Polyphenolic compounds, plant secondary metabolites essential for plant survival, are known for their high antioxidant and anti-inflammatory activity. In addition, several polyphenols, such as phloretin, also have potential antiviral effects, making these compounds potential ingredients of biofunctional foods. A promising source for the extraction of phloretin is a by-product of apple production—apple tree leaves. Focusing on green technologies, the first aim of the present study was to optimize the direct ultrasound-assisted extraction conditions to gain the maximum yield of phloretin from air-dried apple leaves. For the optimization of process parameters, we applied the response surface method with Box–Behnken design. The optimal extraction conditions were extraction time 14.4 min, sonication amplitude 10% and 10 g of sample per 100 mL solvent (70% ethanol, w/w). Using these conditions, we assessed the content of individual and total polyphenolic compounds along with antioxidant activity in the leaves of different autumn and winter apple cultivars grown in Estonia. The analyses were carried out with chromatographic (HPLC-DAD-MS/MS) and spectrophotometric methods. The phloretin concentration ranged from 292 to 726 µg/g and antioxidant activity from 6.06 to 11.42 mg GA eq./g, these being the highest in the local winter cultivars ‘Paide taliõun’ and ‘Tellissaare’, respectively.

The chemical composition of berries and berry pomaces is diverse, containing polyphenolic components that may have both antibacterial and antioxidative properties. In the present study, in vitro antibacterial effect of the extracts of chokeberry, blackcurrant, and rowan berries and berry pomaces against L. monocytogenes, S. aureus, E. coli, and C. jejuni was studied. In addition, the polyphenolic profile and antioxidant activity of these extracts were investigated. The polyphenolic profiles in the aqueous and 30% ethanolic extracts were determined chromatographically by HPLC-MS, and the total polyphenol content was estimated spectrophotometrically by HPLC-DAD-UV. The minimal inhibition concentrations (MICs) of the extracts against tested bacteria were determined by the broth microdilution method. The content of total polyphenols was highest and good antioxidative properties of the extracts were determined for chokeberry and blackcurrant berries and their pomaces. The highest proportions of total quercetin derivatives and anthocyanins were found in the extracts of chokeberry berry/pomace and blackcurrant berry/pomace, respectively. The sensitivity of tested microbes to the extracts of berries and berry pomaces was as follows: S. aureus > L. monocytogenes > E. coli and C. jejuni. In vitro antibacterial activity of tested extracts depended on the extraction solvent, mainly for the ethanolic extracts. Findings suggest that chokeberry and blackcurrant berries and their pomaces can be used as a good source of polyphenols with antioxidative properties, and they also have antibacterial activity against some foodborne pathogenic bacteria. It is important that the valuable compounds are extracted from juice press residues before their disposal.

Inonotus obliquus grows in the Northern Hemisphere on some living broadleaved tree species as a pathogen, causing stem rot. In Estonia, the fungus is well known in the Betula species but can also be found on Alnus. Sterile conks of I. obliquus contain different bioactive compounds, but the quantitative and comparative research of these compounds in conks on different host species is limited. In the current work, I. obliquus was isolated and, evidently, determined from Alnus incana (L.) Moench., Alnus glutinosa (L.) Gaertn., and Betula pendula Roth, and the content of bioactive compounds in conks on these hosts were analysed. All the analysed conks sampled from A. incana and B. pendula contained betulin that varied from 111 to 159 µg/g. A significantly (p < 0.05) higher betulinic acid content was found in conks sampled from A. incana when compared with B. pendula: 474–635 and 20–132 µg/g, respectively. However, the conks from Betula were richer in total polyphenols, flavonols, and glucans. The content of inotodiol was quite similar in the conks from A. incana (7455–8961 µg/g) and B. pendula (7881–9057 µg/g). Also, no significant differences in the lanosterol content were found between the samples from these two tree species. To the best of our knowledge, this study is the first investigation of the chemical composition of I. obliquus parasitizing on Alnus. The results demonstrate that the bioactive compounds are promising in conks of I. obliquus growing not only on Betula but also on the Alnus species. It supports the opportunity to cultivate I. obliquus, also on the Alnus species, thus increasing the economic value of growing this tree species in forestry.

Chaga mushroom (Inonotus obliquus) is a pathogenic fungus that grows mostly on birch species (Betula pendula Roth and B. pubescens Ehrh.) and has traditionally been used as an anticancer medicine. This study aimed to compare the chemical composition and cytotoxic activity of chagas growing on both Betula spp. on various cancer cell lines. The freeze-dried extracts contained triterpenes inotodiol, lanosterol betulin, and betulinic acid typical to conks growing on Betula species. The cytotoxic activity of chaga growing on Betula pendula and B. pubescens 80% ethanolic extracts against 31 human cancer cell lines was evaluated by a sulforhodamine B assay. Chaga extract showed moderate activity against all cancer cell lines examined; it did not result in high cytotoxicity (IC₅₀ ≤ 20 µg/mL). The strongest inhibitions were observed with chaga (growing on B. pendula) extract on the HepG2 and CAL-62 cell line and with chaga (from B. pubescens) extract on the HepG2 cell line, with IC₅₀ values of 37.71, 43.30, and 49.99 μg/mL, respectively. The chaga extracts from B. pendula exert somewhat stronger effects on most cancer cell lines studied than B. pubescens extracts, which can be attributed to a higher content of inotodiol in B. pendula extracts. This study highlights the potential of chaga as a source of bioactive compounds with selective anticancer properties. To the best of our knowledge, this study is the first investigation of the chemical composition of I. obliquus parasitizing on B. pubescens.

Chaga (Inonotus obliquus) is a pathogenic fungus that grows mostly on birch species (Betula pendula and B. pubescens) and has traditionally been used as an anti-cancer medicine. The study aimed to compare the chemical composition and cytotoxic activity of chagas growing on both Betula spp. on various cancer cell lines. The freeze dried extracts contained triterpenes inotodiol, lanosterol betulin and betulinic acid typical to conks growing on Betula species. The cytotoxic activity of chaga growing on Betula pendula and B. pubescens 80% ethanolic extracts against thirty-one human cancer cell lines has been evaluated by a sulforhodamine B assay. The strongest inhibitions were observed with chaga (growing on B. pendula) extract on the HepG2 and CAL-62 cell line, and with chaga (from B. pubescens) extract on HepG2 cell line, with IC50 values of 37.71, 43.30, and 49.99 μg/mL, respectively. The chaga extracts from B. pendula exert somewhat stronger effects on most cancer cell lines studied than B. pubescens extracts. This study highlight the potential of chaga as a source of bioactive compounds with selective anti-cancer properties, encouraging further exploration of its therapeutic mechanisms and potential applications in cancer treatment.