Composition and Antibacterial Effect of Mint Flavorings in Candies and Food Supplements.

Mint flavorings are widely used in confections, beverages, and dairy products. For the first time, mint flavoring composition of mint candies and food supplements (n = 45), originating from 16 countries, as well as their antibacterial properties, was analyzed. The flavorings were isolated by Marcusson's type micro-apparatus and analyzed by GC-MS. The total content of the mint flavoring hydrodistilled extracts was in the range of 0.01 - 0.9%. The most abundant compounds identified in the extracts were limonene, 1,8-cineole, menthone, menthofuran, isomenthone, menthol and its isomers, menthyl acetate. The antimicrobial activity of 13 reference substances and 10 selected mint flavoring hydrodistilled extracts was tested on Escherichia coli and Staphylococcus aureus by broth dilution method. Linalool acetate and (-)-carvone, as most active against both bacteria, had the lowest MIC90 values. (+)-Menthyl acetate, (-)-menthyl acetate, and limonene showed no antimicrobial activity. Three of the tested extracts had antimicrobial activity against E. coli and 8 extracts against S. aureus. Their summary antimicrobial activity was not always in concordance with the activities of respective reference substances.

Essential oil composition of Pimpinella anisum L. fruits from various European countries.

Variations in the essential oil composition of Pimpinella anisum L. fruits obtained from different geographical areas of Europe were determined using capillary GC and GC-MS techniques. The essential oil content of the samples was 10.0-53.6 mL kg(-1) and did not confirm to the European Pharmacopoeia standard in 5 samples out of 14. A total of 21 compounds were identified and significant quantitative differences were observed among the samples. The major component was trans-anethole (76.9-93.7%); the other principal compounds in oils were gamma-himachalene (0.4-8.2%), trans-pseudoisoeugenyl 2-methylbutyrate (0.4-6.4%), p-anisaldehyde (tr-5.4%) and methylchavicol (0.5-2.3%). The highest content of trans-anethole (>90%) was found in the samples from Greece, Hungary, Scotland, Lithuania, Italy, and Germany (2 samples). Essential oil of aniseed from Estonia was rich in gamma-himachalene (8.2%) and trans-pseudoisoeugenyl 2-methylbutyrate (6.4%). The sample from France contained the highest amount of anisaldehyde (5.4%) comparing with other samples (0-3.1%). beta-Bourbonene and alpha-farnesene are determined in anise oil for the first time.

Phytochemical analysis of the essential oil of Thymus serpyllum L. growing wild in Estonia.

Variations in the essential oil composition of Thymus serpyllum L., growing wild in Estonia (33 samples) and in some other countries (Russia, Latvia and Armenia, seven samples) were determined. The oil were obtained from Estonia (46 samples) in yields 0.6-4.4 and 1.9-8.2 mL kg(-1) in other countries. The T. serpyllum herb grown in Estonia usually did not confirmed to the EP standard in the aspect of the essential oil contents (3.0 mL kg(-1)). Variations in the essential oil composition of wild thyme were studied using capillary gas chromatographic methods. A total of 94 components were identified. Thymol and carvacrol, mentioned in literature as principial components, are not the main components of the essential oil of wild thyme growing in Estonia. (E)-nerolidol, caryophyllene oxide, myrcene and borneol chemotypes of wild thyme drug are distinguishable. The chemical composition of samples from Russia, Latvia and Armenia is very variable.

Bioactive components of the hop strobilus: comparison of different extraction methods by capillary electrophoretic and chromatographic methods.

In the present study, the composition of the hop strobilus extract by using different extraction methods under different solvent conditions was analysed and compared. Several separation methods were applied to obtaining detailed information about the hop extract: capillary zone electrophoresis (CZE), high-performance liquid chromatography-mass-spectrometry (HPLC-MS/MS) and gas chromatography-mass-spectrometry (GC-MS). The electropherograms of different extracts varied dramatically. The oxidation reaction of the hop strobilus extract was examined.

