Chemical composition of the volatile extract and antioxidant activities of the volatile and nonvolatile extracts of Egyptian corn silk (Zea mays L.)

A total of 36 compounds, which comprised 99.4% of the extract, were identified by gas chromatography and mass spectrometry (GC-MS) in the volatile dichloromethane extract obtained from Egyptian corn silk. The main constituents of the volatile extract were cis-alpha-terpineol (24.22%), 6,11-oxidoacor-4-ene (18.06%), citronellol (16.18%), trans-pinocamphone (5.86%), eugenol (4.37%), neo-iso-3-thujanol (2.59%), and cis-sabinene hydrate (2.28%). Dried Egyptian corn silk was also directly extracted with petroleum ether, ethanol, and water. All extracts from solvent extraction and the volatile extract described above exhibited clear antioxidant activities at levels of 50-400 microg/mL in the 2,2-diphenyl-1-picrylhydrazyl (DPPH)/linoleic acid assay. The ethanol extract inhibited DPPH activity by 84% at a level of 400 microg/mL. All samples tested via the beta-carotene bleaching assay also exhibited satisfactory antioxidant activity with clear dose responses. This study indicates that corn silk could be used to produce novel natural antioxidants as well as a flavoring agent in various food products.     

Antioxidant activities and volatile constituents of various essential oils

Thirteen essential oils were examined for their antioxidant activity using three different assay systems. Jasmine, parsley seed, rose, and ylang-ylang oils inhibited hexanal oxidation by over 95% after 40 days at a level of 500 microg/mL in the aldehyde/carboxylic acid assay. Scavenging abilities of the oils for the 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical ranged from 39% for angelica seed oil to 90% for jasmine oil at a level of 200 microg/mL. The greatest inhibitory activity toward malonaldehyde (MA) formation from squalene upon UV-irradiation was obtained from parsley seed oil (inhibitory effect, 67%), followed by rose oil (46%), and celery seed oil (23%) at the level of 500 microg/mL. The main compounds of oils showing high antioxidant activity were limonene (composition, 74.6%) in celery seed, benzyl acetate (22.9%) in jasmine, alpha-pinene (33.7%) in juniper berry, myristicin (44%) in parsley seed, patchouli alcohol (28.8%) in patchouli, citronellol (34.2%) in rose, and germacrene (19.1%) in ylang-ylang.     

Determination of ethyl carbamate in distilled alcoholic beverages by gas chromatography with flame ionization or mass spectrometric detection

Quantitative methods are detailed for determination of ethyl carbamate in distilled alcoholic beverages by capillary gas chromatography with flame ionization detection (GC/FID) and by packed-column gas chromatography/mass spectrometry (GC/MS) using selected ion monitoring. Five g samples of distillate of known ethanol concentration are diluted with water to 25% ethanol (v/v), washed with petroleum ether, and extracted with dichloromethane prior to GC/FID or GC/MS analysis. As necessary, sample extracts that exhibit GC/FID interference are passed through alumina for additional cleanup. When internal standards (tert-butyl carbamate and n-butyl carbamate for GC/FID, or ethyl 13C-15N-carbamate for GC/MS) were used for quantitation, the limit of detection for ethyl carbamate was in the range of 5-25 ppb. Coefficients of variation ranged from 3.5 to 6.0% for GC/FID determinations, and from 1.4 to 3.2% for GC/MS. Correlation between methods for 22 random distillate samples ranging in concentration from approximately 40 to 800 ppb gave a correlation coefficient (r) of 0.996.     

