Effect of varying fermentation conditions with ensiling period and inoculum on photosynthetic pigments and phytol content in Italian ryegrass (Lolium multiflorum Lam.) silage.

This study aimed to investigate the effect of an ensiling period (Experiment 1) and adding lactic acid bacteria (LAB, Experiment 2) on the changes in carotenoid, chlorophyll, and phytol in ensiled Italian ryegrass (IR, Lolium multiflorum Lam.). In Experiment 1, the IR herbage ensiled into plastic bags was analyzed for the contents of photosynthetic pigments and phytol over a 5-week period. During the ensiling process, the ß-carotene content decreased (p < .05), whereas the lutein content did not change. Although the chlorophyll content decreased (p < .05) after ensiling, the phytol content barely changed until week 5. In Experiment 2, IR herbage was ensiled without additive, as a Control, or with LAB for 60 days. The pH was lower (p < .05) and lactic acid content was higher (p < .05) for the LAB silage than for the Control. The chlorophyll content in silage was not affected by the LAB; however, the ß-carotene content was higher (p < .05) for the LAB silage than for the Control. Phytol and lutein contents in the herbage did not change after ensiling. These results indicate that phytol and lutein in IR herbage can be preserved well in silage, irrespective of their fermentation condition.

Volatile oil analysis of aerial parts of Boerhavia coccinea (Mill.).

Volatile oil of the aerial part of Boerhavia coccinea was obtained by Clevenger-type apparatus, the chemical components were analysed using gas chromatography (GC) and gas chromatography-mass spectrometry (GC-MS). Twenty constituents were recognised in the oil which accounted for 98.05% of it. This was characterised by high level of ethers (29.85%), while most dominant compound was phytol (24.02%). The essential oil of B. coccinea mainly consisted ethers and alcohols, which is being reported for the first time.

Antioxidant activity and chemical composition of oleoresin from leaves and flowers of Brunfelsia uniflora.

In this study, the temperature and pressure of supercritical CO2 extraction were evaluated to obtain oleoresin of Brunfelsia uniflora leaves and flowers. The oleoresin compounds were identified by gas chromatography-mass spectrometry. The antioxidant activity was evaluated by three different methods. The highest oleoresin yields were 3.32% at 40°C and 200 bar for the leaves, and 1.03% at 60°C and 200 bar for the flowers. The main extracted compounds from leaves were phytol varying from 11.95 to 36.42% and α-tocopherol from 15.53 to 43.10%, and from flowers were geranyl linalool from 11.05 to 21.42% and α-amyrin from 9.66 to 22.12%. Oleoresin obtained at 60°C and 150 bar from leaves presented high antioxidant activity by DPPH (IC50 1.90 mg/mL) and by FRAP (1.8 µmol Fe2+/mg). ß-carotene/linoleic acid co-oxidation oleoresin from leaves at 0.25 mg/mL presented higher antioxidant activity than Trolox. The total phenolic content of the oleoresin from leaves ranged from 66.20 to 83.33 µg/mg and from flowers it was just up to 12.46 µg/mg. The extraction conditions affected yield, chemical composition, and antioxidant activity of oleoresin from leaves and flowers. This is the first report on the antioxidant activity of B. uniflora oleoresin from leaves and flowers and provides subsidies for potential applications in chemical, pharmaceutical, and food industries.

Chemical Compositions and Antioxidant Activities of Essential Oils Extracted from the Petals of Three Wild Tree Peony Species and Eleven Cultivars.

The aim of this study was to investigate the essential oil (EO) compositions and antioxidant activities from petals of three wild tree peony species (Paeonia delavayi, P. lutea, and P. rockii) and eleven P. suffruticosa cultivars from different cultivar groups. The EOs yields varied from 0.63% to 1.25% (v/v) among samples when using supercritical CO2 extraction. One hundred and sixty-three components were detected by GC/MS; and among them, linalool oxide, (Z)-5-dodecen-1-yl acetate, nonadecane, (Z)-5-nonadecene, heneicosane, phytol, and linoleic acid ethyl ester were dominant. According to hierarchical cluster analysis, principal component analysis and correspondence analysis, P. lutea, P. delavayi, and 'High Noon' were clustered in a group described as having a refreshing herbal-like note due to high rates of phytol and linalool oxide. Notably, P. lutea and P. delavayi also had strong DPPH and ABTS radical scavenging activities. These results suggest that P. lutea and P. delavayi are the most promising candidates as useful sources of fragrances and natural antioxidants.

