Chemical composition,antimicrobial, cytotoxic and antioxidant activities of the essential oil of Artemisia indica Willd

Essential oil from the aerial parts of Artemisia indica was analysed by GC-FID and GC–MS. A total of 43 compounds representing 96.8% of the oil were identified and the major components were found to be artemisia ketone (42.1%), germacrene B (8.6%), borneol (6.1%) and cis-chrysanthenyl acetate (4.8%). Antimicrobial activity of the oil was evaluated against seven clinically significant bacterial and two fungal strains. The essential oil and its major constituents exhibited moderate to potent, broad-spectrum antibacterial and antifungal activities targeting both Gram-positive and Gram-negative bacteria. In vitro cytotoxicity evaluation against four human cancer cell lines THP-1 (leukemia), A-549 (lung), HEP-2 (liver) and Caco-2 (colon) showed that the essential oil exhibited concentration dependant growth inhibition in the 10–100 μg/ml dilution range, with IC₅₀ values of 10 μg/ml (THP-1), 25 μg/ml (A-549), 15.5 μg/ml (HEP-2) and 19.5 μg/ml (Caco-2). It was interesting to note that the essential oil also exhibited potent antioxidant activity.     

Composition and stability of phytochemicals in five varieties of black soybeans (Glycine max)

Phytochemical compositions of five varieties of black soybeans (Glycine max) and their stabilities at room temperature, 4 and −80 °C over 14 months were determined by HPLC systems with electrochemical (ECD) and UV detectors. Polyphenol profiling was carried out by a liquid chromatography–electrospray ionisation-mass spectrometry (LC–ESI-MS) with orbitrap as a mass analyser in both positive and negative ion modes, and polyphenols in aglycone forms were quantified by HPLC–ECD. Five different varieties of black soybeans (G. max) contained 249–405 μg/g dry wt of γ-tocopherol and 6.76–14.98 μg/g dry wt of lutein. Major polyphenols in black soybeans (G. max) were daidzein (193–288 μg/g dry wt) and genistein (145–223 μg/g dry wt), mainly present as glucosides and acetyl glucosides. No significant decrease was found in total phenols of black soybeans (G. max) stored at room temperature, 4 or −80 °C for 14 months. On the other hand, lutein and γ-tocopherol degraded significantly within a month of storage at room temperature (p < 0.01), whereas they remained stable up to 6 months at 4 °C and up to 14 months at −80 °C. The current study indicates that black soybeans (G. max) are rich source of γ-tocopherol and phenols (isoflavones, flavonols, flavan-3-ols, proanthocyanidins and anthocyanin) and that the levels vary depending upon varieties. In addition, storage at low temperature is recommended to reduce the loss of fat-soluble phytochemicals in black soybeans (G. max) over an extended period of time.     

Quantification of coumarin derivatives in Noni (Morinda citrifolia) and their contribution of quenching effect on reactive oxygen species

The quantification of coumarin derivatives such as scopoletin, 7-hydroxycoumarin (7-HC) and 4-hydroxycoumarin (4-HC) in Noni (Morinda citrifolia) was described. The coumarin derivatives were determined by HPLC-UV or -fluorescence detection. More than 95% of peak purity for coumarin derivatives in Noni sample was confirmed by a multi-wavelength fluorescence detector. Amounts of scopoletin and 7-HC in Noni juices (A–H) were ranging 5.1–231 μg/ml and 0.04–0.45 μg/ml, respectively (n = 12). No 4-HC was detected in any Noni samples examined.     