Supercritical carbon dioxide extraction of essential oils from Perovskia atriplicifolia Benth

The supercritical fluid extraction (SFE) of the aerial parts of Perovskia atriplicifolia Benth. was studied. The effect of different parameters such as pressure, temperature, modifier identity, and modifier volume on the SFE of the plant was investigated. The extracts were analyzed by GC and GC-MS and compared with the essential oil obtained from P. atriplicifolia Benth by steam distillation. The supercritical extracts and the steam-distilled products had very different compositions. The main constituents of the oil obtained by steam distillation were 1,8-cineole, limonene, camphor, beta-caryophyllene, alpha-pinene, camphene, and alpha-humulene. On the other hand, the major components of SFE extracts were 1,8-cineole, limonene, camphor, beta-caryophyllene, gamma-cadinene, alpha-pinene, and alpha-terpinyl acetate. The results showed that increasing the temperature from 35 to 65 degrees C (at a constant pressure of 100 atm) drastically reduced the number of extracted components. Also, the number of extracted constituents and the percent of main analytes increased when lower pressures were used. Using different modifiers (e.g., methanol, ethanol, dichloromethane, and hexane) for the extraction of the plant at low pressure (100 atm) and temperature (35 degrees C) showed that hexane was more selective than the other modifiers.     

Antimutagenic activity of flavonoids from Pogostemon cablin

A methanol extract from Pogostemon cablin showed a suppressive effect on umu gene expression of SOS response in Salmonella typhimurium TA1535/pSK1002 against the mutagen 2-(2-furyl)-3-(5-nitro-2-furyl)acrylamide (furylfuramide). The methanol extract was re-extracted with hexane, dichloromethane, butanol, and water. A dichloromethane fraction showed a suppressive effect. Suppressive compounds against furylfuramide in the dichloromethane fraction were isolated by SiO(2) column chromatography and identified as 7,4'-di-O-methyleriodictyol (1), 7, 3',4'-tri-O-methyleriodictyol (2), and 3,7,4'-tri-O-methylkaempferol (3). In addition, three flavonoids, ombuine (4), pachypodol (5), and kumatakenin (6), were isolated and identified from the dichrolomethane fraction. Compounds 1 and 3 suppressed >50% of the SOS-inducing activity at <0.6 micromol/mL, and the ID(50) values of both compounds were 0.25 micromol/mL. Compound 2 showed a weakly suppressive effect (17%) at a concentration of 0.6 micromol/mL, and compounds 4-6 did not. These compounds were also assayed with 3-amino-1,4-dimethyl-5H-pyrido[4,3-b]indole (Trp-P-1), which requires liver metabolizing enzymes. Compounds 3-6 suppressed >80% of the SOS-inducing activity of Trp-P-1 at <0.06 micromol/mL, and compounds 1 and 2 suppressed 87 and 63% at a concentration of 0.3 micromol/mL. In addition, these compounds were assayed with activated Trp-P-1, and the suppressed effects of these compounds were further decreased when compared to Trp-P-1. The antimutagenic activities of these compounds against furylfuramide, Trp-P-1, and activated Trp-P-1 were assayed by the Ames test using S. typhimurium TA100.     

2,2-Diphenyl-1-picrylhydrazyl radical-scavenging active components from Polygonum multiflorum thunb

An activity-directed fractionation and purification process was used to identify the antioxidative components of Polygonum multiflorum Thunb. (PM). Dried root of PM was extracted with 95% ethanol and then separated into water, ethyl acetate, and hexane fractions. Among these only the ethyl acetate phase showed strong antioxidant activity by the 2,2-diphenyl-1-picrylhydrazyl (DPPH) test when compared with water and hexane phases. The ethyl acetate fraction was then subjected to separation and purification using silica gel column chromatography and Sephadex LH-20 chromatography. Three compounds showing strong antioxidant activity were identified by spectral methods ((1)H NMR, (13)C NMR, and MS) and by comparison with authentic samples to be gallic acid, catechin, and 2,3,5, 4'-tetrahydroxystilbene 2-O-beta-D-glucopyranoside.     

Antibacterial activity of turmeric oil: a byproduct from curcumin manufacture.

