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Nutmeg is an inexpensive, readily available spice used in a variety of recipes. However, the use of nutmeg powder as a recreational drug for its hallucinogenic effects is resulting in an increase in overdose rates. We encountered a male patient being hospitalized after ingesting 75 g of commercially available nutmeg powder with the intent of committing suicide. There are no available reports documenting the toxic or comatose-fatal blood concentrations or time-course of drug action in cases of nutmeg poisoning. Therefore, to improve patient management, we endeavored to determine the blood serum levels and time-course of the major psychoactive compounds (safrole, myristicin, and elemicin) present in nutmeg. We designed a simple and reliable method using the MonoSpin® extraction kit and gas chromatography-tandem mass spectrometry to detect the presence of these psychoactive compounds in human serum. The method had detection and quantitation limits of 0.14-0.16 and 0.5 ng/mL (lowest calibration points), respectively. The calibration curves displayed excellent linearity (0.996-0.997) for all three compounds at 0.5-300 ng/mL blood concentrations. The intra- and inter-day precision values for quality assurance were in the ranges of 2.4-11 % and 2.5-11 %, respectively; bias ranged from - 2.6 % to 2.1 %. Blood serum levels of safrole, myristicin, and elemicin were measured at admission (approximately 8 h post-ingestion) and approximately 94 h after a post-admission fluid therapy to evaluate their biological half-lives. We developed this method to obtain information on the psychoactive constituents of nutmeg and, thereby, determine the toxicokinetic parameters of nutmeg in a case of nutmeg poisoning., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 Elsevier B.V. All rights reserved.)

Bioinsecticides are regarded as important alternatives for controlling agricultural pests. However, few studies have determined the persistence of these compounds in stored grains. This study aimed at optimizing and validating a fast and effective method for extraction and quantification of residues of safrole (the main component of Piper hispidinervum essential oil) in cowpea beans. It also sought to assess the persistence of this substance in the grains treated by contact and fumigation. The proposed method used headspace solid-phase microextraction (HS-SPME) and gas chromatography with a flame ionization detector (GC/FID). Factors such as temperature, extraction time and type of fiber were assessed to maximize the performance of the extraction technique. The performance of the method was appraised via the parameters selectivity, linearity, limit of detection (LOD), limit of quantification (LOQ), precision, and accuracy. The LOD and LOQ of safrole were 0.0057 and 0.019 μg kg -1 , respectively and the determination coefficient (R 2 ) was >0.99. The relative recovery ranged from 99.26 to 104.85, with a coefficient of variation <15%. The validated method was applied to assess the persistence of safrole residue in grains, where concentrations ranged from 1.095 to 0.052 µg kg -1 (contact) and from 2.16 to 0.12 µg kg -1 (fumigation). The levels measured up from the fifth day represented less than 1% of the initial concentration, proving that safrole have low persistence in cowpea beans, thus being safe for bioinsecticide use. Thus, this work is relevant not only for the extraction method developed, but also for the possible use of a natural insecticide in pest management in stored grains.

Numerous case reports of intoxications with nutmeg seeds (Myristica fragrans, Houtt.) can be found in literature often following their abuse, as psychotropic effects were described after ingestions of large doses. The successful detection of the main ingredients of the nutmeg seeds essential oil elemicin, myristicin, and safrole, as well as their metabolites in human urine by gas chromatography coupled to mass spectrometry (GC-MS) was already described. The aim of this study was to investigate the detectability of the main ingredients of nutmeg seeds and their metabolites in human blood and urine samples using liquid chromatography coupled to linear ion trap mass spectrometry (LC-LIT-MS n ) and liquid chromatography coupled to high-resolution mass spectrometry (LC-HRMS/MS) after nutmeg seed abuse. Sample material of three individuals was retrospectively investigated after a systematic screening approach indicated an intoxication with nutmeg seeds as a likely cause of symptoms. Metabolic patterns in plasma and urine using GC-MS were comparable with those described in earlier publications. Investigations using hyphenated liquid chromatography techniques lead to the detection of myristicin and safrole, as well as further metabolites not described using GC-MS and revealed sulfation as an additional Phase II metabolic pathway. These results might help to detect or confirm future intoxications with nutmeg seeds by using LC-MS techniques., (© 2021 The Authors. Drug Testing and Analysis published by John Wiley & Sons Ltd.)