Phytochemical analysis of the essential oil of Achillea millefolium L. from various European Countries.

Variations in the essential oil composition of Achillea millefolium L. growing in Estonia and in other European countries, were determined. The oils were obtained in yields of 0.9-9.5 mL kg-1. A total of 102 components were identified. The quantitatively most important components of yarrow were sabinene, beta-pinene, 1,8-cineole, artemisia ketone, linalool, alpha-thujone, beta-thujone, camphor, borneol, fenchyl acetate, bornyl acetate, (E)-beta-caryophyllene, germacrene D, caryophyllene oxide, beta-bisabolol, delta-cadinol, chamazulene etc. Samples from Estonia contained high amounts of monoterpenes and chamazulene. High amounts of monoterpenes and chamazulene were also found in samples from Hungary, Greek, Moldavia, Latvia, Lithuania and Germany. The oils from France, Belgium, Russia, Armenia, Spain and Italy were rich in oxygenated monoterpenes and contained a little amount of chamazulene. The drugs from Greece, Estonia, Moldavia and Scotland were rich in sesquiterpenes. The Millefolii herba grown in Estonia conforms to the European Pharmacopoeia (EP) standards in the aspect of the essential oil contents.

Analysis of Carotenoids, Flavonoids and Essential Oil of Calendula officinalis Cultivars Growing in Estonia.

Pot Marigold (Calendula officinalis L.) is an annual ornamental plant with yellow to orange flowers which belongs the the Asteraceae family, its flowers are- used in the pharmaceutical, cosmetic and food industries. The commercial samples of seeds (n=24) of C. officinalis were cultivated in home gardens in Estonia. Total carotenoid contents, determined spectrophotometrically (X=455 nm) in the inflorescences, varied from 0.7 to 2.7%, the cultivar with the highest carotenoid content was 'Dracon'. The total content of flavonoids, determined spectrophotometrically (?=370 nm), varied from 0.8% to 1.7%. The best cultivar contained almost maximum amount of total carotenoids (2.5%) and flavonoids (1.7%) was 'Radio". The oil yields of eight cultivars of C. officinalis obtained were from 0.10 to 0.43%, the highest yield (0.43%) of the oil was obtained from the cultivar 'Double Ball'. A total 70 compounds were identified and analysed by GC-MS in the essential oil of C. officinalis, representing over 95% from total oil. The major components of the oils (>5%) of C. officinalis cultivars were a- cadinole (18.4-32.0%), y-cadinene (9.7-18.9%), viridiflorol (2.0-10.0%), t-cadinole (4.7-7.9%), 8-cadinole (3.3-5.6%), and y-muurolene (0.8-5.1%). There is no clear relation between the colour of the inflorescences of C. officinalis and the total content of flavonoids and carotenoids. The chemotypes of essential oils of C. officinalis were not found among the cultivars studied.

Mint Flavorings from Candies Inhibit the Infectivity of Chlamydia pneumoniae.

The impact of solvent extracts from the distillation water (flavoring extracts) isolated from mint flavored candies on the infectivity of the intracellular bacterium Chlamydia pneumoniae was evaluated by an in vitro model of epithelial cell infections., The mint flavoring extracts were isolated from the candies by simultaneous hydrodistillation and their chemical composition, established by GC-MS, demonstrated menthol and limonene as the most abundant components. Results obtained by treating C. pneumoniae elementary bodies (EBs) with the flavoring extracts or pure reference compounds showed a significant decrease in EB infectivity, achieved with most of the extracts. This antichlamydial activity could be related to the relatively high menthol content of the extracts. Overall, the obtained data indicates that the flavorings present in the candies are able to target the metabolically quiet, non-replicating form of the bacterium and to suppress the spread of this respiratory pathogen from one cell to another.

Selectivity of Pinus sylvestris extract and essential oil to estrogen-insensitive breast cancer cells Pinus sylvestris against cancer cells.