Curcumin, the yellow color pigment of turmeric, is produced industrially from turmeric oleoresin. The mother liquor after isolation of curcumin from oleoresin contains approximately 40% oil. The oil was extracted from the mother liquor using hexane at 60 degrees C, and the hexane extract was separated into three fractions using silica gel column chromatography. These fractions were tested for antibacterial activity by pour plate method against Bacillus cereus, Bacillus coagulans, Bacillus subtilis, Staphylococcus aureus, Escherichia coli, and Pseudomonas aeruginosa. Fraction II eluted with 5% ethyl acetate in hexane was found to be most active fraction. The turmeric oil, fraction I, and fraction II were analyzed by GC and GC-MS. ar-Turmerone, turmerone, and curlone were found to be the major compounds present in these fractions along with other oxygenated compounds.     

Proanthocyanidin profile and ORAC values of Manitoba berries, chokecherries, and seabuckthorn

Six Manitoba fruits were analyzed for their phytochemical content and antioxidant activity in order to increase their production and marketability. The major proanthocyanidins (flavanols) present in whole fruit, juice, and pulp of strawberry, Saskatoon berry, raspberry, wild blueberry, chokecherry, and seabuckthorn were measured. The extraction and purification were facilitated using flash column chromatography, while separation and identification were accomplished by using HPLC and LC-MS techniques. The total proanthocyanidin contents varied from 275.55 to 504.77 mg/100 g in the whole fruit samples. Raspberry contained the highest content, and seabuckthorn showed the lowest content of total flavanols. The highest concentration of proanthocyanidin in juice was found in Saskatoon berry (1363.34 mg/100 mL) and the lowest value in strawberry (622.60 mg/100 mL). HPLC and LC-MS results indicated that epicatechin was the most abundant flavanol followed by B2 in the berry samples, while no catechin or B1 was detected in these fruits. A series of oligomers and polymers were detected in all samples. The recovery percentage was obtained from the ratio of the unspiked samples to the area of spiked samples. Monomers, dimers, oligomers, and polymers gave recovery ranges of 83-99%. The lipophilic and hydrophilic antioxidant capacities of whole fruit, juice, and pulp extracts were measured by the oxygen radical absorbance capacity (ORAC) procedure. In whole fruits, the ORAC values varied from 135 to 479 mg/100 g TE in the MeOH fraction. The corresponding ORAC values varied from 115.30 to 733.15 mg/100 g for the acetone fraction. In juice, all berries showed the same antioxidant capacity (P > 0.05) (133.0-312.0 mg/100 g) in the MeOH fraction, with the exception of raspberry (603.0 mg/100 g). Overall, MeOH fractions mainly contained monomers and dimers with smaller amounts of oligomers and polymers when compared to the acetone fractions. Acetone fractions mainly contained polymers and some oligomers. Although acetone fractions contained a higher quantity of total proanthocyanidins, their antioxidant capacities were lower than MeOH fractions.     

Determination of antioxidant and antimicrobial activities of Rumex crispus L. extracts

The antioxidant activities, reducing powers, 2,2-diphenyl-1-picrylhydrazyl (DPPH) scavenging activities, amount of total phenolic compounds, and antimicrobial activities of ether, ethanol, and hot water extracts of the leaves and seeds of Rumex crispus L. were studied. The antioxidant activities of extracts increase with increasing amount of extracts (50-150 microg). However, the water extracts of both the leaves and seeds have shown the highest antioxidant activities. Thus, addition of 75 microg of each of the above extracts to the linoleic acid emulsion caused the inhibition of peroxide formation by 96 and 94%, respectively. Although the antioxidant activity of the ethanol extract of seed was lower than the water extract, the difference between these was not statistically significant, P > 0.05. Unlike the other extracts, 75 microg of the ether extract of seeds was unable to show statistically significant antioxidant activity, P > 0.05 (between this extract and control in that there is no extract in the test sample). Among all of the extracts, the highest amount of total phenolic compound was found in the ethanol extract of seeds, whereas the lowest amount was found in the ether extract of seeds. Like phenolic compounds, the highest reducing power and the highest DPPH scavenging activity were found in the ethanol extract of seeds. However, the reducing activity of the ethanol extract of seeds was approximately 40% that of ascorbic acid, whereas in the presence of 400 microg of water and ethanol extracts of seeds scavenging activities were about 85 and 90%, respectively. There were statistically significant correlations between amount of phenolic compounds and reducing power and between amount of phenolic compounds and percent DPPH scavenging activities (r = 0.99, P < 0.01, and r = 0.864, P < 0.05, respectively) and also between reducing powers and percent DPPH scavenging activities (r = 0.892, P < 0.05). The ether extracts of both the leaves and seeds and ethanol extract of leaves had shown antimicrobial activities on Staphylococcus aureus and Bacillus subtilis. However, none of the water extracts showed antimicrobial activity on the studied microorganisms.     

