Anti-Candida Activity of Extracts Containing Ellagitannins, Triterpenes and Flavonoids of Terminalia brownii, a Medicinal Plant Growing in Semi-Arid and Savannah Woodland in Sudan.

Various parts of Terminalia brownii (Fresen) are used in Sudanese traditional medicine against fungal infections. The present study aimed to verify these uses by investigating the anti-Candida activity and phytochemistry of T. brownii extracts. Established agar diffusion and microplate dilution methods were used for the antifungal screenings. HPLC-DAD and UHPLC/QTOF-MS were used for the chemical fingerprinting of extracts and for determination of molecular masses. Large inhibition zones and MIC values of 312 µg/mL were obtained with acetone, ethyl acetate and methanol extracts of the leaves and acetone and methanol extracts of the roots. In addition, decoctions and macerations of the leaves and stem bark showed good activity. Sixty compounds were identified from a leaf ethyl acetate extract, showing good antifungal activity. Di-, tri- and tetra-gallotannins, chebulinic acid (eutannin) and ellagitannins, including an isomer of methyl-(S)-flavogallonate, terflavin B and corilagin, were detected in T. brownii leaves for the first time. In addition, genipin, luteolin-7-O-glucoside, apigenin, kaempferol-4'-sulfate, myricetin-3-rhamnoside and sericic acid were also characterized. Amongst the pure compounds present in T. brownii leaves, apigenin and ß-sitosterol gave the strongest growth inhibitory effects. From this study, it was evident that the leaf extracts of T. brownii have considerable anti-Candida activity with MIC values ranging from 312 to 2500 µg/mL.

Hydrolyzable tannins (ellagitannins), flavonoids, pentacyclic triterpenes and their glycosides in antimycobacterial extracts of the ethnopharmacologically selected Sudanese medicinal plant Combretum hartmannianum Schweinf.

In Sudanese traditional medicine, decoctions, macerations, and tonics of the stem and root of Combretum hartmannianum are used for the treatment of persistent cough, a symptom that could be related to tuberculosis (TB). To verify these traditional uses, extracts from the stem wood, stem bark, and roots of C. hartmannianum were screened for their growth inhibitory effects against Mycobacterium smegmatis ATCC 14468. Methanol Soxhlet and ethyl acetate extracts of the root gave the strongest effects (MIC 312.5 and 625 µg/ml, respectively). HPLC-UV/DAD and UHPLC/QTOF-MS analysis of the ethyl acetate extract of the root led to the detection of 54 compounds, of which most were polyphenols and many characterized for the first time in C. hartmannianum. Among the major compounds were terflavin B and its two isomers, castalagin, corilagin, tellimagrandin I and its derivative, (S)-flavogallonic acid dilactone, punicalagin, and methyl-ellagic acid xylopyranoside. In addition, di-, tri- and tetra-galloyl glucose, combregenin, terminolic acid, cordifoliside D, luteolin, and quercetin-3-O-galactoside-7-O-rhamnoside-(2→1)-O-ß-D-arabinopyranoside were characterized. Luteolin gave better growth inhibition against M. smegmatis (MIC 250 µg/ml) than corilagin, ellagic acid, and gallic acid (MIC 500-1000 µg/ml). Our study justifies the use of C. hartmannianum in Sudanese folk medicine against prolonged cough that could be related to TB infection. This study demonstrates that C. hartmannianum should be explored further for new anti-TB drug scaffolds and antibiotic adjuvants.

HPLC-DAD and UHPLC/QTOF-MS Analysis of Polyphenols in Extracts of the African Species Combretum padoides, C. zeyheri and C. psidioides Related to Their Antimycobacterial Activity.