Effects of nitrogen fertilizer and harvesting stage on photosynthetic pigments and phytol contents of Italian ryegrass silage.

This study aimed to investigate the effects of nitrogen (N) fertilizer application and harvesting stage on the contents of chlorophyll, phytol and carotenoids (ß-carotene and lutein) in Italian ryegrass herbage before and after ensiling, and the extent of phytol preservation after ensiling. Three rates of N fertilizer (0, 60 and 120 kg N/ha) were applied by top-dressing as an additional fertilizer. The herbage harvested at booting stage (27 weeks of age) or heading stage (29 weeks of age) were wilted for 1 day, then ensiled for 60 days using a small-scale pouch system. In the pre-ensiled herbages, increasing N fertilizer application increased the contents of crude protein and photosynthetic pigments, and these contents were also higher at the booting stage compared with the heading stage. In the silage, increasing N fertilizer application also increased the contents of crude protein, the photosynthetic pigments and their derivatives (pheophytin and pheophorbide), while harvesting stage did not affect the contents of ß-carotene, chlorophylls or pheophorbide. Nitrogen fertilizer application and early harvesting of herbage increased lutein and phytol contents in Italian ryegrass silage. Lutein and phytol in Italian ryegrass herbage are indicated to be well preserved during ensiling.

Geochemically distinct carbon isotope distributions in Allochromatium vinosum DSM 180T grown photoautotrophically and photoheterotrophically.

Anoxygenic, photosynthetic bacteria are common at redox boundaries. They are of interest in microbial ecology and geosciences through their role in linking the carbon, sulfur, and iron cycles, yet much remains unknown about how their flexible carbon metabolism-permitting either autotrophic or heterotrophic growth-is recorded in the bulk sedimentary and lipid biomarker records. Here, we investigated patterns of carbon isotope fractionation in a model photosynthetic sulfur-oxidizing bacterium, Allochromatium vinosum DSM180T . In one treatment, A. vinosum was grown with CO2 as the sole carbon source, while in a second treatment, it was grown on acetate. Different intracellular isotope patterns were observed for fatty acids, phytol, individual amino acids, intact proteins, and total RNA between the two experiments. Photoautotrophic CO2 fixation yielded typical isotopic ordering for the lipid biomarkers: δ13 C values of phytol > n-alkyl lipids. In contrast, growth on acetate greatly suppressed intracellular isotopic heterogeneity across all molecular classes, except for a marked 13 C-depletion in phytol. This caused isotopic "inversion" in the lipids (δ13 C values of phytol < n-alkyl lipids). The finding suggests that inverse δ13 C patterns of n-alkanes and pristane/phytane in the geologic record may be at least in part a signal for photoheterotrophy. In both experimental scenarios, the relative isotope distributions could be predicted from an isotope flux-balance model, demonstrating that microbial carbon metabolisms can be interrogated by combining compound-specific stable isotope analysis with metabolic modeling. Isotopic differences among molecular classes may be a means of fingerprinting microbial carbon metabolism, both in the modern environment and the geologic record.

Physicochemical and antioxidant characterization of Justicia spicigera.

Extracts with water:ethanol (100:0, 70:30, 50:50, 30:70, 0:100) solutions from fresh (F), just dried (JD), dried and stored for one year (DS) Justicia spicigera leaves were obtained using the stirring and ultrasound techniques. Extracts were analyzed in physicochemical and antioxidant characteristics. Identification of chemical compounds by gas chromatography-mass spectroscopy (GC-MS) was also performed. 2.14±0.91, 5.67±1.70, and 8.52±4.97g Gallic acid equivalents/100g dry weight (d.w.) of phenolic compounds were found, in average, for F, JD, and DS J. spicigera, respectively. 2.22±1.31, 2.58±2.11, and 8.48±3.78g Trolox equivalents/100g d.w. were detected with the ABTS method and 0.49±0.33, 1.23±0.87, and 0.88±0.94g with the DPPH method for F, JD and DS J. spicigera, respectively. Eucalyptol, phytol, and azulene were identified as the main compounds. J. spicigera showed colors (green-iridescent, green-yellow, or pink of different intensities) and antioxidant characteristics depending on the solvent concentration. Extracts could be used in the food and pharmaceutical industries.

