Comparative In Vitro Study: Assessing Phytochemical, Antioxidant, Antimicrobial, and Anticancer Properties of Vaccinium macrocarpon Aiton and Vaccinium oxycoccos L. Fruit Extracts.

The phytochemical diversity and potential health benefits of V. oxycoccos and V. macrocarpon fruits call for further scientific inquiry. Our study aimed to determine the phytochemical composition of extracts from these fruits and assess their antioxidant, antibacterial, and anticancer properties in vitro. It was found that the ethanolic extracts of V. oxycoccos and V. macrocarpon fruits, which contained more lipophilic compounds, had 2-14 times lower antioxidant activity compared to the dry aqueous extracts of cranberry fruit, which contained more hydrophilic compounds. All tested cranberry fruit extracts (OE, OW, ME, and MW) significantly inhibited the growth of bacterial strains S. aureus, S. epidermidis, E. coli, and K. pneumoniae in vitro compared to the control. Cytotoxic activity against the human prostate carcinoma PPC-1 cell line, human renal carcinoma cell line (CaKi-1), and human foreskin fibroblasts (HF) was determined using an MTT assay. Furthermore, the effect of the cranberry fruit extract samples on cell migration activity, cancer spheroid growth, and viability was examined. The ethanolic extract from V. macrocarpon fruits (ME) showed higher selectivity in inhibiting the viability of prostate and renal cancer cell lines compared to fibroblasts. It also effectively hindered the migration of these cancer cell lines. Additionally, the V. macrocarpon fruit extract (ME) demonstrated potent cytotoxicity against PPC-1 and CaKi-1 spheroids, significantly reducing the size of PPC-1 spheroids compared to the control. These findings suggest that cranberry fruit extracts, particularly the ethanolic extract from V. macrocarpon fruits, have promising potential as natural remedies for bacterial infections and cancer therapy.

Edible Gels with Cranberry Extract: Evaluation of Anthocyanin Release Kinetics.

The bioactive compounds found in cranberry fruit are natural antioxidants, and their consumption reduces the risk of diabetes, cardiovascular disease, cancers, and urinary tract infections. Oral gels with cranberry fruit extract are a promising product that can ensure accurate dosage and release of the active compounds and are suitable for people with dysphagia. The aim of this study was to determine the effect of polymeric materials on the dissolution kinetics of cranberry fruit anthocyanins from gel formulations. Gel formulations were prepared using freeze-dried cranberry fruit extract with different gelling excipients: chitosan (G1-G3), sodium carboxymethylcellulose (G4-G6), and sodium carboxymethylcellulose combined with carbomers (G7-G9). The dissolution test showed that the release of anthocyanins from gel formulations G1-G6 and G9 was most intense within the first 10 min, with little change in the anthocyanin content of the acceptor medium afterwards. For the formulations based on carboxymethyl cellulose and carbomers G7 and G8, the amount of anthocyanins released into the acceptor medium gradually increased, which prolonged the release time of the active compounds. The test for the release of anthocyanins from the semi-solid systems through a hydrophilic membrane revealed that within the first hour, the total amount of anthocyanins released from the modeled gel formulations (G1-G9) was within the range of 6.02%-13.50%. The 1% chitosan (G1) gel formulation released the fastest and highest amount of anthocyanins (70% within 6 h). The other formulations showed a slower release of anthocyanins, and after 6 h, the amount of anthocyanins released from formulations G2-G9 was <57%.

Phytochemical Composition of Cranberry (Vaccinium oxycoccos L.) Fruits Growing in Protected Areas of Lithuania.

The fruits of Vaccinium oxycoccos L. are an important source of bioactive compounds with antibacterial, antifungal, and anti-inflammatory effects. Studies on the phytochemical analysis of cranberry fruit samples showed that the qualitative and quantitative composition of biologically active compounds varied in cranberry fruit samples collected from different types of wetland sites: the total anthocyanin content was 698 ± 24-8352 ± 200 µg/g, the total flavonol content-518 ± 16-2811 ± 31 µg/g, the total content of triterpene compounds-4060 ± 122-6542 ± 157 µg/g, the content of chlorogenic acid-17 ± 0.4 µg/g to 1224 ± 41 µg/g, and the total content of proanthocyanidins-919 ± 19 µg EE/g to 3038 ± 137 µg EE/g. The percentage composition of anthocyanins in cranberry fruit varied between the different wetland sites: in some cranberry fruit samples, four anthocyanins (cyanidin-3-galactoside, cyanidin-3-arabinoside, peonidin-3-galactoside, and peonidin-3-arabinoside) were predominant, while in other samples, six anthocyanins (cyanidin-3-galactoside, cyanidin-3-arabinoside, peonidin-3-galactoside, peonidin-3-arabinoside, cyanidin-3-glucoside, and peonidin-3-glucoside) predominated. The results of these studies showed the differences in the composition of secondary metabolites in the studied cranberry samples and prove that the standardization of the qualitative and quantitative composition of cranberry fruit raw materials and the application of routine tests are necessary for the expansion of the use of botanical raw materials in the production of functional foods and phytopreparations.