Simultaneous quantification of selected compounds from Salvia herbs by HPLC method and their application

A simple and reliable reversed-phase high performance liquid chromatographic method was developed and validated for simultaneous determination of danshensu, protocatechuic aldehyde, rosemarinic acid, salvianolic acid B, dihydrotanshinone I, tanshinone I, cryptotanshinone and tanshinone IIA in Salvia herbs. The contents of these eight main components were compared between Salvia miltiorrhiza Bunge and Salvia przewalskii Maxim, which were used for treating coronary heart diseases, cerebrovascular disease, bone loss, hepatitis, hepatocirrhosis and chronic renal failure. The samples were successfully separated by the HPLC method. All the standard compounds showed a good linearity (R² > 0.9994) in the relatively wide concentration range. The limit of detection of the eight compounds was in the range of 0.05–0.5 μg/ml and the limit of quantification was in the range of 0.25–1.50 μg/ml. The intra-day variability was in the range of 0.03–8.08% and the inter-day variability was in the range of 0.18–12.36%. The recoveries of the selected compounds were in the range of 96.2–108.1%. This method was accurate, precise and reproducible, and could be successfully applied to the quality control and stable experiment for the preparations consisted of these active components, and the content comparison of the herbs from Salvia species.     

Carotenoid composition of Algerian date varieties (Phoenix dactylifera) at different edible maturation stages

The aim of this study was to investigate the carotenoid composition and the provitamin A value of three palm date (Phoenix dactylifera) varieties (Deglet-Nour, Hamraya and Tantebouchte) from Algeria at three different ripening stages (khallal, rutab and tamr). Chromatographic analysis showed that the major carotenoid pigment present in dates is lutein followed by β-carotene, with an evident carotenoid disappearance during ripening from the khallal to the tamr stage. The different date fruits present a total carotenoid content in the range of 61.7–167, 32.6–672, and 37.3–773 μg/100 g fresh weight (FW) in Deglet-nour, Tantebouchte and Hamraya varieties, respectively. The rutab stage of Tantebouchte showed the lowest carotenoid content of 32.6 μg/100 g FW, whereas the khallal stage of Hamraya presented the highest value, 773 μg/100 g FW, followed by Tantebouchte with 672 μg/100 g FW. Provitamin A value (due exclusively to β-carotene) increased from 0.4 to 0.5 RE/100 g in Deglet-Nour fruits, but decreased from 11.7 to 1.6 RE/100 g and from 3.9 to 0.5 RE/100 g in Tantebouchte and Hamraya fruits, respectively, during ripening. The lowest value was found at the tamr stage of the Deglet-Nour variety (0.5 RE/100 g) whereas the highest provitamin A content was found at the khallal stage of the Tantebouchte variety (11.7 RE/100 g).     

Composition,antimicrobial, antiradical and spasmolytic activity of Ferula heuffelii Griseb. ex Heuffel (Apiaceae) essential oil

The essential oil from underground parts of Ferula heuffelii from N.E. Serbia, was analysed using GC and GC–MS. The main compounds of the essential oil were elemicin (35.4%) and myristicin (20.6%). The essential oil exhibited the best antimicrobial activity against two strains of Candida albicans (MIC = 7.0 and 13.7 μg/ml), as well as against Micrococcus luteus (MIC = 13.7 μg/ml), Staphylococcus epidermidis (MIC = 17.6 μg/ml), Bacillus subtilis (MIC = 21.1 μg/ml) and Micrococcus flavus (MIC = 28.2 μg/ml). In the DPPH radical scavenging assay, essential oil showed substantial activity with SC₅₀ = 22.43 μl/ml. The essential oil was also tested for antispasmodic activity. It inhibited spontaneous contraction of isolated rat ileum dose-dependently, and at the concentration of 86.64 μg/ml exhibited 50% of the maximum effect of atropine. After incubation with 75.00 μg/ml of essential oil, acetylcholine did not induce contractions of ileum, and at 250.00 μg/ml, the essential oil almost completely abolished the spasmodic effect of potassium chloride (80 mM).     

The determination of vitamin D3 in bovine milk by liquid chromatography mass spectrometry