BACKGROUND: So far, the anticancer action of pine tree extracts has mainly been shown for the species distributed widely around the Asian countries. OBJECTIVE: Therefore, this study was performed to examine the potential cytotoxicity of Scots pine (Pinus sylvestris L.) native also to the European region and growing widely in Estonia. MATERIALS AND METHODS: The cytotoxic activity of methanol extract and essential oil of Scots pine needles was determined by sulforhodamine B assay in different human cancer cell lines. RESULTS: This needle extract was found to suppress the viability of several human cancer cell lines showing some selectivity to estrogen receptor negative breast cancer cells, MDA-MB-231(half maximal inhibitory concentration [IC50] 35 µg/ml) in comparison with estrogen receptor-positive breast cancer cells, MCF-7 (IC50 86 µg/ml). It is the strongest cytotoxic effect at all measured, thus far for the needles and leaves extracts derived from various pine species, and is also the first study comparing the anticancer effects of pine tree extracts on molecularly different human breast cancer cells. The essential oil showed the stronger cytotoxic effect to both negative and positive breast cancer cell lines (both IC50 29 µg/ml) than pine extract (IC50 42 and 80 µg/ml, respectively). CONCLUSION: The data from this report indicate that Scots pine needles extract and essential oil exhibits some potential as chemopreventive or chemotherapeutic agent for mammary tumors unresponsive to endocrine treatment.

Effect of storage on the essential oil composition of Piper nigrum L. fruits of different ripening states.

The qualitative and quantitative composition of the essential oil from black, green, and white pepper was determined by using a simultaneous distillation and extraction micromethod for oil isolation and gas chromatography (GC)/flame ionization detection (FID) and GC/mass spectrometry (MS) analysis techniques. The most abundant compounds in pepper oils were (E)-beta-caryophyllene (1.4-70.4%), limonene (2.9-38.4%), beta-pinene (0.7-25.6%), Delta-3-carene (1.7-19.0%), sabinene (0-12.2%), alpha-pinene (0.3-10.4%), eugenol (0.1-41.0%), terpinen-4-ol (0-13.2%), hedycaryol (0-9.1%), beta-eudesmol (0-9.7%), and caryophyllene oxide (0.1-7.2%). Green pepper corn obtained by a sublimation drying method gave more oil (12.1 mg/g) and a much higher content of monoterpenes (84.2%) in the oil than air-dried green pepper corn (0.8 mg/g and 26.8%, respectively). The oil from ground black pepper contained more monoterpenes and less sesquiterprnes and oxygenated terpenoids as compared to green and white pepper oils. After 1 year of storage of pepper samples in a glass vessel at room temperature, the amount of the oils isolated decreased, the content of terpenes decreased, and the amount of oxygenated terpenoids increased. Differently from other pepper samples, 1 year storage of green pepper corn raised the oil amount more than twice of both drying methods.

Content and composition of the essential oil of Thymus serpyllum L. growing wild in Estonia.

OBJECTIVE: The aim of this work was to analyze the essential oil content and its composition in the drug (Serpylli herba) of wild thyme (Thymus serpyllum L.) originating from 20 different natural places of growth in Estonia. MATERIAL AND METHODS: The quantitative content of essential oil was determined according to the method of European Pharmacopoeia. Gas chromatographic analysis of essential oils was carried out using a gas chromatography with flame ionization detector on two fused silica capillary columns with bonded stationary phases NB-30 and SW-10. The identification of the oil components was accomplished by comparing their retention indices on two columns with the retention indices values of standard compounds, with our retention indices data bank and with literature data. The results obtained were confirmed by gas chromatography / mass spectrometry. RESULTS: The content of essential oil is between 0.6 and 4 ml/kg and usually is not in conformity with European Pharmacopoeia standard (3 ml/kg). There were 55 components identified in the essential oil of wild thyme of Estonian origin. Differently from the data in the literature of foreign countries, thymol and carvacrol are not the main components of the essential oil of wild thyme growing in Estonia. The main components here are (E)-nerolidol, caryophyllene oxide, myrcene, (E)-beta-caryophyllene and germacrene D. CONCLUSION: In Estonia, the (E)-nerolidol-caryophyllene oxide, (E)-nerolidol-myrcene and myrcene chemotypes of wild thyme drug are distinguishable.