Peculiarities of the Variation of Biologically Active Compounds in Fruit of Vaccinium oxycoccos L. Growing in the Cepkeliai State Strict Nature Reserve.

This study was carried out to analyze the accumulation patterns of anthocyanins, proanthocyanidins, flavonols, chlorogenic acid, and triterpene compounds in fruit samples of Vaccinium oxycoccos L. berries growing in the Cepkeliai State Strict Nature Reserve in Lithuania. Studies were carried out on the phytochemical composition of cranberry fruit samples during the period of 2020-2022. Anthocyanins, flavonols, chlorogenic acid and triterpene compounds were identified and quantified using UPLC-DAD methods, and proanthocyanins were determined using spectrophotometric methods. The content of identified compounds varied, as reflected in the total amounts of anthocyanins (710.3 ± 40 µg/g to 6993.8 ± 119 µg/g), proanthocyanidins (378.4 ± 10 µg EE/g to 3557. 3 ± 75 µg EE/g), flavonols (479.6 ± 9 µg/g to 7291.2 ± 226 µg/g), chlorogenic acid (68.0 ± 1 µg/g to 3858.2 ± 119 µg/g), and triterpenoids (3780.8 ± 98 µg/g to 7226.9 ± 224 µg/g). Cranberry fruit samples harvested from open oligotrophic wetland habitats contained higher levels of anthocyanins, anthocyanidins, flavonol glycosides, and proanthocyanidins. The highest levels of triterpene compounds were found in the cranberry fruits harvested in the spring of the following year after the snowmelt. The use of principal component analysis showed that cranberry plant material harvested in October and November had higher levels of bioactive compounds.

Formulation and Biopharmaceutical Evaluation of Capsules Containing Freeze-Dried Cranberry Fruit Powder.

Cranberry fruits are an important source of anthocyanins and anthocyanidins. The aim of the present study was to investigate the effect of excipients on the solubility of cranberry anthocyanins and their dissolution kinetics as well as on the disintegration time of the capsules. Selected excipients (sodium carboxymethyl cellulose, beta-cyclodextrin and chitosan) were found to affect the solubility and release kinetics of anthocyanins in freeze-dried cranberry powder. Capsule formulations N1-N9 had a disintegration time of less than 10 min, and capsule formulation N10 containing 0.200 g of freeze-dried cranberry powder, 0.100 g of Prosolv (combination of microcrystalline cellulose and colloidal silicon dioxide), and 0.100 g of chitosan had a capsule disintegration time of over 30 min. The total amount of anthocyanins released into the acceptor medium ranged from 1.26 ± 0.06 mg to 1.56 ± 0.03 mg. Capsule dissolution test data showed that the time to release into the acceptor medium was statistically significantly longer for the chitosan-containing capsule formulations compared to the control capsules (p < 0.05). Freeze-dried cranberry fruit powder is a potential source of anthocyanin-rich dietary supplements, and the choice of excipient chitosan could be a suitable solution in capsule formulations providing greater anthocyanin stability and modified release in the gastrointestinal tract.

Phytochemical Composition of the Fruit of Large Cranberry (Vaccinium macrocarpon Aiton) Cultivars Grown in the Collection of the National Botanic Garden of Latvia.