A renewed international interest in vitamin D status has revealed significant deficiencies in several populations, including Australia. Vitamin D exists in two forms, cholcalciferol (D₃) and ergocalciferol (D₂). The main source of vitamin D₃ is from exposure of 7-dehydrocholesterol present in the skin to UV irradiation. However, there is an absolute requirement for vitamin D through proper dietary intake if humans live in the absence of sunlight or exclusively indoors. Bovine milk is considered to be a good dietary source of vitamin D₃, even though the levels are quite low. This paper describes robust methods using liquid chromatography–linear ion trap mass spectrometry (LC–MSⁿ) and liquid chromatography–tandem mass spectrometry (LC–MS/MS) to measure the levels of vitamin D₃ in fresh bovine milk (0.05 μg/100 ml), commercial (natural and fortified) milk samples (0.01–2 μg/100 ml) and a dairy based infant formula (8 μg/100 g), without the need for extensive clean-up procedures. The limits of quantification (LOQ) are 0.01 μg/100 ml and 0.02 μg/100 ml for LC–MSⁿ and LC–MS/MS, respectively. Recoveries of vitamin D₃ added to the samples prior to saponification were satisfactory (range 60–90%). 25-Hydroxyvitamin D₃ was not present in any of the samples analysed (LOQ = 0.01 μg/100 ml, recovery range 30–40%).     

Organic acids,antioxidant capacity,phenolic content and lipid characterisation of Georgia-grown underutilized fruit crops

Four underutilized Georgia-grown fruit crops, namely loquat (Eriobotrya japonica), mayhaw (Crataegus sp.), fig (Ficus carica), and pawpaw (Asimina triloba), and their leaves were analysed for total polyphenols by Folin–Ciocalteau method, and antioxidant capacity by ferric-reducing antioxidant power (FRAP) and Trolox-equivalent antioxidant capacity (TEAC) assays. Organic acids and phenolic compounds were identified by RP-HPLC. For lipid profile, fruits were separated into two fractions – seed and fruit (i.e., without seed); lipid was extracted using the Folch method and analysed for fatty acids, phytosterols, tocopherols, and phospholipids. The major organic acid identified in all samples was malic acid (177–1918 mg/100 g FW). The predominant phenolic acids in all the fruits were gallic (1.5–6.4 mg/100 g FW) and ellagic (0.2–33.8 mg/100 g FW), and the most abundant flavonoid was catechin (12.2–37.8 mg/100 g FW). Total lipid content varied from 0.1% in mayhaw fruit to 21.5% in pawpaw seed. Linoleic acid was the predominant fatty acid in all of the samples (28.2–55.7%).     

Effect of canned tuna fish processing steps on lead and cadmium contents of Iranian tuna fish

The contents of Pb and Cd in two species of Iranian tuna fish (yellowfin and skipjack), which were caught from the Persian Gulf and Oman Sea, and the effects of canning processing steps on their contents were assessed by electrothermal atomic absorption spectrometry. The results revealed that the levels of lead and cadmium throughout the processing steps in yellowfin were in range of 0.154 ± 0.019–0.441 ± 0.025 μg/g and 0.029 ± 0.002–0.084 ± 0.0005 μg/g, respectively. Pb and Cd concentrations from received fish to final product in skipjack were found to be in range of 0.072 ± 0.031–0.218 ± 0.031 μg/g and 0.016 ± 0.001–0.062 ± 0.002 μg/g, respectively. The limit of detection for lead and cadmium were 0.058 μg/g (11.6022 μg/l) and 0.0007 μg/g (0.1485 μg/l), respectively. Results from paired sample t-test analysis showed that defrosting, cooking, and sterilisation by autoclave would reduce the contents of lead and cadmium, considerably.     

Antioxidant properties and chemical composition of technical Cashew Nut Shell Liquid (tCNSL)

The present study analysed the antioxidant activity of the technical Cashew Nut Shell Liquid (tCNSL) using the 2,2-diphenyl-1-picrylhydrazyl (DPPH) scavenging assay and the xanthine oxidase assay, as well as in vivo evaluation by Saccharomycescerevisiae assay. Also, the chemical composition of tCNSL was determined by gas chromatography–mass spectrometry (GC–MS), revealing the presence of cardanols (40.26%), cardols (29.95%), phytosterol (10.68%), triacontanes (4.66%) and anacardic acid (1.79%). The DPPH-based assay results showed that tCNSL 1000 μg/ml reduced the radical level by 88.9%, and the scavenging of hydroxyl radicals in the xanthine oxidase assay indicated significant antioxidant activity (IC₅₀ = 702 μg/ml). In addition, tCNSL exerts an important protective effect against oxidative stress in yeast when used in 100–500 μg/ml concentration range, when exposed to paraquat or H₂O₂, indicating an in vivo antioxidant effect.     