Composition of the essential oil of the Rhododendron tomentosum Harmaja from Estonia.

Wild Rhododendron tomentosum Harmaja shoots were collected from four localities of Estonia. Essential oils, isolated from dried samples by simultaneous distillation and extraction, were analysed using GC-FID and Gas chromatography-mass spectrometry methods. The yields of oils were in the range 0.14-0.87%. A total of 72 constituents, accounting for over 95% of the total oil yield, were identified. The major components in the all four oils studied were palustrol (15.9-53.5%), ledol (11.8-18.3%), γ-terpineol (0-31.2%), p-cymene (0.1-13.9%), lepalone (0.7-6.5%), lepalol (1.0-6.5%) and cyclocolorenone (1.0-6.4%). Two different chemotypes of R. tomentosum were found in Estonia and one of them was rich in palustrol (41.0-53.5%) and ledol (14.6-18.3%). The second chemotype, for the first time, was rich in γ-terpineol (24.7-31.2%) and contained less of palustrol (15.9-16.7%) and ledol (11.8-12.8%), but more p-cymene (12.5-13.9%). Also, γ-terpineol was identified for the first time in the oils of R. tomentosum.

Essential oil composition of Foeniculum vulgare Mill. fruits from pharmacies in different countries.

Variations in the essential oil composition of Foeniculum vulgare Mill. commercial fruits obtained from retail pharmacies in Estonia, Norway, Austria and Moldova and from a spice shop in Turkey were determined using capillary GC techniques. The essential oil content of all the samples was 5-51 mL kg(-1) and between 22 and 51 mL kg(-1) in fennel fruits bought from pharmacies. A total of 34 compounds were identified. The major component was trans-anethole (34.8-82.0%); the other principal compounds in oils were fenchone (1.6-22.8%), estragole (2.4-17.0%), limonene (0.8-16.5%), and cis-anethole (0.1-8.6%). The yield of essential oil (5.0 mL kg(-1)) and content of trans-anethole was very low (34.8%) in the Turkish spice sample. Maximum yield of essential oil was found in fennel from Norway and Austria (50.7 and 50.5 mL kg(-1), respectively); these samples were rich in fenchone (21.2% and 22.8%), but contained less trans-anethole (64.6-63.7) than samples from Estonia and Moldova (82.0% and 80.9%). The typical samples of sweet fennel (bought from Estonia and Moldova) and bitter fennel (from Norway and Austria) were found to conform completely or partially to EP standards, although fennel type was always not marked on the packages.

Variation in the composition of the essential oils, phenolic compounds and mineral elements of Hypericum perforatum L. growing in Estonia.

A comprehensive investigation of the chemical composition of the aerial parts of Hypericum perforatum L. collected in three habitations in Estonia was carried out. An analysis by gas chromatography-mass spectrometry and gas chromatography-flame ionisation detection established the main components of the essential oils. The phenolic compounds both in ethanol and water extracts of the plant were analysed using liquid chromatography-mass spectrometry (LC-MS) and capillary zone electrophoresis. In addition to the earlier published polyphenols, several new phenolic acids and flavonoids, such as quercetin hexoside malonates and an A-type catechin-epicatechin trimer were identified in this Hypericum for the first time. The contents of the pharmaceutically important antidepressants hyperforin and hypericin were also estimated by LC-MS and compared with the data in the literature. The composition of the mineral elements was determined by atomic absorption spectroscopy. The results of the study demonstrate a rather high variability in the content of different substance groups in H. perforatum L. and, hence, the need for a survey of the raw material in the course of selection of raw materials for pharmaceutical preparations.

Chemical investigation of the essential oil from berries and needles of common juniper (Juniperus communis L.) growing wild in Estonia.