Background: Safrole is a natural compound extracted from various plants, and has shown various biological activities. The current study aimed to investigate the antioxidant, antidiabetic, antimicrobial, and anticancer activity of safrole oil and to study the influence of safrole nanoemulgel on these activities., Methods: The antioxidant and antidiabetic in-vitro assays were conducted using standard biomedical methods. The safrole oil nanoemulgel was developed using a self-emulsifying technique. Then the antimicrobial activity of the safrole oil and safrole nanoemulgel were performed on different microbial species, and cytotoxicity was determined against Hep3B cancer cell lines using the MTS assay., Results: Safrole oil showed moderate antioxidant activity compared with standard Trolox, with IC 50 value 50.28 ± 0.44 and 1.55 ± 0.32 μg/ml, respectively. Moreover, it had potent α-amylase inhibitory activity (IC 50 11.36 ± 0.67 μg/ml) compared with Acarbose (IC 50 value 5.88 ± 0.63). The safrole nanoemulgel had pseudo-plastic behaviour, droplet sizes below 200 nm, a polydispersity index (PDI) below 0.3, and a zeta potential of less than - 30 mV. Safrole oil has potential antimicrobial and anticancer activities, and these activities were improved with safrole nanoemulgel., Conclusion: The safrole oil may be applied for the prevention and treatment of oxidative stress, diabetes, different microbial species and cancer, and these activities could be improved by nano-carriers.

The application of reclaimed asphalt pavement has been widely encouraged due to its significant economic and environmental benefits. However, it is necessary to add rejuvenators to ensure its performance. Currently, bio-oil-based regenerants have attracted attention owing to their advantages of renewability and cost savings. The purpose of this paper is to study the use of recycled vegetable oil waste (R-oil) and recycled polyethylene particles for the regeneration and modification of aged asphalt. Physical, rheological, and chemical tests were used to figure out their influence on the pavement performance of aged asphalt. According to the physical test indices (penetration, softening point, and ductility), the performance of the rejuvenated asphalt was better than that of virgin asphalt. The workability and low-temperature performance of the rejuvenated asphalt were basically the same as those of virgin asphalt, and its fatigue and high-temperature performance were better. Infrared spectroscopy showed that R-oil diluted the high-polarity sulfoxide base of aged asphalt. Gel permeation chromatography showed that its molecular weight dispersion was better than that of aged asphalt. Therefore, R-oil and polyethylene can improve the pavement performance and chemical properties of aged asphalt., Competing Interests: The authors have declared that no competing interests exist.

Alkenylbenzenes are natural toxins with genotoxic and carcinogenic effects in rodents, which are highly present in condiments frequently consumed. The aim of this study was the development of the first multi-analyte method for the determination of eight alkenylbenzenes (eugenol, methyl eugenol, acetyl eugenol, trans-isoeugenol, safrole, estragole, myristicin and trans-anethole) in different pepper varieties by gas chromatography coupled to high-resolution mass spectrometry (GC-HRMS-Q-Orbitrap) in combination with a simple ultrasound-assisted extraction method (UAE). The method was successfully validated, and it was applied for studying the presence of these analytes in peppers as well as to elucidate the effects of the berries' maturity and the geographical origin on alkenylbenzene contents. The analysis of the pepper samples showed that eugenol (10.5-120 mg/kg), trans-anethole (10.7-42.7 mg/kg) and estragole (2.2-45.7 mg/kg) tended to be the most detected alkenylbenzenes at high levels, whereas trans-isoeugenol (0.69-3.6 mg/kg) and safrole (0.20-3.0 mg/kg) were minor components. Estragole (PubChem CID: 8815); trans-anethole (PubChem CID: 637563); Myristicin (PubChem CID: 4276); Safrole (PubChem CID: 5144); Eugenol (PubChem CID: 3314); Methyl eugenol (PubChem CID: 7127); Acetyl eugenol (PubChem CID: 7136); trans-Isoeugenol (PubChem CID: 853433); Caffeine (PubChem CID: 2519); Dicyclohexylmethanol (PubChem CID: 78197)., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2020 Elsevier Ltd. All rights reserved.)