Phytyl Fatty Acid Esters in the Pulp of Bell Pepper (Capsicum annuum).

Phytyl fatty acid esters (PFAE) are esters of fatty acids with the isoprenoid alcohol phytol (3,7R,11R,15-tetramethylhexadec-2E-enol). In this study, PFAE were identified and quantified in bell pepper using gas chromatography with mass spectrometry (GC-MS). All red (n = 14) and yellow (n = 6) samples contained six or seven PFAE at 0.9-11.2 mg/100 g fresh weight. By contrast, PFAE were not detected in green bell pepper samples (n = 3). PFAE might eventually be a source for bioavailable phytol, which can be transformed into phytanic acid by humans. Phytanic acid cannot be properly degraded by patients who suffer from Refsum's disease (tolerable daily intake (TDI) ≤ 10 mg of phytanic acid). The phytol moiety of the PFAE (0.4-5.4 mg/100 g fresh weight) would contribute up to ∼50% to the TDI with the consumption of only one portion of bell pepper fruit pulp.

Characteristic Chemical Components and Aroma-active Compounds of the Essential Oils from Ranunculus nipponicus var. submersus Used in Japanese Traditional Food.

Ranunculus nipponicus var. submersus is an aquatic macrophyte; it is known as a wild edible plant in Japan for a long time. In this study, the essential oils from the fresh and dried aerial parts of R. nipponicus var. submersus were extracted by hydrodistillation and analyzed by gas chromatography (GC) and GC-mass spectrometry (GC-MS). Moreover, important aroma-active compounds were also detected in the oil using GC-olfactometry (GC-O) and aroma extract dilution analysis (AEDA). Thus, 98 compounds (accounting for 93.86%) of the oil were identified. The major compounds in fresh plant oil were phytol (41.94%), heptadecane (5.92%), and geranyl propionate (5.76%), while those of. Dried plant oil were ß-ionone (23.54%), 2-hexenal (8.75%), and dihydrobovolide (4.81%). The fresh and dried oils had the green-floral and citrus-floral odor, respectively. The GC-O and AEDA results show that phenylacetaldehyde (green, floral odor, FD-factor = 8) and ß-ionone (violet-floral odor, FD-factor = 8) were the most characteristic odor compounds of the fresh oils. ß-Cyclocitral (citrus odor, FD-factor = 64) and ß-ionone (violet-floral odor, FD-factor = 64) were the most characteristic odor compounds of the dried oil. These compounds are thought to contribute to the flavor of R. nipponicus var. submersus.

2-Methyl-3-buten-2-ol (MBO) synthase expression in Nostoc punctiforme leads to over production of phytols.

Phytol is a diterpene alcohol of medicinal importance and it also has potential to be used as biofuel. We found over production of phytol in Nostoc punctiforme by expressing a 2-Methyl-3-buten-2-ol (MBO) synthase gene. MBO synthase catalyzes the conversion of dimethylallyl pyrophosphate (DMAPP) into MBO, a volatile hemiterpene alcohol, in Pinus sabiniana. The result of enhanced phytol production in N. punctiforme, instead of MBO, could be explained by one of the 2 models: either the presence of a native prenyltransferase enzyme with a broad substrate specificity, or appropriation of a MBO synthase metabolic intermediate by a native geranyl diphosphate (GDP) synthase. In this work, an expression vector with an indigenous petE promoter for gene expression in the cyanobacterium N. punctiforme was constructed and MBO synthase gene expression was successfully shown using reverse transcriptase (RT)-PCR and SDS-PAGE. Gas chromatography--mass spectrophotometry (GC-MS) was performed to confirm phytol production from the transgenic N. punctiforme strains. We conclude that the expression of MBO synthase in N. punctiforme leads to overproduction of an economically important compound, phytol. This study provides insights about metabolic channeling of isoprenoids in cyanobacteria and also illustrates the challenges of bioengineering non-native hosts to produce economically important compounds.