Chemical and biological variability of hot pepper fruits (Capsicum annuum var. acuminatum L.) in relation to maturity stage

The aim of the present work was to evaluate the chemical composition and the radical-scavenging and antioxidant activities of hot pepper fruits (Capsicum annuum L. var. acuminatum) at three maturity stages (small green, green and red). GC–MS analysis of n-hexane and chloroform fractions showed a different composition between the three stages of ripening. The first stage of maturation (small green) showed the highest radical-scavenging activity (IC₅₀ of 129 μg/ml). Using the bovine brain peroxidation assay, the methanolic extract of green pepper showed significant antioxidant activity (IC₅₀ of 522 μg/ml). Addition of methanolic extract of red and green pepper inhibited oxidation of linoleic acid. Methanolic extract of red pepper showed greater antioxidative potency than the others (IC₅₀ of 3 μg/ml). The different composition of lipophilic compounds and the various amount of phenolics, showed in the three stage of ripening of C. annuum var. acuminatum fruits, modifies the antioxidant activity.     

GC–MS quantification and sensory thresholds of guaiacol in orange juice and its correlation with Alicyclobacillus spp.

Guaiacol is a profoundly negative taint/off-flavour produced by the increasingly common microbial contamination of fruit juices with Alicyclobacillus spp. The objectives of this study included: determining sensory thresholds for guaiacol in orange juice (OJ), developing an analytical method whose detection limits were equivalent to sensory thresholds and determining levels of Alicyclobacillus spp. that would produce detectable levels of guaiacol. A 12 member trained panel was used to establish guaiacol detection and recognition thresholds. Guaiacol ortho and retro nasal detection thresholds in OJ were 0.70 and 0.53 μg/l respectively. Odour recognition threshold was 2.0 μg/l. A SPME GC–MS Selected Ion Monitoring (SIM) procedure was optimised to achieve analytical detection limits of 0.5 μg/l. Optimum guaiacol detection limit was achieved using only responses from m/z 109 and 124. Ion ratios (m/z 109/124) and linear retention index value matching were used to confirm the identification of guaiacol. Quantification was achieved using peak areas from standard guaiacol additions in orange juice between 0.5 and 100 μg/l. Alicyclobacillus growth of 2.2 log CFU/ml in OJ produced just detectable levels (0.7 μg/l) of guaiacol.     

Anti-oxidant,anti-proliferative and anti-inflammatory activities of the extracts from black raspberry fruits and wine

The purpose of the study was to determine polyphenols, anthocyanins and ascorbic acid in the extracts of black raspberry fruits and wine, along with their anti-oxidant, anti-proliferative and anti-inflammatory activities. Black raspberry fruits without or with seeds crushed were blended in 60% ethanol (FE and FES, respectively) or in water (FW and FWS, respectively). Black raspberry wine without or with seeds crushed (W and WS, respectively) were prepared. Polyphenol content was the highest in the FES (8.25 mg/g fruit). Generally the ethanol extracts with seeds crushed showed higher anti-oxidant activities with the lowest DPPH IC₅₀ (130 μg/ml (freeze-dried extract/reaction solution)) for the FES and the lowest ABTS IC₅₀ (198 μg/ml) for the WS. Cell viabilities were reduced by 13–70% when treated with 100 μg/ml (freeze-dried extract/medium) for HT-29 cells and 1000 μg/ml for LNCaP cells. The FES most actively suppressed nitric oxide production in LPS-stimulated RAW264.7 cells (p < 0.05). Superoxide dismutase and glutathione peroxidase activities treated with the extracts were higher than the control (p < 0.05).     