High doses of nutmeg (seeds from Myristica fragrans Houtt.) can be abused as a psychoactive drug due to phenylpropene ingredients. During controlled abstinence, e.g., in forensic psychiatric clinics, nutmeg abuse has to be distinguished from an ingestion of other spices having phenylpropene ingredients (e.g., black pepper or garden lovage) or unintentional low-dose nutmeg intake. The aim of this study was to develop an evaluation model for the estimation of time point and amount of nutmeg abuse and differentiation from ingestion of other spices or low doses of nutmeg based on the gas chromatographic-mass spectrometric (GC-MS) analysis of urine samples. A total of 3 volunteers ingested 1.5 g of freshly ground nutmeg. No symptoms were reported. Urine samples were collected for up to 3 days. In addition, 18 blank samples from volunteers with regular diet and 2 authentic samples from forensic psychiatry patients with supposed nutmeg abuse were analyzed. All samples were analyzed by GC-MS in full scan mode. Metabolites of the nutmeg ingredients safrole, myristicin and elemicin were identified via a library search. For semi-quantitative estimations, the area ratios of the analytes to the internal standard (MDMA-d5) were normalized to the creatinine concentration. Up to 8 different metabolites were detected for at least 18 hours after intake of 1.5 g of nutmeg. In the two authentic samples, the normalized area ratios of those metabolites were 0.5-14 times the maximum reached in the intake study. Two additional metabolites could be detected in authentic samples. Probably due to ingestion of other spices, 5 of the 8 metabolites after intake of 1.5 g of nutmeg were detected in blank urine samples as well. The intake of high doses of nutmeg can be differentiated from the ingestion of other spices or low doses of nutmeg via standard GC-MS analysis of urine and application of the proposed evaluation model., (© The Author(s) 2019. Published by Oxford University Press. All rights reserved. For Permissions, please email: journals.permissions@oup.com.)

Safrole is a well-known carcinogenic agent that is present in camphor trees. In this study, a gas chromatographic method was established to quantitate the levels of safrole in essential oils using n-decyl alcohol as an internal standard. The method used a nonpolar column and was able to detect concentrations of safrole as low as 5 µg/ml in the samples. Following addition of 2-10 mg of safrole into 1 g of essential oil extracted from Stout Camphor wood ( Cinnamomum kanehirai Hayata) or 1-10 mg of safrole into 1 g of essential oil extracted from Small-flower Camphor wood ( Cinnamomum micranthum Hayat), the recovery rates of safrole were determined. With direct injection of samples into the gas chromatograph, the results showed that the recovery was more than 96.1%, with a coefficient of variation below 5.6%. We then analyzed 23 commercially available Stout Camphor and other essential oil samples and found that 21 of them contained safrole in the range of 37.65-355.07 mg/g. In addition, in the heavier essential oil distilled from Small-flower Camphor wood, the safrole level was up to 642.98 mg/g. Our results demonstrated that most camphor essential oils on the market have a carcinogenic potential due to their high safrole levels.

Previous studies have demonstrated that the widely distributed desert plant Eremophila longifolia has at least six geographically defined essential oil chemotypes. The focus of the present study is to extend and enhance information concerning known chemotypes and to investigate the involvement of cell nuclei ploidy in this variation. Forty field collected specimens of E. longifolia were taken from most of the mainland states of Australia then subjected to hydrodistillation to produce essential oils, which were then chemically characterised. Ploidy was determined using relative fluorescence of cell nuclei stained with propidium iodide, measured in a flow cytometer. Using principal component analysis (PCA), at least three essential oil chemotypes, in addition to the six already described, were identified in the present study. Previously described high yielding essential oil chemotypes were also characterised in terms of diploidy. For the first time diploid populations were identified in New South Wales, correlating with high yielding isomenthone/menthone and karahanaenone chemotypes. Furthermore, the separate diploid population previously described from Western Australia was demonstrated to be the safrole/methyl eugenol type, which is restricted to a small geographic range in far north-west Western Australia (Murchison District). All other chemotypes were shown to be tetraploid, including apparently randomly emerging individuals, representative of chemotypes producing low yields of isomenthone/menthone and karahanaenone similar in composition to the high yielding diploid types., (Copyright © 2014 Elsevier Ltd. All rights reserved.)