Chemical composition profiling and antifungal activity of the essential oil and plant extracts of Mesembryanthemum edule (L.) bolus leaves.

BACKGROUND: Essential oil from Mesembryanthemum edule leaves have been used by the Eastern Cape traditional healers for the treatment of respiratory tract infections, tuberculosis, dysentery, diabetic mellitus, laryngitis and vaginal infections. The investigation of bioactive compounds in the essential oil of this plant could help to verify the efficacy of the plant in the management or treatment of these illnesses. MATERIALS AND METHODS: Various concentrations of the hydro-distilled essential oil, ranging from 0.005-5 mg/ml, were tested against some fungal strains, using the micro-dilution method. Minimum inhibitory activity was compared with four other different crude extracts of hexane, acetone, ethanol and aqueous samples from the same plant. The chemical composition of the essential oil, hexane, acetone and ethanol extracts was determined using GC-MS. RESULTS: GC/MS analysis of the essential oil resulted in the identification of 28 compounds, representing 99.99% of the total oil. Phytoconstituents of hexane, acetone and ethanol extracts yielded a total peak chromatogram of fifty nine compounds. A total amount of 10.6% and 36.61% of the constituents were obtained as monoterpenes and oxygenated monoterpenes. Sesquiterpene hydrocarbons (3.58%) were relatively low compared to the oxygenated sesquiterpenes (9.28%), while the major concentrated diterpenes and oxygenated diterpenes were 1.43% and 19.24 %, respectively and phytol 12.41%. Total amount of fatty acids and their methyl esters content, present in the oil extract, were found to be 19.25 %. Antifungal activity of the oil extract and four solvent extracts were tested against five pathogenic fungal strains. The oil extract showed antifungal activity against Candida albican, Candida krusei, Candida rugosa, Candida glabrata and Cryptococcus neoformans with MIC ranges of 0.02 0.31 mg/ml. Hexane extract was active against the five fungal strains with MICs ranging between 0.02-1.25 mg/ml. Acetone extracts were active against C. krusei only at 0.04mg/ml. No appreciable antifungal activity was found in either ethanol or water extracts when compared with commercial antibiotics. CONCLUSION: The profile of chemical constituents found in M. edule essential oil and its antifungal properties support the use of M. edule by traditional healers as well as in the pharmaceutical and food industries as a natural antibiotic and food preservative.

[Chemical components from essential oil of Pandanus amaryllifolius leaves].

OBJECTIVE: To analyze the chemical compositions of Pandanus amaryllifolius leaves essential oil extracted by steam distillation. METHODS: The essential oil of Pandanus amaryllifolius leaves was analyzed by gas chromatography-mass spectrum, and the relative content of each component was determined by area normalization method. RESULTS: 128 peaks were separated and 95 compounds were identified, which weighed 97.75%. The main chemical components of the essential oil were phytol (42.15%), squalene (16.81%), what's more pentadecanal (6.17%), pentadecanoic acid (4.49%), 3, 7, 11, 15-tetramethyl-2-hexadecen-1-ol (3.83%), phytone (2.05%) and the other 74 chemical compositions were firstly identified from the essential oil of Pandanus amaryllifolius leaves. CONCLUSION: The chemical compositions of Pandanu samaryllifolius leaves essential oil was systematically, deeply isolated and identified for the first time. This experiment has provided scientific foundation for further utilization of Pandanus amaryllifolius leaves.

Pachira glabra Pasq. essential oil: chemical constituents, antimicrobial and insecticidal activities.