骆驼蓬种子提取物对斜纹夜蛾的生物活性效应

用甲醇、乙醇、丙酮、氯仿、乙酸乙酯、苯、石油醚等7种溶剂对骆驼蓬种子进行冷浸提取。提取率分别为23．91％。18．68％．17．80％，18．49％，14．31％．15．13％，17．96％。7种提取物中．以种子乙醇提取物(ESPH)对斜纹夜蛾3龄幼虫的非选择性拒食活性最高。24h拒食率为70．15％。采用酸水总生物碱提取法对ESPH进行初步分离。得到氯仿、正丁醇、水及总生物碱萃取物。生物测定结果表明．总生物碱萃取物对斜纹夜蛾幼虫的生长发育抑制作用最为强烈，其次为氯仿萃取物。     

In vitro antioxidant and ex vivo protective activities of green and roasted coffee

The antioxidant properties of green and roasted coffee, in relation to species (Coffea arabica and Coffea robusta) and degree of roasting (light, medium, dark), were investigated. These properties were evaluated by determining the reducing substances (RS) of coffee and its antioxidant activity (AA) in vitro (model system beta-carotene-linoleic acid) and ex vivo as protective activity (PA) against rat liver cell microsome lipid peroxidation measured as TBA-reacting substances. RS of C. robustasamples were found to be significantly higher when compared to those of C. arabica samples (p < 0.001). AA for green coffee samples were slightly higher than for the corresponding roasted samples while PA was significantly lower in green coffee compared to that of all roasted samples (p < 0.001). Extraction with three different organic solvents (ethyl acetate, ethyl ether, and dichloromethane) showed that the most protective compounds are extracted from acidified dark roasted coffee solutions with ethyl acetate. The analysis of acidic extract by gel filtration chromatography (GFC) gave five fractions. Higher molecular mass fractions were found to possess antioxidant activity while the lower molecular mass fractions showed protective activity. The small amounts of these acidic, low molecular mass protective fractions isolated indicate that they contain very strong protective agents.     

Suppression of chemical mutagen-induced SOS response by alkylphenols from clove (Syzygium aromaticum) in the Salmonella typhimurium TA1535/pSK1002 umu test