Combretum padoides Engl. & Diels, C. psidioides Welv. and C. zeyheri Sond. are used forthe treatment of infections and tuberculosis related symptoms in African traditional medicine. In orderto verify these uses, extracts were screened for their growth inhibitory eects against M. smegmatisATCC 14468. Ultra-high pressure liquid chromatography coupled to quadrupole time-of-flightmass spectrometry (UHPLC/QTOF-MS) and GC-MS were used to investigate the polyphenoliccomposition in the active extracts. The lowest minimum inhibitory concentration (MIC), 625 g/mL,was shown by a methanol extract of the stem bark of C. psidioides. A butanol extract of C. psidioidesgave large inhibition zone diameters (IZD 21 mm) and inhibited 84% of the mycobacterial growthat 312 g/mL. Combretastatin B-2 and dihydrostilbene derivatives were present in the methanolextract of C. psidioides, whereas the butanol extract of this species contained punicalagin, corilagin,and sanguiin H-4. Methanol and butanol extracts of the stem bark of C. padoides gave large inhibitionzone diameters (IZD 26.5 mm) and MIC values of 1250 and 2500 g/mL, respectively. C. padoidescontained an ellagitannin with a mass identical to punicalagin ([M-H]- 1083.0587) and a corilaginlike derivative ([M-H]- 633.0750) as well as ellagic acid arabinoside and methyl ellagic acid xyloside.A butanol extract of the roots of C. zeyheri showed mild antimycobacterial activity and containeda gallotannin at m/z [M-H]- 647.0894 as the main compound along with punicalagin and threeunknown ellagitannins at m/z [M-H]- 763.0788, 765.0566, and 817.4212. Our results indicate thatthe studied species of Combretum contain phenolic and polyphenolic compounds with possiblepotential as leads for antimycobacterial drugs or as adjuvants for conventional anti-TB drugs.

Potential Anti-Tuberculosis Activity of the Extracts and Their Active Components of Anogeissus Leiocarpa (DC.) Guill. and Perr. with Special Emphasis on Polyphenols.

In Sudanese traditional medicine, decoctions of the stem bark of Anogeissus leiocarpa are used for the treatment of tuberculosis (TB). However, this plant has not been investigated before for its antimycobacterial effects. Our screening results show, for the first time, that many extracts of various parts of A. leiocarpa exhibit growth inhibitory activity against Mycobacterium smegmatis. Minimum inhibitory concentration (MIC) values ranged between 625 and 5000 µg/mL, with an ethyl acetate extract of the root showing the lowest MIC value. The good antimycobacterial effects of the root part could be due to its high concentration of ellagic acid derivatives, ellagitannins, and flavonoids. Thin layer chromatography (TLC) fractionation resulted in some fractions with better activity than the starting point crude methanol extract (MIC 2500 µg/mL). Those fractions with the lowest MIC values contained a high number of antioxidant compounds. Fractions 3 and 4 (MIC 1500 and 1000 µg/mL, respectively) contained high concentrations of di-methyl ellagic acid ([M-H] - 329.0318). Fraction 6 (MIC 2000 µg/mL) contained a lower concentration of di-methyl ellagic acid and was not as growth inhibitory as fractions 3 and 4. Moreover, in fraction 3, an acetylated ellagic acid derivative ([M-H] - 343.0477) and di-methyl-ellagic acid xyloside ([M-H]- 461.0739) were tentatively characterized. Di-methyl ellagic acid xyloside was also present in fraction 4 and could strongly contribute to the antimycobacterial effect of this fraction. Additionally, protocatechuic acid ([M-H]-at m/z 153.0196) was present in fraction 4. Our antimycobacterial results obtained from this research justify the use of A. leiocarpa in Sudanese folk medicine against cough related to TB. Roots, stem bark, and leaves of A. leiocarpa are sources for new potent anti-TB drug lead compounds.

Alkaloid-Rich Crude Extracts, Fractions and Piperamide Alkaloids of Piper guineense Possess Promising Antibacterial Effects.

Piper guineense is a food and medicinal plant commonly used to treat infectious diseases in West-African traditional medicine. In a bid to identify new antibacterial compounds due to bacterial resistance to antibiotics, twelve extracts of P. guineense fruits and leaves, obtained by sequential extraction, as well as the piperine and piperlongumine commercial compounds were evaluated for antibacterial activity against human pathogenic bacteria. HPLC-DAD and UHPLC/Q-TOF MS analysis were conducted to characterize and identify the compounds present in the extracts with promising antibacterial activity. The extracts, with the exception of the hot water decoctions and macerations, contained piperamide alkaloids as their main constituents. Piperine, dihydropiperine, piperylin, dihydropiperylin or piperlonguminine, dihydropiperlonguminine, wisanine, dihydrowisanine and derivatives of piperine and piperidine were identified in a hexane extract of the leaf. In addition, some new piperamide alkaloids were identified, such as a piperine and a piperidine alkaloid derivative and two unknown piperamide alkaloids. To the best of our knowledge, there are no piperamides reported in the literature with similar UVλ absorption maxima and masses. A piperamide alkaloid-rich hexane leaf extract recorded the lowest MIC of 19 µg/mL against Sarcina sp. and gave promising growth inhibitory effects against S. aureus and E. aerogenes as well, inhibiting the growth of both bacteria with a MIC of 78 µg/mL. Moreover, this is the first report of the antibacterial activity of P. guineense extracts against Sarcina sp. and E. aerogenes. Marked growth inhibition was also obtained for chloroform extracts of the leaves and fruits against P. aeruginosa with a MIC value of 78 µg/mL. Piperine and piperlongumine were active against E. aerogenes, S. aureus, E. coli, S. enterica, P. mirabilis and B. cereus with MIC values ranging from 39⁻1250 µg/mL. Notably, the water extracts, which were almost devoid of piperamide alkaloids, were not active against the bacterial strains. Our results demonstrate that P. guineense contains antibacterial alkaloids that could be relevant for the discovery of new natural antibiotics.