The chemical composition of essential oil obtained by hydrodistillation of the leaves of Pachira glabra Pasq., (PgEO) has been studied by Gas Chromatography (GC) and Gas Chromatography coupled with Mass Spectrometry (GC/MS). Thirty three constituents representing 98.4% of total contents were identified from the essential oil. The major constituents of oil were limonene (23.2%), ß-caryophyllene (14.5%), phtyol (8.5%) and ß-bisabolene (6.3%). The antimicrobial activity of the PgEO was evaluated against a panel of ten bacteria and three fungal strain using agar diffusion and broth microdilution methods. Results have shown that the PgEO exhibited moderate to strong antimicrobial activity against the tested microorganisms except Citrobacter youagae, Micrococcus spp. and Proteus spp. The activity zones of inhibition (ZI) and minimum inhibitory concentrations (MIC) ranged between 13.7 mm-24.0 mm and 0.3 mg/mL-2.5 mg/mL, respectively. The insecticidal activity of PgEO was assayed against the adult Sitophilus zeamais. The lethal concentrations (LC50 and LC90) of the PgEO showed it to be toxic against adult S. zeamais at 32.2 and 53.7 mg/mL, respectively. This is the first report on the chemical composition and in vitro biological activities of essential oil of P. glabra growing in Nigeria.

[GC-MS analysis of chemical constituents of volatile oil from flowers of Rhododendron mucronatum].

OBJECTIVE: To analyze the chemical constituents of volatile oil from flowers of Rhododendron mucronatum. METHODS: The volatile oil was extracted by water-steam distillation and analyzed by GC-MS. RESULTS: Forty-nine compounds, which occupied 79.55% of total constituents, were identified. The major constituents were linalool, beta-eudesmene, phytol, benzyl benzoate, benzyl salicylate and nonyl aldehyde. CONCLUSION: The chemical constituents of volatile oil, which contain many bioactive constituents, are mainly composed of terpenes, esters, and alkanes.

[Study on chemical constituents of volatile oil from rhizomes and leaves of Pileostegia viburnoides var. glabrescens by GC-MS].

OBJECTIVE: To analyze the chemical constituents of volatile oil from the rhizomes and leaves of Pileostegia viburnoides var. glabrescens by GC-MS. METHODS: The volatile oil was extracted from the rhizomes and leaves of Pileostegia viburnoides var. glabrescens by steam distillation. The constituents of volatile oil were identified by GC-MS technology. RESULTS: 37 compounds were identified from the oil of rhizomes. 36 compounds were identified from the oil of leaves. The rhizomes and leaves volatile oil had 18 compounds in common. CONCLUSION: This study is the first one to report the volatile components of Pileostegia viburnoides var. glabrescens. It can provide a scientific basis for rational use of the rhizomes and leaves of Pileostegia viburnoides var. glabrescens.

Differentiation of refined and virgin edible oils by means of the trans- and cis-phytol isomer distribution.

The differentiation of nonrefined (e.g., cold-pressed) and refined edible oils is an important task in food control because of the higher commercial value of the former. Here, we explored the suitability of the relative abundance of cis-phytol as a marker for authentication of nonrefined edible oils. Phytol, the tetramethyl-branched, monoenoic alcohol, is found widespread in nature as a part of chlorophyll. In chlorophyll, only trans-phytol is found. In this study, we present a method for the analysis of the phytol isomers, considering that traces of cis-phytol (contributing 0.1% to the phytol content) can be determined next to trans-phytol. For this purpose, phytol was gathered with the unsaponifiable matter from the oil, trimethylsilylated, and analyzed by gas chromatography coupled to mass spectrometry. With this method, 27 samples of edible oils (16 refined and 11 nonrefined edible oils) were analyzed for the abundance of cis-phytol relative to trans-phytol. In the nonrefined oils (e.g., olive oil, rapeseed oil, maize oil, and sunflower oil), cis-phytol contributed 0.1% (n = 3) or less (n = 8) to the phytol content. In contrast, the refined olive oils (n = 4) contained a share of 1.3-3% cis-phytol; the refined rapeseed oil (n = 3) contained a share of 0.7-1.0% cis-phytol; and the refined sunflower oil (n = 4) contained a share of 0.3-0.9% cis-phytol. Only one refined pomegranate kernel did not contain cis-phytol. The phytol concentration was not suited to distinguish nonrefined from refined oils. In contrast, our data suggest that the virtual absence of cis-phytol can be used as a marker for nonrefined (e.g., cold-pressed) edible oils.

[Analysis of the chemical constituents of essential oil from Ligustrum quihoui by GC-MS].