A methanol extract from clove (Syzygium aromaticum) showed a suppressive effect of the SOS-inducing activity on the mutagen 2-(2-furyl)-3-(5-nitro-2-furyl)acrylamide (furylfuramide) in the Salmonella typhimurium TA1535/pSK1002 umu test. The methanol extract was re-extracted with hexane, dichloromethane, ethyl acetate, butanol, and water. The hexane fraction showed a suppressive effect. Suppressive compounds in the hexane fraction were isolated by silica gel column chromatography and identified as trans-isoeugenol (1) and eugenol (2) by GC, GC-MS, IR, and (1)H and (13)C NMR spectroscopy. Compounds 1 and 2 suppressed the furylfuramide-induced SOS response in the umu test. Compounds 1 and 2 suppressed 42.3 and 29.9% of the SOS-inducing activity at a concentration of 0.60 micromol/mL. These compounds were assayed with other mutagens, 4-nitroquinolin 1-oxide (4NQO) and N-methyl-N'-nitro-N-nitrosoguanidine (MNNG). In addition, compounds 1 and 2 were assayed with aflatoxin B(1) (AfB(1)) and 3-amino-1,4-dimethyl-5H-pyrido[4,3-b]indole (Trp-P-1), which require liver metabolizing enzymes. These compounds showed suppressive effects of the SOS-inducing activity against furylfuramide, 4NQO, AfB(1), and Trp-P-1. To research the structure-activity relationship, methyl esters of 1 and 2 (1Me and 2Me) and o-eugenol (3), as compounds similar to 2, were also assayed with all mutagens. Compounds 1Me, 2Me, and 3 showed weak suppressive effects of the SOS-inducing activity against furylfuramide.     

竹黄无性型菌株液体发酵菌丝提取物的生物活性研究

测定竹黄无性型菌株ZHLS-03发酵菌丝体中竹红菌甲素的含量,并对其菌丝提取物的清除自由基活性和抑菌活性进行了研究。按50μg/ml的浓度测定了菌丝体石油醚、乙酸乙酯和甲醇等3种提取物对水稻纹枯病菌、稻瘟病菌、棉花枯萎病菌3种植物病原菌以及白色假丝酵母、金黄色葡萄球菌2种病原细菌的抑制能力;同时测定菌丝体的石油醚、乙酸乙酯和饱和正丁醇等3种提取物对DPPH自由基的清除率。结果表明：菌株ZHLS-03液体培养至6d,菌丝体中竹红菌甲素含量最高,可达到28029mg/g;菌丝体甲醇和正丁醇提取物分别具有较强的抑菌活性和清除自由基活性,甲醇相对棉花枯萎病菌的抑制率为7478%,抗白色假丝酵母的抑菌圈直径达到2069mm,且反应15min后正丁醇提取物对DPPH自由基的清除率可达9324%。     

Antioxidative activities of volatile extracts from green tea, oolong tea, and black tea

Antioxidative activities of volatile extracts from six teas (one green tea, one oolong tea, one roasted green tea, and three black teas) were investigated using an aldehyde/carboxylic acid assay and a conjugated diene assay. The samples were tested at levels of 20, 50, 100, and 200 micrograms/mL of dichloromethane. The results obtained from the two assays were consistent. All extracts except roasted green tea exhibited dose-dependent inhibitory activity in the aldehyde/carboxylic acid assay. A volatile extract from green tea exhibited the most potent activity in both assays among the six extracts. It inhibited hexanal oxidation by almost 100% over 40 days at the level of 200 micrograms/mL. The extract from oolong tea inhibited hexanal oxidation by 50% in 15 days. In the case of the extract from roasted green tea, the lowest antioxidative activity was obtained at the level of 200 micrograms/mL, suggesting that the extract from roasted green tea contained some pro-oxidants. The extracts from the three black teas showed slight anti- or proactivities in both assays. The major volatile constituents of green tea and roasted green tea extracts, which exhibited significant antioxidative activities, were analyzed using gas chromatography-mass spectrometry. The major volatile chemicals with possible antioxidative activity identified were alkyl compounds with double bond(s), such as 3,7-dimethyl-1,6-octadien-3-ol (8.04 mg/kg), in the extract from green tea and heterocyclic compounds, such as furfural (7.67 mg/kg), in the extract from roasted green tea. Benzyl alcohol, which was proved to be an antioxidant, was identified both in a green tea extract (4.67 mg/kg) and in a roasted tea extract (1.35 mg/kg).     