Terminalia laxiflora and Terminalia brownii contain a broad spectrum of antimycobacterial compounds including ellagitannins, ellagic acid derivatives, triterpenes, fatty acids and fatty alcohols.

ETHNOPHARMACOLOGICAL RELEVANCE: Terminalia laxiflora Engl. & Diels, (Sudanese Arabic name: Darout الدروت) and Terminalia brownii Fresen (Sudanese Arabic name: Alshaf ألشاف) (Combretaceae) are used in Sudanese traditional folk medicine and in other African countries for treatment of infectious diseases, TB and its symptoms, such as cough, bronchitis and chest pain. AIM OF STUDY: Because of the frequent use of T. laxiflora and T. brownii in African traditional medicine and due to the absence of studies regarding their antimycobacterial potential there was a need to screen extracts of T. laxiflora and T. brownii for their growth inhibitory potential and to study the chemical composition and compounds in growth inhibitory extracts. MATERIALS AND METHODS: The plant species were collected in Sudan (Blue Nile Forest, Ed Damazin Forestry areas) and selected according to their uses in traditional medicine for the treatment of bacterial infections, including TB. Eighty extracts and fractions of the stem bark, stem wood, roots, leaves and fruits of T. laxiflora and T. brownii and nine pure compounds present in the active extracts were screened against Mycobacterium smegmatis ATCC 14468 using agar diffusion and microplate dilution methods. Inhibition zones and MIC values were estimated and compared to rifampicin. HPLC-UV/DAD, GC/MS and UHPLC/Q-TOF MS were employed to identify the compounds in the growth inhibitory extracts. RESULTS: The roots of T. laxiflora and T. brownii gave the best antimycobacterial effects (IZ 22-27 mm) against Mycobacterium smegmatis. The lowest MIC of 625 µg/ml was observed for an acetone extract of the root of T. laxiflora followed by methanol and ethyl acetate extracts, both giving MIC values of 1250 µg/ml. Sephadex LH-20 column chromatography purification of T. brownii roots resulted in low MIC values of 62.5 µg/ml and 125 µg/ml for acetone and ethanol fractions, respectively, compared to 5000 µg/ml for the crude methanol extract. Methyl (S)-flavogallonate is suggested to be the main active compound in the Sephadex LH- 20 acetone fraction, while ellagic acid xyloside and methyl ellagic acid xyloside are suggested to give good antimycobacterial activity in the Sephadex LH-20 ethanol fraction. RP-18 TLC purifications of an ethyl acetate extract of T. laxiflora roots resulted in the enrichment of punicalagin in one of the fractions (Fr5). This fraction gave a five times smaller MIC (500 µg/ml) than the crude ethyl acetate extract (2500 µg/ml) and this improved activity is suggested to be mostly due to punicalagin. 1,18-octadec-9-ene-dioate, stigmast-4-en-3-one, 5α-stigmastan-3,6-dione, triacontanol, sitostenone and ß-sitosterol were found in antimycobacterial hexane extracts of the stem bark of both studied species. Of these compounds, 1,18-octadec-9-ene-dioate, stigmast-4-en-3-one, 5α-stigmastan-3,6-dione, triacontanol, sitostenone have not been previously identified in T. brownii and T. laxiflora. Moreover, both plant species contained friedelin, betulinic acid, ß-amyrine and two unknown oleanane-type triterpenoids. Of the listed compounds, friedelin, triacontanol and sitostenone gave a MIC of 250 µg/ml against M. smegmatis, whereas stigmasterol and ß-sitosterol gave MIC values of 500 µg/ml. CONCLUSIONS: Our results show that T. laxiflora and T. brownii contain antimycobacterial compounds of diverse polarities and support the traditional uses of various parts of T. laxiflora and T.brownii as decoctions for treatment of tuberculosis. Further investigations are warranted to explore additional (new) antimycobacterial compounds in the active extracts of T. laxiflora and T. brownii.

