Development and Validation of the UPLC-DAD Methodology for the Detection of Triterpenoids and Phytosterols in Fruit Samples of Vaccinium macrocarpon Aiton and Vaccinium oxycoccos L.

Cranberries are used in the production of medicinal preparations and food supplements, which highlights the importance of triterpene compounds determination in cranberry fruit raw material. The aim of our study was to develop and validate for routine testing suitable UPLC-DAD methodology for the evaluation of triterpene acids, neutral triterpenoids, phytosterols, and squalene content in cranberry samples. The developed and optimized UPLC-DAD methodology was validated according to the guidelines of the International Council for Harmonization (ICH), evaluating the following parameters: range, specificity, linearity (R2 > 0.999), precision, LOD (0.27−1.86 µg/mL), LOQ (0.90−6.18 µg/mL), and recovery (80−110%). The developed and validated technique was used for the evaluation of triterpenic compounds in samples of Vaccinium macrocarpon and Vaccinium oxycoccos fruits, and their peels, pulp and seeds. The studied chromatogram profiles of Vaccinium macrocarpon and Vaccinium oxycoccos were identical but differed in the areas of the analytical peaks. Ursolic acid was the dominant compound in fruit samples of Vaccinium macrocarpon and Vaccinium oxycoccos. The highest amounts of triterpenic compounds were detected in the cranberry peels samples. The developed method for the detection of triterpene compounds can be applied in further studies for routine testing on the qualitative and quantitative composition of fruit samples of Vaccinium macrocarpon and Vaccinium oxycoccos species and cultivars.

Bioactive Compounds, Antioxidant Activity, and Biological Effects of European Cranberry (Vaccinium oxycoccos).

Lesser known fruits or underutilized fruit species are recently of great research interest due to the presence of phytochemicals that manifest many biological effects. European cranberry, Vaccinium oxycoccos fruit, as an important representative of this group, is a valuable source of antioxidants and other biologically active substances, similar to American cranberry (V. macrocarpon) which is well known and studied. European cranberry fruit is rich especially in polyphenolic compounds anthocyanins (12.4⁻207.3 mg/100 g fw), proanthocyanins (1.5⁻5.3 mg/100 g fw), and flavonols, especially quercetin (0.52⁻15.4 mg/100 g fw), which mostly contribute to the antioxidant activity of the fruit. Small cranberry is also important due to its various biological effects such as urinary tract protection (proanthocyanidins), antibacterial and antifungal properties (quercetin, proanthocyanidins, anthocyanins), cardioprotective (proanthocyanidins) and anticancer activities (proanthocyanidins), and utilization in food (juice drinks, jams, jellies, sauces, additive to meat products) and pharmacological industries, and in folk medicine.

Antimicrobial protection of minced pork meat with the use of Swamp Cranberry (Vaccinium oxycoccos L.) fruit and pomace extracts.

The objective of the study was to determine the antimicrobial activity of Swamp Cranberry (Vaccinium oxycoccos) fruit and pomace extracts (FSCE and PSCE) and their efficiency in minced pork meat. Ethanol (96 and 40%) and water were used for raw material extraction. Organic acids, flavonols, terpenes and stilbenes composition of the extracts was determined using HPLC. Minimal inhibitory concentration and minimum bactericidal/fungicidal concentration were determined for bacteria and fungi strains using the broth macrodilution method. The growth inhibition of Staphylococcus aureus, Listeria monocytogenes, Salmonella Enteritidis, and Escherichia coli in inoculated fresh minced pork meat containing 2.5% we-PSCE or we-FSCE (prepared by using 40% ethanol) were evaluated within 6 days of refrigeration storage. Swamp Cranberry pomace extracts contained stilbenes and more organics acids and flavonols than fruit extracts. Extracts inhibited Gram-positive bacteria strains stronger than Gram-negative, regardless of used raw material. The extracts did not show antifungal activity. Water-ethanol extracts (we-FSCE and we-PSCE) had stronger antibacterial properties than ethanolic extracts (e-FSCE and e-PSCE) and aqueous extracts (w-FSCE and w-PSCE). A 2.5% addition of we-PSCE or we-FSCE to minced pork meat resulted in a reduction of the number of pathogenic cells by 4 log cycles after 4 days of refrigeration storage. Baked burgers containing 2.5% of these extracts obtained high ratings for color, taste, odor, juiciness, and overall acceptability that did not differ statistically from control samples. Extracts from Swamp Cranberry constitute interesting candidates for natural preservatives of minced pork meat.

Nephroprotective effect of cranberry (Vaccinium oxycoccos) in streptozocin-induced diabetic nephropathy in mice.

