HS-SPME analysis of the volatiles profile of water celery (Apium nodiflorum), a wild vegetable with increasing culinary interest.

Water celery (Apium nodiflorum) is a wild plant traditionally harvested in some Mediterranean areas for being consumed raw. Despite its appreciated organoleptic properties, the aromatic profile of the fresh vegetable remains to be studied. In the present study, volatile compounds from five wild populations were extracted by the headspace-solid phase microextraction technique, analysed by gas cromatography-mass spectrometry, and compared to related crops. The wild species had a high number of aromatic compounds. It was rich in monoterpenes (49.2%), sesquiterpenes (39.4%) and phenylpropanoids (9.6%), with quantitative differences among populations, in absolute terms and relative abundance. On average, germacrene D was the main compound (16.6%), followed by allo-ocimene (11.9%) and limonene (11.1%). Only in one population, the levels of limonene were greater than those of germacrene D. Among phenylpropanoids, dillapiol displayed the highest levels, and co-occurred with myristicin in all populations except one. These differences may have a genetic component, which would indicate the possibility of establishing selection programmes for the development of water celery as a crop adapted to different market preferences. On the other hand, comparison with related crops revealed some similarities among individual volatiles present in the different crops, which would be responsible of the common aroma notes. However, water celery displayed a unique profile, which was in addition quantitatively richer than others. Thus, this differentiation may promote the use of water celery as a new crop.

[Determination of myclobutanil and difenoconazole residues in pollen and honey of litchi by high performance liquid chromatography-tandem mass spectrometry].

An effective method was developed for the determination of two major fungicides including myclobutanil and difenoconazole residues in pollen and honey of litchi by modified QuEChERS-high performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS). The pollen and honey samples were all extracted by acetonitrile, the pollen samples were cleaned-up by 0.9 g anhydrous magnesium sulfate (MgSO4), 0.15 g primary secondary amine (PSA) and 0.15 g C18; the honey samples were cleaned-up by 0.9 g MgSO4 and 0.15 g PSA. The 0.1% (v/v) formic acid aqueous solution-acetonitrile (25:75, v/v) were used as the mobile phases. The extracts were separated on a Poroshell-120 EC-C18 chromatographic column, the positive electrospray ion (ESI+) source and selected ion monitoring (SIM) mode were used. The analytes were quantified by the matrix matching standard solutions. The matrix matched standard solutions of myclobutanil and difenoconazole showed good linearities in the range of 1-100 µg/L, and the correlation coefficients (r2) were all above 0.9990. The limits of detection (LODs) of myclobutanil and difenoconazole were 0.25 µg/kg and 0.50 µg/kg, respectively. The limits of quantification (LOQs) of myclobutanil and difenoconazole were 0.83 µg/kg and 1.7 µg/kg, respectively. The average recoveries of myclobutanil and difenoconazole in pollen and honey samples were 87.0%-95.2% and 90.1%-96.4% with the relative standard deviations of 1.2%-3.6% and 0.7%-4.1%, respectively. The method is quick, easy and sensitive, and it is suitable for the rapid determination and trace analysis of myclobutanil and difenoconazole in pollens and honeys of litchi. The method can provide data support for the exposure risk assessment of bees and other pollination insects.

Essential oil constituents and antimicrobial activity of Pycnocycla bashagardiana Mozaff. from Iran.

Pycnocycla bashagardiana is a rare endemic and endangered species that has been used in folkloric medicine in Southern Iran. This study aimed to evaluate the essential oil constituents and antimicrobial activity of wild and cultivated p. bashagardiana. The aerial parts of wild and cultivated plants were collected from two provinces of Iran. The essential oil was isolated by hydrodistillation and analyzed by a combination of capillary GC and GC-MS. The main components in wild plants were myristicin (39.12%), (E)-ß-ocimene (21.97%), sabinene (15.0%) and cis-iso-miristicin (2.67%) and in cultivated plants, (E)-ß-ocimene (55.40%), myristicin (18.27%), (Z)-ß-ocimene (12.47%) and cis-iso-miristicin (2.94%) were the main constituents in essential oil. The in vitro antimicrobial activity of the essential oil of P. bashagardiana were studied against Staphylococcus aureus, Staphylococcus epidermidis, Escherichia coli and Candida albicans for the first time. The results showed that the oil exhibited strong antimicrobial activity against all the tested pathogens.