Dry-air drying at room temperature - a practical pre-treatment method of tree leaves for quantitative analyses of phenolics?

INTRODUCTION: In ecological experiments, storage of plant material is often needed between harvesting and laboratory analyses when the number of samples is too large for immediate, fresh analyses. Thus, accuracy and comparability of the results call for pre-treatment methods where the chemical composition remains unaltered and large number of samples can be treated efficiently. OBJECTIVE: To study if a fast dry-air drying provides an efficient pre-treatment method for quantitative analyses of phenolics. METHODOLOGY: Dry-air drying of mature leaves was done in a drying room equipped with dehumifier (10% relative humidity, room temperature) and results were compared to freeze-drying or freeze-drying after pre-freezing in liquid nitrogen. The quantities of methanol-soluble phenolics of Betula pendula Roth, Betula pubescens Ehrh., Salix myrsinifolia Salisb., Picea abies L. Karsten and Pinus sylvestris L. were analysed with HPLC and condensed tannins were analysed using the acid-butanol test. RESULTS: In deciduous tree leaves (Betula, Salix), the yield of most of the phenolic compounds was equal or higher in samples dried in dry-air room than the yield from freeze-dried samples. In Picea abies needles, however, dry-air drying caused severe reductions in picein, stilbenes, condensed tannin and (+)-catechin concentrations compared to freeze-drying. In Pinus sylvestris highest yields of neolignans but lowest yields of acetylated flavonoids were obtained from samples freeze-dried after pre-freezing. CONCLUSION: Results show that dry-air drying provides effective pre-treatment method for quantifying the soluble phenolics for deciduous tree leaves, but when analysing coniferous species, the different responses between structural classes of phenolics should be taken into account.

A Seven-Year Study of Phenolic Concentrations of the Dioecious Salix myrsinifolia.

In boreal woody plants, concentrations of defensive phenolic compounds are expected to be at a high level during the juvenile phase and decrease in maturity, although there is variation between plant species. Females of dioecious species, like most of the Salicaceae, are expected to invest their resources in defense and reproduction, while males are expected to be more growth-oriented. We studied age- and sex-dependent changes in leaf and stem phenolics, and in height and diameter growth in a dioecious Salix myrsinifolia plants over a seven-year time period. In addition, we registered flowering as well as rust damage in the leaves. From the first year and throughout ontogenetic development from juvenile to adult phases, there was no significant change in the concentrations of any of the studied compounds in the leaves of S. myrsinifolia. In the stems, the concentrations of six out of 43 identified compounds decreased slightly with age, which may be partly explained by dilution caused by the increment in stem diameter with age. The fairly steady chemistry level over seven years, accompanied by moderate genotypic phenolic variation, indicates important roles of chemical defenses against herbivory for this early-successional species.

Epidihydropinidine, the main piperidine alkaloid compound of Norway spruce (Picea abies) shows promising antibacterial and anti-Candida activity.

This study reports for the first time promising antibacterial and antifungal effects of epidihydropinidine, the major piperidine alkaloid in the needles and bark of Norway spruce, Picea abies (L.) Karsten. Epidihydropinidine was growth inhibitory against all bacterial and fungal strains used in our investigation, showing the lowest MIC value of 5.37µg/mL against Pseudomonas aeruginosa, Enterococcus faecalis, Candida glabrata and C. albicans. Epidihydropinidine was nearly three times more active than tetracycline against P. aeruginosa and E. faecalis. Promising antibacterial effects were also recorded against Staphylococcus aureus and Bacillus cereus (MIC 10.75µg/mL) as well as against Salmonella enterica (MIC and MBC 43µg/mL). Our preliminary results suggest that epidihydropinidine as well related alkaloids of Norway spruce could be powerful candidates for new antibiotics and for preventing food spoilage.

Quantitative metabolite profiling of edible onion species by NMR and HPLC-MS.

Allium genus is a treasure trove of valuable bioactive compounds with potentially therapeutically important properties. This work utilises HPLC-MS and a constrained total-line-shape (CTLS) approach applied to (1)H NMR spectra to quantify metabolites present in onion species to reveal important inter-species differences. Extensive differences were detected between the sugar concentrations in onion species. Yellow onion contained the highest and red onion the lowest amounts of amino acids. The main flavonol-glucosides were quercetin 3,4'-diglucoside and quercetin 4'-glucoside. In general, the levels of flavonols were, higher in yellow onions than in red onions, and garlic and leek contained a lower amount of flavonols than the other Allium species. Our results highlight how (1)H NMR together with HPLC-MS can be useful in the quantification and the identification of the most abundant metabolites, representing an efficient means to pinpoint important functional food ingredients from Allium species.