In this study, we conducted a qualitative and quantitative analysis of anthocyanins, proanthocyanidins, flavonols and triterpenoids in samples of introduced and bred large cranberry fruits from the collection of the National Botanic Garden of Latvia. The highest total anthocyanin levels (8638-9316 µg/g) were detected in the fruit samples of the cranberry cultivars 'Black Veil', 'Franclin' and 'Early Black'. The highest total proanthocyanidin levels (2775-3389 µg/g) were found in cranberries of cultivars 'Kalnciema Agra', 'Kalnciema Tumsa', 'Searles', 'Howes', and 'Kalnciema Raziga'. The highest levels of flavonol compounds (1373-1402 µg/g) were detected in cranberries of cultivars 'Howes', 'Black Veil' and 'Salaspils Melnas'. The highest levels of triterpenoids (5292-5792 µg/g) were determined in cranberries of cultivars 'Kalnciema Agra', 'Septembra', 'Dzbruklene' and 'Early Black'. The results of our study allow for the estimation of differences in the content of secondary metabolites in the fruit samples of the studied cranberry cultivars and for the selection of promising cultivars for further introduction and cultivation in the climatic conditions of the Baltic countries. These results are also important for the selection of the most promising cranberry cultivars for the preparation of cranberry raw material, and the high-quality composition of plant material ensures the effectiveness of cranberry supplements and other cranberry preparations.

Changes in the Composition of Biologically Active Compounds during the Ripening Period in Fruit of Different Large Cranberry (Vaccinium macrocarpon Aiton) Cultivars Grown in the Lithuanian Collection.

In our investigation, we evaluated the content of chlorogenic acid, proanthocyanidins, anthocyanins, flavonols, triterpenoids, and phytosterols in cranberry fruit extracts of the cultivars 'Baifay', 'Early Black', 'Howes', 'Pilgrim', 'Red Star', and 'Stevens' grown in Lithuania, as well as changes in the antioxidant activity in extracts of fruit samples of these cultivars during the period of berry maturation. The highest amount of proanthocyanidins (8.87 ± 0.57 mg EE/g EE) and flavonols (3688.52 ± 22.85 µg/g) was determined in cranberries of the cultivar 'Howes' harvested on 12 August. Remarkably, the highest anthocyanins content (9628.62 ± 266 µg/g) was determined in cranberries of the cultivar 'Howes' harvested on 22 October. The study showed that the content of phytochemical compounds in cranberries varied between 12 August and 22 October; the content of proanthocyanidins decreased by a factor of about 2, the content of chlorogenic acid decreased by a factor of about 1.3, the content of flavonols decreased by a factor of about 2, and the content of anthocyanins increased by 27 to 450 times. A strong correlation was found between the total proanthocyanidin content of cranberry fruit extracts and their in vitro antiradical and reducing activity (r = 0.781 and 0.726, respectively, p < 0.001). The data of our study detail the accumulation of the phytochemical composition of biologically active compounds in cranberry samples during the stages of maturity, therefore these data are significant for the assessment of harvest time of cranberry and can be applied to select cranberry cultivars for further cultivation in Lithuanian climatic conditions.

Evaluation of the Phytochemical Composition of Phenolic and Triterpene Compounds in Fruit of Large Cranberries (Vaccinium macrocarpon Aiton) Grown in Latvia.

We carried out a qualitative and quantitative analysis of the phytochemical composition of the fruits of large cranberry cultivars 'Ben Lear', 'Bergman', 'Kalnciema agra', 'Lemunyon', 'Pilgrim', 'Stevens', and 'Tina' grown in Latvian climatic conditions. The following predominant compounds were found in cranberry fruit samples: peonidin-3-O-galactoside, peonidin-3-O-arabinoside, cyanidin-3-O-galactoside, cyanidin-3-O-arabinoside, myricetin-3-galactoside, quercetin-3-galactoside, quercetin-3-α-L-arabinofuranoside, quercetin 3-rhamnoside, ursolic acid, and oleanolic acid. During the berry ripening period (from 16 August until 15 September), a trend of decreasing amounts of compounds was found in the fruit samples of the studied cranberry cultivars: the total amount of proanthocyanidins decreased by 1.3 times, the total amount of the identified flavonols decreased by 1.3 times, the total amount of triterpenoids decreased by 1.2 times, and the total amount of chlorogenic acid decreased by 1.7 times. During the period from 16 August until 15 September, the total amount of anthocyanins in the cranberry fruit samples increased by 2.6 to 17 times. The highest total amount of anthocyanins (5305.80 ± 27 µg/g) was detected in fruit samples of the cranberry cultivar 'Kalnciema agra' collected on 15 September. The amount of biologically active compounds in cranberry fruit samples varies during berry ripening. Thus, the choice of the picking time is one of the factors that determines the phytochemical composition of raw cranberry material.

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.