Antimutagenic activity of isoflavone from Pueraria lobata

A methanol extract from Pueraria lobata showed a suppressive effect on umu gene expression of the SOS response in Salmonella typhimurium TA1535/pSK1002 against the mutagen 2-(2-furyl)-3-(5-nitro-2-furyl)acrylamide (furylfuramide). The methanol extract from P. lobata was re-extracted with hexane, dichloromethane, ethyl acetate, butanol, and water, respectively. A suppressive compound in the dichloromethane and ethyl acetate extract fractions was isolated by SiO(2) column chromatography and identified as tectorigenin (1) by EI-MS and (1)H and (13)C NMR spectroscopy. Compound 1 and its methylated derivative [7,4'-di-O-methyltectorigenin (2)] had the suppressive effects on umu gene expression of the SOS response in Salmonella typhimurium TA1535/pSK1002 against furylfuramide, 4-nitroquinoline-1-oxide, N-methyl-N'-nitrosoguanidine, and activated Trp-P-1, which do not require live metabolic activation by S9. These compounds also showed suppression of SOS-inducing activity against Trp-P-1 and AfB(1), which requires liver metabolizing enzymes. In addition to the antimutagenic activities of these compounds against furylfuramide, Trp-P-1 and activated Trp-P-1 were also assayed by an Ames test using S. typhimurium TA100.     

Antimutagenic activity of polymethoxyflavonoids from Citrus aurantium

The methanol extract from Citrus aurantium showed a suppressive effect on umu gene expression of SOS response in Salmonella typhimurium TA1535/pSK1002 against the mutagen 2-(2-furyl)-3-(5-nitro-2-furyl)acrylamide (furylfuramide). The methanol extract from C. aurantium was successively re-extracted with hexane, dichloromethane, butanol, and water. A dichloromethane fraction showed a suppressive effect. The suppressive compounds in the dichloromethane fraction were isolated by SiO(2) column chromatography and identified as tetra-O-methylscutellarein (1), sinensetin (2), and nobiletin (3) by EI-MS and (1)H- and (13)C NMR spectroscopy. These compounds suppressed the furylfuramide-induced SOS response in the umu test. Gene expression was suppressed 67%, 45%, and 25% at a concentration of 0.6 micromol/mL, respectively. The ID(50) value (50% inhibition dose) of compound 1 was 0. 19 micromol/mL. These compounds were assayed with other mutagens, 3-amino-1,4-dimethyl-5H-pyrido[4,3-b]indole (Trp-P-1), which requires liver metabolizing enzymes, activated Trp-P-1, and UV irradiation. These compounds showed of all mutagen-induced SOS response in the umu test. In addition, compounds 1-3 exhibited antimutagenic activity in the S. typhimurium TA100 Ames test.     

Cyclooxygenase inhibitory and antioxidant compounds from the mycelia of the edible mushroom Grifola frondosa

The bioassay-guided isolation and purification of the hexane extract of the cultured mycelia of Grifola frondosa led to the characterization of a fatty acid fraction and three compounds, ergosterol (1), ergostra-4,6,8(14),22-tetraen-3-one (2), and 1-oleoyl-2-linoleoyl-3-palmitoylglycerol (3). The composition of fatty acid fraction was confirmed as palmitic, oleic, and linoleic acids by GC-MS and by comparison with the retention values of authentic samples. The structures of compounds 1-3 were established by spectroscopic methods. The fatty acid fraction and compounds 1-3 showed cyclooxygenase (COX) enzyme inhibitory and antioxidant activities. The inhibition of COX-1 enzyme by the fatty acid fraction and compounds 1-3 at 250 microg/mL were 98, 37, 55, and 67%, respectively. Similarly, COX-2 enzyme activity was reduced by fatty acid fraction and compounds 1-3 at 250 microg/mL by 99, 37, 70, and 4%, respectively. The inhibitions of liposome peroxidation by the fatty acid fraction and compounds 1 and 2 at 100 microg/mL were 79, 48, and 42%, respectively. This is the first report of compounds 2 and 3 from the cultured mycelia of G. frondosa. The COX inhibitory activities of compounds 1-3 are reported here for the first time.     