Ultra high performance liquid chromatography-electrospray ionization-tandem mass spectrometry and pharmacokinetic analysis of justicidin B and 6'-hydroxy justicidin C in rats.

Arylnaphthalene lignans have attracted considerable interest with the discovery of their antineoplastic activities. Two such compounds are justicidin B and 6'-hydroxy justicidin C, both of which have been isolated from the herb Justicia procumbens. We sought to develop and validate a sensitive and accurate, ultra high performance liquid chromatography with electrospray ionization tandem mass spectrometry method for the structural determination and pharmacokinetics of justicidin B and 6'-hydroxy justicidin C. Chromatographic separation was achieved on an Agilent 300SB-C18 column using water (0.5% formic acid, 10 mM NH4 COOH) methanol as the mobile phase. The plasma samples obtained after oral administration of the active extract of Justicia procumbens were successfully analyzed with our novel method, thereby demonstrating its sound applicability and reliability. The lower limit of quantification for justicidin B and 6'-hydroxy justicidin C was 0.50 and 1.00 ng/mL in 50 µL rat plasma, respectively. The elimination half-life and clearance of justicidin B was estimated to be 1.27 ± 0.61 h and 5.40 ± 0.22 L/h/kg while that of 6'-hydroxy justicidin C was 2.07 ± 0.70 h and 11.84 ± 1.06 L/h/kg. This newly developed and validated method was successfully applied to the quantification and pharmacokinetic study of justicidin B and 6'-hydroxy justicidin C in rats.

Occurrence of piperidine alkaloids in Piper species collected in different areas.

A simple and convenient method was established for simultaneous quantitative determination of piperine and piperlonguminine in dried fruits of Piper longum and allied plants. The average content of piperine in P. longum (18.26 mg/g, range 12.05-33.23 mg/g) was about one half that of P. nigrum (40.09 mg/g, range 29.57-54.23 mg/g), but the content of piperlonguminine in P. longum was in the range of 0.42-1.82 mg/g, and the average content of piperlonguminne (0.91 mg/g) was about seven times higher than that in P. nigrum (0.13 mg/g). A sample of P. longum from Vietnam and a sample of P. retrofractum collected in Ishigaki, Japan, showed high contents of piperine and piperlonguminine. On the other hand, a sample of P. betle collected in Taiwan showed low content of piperine, and piperlonguminine was not detected.

Effect of growth stage on essential-oil yield and composition of Daucus sahariensis.

The essential oils obtained by hydrodistillation from Daucus sahariensis Murb. harvested at three different growth stages were characterized by GC/MS analysis. In total, 88 compounds were identified, with myristicin (29.8-51.7%), myrcene (6.7-31.1%), α-pinene (11.6-14.8%), and limonene (5.3-11.5%) as main constituents. Monoterpene hydrocarbons were the most represented compounds in the oils of the plant samples collected during the flower-budding and full-flowering periods. On the contrary, during the fruiting stage, the oils were dominated by phenylpropanoids. The essential oils were subject of considerable variation in their composition during the various developmental stages, particularly concerning the content of myrcene that decreased significantly passing from the vegetative to the fruiting stage. Conversely, for myristicin, the opposite trend was observed. Furthermore, the essential-oil yields were quite low during the flower-budding phase (0.27%), but rapidly increased during plant development (0.63 and 0.68% for the flowering and fruiting phases, resp.).

Comparison of greenhouse and field degradation behaviour of isoprocarb, hexaflumuron and difenoconazole in Perilla frutescens.