The chemical composition of the essential oil from PINELLIA TUBER (Japanese name: Hange), the dried rhizome of Pinellia ternata, was investigated by capillary gas chromatography (GC) and GC-mass spectrometry (MS) analyses. The oil obtained from Pinellia tuber was revealed the presence of 114 compounds, representing 90.6% of the total oil identified. This colorless oil had a spicy and woody odor. The main components of the oil were β-cubebene (8.8%), atractylon (7.8%), methyl eugenol (6.2%), and δ-cadinene (5.3%). Fifteen major odor-active compounds were identified in the essential oil from PINELLIA TUBER by the GC-olfactometry (GC-O) and aroma extract dilution analysis (AEDA). Among these, safrole (spicy) and β-vatirenene (woody) showed the highest flavor dilution (FD) factor (128), followed by paeonol (FD = 64; woody, spicy), α-humulene (FD = 64; woody), and β-phenylnaphthalene (FD = 64; spicy).

Quantitation of tr-cinnamaldehyde, safrole and myristicin was carried out in 70 samples of cola-flavoured soft drinks purchased in eight European countries with the purpose of assessing the variability in the levels of these substances. Results indicated a limited variability in the content of the three substances: the ratio between the 90th and the 10th percentile concentration amounted to 21, 6 and 13 for tr-cinnamaldehyde, safrole and myristicin, respectively. The uncertainty in the assessment of dietary exposure to these substances due to the variability of their level in cola-flavoured drinks was low. Based on these analytical data and on refined food consumption data, estimates of exposure to safrole associated to cola drink consumption, along with Margin of Exposure (MOE) values, were obtained. For high consumers of cola-flavoured soft drinks in certain age groups, within some European countries, MOE values lower than 10,000 resulted, MOE values of 10,000 or higher having been stated by the EFSA as a quantitative criterion to identify low concern from a public health point of view and low priority for risk management actions. The lowest MOE values, from 1900 to 3000, were observed for children and teen agers in the United Kingdom and Ireland., (Copyright © 2013 Elsevier Ltd. All rights reserved.)

A high performance liquid chromatography-linear ion trap triple stage mass spectrometry (HPLC-IT/MS3) method was established to detect aldicarb, aldicarb sulfoxide, and aldicarb sulfone in peanuts. The samples were extracted by acetonitrile saturated with cyclohexane, followed by clean-up with gel permeation chromatography (GPC). The determination was performed using HPLC-IT/MS3 for the identification and quantification of the compounds. The separation was carried on a Capcell PAK CR column with gradient elution using 5 mmol/L acetic acid/ammonium acetate/acetonitrile as mobile phase. The ionization of molecules was performed by electrospray mode. Selective reaction monitoring (SRM) was the acquisition mode used for the monitoring of MS3 transitions for each compound using aldicarb-d3 as internal standard for three analytes. Matrix effects were evaluated by comparing the recovery of matrix-matched and solvent-based calibration curves. The calibration graphs were linear in the ranges of 10 - 500 microg/L and the detection limits ranged from 4 to 5 microg/kg. The average recoveries ranged between 81.5% and 115% at three different spiked levels (10, 20 and 40 microg/kg). Satisfactory results were obtained in the determination of real peanut samples by this method.

Objective: To analyze the genetic diversity and the volatile components of Asarum sieboldii from seven habitats in Qin-ba region., Methods: The genetic diversity of the herb was analyzed by ISSR (inter simple sequence repeat) markers; The relative content volatile components of the herb were dectected by head space solid-phase microextraction gas chromatogrphy-mass spectrometry (HS-SPME-GC-MS). The contents of the 3 main components were analyzed by steam distillation gas chromatography-mass spectrometry (GC-MS)., Results: 57 bands were amplified from 7 populations by 6 reliable primers, 51 of which were polymorphic (89.47% of the total). The cluster analysis presented that these resources were divided into two main groups. There were differences in the chemical components and the contents of Asarum sieboldii from the 6 wild habitats. Except for some same components, many unique components were identified in them respectively. In addition, some components could be detected only in some populations which had smaller genetic distance., Conclusion: Cluster analysis shows no direct correlation between genetic distance and geographic distance of Asarum sieboldii in Qin-ba region. The accumulations of some volatile components of Asarum sieboldii are possibly related to genetic diversity.

