Jaana Liimatainen

Phenolic compounds extracted by acidic aqueous ethanol from berries and leaves of different berry plants.

Phenolic compounds of berries and leaves of thirteen various plant species were extracted with aqueous ethanol and analyzed with UPLC-DAD-ESI-MS, HPLC-DAD, and NMR. The total content of phenolics was consistently higher in leaves than in berries (25-7856 vs. 28-711mg/100g fresh weight). Sea buckthorn leaves were richest in phenolic compounds (7856mg/100g f.w.) with ellagitannins as the dominant compound class. Sea buckthorn berries contained mostly isorhamnetin glycosides, whereas quercetin glycosides were typically abundant in most samples investigated. Anthocyanins formed the dominating group of phenolics in most dark-colored berries but phenolic acid derivatives were equally abundant in saskatoon and chokeberry berries. Caffeoylquinic acids constituted 80% of the total phenolic content (1664mg/100g f.w.) in bilberry leaves. B-type procyanidins and caffeoylquinic acids were the major phenolic compounds in hawthorn and rowanberry, respectively. Use of leaves of some species with prunasin, tyramine and β-p-arbutin, may be limited in food applications.

Birch inner bark procyanidins can be resolved with enhanced sensitivity by hydrophilic interaction HPLC-MS

A complex mixture of procyanidin aglycones was isolated by Sephadex LH-20 column chromatography from the silver birch inner bark, which is a polyphenol-rich source of natural antioxidants. Procyanidins were studied by using hydrophilic interaction liquid chromatography combined with high-resolution ESI-TOF-MS. A good chromatographic separation was achieved and procyanidins eluted according to their increasing degree of polymerization. Individual procyanidins were detected from dimers up to the degree of polymerization of 22 by their negative-ion mass spectra. The results showed that hydrophilic interaction liquid chromatography can be successfully applied for the analysis of high-molecular-weight procyanidins with enhanced sensitivity in electrospray mass spectrometry.

Characterization of phenolic compounds from inner bark of Betula pendula

Abstract A method has been developed for the characterization of biologically active silver birch (Betula pendula) inner bark phenolics based on high-performance liquid chromatography/diode array detector (HPLC-DAD)/electrospray ionization-mass spectrometry (ESI-MS). It was demonstrated that the inner bark contains high amounts of flavonoids, arylbutanoids, diarylheptanoids, simple phenolic compounds, phenolic acids, lignans, and procyanidins. Altogether, 30 individual compounds were characterized based on their ultraviolet (UV) and MS data. Structures of 22 compounds were confirmed by nuclear magnetic resonance (NMR) spectroscopy. In addition to previously reported phenolic compounds, 12 compounds were identified in silver birch inner bark for the first time; two of them are novel compounds: 3-β-glucopyranosyloxy-2-hydroxy-1-(4-hydroxy-3-methoxy-phenyl)-propan-1-one and 1,7-bis-(4-hydroxyphenyl)-3-heptanol 3-O-β-ap-iofuranosyl-(1→2)-β-glucopyranoside.

Phenolic Compounds of the Inner Bark of Betula pendula: Seasonal and Genetic Variation and Induction by Wounding

The contents of individual phenolic compounds in the inner bark of silver birch (Betula pendula Roth) were analyzed by HPLC-DAD. Samples from 21 mature trees originating from three micropropagated parent trees were collected six times over a 1-year period. Significant seasonal variation in the quantities of ten compounds and four chromatographically unresolved compound pairs was found. A majority of the compounds also exhibited significant quantitative variation among birch clones. There were no qualitative differences associated with the season or among the clones. However, wounding of the bark induced the production of new types of bark phenolics: several ellagitannins were detected in the callus tissues of birch for the first time.

Procyanidin xylosides from the bark of Betula pendula

A procyanidin dimer xyloside, catechin-(4α→8)-7-O-β-xylopyranosyl-catechin, was isolated from the inner bark of Betula pendula and its structure was determined using 1D and 2D NMR, CD and high-resolution ESIMS. Interestingly, the 7-O-β-xylopyranose unit was found to be present in the lower terminal unit of the dimer. In addition to this procyanidin dimer xyloside, an entire series of oligomeric and polymeric procyanidin xylosides was detected. Their structures were investigated by hydrophilic interaction HPLC-HRESIMS. Procyanidin glycosides are still rarely found in nature.

Online Measurement of Real-Time Cytotoxic Responses Induced by Multi-Component Matrices, such as Natural Products, through Electric Cell-Substrate Impedance Sensing (ECIS)

Natural products are complex matrices of compounds that are prone to interfere with the label-dependent methods that are typically used for cytotoxicity screenings. Here, we developed a label-free Electric Cell-substrate Impedance Sensing (ECIS)-based cytotoxicity assay that can be applied in the assessment of the cytotoxicity of natural extracts. The conditions to measure the impedance using ECIS were first optimized in mice immortalized hypothalamic neurons GT1-7 cells. The performance of four natural extracts when tested using three conventional cytotoxicity assays in GT1-7 cells, was studied. Betula pendula (silver birch tree) was found to interfere with all of the cytotoxicity assays in which labels were applied. The silver birch extract was also proven to be cytotoxic and, thus, served as a proof-of-concept for the use of ECIS. The extract was fractionated and the ECIS method permitted the distinction of specific kinetic patterns of cytotoxicity on the fractions as well as the extract’s pure constituents. This study offers evidence that ECIS is an excellent tool for real-time monitoring of the cytotoxicity of complex extracts that are difficult to work with using conventional (label-based) assays. Altogether, it offers a very suitable cytotoxicity-screening assay making the work with natural products less challenging within the drug discovery workflow.

Sephadex LH-20 fractionation and bioactivities of phenolic compounds from extracts of Finnish berry plants

In order to assist developing a natural, safe food-preservative, aqueous ethanolic extracts of leaves and berries of eight Finnish berry plants were fractionated with Sephadex LH-20 column chromatography. For each fraction, phenolic compounds were analyzed with NMR, UPLC-DAD-ESI-MS and HPLC-DAD. The antioxidant activities of the fractions were investigated using oxygen radical absorbance capacity (ORAC) assay, and the antibacterial activities were evaluated against foodborne pathogens Staphylococcus aureus and Escherichia coli. Antioxidant activities of the fractions correlated highly with both the total concentration and structural feature of phenolic compounds, including both flavonoids and non-flavonoid phenolics. ORAC value correlated strongly with the concentration of (+)-catechin, (-)-epicatecin, quercetin glycosides, and anthocyanins. Increase in size and number of sugar moieties may reduce the antioxidative activities of quercetin glycosides. Type of sugar moieties may have a significant role in influencing peroxyl-radicals scavenging ability of quercetin glycosides with monosaccharides as a single sugar moieties. Most of the fractions inhibited the target microbes. S. aureus strains expressed a higher sensitivity to phenolic compounds than E. coli strains.