Carotenoid composition from the Brazilian tropical fruit camu–camu (Myrciaria dubia)

Camu–camu (Myrciaria dubia) is a small berry, native to the Amazon, known as a rich source of ascorbic acid. The carotenoid composition of this fruit was determined using high performance liquid chromatography-diode array detection on C₁₈ and C₃₀ columns. Fruits produced in two different regions of São Paulo State, Iguape and Mirandópolis, were analysed. All-trans-lutein was the major carotenoid in camu–camu fruits from both regions, ranging from 45% to 55% of the total carotenoid content (160.5 ± 93.1 μg/100 g for Iguape and 601.9 ± 75.6 μg/100 g for Mirandópolis fruits), followed by β-carotene, violaxanthin and luteoxanthin. The levels of lutein, β-carotene, violaxanthin, luteoxanthin and other minor carotenoids were significantly higher in the camu–camu produced in Mirandópolis region, most probably due to the higher temperature and light exposure found in this region, in comparison to those from Iguape. Maturation was also an important feature affecting batches from the same region.     

Antimicrobial properties and analytical profile of traditional Eruca sativa seed oil: Comparison with various aerial and root plant extracts

The present study investigates the antimicrobial activity of various solvent extracts of Eruca sativa (aerial and root) and seed oil against-antibiotic resistant Gram-negative (Escherichia coli, Pseudomoms aeruginosa and Shigella flexneri) and Gram-positive (Staphylococcus aureus and Bacillus subtilis) bacteria. Among the various preparations, seed oil was the most active, exhibiting a maximum zone inhibition of 97% for Gram-positive bacteria and of 74–97% for Gram-negative bacteria. The MIC of the seed oil was found to be 65–75 and 60–70 μg/ml for Gram-negative and Gram-positive bacteria, respectively. Analytical investigation on main volatile and non-volatile components was performed on seed oil. Among the formers allyl isothiocyanate (40 μg/g), 3-butenyl isothiocyanate (260 μg/g), 4-methylsulfinybutyl isothiocyanate (sulforaphane 743 μg/g), 2-phenylethyl isothiocyanate (159 μg/g) and bis(isothiocyanatobutyl)disulphide (∼5000 μg/g) were determined by head space/SPME/GC–MS analysis. Free fatty acids were 1.6% w/w of the oil and overall 25 fatty acids were identified. Erucic and oleic acids were the main fatty acids both in the free (7.8 and 2.1 mg/ml) and esterified forms (50.6% w/w and 14.9% w/w of total fatty acids). Unsaponifiable fraction was 1.8% w/w.     

Quantitative analysis of phenolic compounds in Chinese hawthorn (Crataegus spp.) fruits by high performance liquid chromatography–electrospray ionisation mass spectrometry

Eleven major phenolic compounds (hyperoside, isoquercitrin, chlorogenic acid, ideain, epicatechin, two procyanidin (PA) dimers, three PA trimers and a PA dimer-hexoside) were quantified in the fruits of 22 cultivars/origins of three species of the Chinese hawthorn (Crataegus spp.) by HPLC–ESI-MS-SIR. Hyperoside (0.1–0.8 mg/g dry mass [DM]), isoquercitrin (0.1–0.3 mg/g DM), chlorogenic acid (0.2–1.6 mg/g DM), epicatechin (0.9–11.7 mg/g DM), PA B2 (0.7–12.4 mg/g DM), PA dimer II (0.1–1.5 mg/g DM), PA trimer I (0.1–2.7 mg/g DM), PA trimer II (0.7–6.9 mg/g DM), PA trimer III (0.01–1.2 mg/g DM) and a PA dimer-hexoside (trace–1.1 mg/g DM) were detected in all the samples. Ideain (0.0–0.7 mg/g DM) was found in all the samples except Crataegus scabrifolia. Significant correlations between the contents of individual PA aglycons were observed (r > 0.9, P < 0.01). A strong correlation between flavonols was also shown (r = 0.71, P < 0.01). Fruits of Crataegus pinnatifida var. major had higher contents of PAs but lower contents of flavonols compared with Crataegus brettschneideri. The fruits of C. scabrifolia contained the highest level of PA dimer-hexoside, which was present in trace amounts in the fruits of C. pinnatifida.     