OBJECTIVES: Diabetic nephropathy is a chief reason of mortality particularly in individuals with renal dysfunction. The current research was aimed to assess the nephroprotective portion of Vaccinium oxycoccos toward mice diabetic nephropathy induced by streptozotocin (STZ). V. oxycoccos was purchased and used for hydroalcoholic extraction. METHODS: Sixty male mice were subjected to STZ-intraperitoneal injection (45 mg/kg). After diabetes induction, mice were divided into five groups of diabetic control (received only STZ), non-diabetic control (received only citrate buffer), two V. oxycoccos treatment (received V. oxycoccos extract (200 and 400 mg/kg) oral daily by gavage), and metformin treatment (received metformin (500 mg/kg) oral daily by gavage). Glucose and weight of mice were checked weekly. RESULTS: After 28 days, the effect of V. oxycoccos extract on serum and urine parameters were assessed. STZ caused significant decreased in the mice body weight. Mice treated with the V. oxycoccos (400 mg/kg) harbored the lowest weight loss at day 28 (70.2±1.38 g). STZ caused significant increase in the mice FBS. Mice treated with the V. oxycoccos (400 mg/kg) harbored the lowest FBS at day 28 (189.2±1.20 mg/dL). Treatment of mice with V. oxycoccos (400 mg/kg) caused the lowest increase in the levels of cholesterol, HbA1c and triglycerides compared to the diabetic control mice. Compared to the diabetic control group, mice treated with V. oxycoccos (400 mg/kg) had the highest HDL, insulin, SOD, and GSH (p<0.05). The lowest serum BUN, CR, and UR were found in mice treated with V. oxycoccos (400 mg/kg). Anti-inflammatory effects of V. oxycoccos (400 mg/kg) was shown by the lowest TNF-α, IL-6, and TGF-ß1 concentration in mice treated with V. oxycoccos (400 mg/kg). CONCLUSIONS: The current study disclosed that treatment with V. oxycoccos resulted in substantial development in the serum and urine parameters and also antioxidant and anti-inflammatory response of STZ-induced diabetic mice.

Fungi Present in the Clones and Cultivars of European Cranberry (Vaccinium oxycoccos) Grown in Lithuania.

Fungi are associated with the European cranberry (Vaccinium oxycoccos L.) and play important roles in plant growth and disease control, especially in cranberry yields. This article presents the results of a study which was aimed to investigate the diversity of fungi found on different clones and cultivars of the European cranberry grown in Lithuania, causing twigs, leaf diseases and fruit rots. In this study seventeen clones and five cultivars of V. oxycoccos were selected for investigation. Fungi were isolated via the incubation of twigs, leaves and fruit on a PDA medium and identified according to their cultural and morphological characteristics. Microscopic fungi belonging to 14 genera were isolated from cranberry leaves and twigs, with Physalospora vaccinii, Fusarium spp., Mycosphaerella nigromaculans and Monilinia oxycocci being the most frequently isolated fungi. 'Vaiva' and 'Zuvinta' cultivars were the most susceptible to pathogenic fungi during the growing season. Among the clones, 95-A-07 was the most susceptible to Phys. vaccinii, 95-A-08 to M. nigromaculans, 99-Z-05 to Fusarium spp. and 95-A-03 to M. oxycocci. Microscopic fungi belonging to 12 genera were isolated from cranberry berries. The most prevalent pathogenic fungi M. oxycocci were isolated from the berries of the cultivars 'Vaiva' and 'Zuvinta' and clones 95-A-03 and 96-K-05.

Formulation and evaluation of oral disintegrating films using a natural ingredient against Streptococcus mutans.

Background: Oral disintegrating films (ODFs) are one of the forms of drug delivery system with better patient compliance. The advantage is that it disintegrates quickly when placed on the tongue and has better bioavailability. Aim: The aim of this study is to develop an ODF using Vaccinium oxycoccos and Plectranthus amboinicus targeting Streptococcus mutans. Setting and Design: This in vitro study was conducted at an institutional laboratory. Materials and Methods: The chemical composition of aqueous extracts of Vaccinium oxycoccos and Plectranthus amboinicus was examined using gas chromatography (GC)-mass spectrometry (MS). Extracts were added at its minimal inhibitory concentration (MIC) into the hydroxy propylmethyl cellulose (HPMC) polymer matrix solution to develop active ODF. The study concentrated on assessing the physical properties such as thickness of film, PH of the film, folding endurance, swelling test, disintegration, and dissolution test. Color analysis, Fourier transform infrared (FTIR), spectroscopy, and scanning electron microscope (SEM) were the mechanical properties of the film assessed. Statistical Analysis: Data were analyzed statistically, one-way analysis of variance followed by post hoc analysis for the assessment of MIC. Descriptive statistics were performed for the analysis of film properties. Results: MIC was 25 µg/ml for Vaccinium oxycoccos and 50 µg/ml for Plectranthus amboinicus. Three percentage HPMC with 1% citric acid and 1% aspartame was used to develop a polymer matrix. Films pH was between 6 and 7. FTIR and SEM analysis were done to confirm the attachment of active compound in a polymer matrix. Conclusion: Vaccinium oxycoccos and Plectranthus amboinicus showed good antibacterial activity, therefore could be a potent source to minimize the incidence of S. mutans.