspheres were characterized comparatively and the bioactives were monitored by UV-Vis spectrometry.  In the lipophilic extract there were identified, before and after encapsulation, different classes of compounds, from lipids, to phenolic acid derivatives, flavonoids and carotenoids. Carotenoids were the major components having concentrations from 9.16 up to 19.71 mg/100 g sample. The viscosity of  each emulsion including juices, oil and natrium alginate 2%, before encapsulation, showed differences, dependent on the oil addition and speed of homogenization. The macroscopic and microscopic structure of microspheres and microcapsules were comparatively evaluated. Both microspheres and microcapsules had external diameters  ranging from 750 to 900 μm and the microcapsules’ oily core of 150-180 μm. The results obtained from emulsion’s viscosity will be correlated with the rigidity and optimal release rate of bioactive molecules from microcapsules and microspheres.  Further studies are directed towards these aspects

HPLC-UV Analysis Coupled with Chemometry to Identify Phenolic Biomarkers from Medicinal Plants, used as Ingredients in Two Food Supplement Formulas

. High performance liquid chromatography (HPLC) with UV detection is nowadays the reference method to identify and quantify the biomarkers of quality and authenticity of plants and food supplements. Seven medicinal plants were collected from wild flora: Taraxacum officinalis (1), Cynara scolimus (2), Silybum marianum (3), Hypericum perforatum (4),  Chelidonium majus (5), Lycopodium clavatum (6) and  Hippophae rhamnoides (7)  leaves and fruits.  Two products (A and B) were obtained by mixing individual plant powders. Therefore product A was obtained by mixing dandelion, artichoke and milk thistle, 1:1:1 while product B by mixing St John’s wort, Celandine and Wolf’s claw, 1:1:1. The methanolic extracts of individual plants as well as three different extracts of products A and B (using acidulated water, neutral water and acidulated methanol) were analyzed using HPLC-UV for their phenolics’ fingerprint and composition. The qualitative (untargeted analysis) and quantitative (targeted analysis) results were further compared using Principal Component Analysis (PCA) in order to identify their specific biomarkers. Thus, quantitative evaluation of individual phenolics in case of individual plants and products A and B extracts, showed specific and significant differences of composition. Both products A and B contained elagic acid as major compound. For product A, good biomarkers were trans-cinnamic, chlorogenic, caffeic and p-coumaric acids, as well silymarin and silibine originating from milk thistle. For product B, good biomarkers were quercetin and kaempherol, gallic and protocatecuic acids, this product being rich in flavonoids. In conclusion, HPLC-UV coupled with PCA analysis proved to be a rapid and useful way to identify the main biomarkers of plants’ authentication, as well of final products’ quality and safety