The essential oils obtained by simultaneous distillation and extraction (SDE) from the fresh and dried needles and dried berries of Juniperus communis L. of Estonian origin were subjected to GC-FID and GC-MS analyses. The yields of the oils ranged between 0.2% and 0.6% from juniper berries and between 0.5% and 1.0% from needles (dried weight). A total of 87 compounds were identified, representing over 95% of the oil. The major compounds in the needle oil were monoterpenes α-pinene (33.3-45.6%), sabinene (0.2-15.4%), limonene (2.8-4.6%) and sesquiterpenes (E)-ß-caryophyllene (0.8-10.3%), α-humulene (0.8-6.2%) and germacrene D (3.0-7.8%). The juniper berry oil was rich in α-pinene (53.6-62.3%), ß-myrcene (6.5-6.9%) and germacrene D (4.5-6.1%). The main oxygenated terpenoids found in the needle oil were germacrene D-4-ol (0.4-4.0%) and α-cadinol (to 2.7%). The oil from fresh needles contained high amounts of (E)-2-hexenal (3.7-11.7%).

Content and composition of the essential oil of Chamomilla recutita (L.) Rauschert from some European countries.

Variations in the essential oil composition of Chamomilla recutita (L.) Rauschert from different European countries were determined. A total of 39 components were identified, representing over 92% of the total oil yield. The principal biologically active compounds in chamomile oils were bisabolol oxide A (3.1-56.0%), alpha-bisabolol (0.1-44.2%), bisabolol oxide B (3.9-27.2%), cis-enyne-bicycloether (8.8-26.1%), bisabolon oxide A (0.5-24.8%), chamazulene (0.7-15.3%), spathulenol (1.7-4.8%) and (E)-beta-farnesene (2.3-6.6%). In 8 chamomile samples from 13, bisabolol oxide A (27.5-56.0%) was predominant (among them in three Estonian samples). alpha-Bisabolol (23.9-44.2%) was predominant in the samples from Moldova, Russia and the Czech Republic. The sample from Armenia was rich in bisabolol oxide B (27.2%) and chamazulene (15.3%). The oils were obtained in yields of 0.7-6.7 mL kg(-1) and the minimum limit of 4 mL kg(-1) stated by the European Pharmacopoeia was exceeded only in 13 samples from 13 analysed drugs.

Composition of essential oil of aerial parts of Chamomilla suaveolens from Estonia.

Variations in the essential oil composition of aerial parts of pineapple weed (Chamomilla suaveolens (Pursh) Rydb.) growing wild in Estonia, were determined using GC/FID and GC/MS. Forty-four components were identified, representing over 90% of the total oil. Nine compounds have not been mentioned in the literature before. The principal biologically active compounds in C. suaveolens oils were (Z)-en-yne-dicycloether (17.0-40.7%), (E)-beta-farnesene (19.5-32.2%), geranyl isovaleriate (8.4-18.4%), palmitic acid (0.3-9.4%) and myrcene (1.1-7.9%). The investigation seems to approve the benefit of using aerial parts of pineapple weed as the substitute for flowers.

Composition of the essential oil of Salvia officinalis L. from various European countries.

Variations in the essential oil composition of Salvia officinalis L. growing in Estonia and in other European countries were determined. The oils were obtained in yields of 2.2-24.8 mL kg(-1). In three samples, the content of essential oil did not conform to the EP standard (10 mL kg(-1)). Variations in the essential oil composition of sage were studied using capillary gas chromatographic methods. A total of 40 components were identified. The principal components in the sage oils were 1,8-cineole, camphor, alpha-thujone, beta-thujone, borneol, and viridiflorol. The chemotypes of sage were not determined in investigated samples. The concentration of the main compounds in the drugs cultivated in Estonia varied in about the same range as the concentrations of these compounds in the oils of drugs obtained from other countries. The comparatively high concentration of toxic thujones seem to be characteristic to sage leaves cultivated in Estonia.