Antibacterial labdane diterpenoids of Ulva fasciata Delile from southwestern coast of the Indian Peninsula

Chromatographic purification of the dichloromethane-soluble fraction of alga, on neutral alumina, using increasing concentrations of ethylacetate/n-hexane as eluents, yielded seven labdane diterpenoids (1–7) as major constituents of green alga Ulva fasciata. Structures of these diterpenoids were established using extensive spectroscopic techniques. Antimicrobial assay showed that the compounds labda-14-ene-3α,8α-diol (2) and labda-14-ene-8α-hydroxy-3-one (4) were inhibitory to the growth of Vibrio parahaemolyticus and Vibrio alginolyticus with minimum inhibitory concentrations of 30 μg/ml by 2, and 40 μg/ml by 4, respectively against the former and 30 μg/ml by 2, and 80 μg/ml by 4, respectively, against the latter. Structure–activity relationship analyses revealed that the compounds with electronegative hydroxyl or carbonyl group(s) exhibit greater activities, apparently by proton exchange reaction with the basic aminoacyl residue at the macromolecular receptor site of virulent enzymes of pathogenic bacteria. These might provide promising therapeutic agents against infections with multi-resistant Gram-negative fish pathogenic bacteria.     

Liquid chromatography–tandem mass spectrometry for the determination of chrysoidine in yellow-fin tuna

A high performance liquid chromatography (HPLC) coupled to electrospray ionisation tandem mass spectrometry (MS/MS) method was described for the residue detection of chrysoidine in yellow-fin tuna in the present study. Samples were cleaned up with solid phase extraction (SPE) cartridge, and then injected into HPLC for separation. Multiple-reaction monitoring (MRM) was applied for quantitative determination. Results showed that the low limit of detection (LOD) of the method was 1.25 × 10⁻¹² g, and the low limit of quantification (LOQ) was 0.42 μg/L. The standard calibration curve was y = 2333.9x −845 (r² > 0.99) with the linear range of 0.63–100 μg/L. The average recoveries of chrysoidine ranged from 86.0% to 108.0% when the spiked concentration was from 0.5 μg/kg to 20 μg/kg. And the developed method also showed the good test precisions (RSD%: 4.38–14.27%).     

Antioxidant and free radical scavenging activities of polyphenol-enriched curry leaf (Murraya koenigii L.) extracts

The in vitro antioxidant properties of different extracts (water, alcohol, alcohol:water, hexane or chloroform extract) of curry leaves (Murraya koenigii L.) were evaluated using various assays. The alcohol:water (1:1) extract of curry leaves (AWEC) showed the highest antioxidant and free radical scavenging activity. It inhibited membrane lipid peroxidation by 76%, at 50 μg/ml, scavenged 93% of superoxides at 200 μg/3 ml and scavenged approximately 90% of hydroxyl and 1,1-diphenyl-2-picrylhydrazayl radicals at 4–5-fold lower concentrations compared to the other tested extracts. In addition, the alcohol:water extract reduced cytochrome c and ferric ion levels, chelated ferrous ions and inhibited ferrous sulfate:ascorbate-induced fragmentation and sugar oxidation of DNA. These results establish the antioxidant potential of AWEC, which could be used as natural antioxidant source.     

Cytoprotective effect of a bilberry extract against oxidative damage of rat hepatocytes

The effect of a bilberry extract (BE, 25% anthocyanins) against oxidative damage in primary cultures of rat hepatocytes, induced by tert-butyl hydroperoxide and allyl alcohol, was investigated. BE displayed cytoprotective effects at 100 and 500 μg/ml in the MTT viability test. It protected the cells against lactate dehydrogenase leakage and lipoperoxidation products formation. Maximum protection (58%) was noted using 500 μg/ml of BE and intoxication by allyl alcohol. The observed cytoprotective effect is probably due to the antioxidant properties of its constituents, mainly anthocyanins. BE scavenged DPPH (IC₅₀ 3.99 ± 0.14 μg/ml) and enzymatically generated superoxide radical with an activity equivalent to 108 ± 7.2 U of superoxide dismutase per mg of extract. Our results support the use of bilberry and bilberry extracts in functional foods and food supplements designed for the prevention of chronic diseases associated with oxidative stress.