In the current investigation, the influence of the plant growth stages (prior to emergence of flower heads or vegetative, anthesis and fruiting) on the essential oil content and composition in the aerial parts of Prangos uloptera DC. was studied. Quantitative and qualitative differences were found among oil of aerial parts at three phenological stages. The principle compounds of the oil before emergence of flower heads were saferole (21.6%) and α-pinene (20%), while the oil at anthesis was composed of α-bisabolol (30.5%), saferole (19.11%) and (+)-spathulenol (12.9%). The oil at fruiting stage displayed γ-terpinene (35.5%) and trans-anethole (23.5%) as major constituents. These findings may confer valuable information regarding chemical composition of P. uloptera DC., which could be beneficial in the fields of herbal medicine.

Chemical composition, antioxidant and antimicrobial activities of the fresh leaves and stems oils of Piper caninum were investigated. A total of forty eight constituents were identified in the leaves (77.9%) and stems (87.0%) oil which were characterized by high proportions of phenylpropanoid, safrole with 17.1% for leaves and 25.5% for stems oil. Antioxidant activities were evaluated by using β-carotene/linoleic acid bleaching, DPPH radical scavenging and total phenolic content. Stems oil showed the highest inhibitory activity towards lipid peroxidation (114.9 ± 0.9%), compared to BHT (95.5 ± 0.5%), while leaves oil showed significant total phenolic content (27.4 ± 0.5 mg GA/g) equivalent to gallic acid. However, the essential oils showed weak activity towards DPPH free-radical scavenging. Evaluation of antimicrobial activity revealed that both oils exhibited strong activity against all bacteria strains with MIC values in the range 62.5 to 250 μg/mL, but weak activity against fungal strains. These findings suggest that the essential oils can be used as antioxidant and antimicrobial agents for therapeutic, nutraceutical industries and food manufactures.

A pressurized liquid extraction and GC-MS method was developed for simultaneous quantitative determination of the seven components, including cinnamaldehyde, copaene, cinnamic acid, coumarin, 2-methoxycinnamaldehyde, 2-methoxycinnamic acid and safrole in Cinnamomum cassia. The results showed that methanol and ethanol was not available for extraction of cinnamaldehyde and 2-methoxycinnamaldehyde due to aldol reaction. The developed method was validated to be sensitive, accurate and simple, and was successfully employed for the analysis of 15 samples of C. cassia. The contents of the investigated components were significantly variant and cinnamaldehyde is the most abundant compound, but safrole was not detected in all samples.

X-ray photoelectron spectroscopy was used to investigate the characteristics and evolution of sulfur (S) in mixtures of bituminous coal and sewage sludge (SS) and their chars during isothermal combustion. Five groups of mixtures with SS content of 0%, 10%, 20%, 30% and 100%, were examined at different burn-off ratios (beta) of 0, 30%, 50%, 70% and 100%. The S in the coal mainly exist as the forms of mercaptan (S1), sulfide (S2), thiophene (S3), sulfoxide (S4), sulfone (S5) and sulfate (S6). During the coal combustion process, the content of S1 and S2 decreased, while that of S3 and S5 increased in the early stage and decreased in the late stage. The S4 content increased throughout the entire process of combustion. Small amount of S6 was detected, showing a fluctuated pattern. The trend of S1, S2, S5 and S6 in SS was alike with that in coal, whereas S4 decreased at the end of combustion. The changing process of S3 in SS was opposite to that of coal, while the composition of S in the mixtures resulted from the mixing of coal and SS. The transformation of each functional group during co-combustion were correlated with their transformation characteristics during the mono-combustion of coal and SS, and no obvious interaction was observed, which demonstrated that the coal-origin and SS-origin sulfur in mixtures kept their own characteristics in the combustion. SS may accumulate on the solid surface as alpha increase, resulting its significant influence on the evolution of each form of S. When alpha was low, most of the S-contained functional groups presented the characteristics of coal. The percentage of coal-origin functional groups declined as alpha increased. The transforming trends of most functional groups were similar with that of SS when alpha reached 30%.