Sex-related differences in growth and carbon allocation to defence in Populus tremula as explained by current plant defence theories.

Plant defence theories have recently evolved in such a way that not only the quantity but also the quality of mineral nutrients is expected to influence plant constitutive defence. Recently, an extended prediction derived from the protein competition model (PCM) suggested that nitrogen (N) limitation is more important for the production of phenolic compounds than phosphorus (P). We aimed at studying sexual differences in the patterns of carbon allocation to growth and constitutive defence in relation to N and P availability in Populus tremula L. seedlings. We compared the gender responses in photosynthesis, growth and whole-plant allocation to phenolic compounds at different combination levels of N and P, and studied how they are explained by the main plant defence theories. We found no sexual differences in phenolic concentrations, but interestingly, slow-growing females had higher leaf N concentration than did males, and genders differed in their allocation priority. There was a trade-off between growth and the production of flavonoid-derived phenylpropanoids on one hand, and between the production of salicylates and flavonoid-derived phenylpropanoids on the other. Under limited nutrient conditions, females prioritized mineral nutrient acquisition, flavonoid and condensed tannin (CT) production, while males invested more in above-ground biomass. Salicylate accumulation followed the growth differentiation balance hypothesis as low N mainly decreased the production of leaf and stem salicylate content while the combination of both low N and low P increased the amount of flavonoids and CTs allocated to leaves and to a lesser extent stems, which agrees with the PCM. We suggest that such a discrepancy in the responses of salicylates and flavonoid-derived CTs is linked to their clearly distinct biosynthetic origins and/or their metabolic costs.

Metabolite profiling of leek (Allium porrum L) cultivars by (1) H NMR and HPLC-MS.

INTRODUCTION: Leek (Allium ampeloprasum var. porrum) is consumed as a vegetable throughout the world. However, little is known about the metabolites of leek cultivars, especially those with potentially important beneficial properties for human health. OBJECTIVE: We provide new information for the overall metabolite composition of several leek cultivars grown in Europe by using HPLC-MS and (1) H NMR. METHODS: The use of a novel CTLS/NMR (constrained total-line-shape nuclear magnetic resonance) approach was found to be capable of reliable quantification, even with overlapping metabolite signals in the (1) H NMR of plant metabolites. Additionally, a new application for leek flavonoids was optimised for HPLC-MS. RESULTS: The total concentration of carbohydrates (glucose, fructose, kestose/nystose and sucrose) and nine amino acids varied by fourfold in leek juice from different cultivars, while the total concentrations of four organic acids were similar in all cultivars. All the quantified flavonols were kaempferol derivatives or quercetin derivatives and threefold differences in flavonol concentrations were detected between cultivars. CONCLUSION: In this study, various phytochemical profiles were determined for several leek cultivars by (1) H NMR spectroscopy with CTLS combined with HPLC-MS. The wide variation in bioactive compounds among commercial leek cultivars offers promising opportunities for breeders to raise the levels of important biochemical compounds in leek breeding lines, and also provides some objective measure for quality assurance for the leek industry.

Variation in the anthocyanin concentration of wild populations of crowberries (Empetrum nigrum L subsp. hermaphroditum).

Crowberry (Empetrum nigrum L.) is a relatively under-utilized wild berry that occurs widely throughout the northern hemisphere such as in Canada, Eurasia, and northern Europe. In this work, the anthocyanins of crowberries were analyzed from four geographically distinct crowberry populations in Finland using HPLC-DAD and HPLC-ESI/MS/MS. A total number of 15 anthocyanins were detected; 15 (11 structure elucidated) in all samples in order to profile-specific anthocyanin compositions throughout Finland. The major anthocyanin found in the samples collected from central and eastern Finland was delphinidin 3-galactoside accounting for more than 24% of the total anthocyanin content, while the cyanidin 3-galactoside was the major anthocyanin in the northernmost and in the western samples. Significant variation in the concentrations of different anthocyanins between and within crowberry populations were found suggesting that the synthesis of anthocyanins is modified by site-specific environmental conditions. The suitability of the crowberries as a potential source of health-promoting ingredients for incorporation into pharmaceutical and food industrial products is highlighted in this work due to the diverse anthocyanin profile.