Tyrosinase inhibitor from black rice bran

The inhibitor of tyrosinase activity in black rice bran was investigated. The methanol extract from black rice bran was re-extracted with hexane, chloroform, ethyl acetate, or water. The ethyl acetate extract had the most potent inhibition against tyrosinase activity by 80.5% at a concentration of 0.4 mg/mL. Inhibitory compound in the ethyl acetate fraction was isolated by silica gel column chromatography, and identified as protocatechuic acid methyl ester (compound 1) by GC, GC-MS, IR, and 1H and 13C NMR spectroscopy. Compound 1 inhibited 75.4% of tyrosinase activity at a concentration of 0.50 micromol/mL. ID(50) (50% inhibition dose) value of compound 1 was 0.28 micromol/mL. To study the structure-activity relationship, protocatechuic acid (2), vanillic acid (3), vanillic acid methyl ester (4), isovanillic acid (5), isovanillic acid methyl ester (6), veratric acid (7), and veratric acid methyl ester (8) were also assayed.     

Rapid quantitation and confirmation of aflatoxins in corn and peanut butter, using a disposable silica gel column, thin layer chromatography, and gas chromatography/mass spectrometry

A simple, rapid, and solvent-efficient method for determining aflatoxins in corn and peanut butter is described. Aflatoxins B1, B2, G1, and G2 were extracted from 50 g sample with 200 mL methanol-water (85 + 15). A portion of the extract was diluted with 10% NaCl solution to a final concentration of 50% methanol, and then defatted with hexane. The aflatoxins were partitioned into chloroform. The chloroform solution was evaporated, and the residue was placed on a 0.5 g disposable silica gel column. The column was washed with 3 mL each of hexane, ethyl ether, and methylene chloride. Aflatoxins were eluted with 6 mL chloroform-acetone (9 + 1). The solvent was removed by evaporation on a steam bath, and the aflatoxins were determined using thin layer chromatography (TLC) with silica gel plates and a chloroform-acetone (9 + 1) developing solvent. Overall average recovery of aflatoxin B1 from corn was 82%, and the limit of determination was 2 ng/g. For mass spectrometric (MS) confirmation, aflatoxin B1 in the extract from 3 g sample (20 ng/g) was purified by TLC and applied by direct on-column injection at 40 degrees C into a 6 m fused silica capillary gas chromatographic column. The column was connected directly to the ion source. After injection, the temperature was rapidly raised to 250 degrees C, and the purified extract was analyzed by negative ion chemical ionization MS.     

Isolation and identification of insecticidal components from Citrus aurantium fruit peel extract

Three active components were identified by bioassay-guided fractionation of bitter orange ( Citrus aurantium L.) fruit peel petroleum ether extract. Silica gel fractionation of the extract yielded a fraction that inflicted up to 96% mortality to adults of the olive fruit fly Bactrocera oleae (Gmelin) three days post-treatment. Subsequent HPLC purification of the active fraction resulted in the isolation of three components, eluted in fractions F 222, F 224, and F 226, that induced adult mortality. Considering the data obtained from UV, FTIR, MS, and (1)H NMR spectra, they were identified as 7-methoxy-8-(3'-methyl-2'-butenyl)-2 H-1-benzopyran-2-one (osthol), 4-methoxy-7 H-furo[3,2- g]benzopyran-7-one (bergapten), and 4-(( E)-3'-methyl-5'-(3',3'-dimethyloxiran-2'-yl)pent-2'-enyloxy)-7 H-furo[3,2- g][1]benzopyran-7-one (6',7'-epoxybergamottin). Our results are in concordance with those reported in the literature and were further verified by direct comparison to authentic components. 6',7'-Epoxybergamottin was toxic when tested individually, while bergapten and osthol were found to act synergistically to 6',7'-epoxybergamottin.