Isoprocarb, hexaflumuron and difenoconazole were used in Perilla frutescens at 600, 60 and 75 g a.i./ha respectively. High performance liquid chromatography-tandem mass spectrometry was used for residue determination because of high selectivity and simple treatment. The results showed that the half-lives of isoprocarb, hexaflumuron and difenoconazole at greenhouse condition were 0.71, 1.63 and 1.21 days respectively, and at field condition, the values were 1.13, 1.07 and 0.92 days respectively.

Extraction and separation of volatile and fixed oils from seeds of Myristica fragrans by supercritical CO2: chemical composition and cytotoxic activity on Caco-2 cancer cells.

UNLABELLED: Isolation of volatile and fixed oils from nutmeg have been obtained by supercritical fractioned extraction with carbon dioxide. Extraction experiments were carried out at pressures of 90 and 250 bar and temperature of 40 °C. The extraction step performed at 90 bar produced a volatile fraction mainly formed by myristicin (32.8%), sabinene (16.1%), α-pinene (9.8%), ß-pinene (9.4%), ß-phellandrene (4.9%), safrole (4.1%) and terpinen-4-ol (3.6%). The oil yield relative to this step of the process was 1.4% by weight of the charge. The last extraction step at 250 bar produced a butter-like material (nutmeg butter). The yield of this step was 14.4% by weight. The most represented fatty acids of fixed oil from nutmeg were 14:0 (79.2%), 18:1 n-9 (7.4%) and 16:0 (6.1%), and in particular the unsaturated fatty acids 18:1 n-9 averaged 32.96 µg/mg of oil. The level of myristicin in the nutmeg essential and fixed oils was also directly quantified by reversed HPLC-DAD. Moreover, the essential oil obtained from nutmeg, as well as myristicin, showed a significant in vitro inhibitory effect on the growth of a colon cancer cell line (undifferentiated Caco-2 cells). PRACTICAL APPLICATION: In this study, the chemical characterization and the anticancer activity of nutmeg oils obtained by supercritical extraction with carbon dioxide were investigated. This is important for their potential application in food and pharmaceutical industries.

Optimized ultrasonic assisted extraction-dispersive liquid-liquid microextraction coupled with gas chromatography for determination of essential oil of Oliveria decumbens Vent.

Ultrasonic assisted extraction-dispersive liquid-liquid microextraction (UAE-DLLME) coupled with gas chromatography (GC) was applied for extraction and determination of essential oil constituents of the plant Oliveria decumbens Vent. Scanning electron microscopy (SEM) was used to see the effect of ultrasonic radiation on the extraction efficiency. By comparison with hydrodistillation, UAE-DLLME is fast, low cost, simple, efficient and consuming small amount of plant materials (∼1.0 g). The effects of various parameters such as temperature, ultrasonication time, volume of disperser and extraction solvents were investigated by a full factorial design to identify significant variables and their interactions. The results demonstrated that temperature and ultrasonication time had no considerable effect on the results. In the next step, a central composite design (CCD) was performed to obtain the optimum levels of significant parameters. The obtained optimal conditions were: 0.45 mL for disperser solvent (acetonitrile) and 94.84 µL for extraction solvent (chlorobenzene). The limits of detection (LODs), linear dynamic range and determination coefficients (R(2)) were 0.2-29 ng mL(-1), 1-2100 ng mL(-1) and 0.995-0.998, respectively. The main components of the essential oil were: thymol (47.06%), carvacrol (23.31%), gamma-terpinene (18.94%), p-cymene (8.71%), limonene (0.76%) and myristicin (0.63%).

[Analysis of the chemical constituents of volatile oil from Asarum insigne by GC-MS].

OBJECTIVE: To analyse the chemical constituents of volatile oil from Asarum insigne. METHODS: The volatile oil from Asarum insigne was isolated with steam distillation and identified by capillary GC/MS method. RESULTS: 68 Volatile components were identified and determined, accounting for 92.18% of the total peak area. The main volatile compounds and their relative contents are camphene (13.48%), alpha-pinene (12.44%), beta-pinene (11.07%), borneol (8.12%), trans-beta-farnesene (5.91%), elemicin (5.38%), 1,3-benzodioxole-5-(2-propenyl) (3.06%), myristicin (2.95%), ledene (2.47%), eucalyptol (2.33%), patchouli alcohol (2.25%), alpha-bisabolene (2.04%) and bornyl acetate (1.36%) etc. CONCLUSION: The study provided solid and scientific proof for the exploitation and utilization of Asarum insigne.