Effect of latitude and altitude on the terpenoid and soluble phenolic composition of juniper (Juniperus communis) needles and evaluation of their antibacterial activity in the boreal zone.

The demand for dry juniper (Juniperus communis) needles as a raw material for the food, pharmaceutical, and cosmetic industries has increased rapidly in recent years. Juniper needles are known to be rich in terpenoids and phenolics, but their chemical composition and antibacterial properties have not been well-characterized. In this study, we describe the soluble phenolic and terpenoid composition of juniper needles collected in Finland (n = 125) and demonstrate that the concentration of these compounds clearly increased with latitude and altitude with, however, a stronger latitudinal effect (a higher content of monoterpenoids, proanthocyanidins, and flavonols in northern latitudes). Analysis of methanolic extracts showed quite good activity against both antibiotic-sensitive and -resistant Staphylococcus aureus strains and suggested an important role of the soluble phenolic fraction. Finally, we demonstrate the relative lack of toxicity of juniper extracts on keratinocytes and fibroblastic cells, raising the possibility of their use in preventing bacterial skin infection.

Effect of the temperature and the exclusion of UVB radiation on the phenolics and iridoids in Menyanthes trifoliata L. leaves in the subarctic.

The long-term effects of UVB exclusion and temperature on the methanol extractable (ME) phenolics (flavonoids, phenolic acids) and iridoids of Menyanthes trifoliata L. (Mt) leaves were studied in northern Finland (68 degrees N) using wooden frames covered with filters for UVB exclusion (polyester filter), control (cellulose acetate filter) and ambient (no filter) conditions. Analysis of ambient plots showed no effect of the daily mean temperature (2sigma = 1.58 degrees C) on the leaf ME compound content and composition, but minimum temperatures decreased the flavonol content. UVB exclusion did not affect the total ME compound content but significantly decreased the proportion of flavonols concomitantly with an increase in iridoids. Due to its high iridoid content, Mt appears as an interesting model plant for studying the iridoid biosynthesis and its regulation under stress conditions.

Medicinal properties, in vitro protocols and secondary metabolite analyses of scots pine.

Scots pine (Pinus sylvestris L.) is known as an economically important forest tree with a wide distribution throughout the Northern hemisphere. Recently, the species has also become recognized as a novel source of functional food and bioactive compounds with medicinal properties. The present paper provides up-to-date information on protocols for somatic embryogenesis (i.e., the most promising in vitro method for vegetative propagation of Scots pine). Endophyte protocols cover the topics of endophyte isolation, identification and elimination from in vitro cultures. Moreover, the protocols for secondary metabolite analyses are described in order to emphasize the emerging role of Scots pine as a medicinal plant.

Effect of different organic farming methods on the phenolic composition of sea buckthorn berries.

The effects of different organic cultivation methods on the berry phenolics of two Finnish sea buckthorn (Hippophae rhamnoides L. ssp. rhamnoides ) cultivars, 'Terhi' and 'Tytti', were studied in an experimental field at a coastal area in Merikarvia, western Finland. Cultivation methods included different fertilizers (designed for organic cultivation), mulches (organic and plastic), and land contours (flat land versus ridged beds). Two experiments were conducted: The first, a fertilization experiment, allowed for the estimation of the effects of cultivar, fertilizer, land contour, and all of their interactions. The second experiment, a mulch experiment, allowed for the estimation of the effects of mulch, land contours, and their interactions for the cultivar 'Tytti'. Berry phenolics were analyzed by high-performance liquid chromatography (HPLC) with ultraviolet (UV) detection. The results suggest that there are significant differences between the cultivars and cultivation methods. The concentrations of quercetin derivatives 1-3, isorhamnetin 3,7-diglucoside, quercetin-3-glucoside-7-rhamnoside, quercetin 3-glucoside, isorhamnetin 3-glucoside, and flavonoid derivative 3 were higher in 'Tytti' than in 'Terhi', while concentrations of isorhamnetin-glucoside 2 and 3 were higher in 'Terhi' than in 'Tytti'. Flat land increased the concentrations of isorhamnetin 3,7-diglucoside, isorhamnetin-glucoside 1, quercetin derivatives 2 and 4, and condensed tannins. Mulch did not have any significant effect on the concentrations of phenolic compounds. These results indicate that the phenolic accumulation in berries of studied sea buckthorn cultivars seems to be mainly dependent upon cultivar selection and soil structure.