OBJECTIVE: [corrected] To study the chemical constituents of essential oil from Ligustrum quihoui. METHODS: Essential oil was extracted by steam distillation (SD). The chemical constituents of essential oil was analyzed by GC-MS. RESULTS: The chemical components in the oil were qualitatively and quantitatively analyzed by GC-MS, 76 components were seperated and 35 components were identified. The main components are n-Hexadecanoic acid (17.28%), (Z, Z, Z)-9, 12, 15-Octadecatrienoic acid, ethyl ester (12.13%), Phytol (5.80%). CONCLUSION: The method is simple, reliable and with good reprodutivity.

Identification and quantification of metallo-chlorophyll complexes in bright green table olives by high-performance liquid chromatrography-mass spectrometry quadrupole/time-of-flight.

Five different samples of table olives, two regular Spanish table olives and three "bright green table olives", have been analyzed by HPLC-MS/MS to determine their pigment profile. Typical pigment profiles of almost all table olives show primarily chlorophyll derivatives lacking metals (e.g., pheophytin a/b and 15(2)-Me-phytol-chlorin e(6)). Bright green table olives have a unique profile including metallo-chlorophyll complexes (Cu-15(2)-Me-phytol-chlorin e(6) with 26-48% and Cu-pheophytin a with 3-18%) as their major pigments. New tentative structures have been identified by MS such as 15(2)-Me-phytol-rhodin g(7), 15(2)-Me-phytol-chlorin e(6), 15(2)-Me-phytol-isochlorin e(4), Cu-15(2)-Me-phytol-rhodin g(7), Cu-15(2)-Me-phytol-chlorin e(6), and Cu-15(2)-Me-phytol-isochlorin e(4), and new MS/MS fragmentation patterns are reported for Cu-15(2)-Me-phytol-rhodin g(7), Cu-15(2)-Me-phytol-chlorin e(6), Cu-pheophytin b, Cu-pheophytin a, Cu-pyropheophytin b, and Cu-pyropheophytin a. The presence of metallo-chlorophyll derivatives is responsible for the intense color of bright green table olives, but these metallo-chlorophyll complexes may be regarded as a "green staining" defect that is unacceptable to consumers.

[Study on the volatile oil of leaf of red gland Lonicera in guangxi].

OBJECTIVE: To compare volatile oil of Lonicera confusa (red gland Lonicera) fresh leaves with dry leaves and provide evidence for further study of Lonicera confusa leaves. METHODS: GC-MS was used to identify chemical composition in volatile oil in red gland Lonicera leaves. RESULTS: 93 chemical components were separated in fresh leaves in which 39 chemical components were identified; 88 chemical components were separated in dry leaves in which 51 chemical components were identified. The main components of fresh leaves of red gland Lonicera were n-Hexadecanoic acid (11.90%), Phytol (11.79) and 9,12,15-Octadecatrienoic acid, methyl ester, (Z, Z, Z)(7.08); The main components of dry leaves of red gland Lonicera were 1,6-Octadien-3-ol, 3,7-dimethyl-(27.62%), Phytol (7.57%) and 2, 6, 10, 14, 18, 22-Tetracosahexaene, 2, 6, 10, 15, 19, 23-hexamethyl-, (all-E) (4.70%). CONCLUSION: There are significant differences between volatile oil between fresh and dry leaves.

[Analysis of volatile oil from Ardisia brevicaulis].

OBJECTIVE: To analyze and compare the compounds in the essential oil from the leaves and roots of Ardisia brevicaulis. METHODS: The essential oil were obtained by steam distillation. The chemical components were separated and identified by gas chromatography-mass spectrometry (GC-MS). RESULTS: 38 compounds were identified from the leaves (65.952% of the total essential oil) and 46 compounds were identified from the roots (54.890% of the total essential oil). The main constituents of the leaves essential oil were Palmitic acid (43.329%), Fitone (2.430%), Phytol (3.142%), and so on. The main constituents of the roots essential oil were Calamenene (2.913%), cis-alpha-Bisabolene (5.222%), gamma-Muurolene (14.227%), Caryophyllene (11.592%), and so on. CONCLUSION: The constituents of volatile oil extracted from the leaves and roots of Ardisia brevicaulis were different, so the leaves and roots of Ardisia brevicaulis should be utilized differently in clinical application.