Residues of Pesticides in honeybee (Apis mellifera carnica) bee bread and in pollen loads from treated apple orchards.

Honey bee (Apis mellifera carnica) colonies were placed in two apple orchards treated with the insecticides diazinon and thiacloprid and the fungicide difenoconazole in accordance with a Protection Treatment Plan in the spring of 2007. Pollen and bee bread were collected from combs inside the hives. The residue of diazinon in pollen loads 10 days after orchard treatment was 0.09 mg/kg, and the same amount of residue was found in bee bread 16 days after treatment. In pollen loads 6 days after application 0.03 mg/kg of thiacloprid residues and 0.01 mg/kg of difenoconazole were found on the first day after application. Possible sub-lethal effects on individual honey bees and brood are discussed.

Inhibitory effect of ethanol extract of Piper longum L. on rabbit platelet aggregation through antagonizing thromboxane A2 receptor.

Piper longum L. has been used as a crude drug for the treatment of disorders of poor peripheral blood circulation in Asia. However, the detailed mechanism of its action has not been clarified as yet. In the present study, we examined the effects of several extracts of Piper longum L. on rabbit platelet function. Thromboxane A(2) receptor agonist U46619 caused rabbit platelet aggregation, which was potently inhibited by the ethanol or butanol extract of Piper longum L. The ethanol extract inhibited U46619-induced platelet aggregation in a concentration-dependent manner, but only weakly inhibited that induced by thrombin. The maximum response to U46619 was reduced by 100% ethanol extract concentration dependently, suggesting that the inhibitory mode of U46619-induced platelet aggregation by the ethanol extract was non-competitive. The extract also inhibited U46619-induced phosphoinositide hydrolysis with a similar concentration dependency to the platelet aggregation. Furthermore, the extract inhibited binding of [(3)H]SQ29548 to thromboxane A(2) receptor in intact platelets in a concentration-dependent manner. These results suggest that Piper longum L. contains a constituent(s) that inhibits platelet aggregation as a non-competitive thromboxane A(2) receptor antagonist.

Comparative analysis of the oil and supercritical CO2 extract of Ridolfia segetum (L.) Moris.

Supercritical carbon dioxide extraction allowed to obtain the volatile oil of different aerial parts of Ridolfia segetum (L.) Moris. Extraction conditions were as follows: pressure, 90 bar; temperature, 50 degrees C and carbon dioxide flow, Phi = 1.0 kg h(-1). Waxes were entrapped in the first separator set at 90 bar and -10 degrees C. The oil was recovered in the second separator working at 15 bar and 10 degrees C. The main components of the flower oil were alpha-phellandrene (19.4%), terpinolene (20.5%), piperitenone oxide (11.6%), beta-phellandrene (8.2%), (Z)-beta-ocimene (7.8%), myristicin (7.5%) and p-cymene (4.4%). The comparison with the hydrodistilled (HD) oil reveal that the significative difference was the content of sesquiterpenes which are higher in the supercritical fluid extraction (SFE) products. Collection of samples at different extraction times during supercritical extraction, allowed to monitor the change of the oil composition. Lighter compounds, as hydrocarbon monoterpenes, were extracted in shorter times than the heavier hydrocarbon and oxygenated sesquiterpenes. The oil from the steams was characterized by a high content of alpha-phellandrene (12.9%), terpinolene (11.6%), myristicin (11.0%), p-cymene (9.9%), beta-phellandrene (8.2%) and (Z)-beta-ocimene (6.0%) while the main components of the fruits were found to be myristicin (70.8%), piperitenone oxide (19.9%) and dill apiole (4.2%).

[Comparing analysis of components in volatile oils of nutmeg and prepared nutmeg by GC-MS].

OBJECTIVE: To compare components in volatile oils of nutmeg and prepared nutmeg. METHOD: Volatile oil from nutmeg and prepared nutmeg were extracted by vapor distillation. The chemical components in two kinds of volatile oils were determined and indentified by GC-MS. RESULT: The change in quantity and quality of components in volatile oils were observed after processing. 13 new components occurred and 4 components disappeared in volatile oils after processing. The contents of methyleugenol and methylisoeugenol that are active ingredients were increased. The contents of myristicin and safrol that are toxic ingredients in volatile oils were decreased. CONCLUSION: The processing method of nutmeg by soaking with water and roasting with bran is scientific.

A new lignan from the Jian-er syrup and its content determination by RP-HPLC.

The Jian-er syrup is a commonly used traditional Chinese medicinal preparation refined from two herbs, Metaplexis japonica (Thunb.) Makino and Justicia procumbens L. A new lignan, named 6'-hydroxy-Justicidin B, was isolated from the ethyl acetate extract of the Jian-er syrup and its structure was established on the basis of spectral analysis. The content of 6'-hydroxy-Justicidin B in the Jian-er syrup was determined by RP-HPLC for the first time. Chromatographic separation was achieved on an Agilent Zorbax SB-C18 column (150 mmx4.6 mm i.d., 5 microm) at 25 degrees C and the mobile phase was a mixture of acetonitrile and water (31:69 v/v) detected at 256 nm at a flow rate of 1.0 ml/min. This method was validated in terms of selectivity, linearity, precision, accuracy, limit of detection, limit of quantitation and solution stability. It can be used to control the quality of this preparation effectively.

[Study on chemical constituents of the essential oil from Myristica fragrans Houtt. by supercritical fluid extraction and steam distillation].

Essential oils were extracted from Myristica fragrans Houtt. by supercritical fluid extraction (SFE) and steam distillation (SD). Their components were analyzed by gas chromatography-mass spectrometry and compared. 48 compounds were identified for the essential oil extracted by supercritical carbon dioxide, and its main components have been found to be myristic acid, myristicin, terpinen-4-ol, alpha-pinene and safrole. 38 compounds were identified for the essential oil obtained by SD, and its main components have been found to be beta-pinene, terpinen-4-ol, alpha-pinene, gamma-terpinene and beta-phellandrene.

[GC-MS analysis of essential oils from seeds of Myristica fragrans in Chinese market].

OBJECTIVE: To analyze the constituents of the essential oils extracted from nine samples of commercial seeds of Myristica fragrans Houtt respectively and to provide qualities control foundations. METHOD: Water steam distillation and GC-MS were used. RESULT: 95-118 compounds were separated respectively. 79 Compounds being identified which were 95.18%-98.70% of the total essential oil. CONCLUSION: Myristicin (39.63%) and terpene series were the main compounds.

[Effect of different processing conditions on content of myrisiticin, volatile oil and fatty lipid in semen Myristicae].

The effect of different processing conditions on contents of myrisiticin, volatile oil and fatty lipid in Semen Myristicae was studied by orthogonal design. The result shows that these processing conditions could not influence the contents of myrisiticin, volatile oil and fatty lipid in the processed products of Semen Myristicae, suggesting that processing does not necessarily lower toxicity.

[Determination of myristicin and safrol in volatile oil of nutmeg and its processed products by HPLC].

The contents of myristicin and safrol in the volatile oil of nutmeg and its processed products were determined by HPLC. This method is fast, accurate and gives good resolution. It has also been confirmed that myristicin tends to decrease in varying degrees after the nutmeg has been processed.

[Comparative study on nutmeg, mace and their processed products].

The contents of volatile oil, myristicin, safrol and methyleugenol, as well as the composition and content of fatty oil in nutmeg, mace and their processed products were comparatively studied. The composition of volatile oil and the detection of trimyristin were compared by TLC. A scientific basis for the development and application of mace has thus been provided.