Introduction: Asari radix et rhizoma (Xixin, Manchurian Wildginger, Asarum spp) is a herbal drug commonly used as an ingredient in traditional Chinese medicine (TCM). Many species of Asarum contain safrole and methyleugenol as the main components of their volatile oils. However, toxicological studies have shown that safrole and methyleugenol may be a hepatocarcinogen and/or genotoxic leading to concerns regarding the habitual consumption of this herbal drug., Materials and Methods: An HPLC method was established to assess the levels of safrole and methyleugenol in five batches of Asari radix et rhizoma and two TCM formulae containing this herbal drug as an ingredient. Analyses showed that the content of safrole in the dried herbal drugs tested ranged from 0.14-2.78 mg/ g whilst the content of methyleugenol ranged from 1.94-16.04 mg/g., Results: The present study demonstrated that following a 1-hour decoction, the amount of safrole was decreased by more than 92% resulting in the equivalent of no more than 0.20 mg/g safrole remaining in the aqueous extract. Similarly, the content of methyleugenol was decreased to the equivalent of 0.30-2.70 mg/g. Furthermore, both TCM formulae, after decoction, showed negligible amounts of safrole (maximum, the equivalent of 0.06 mg/ g), and only 1.38-2.71 mg/g of methyleugenol., Conclusions: The present study shows that a decoction procedure, similar to that traditionally used for Chinese herbal preparations, is able to effectively reduce the amount of safrole and methyleugenol effectively. Such a reduction in the content of safrole should be acceptable for therapeutic use., (Copyright 2009 S. Karger AG, Basel.)

Betel quid chewing, which contributes high concentration of safrole in saliva, is a popular oral habit in Taiwan. Safrole is a documented rodent hepatocarcinogen, yet its hepatocarcinogenic potential in human is not known. Here, we used LC/ESI-ITMS(n) and LC/QTOF-MS confirmed safrole-dGMP as reference standard to detect the safrole-DNA adduct in hepatic tissues from HBsAg-/HCV-seronegative hepatocellular carcinoma patients by (32)P-postlabeling. We first synthesized and confirmed safrole-dGMP by LC/MS. Two isomeric safrole-dGMPs were characterized as N(2)-(trans-isosafrol-3'-yl) deoxyguanosine and N(2)-(safrol-1'-yl) deoxyguanosine. This technique was able to detect hepatic safrole-DNA adduct in mice that were treated with safrole but not sensitive enough to detect safrole-DNA adduct in human samples. Using the nuclease P1 version of the (32)P-postlabeling technique, we detected the presence of safrole-DNA adduct in two out of 28 hepatic tissues from hepatocellular carcinoma patients, and only these two patients had a history of betel quid chewing lasting more than 10 years. From co-chromatography with the mass confirmed safrole-dGMPs, this safrole-DNA adduct was identified as N(2)-(trans-isosafrol-3'-yl) deoxyguanosine. These results suggest that betel quid-containing safrole might be involved in the pathogenesis of hepatocellular carcinoma in human beings and LC/MS has the potential to identify DNA adducts in clinical samples.

Square-wave adsorptive-stripping voltammetry technique has been used to develop a method for the determination of fenthion in olive oil. Due to the fact that fenthion does not give any electrochemical signal at mercury electrode, the method has been based on a previous oxidation of fenthion to its metabolite, fenthion-sulfoxide, by using KMnO(4). The metabolite gives rise to a peak due to an adsorptive-reductive process at -0.786 V. Fenthion is isolated from olive oil by carrying out a solid-liquid extraction procedure using silica cartridge, followed by a liquid-liquid partitioning with acetonitrile. The detection limit in olive oil is 78.8 ng g(-1) and recoveries for four levels of fortification are ranged from 85% to 109%. On the other hand, it has been developed a method for the simultaneous determination of fenthion and its metabolite fenthion-sulfoxide, in river water. Pesticides are isolated from water by carrying out a liquid-liquid partitioning with trichloromethane. The detection limits are 0.41 ng g(-1) and 0.44 ng g(-1), for fenthion and fenthion-sulfoxide, respectively. Recoveries for three levels of fortification are ranged from 96% to 103% for fenthion and 94% to 104% for fenthion-sulfoxide.

Various studies have been drawn toward the beneficial properties of fruit juices because they have several components, such as phenols, vitamins, and flavonoids, with antioxidant effects. However, fruit juices can also contain residues of pesticides used as standard pest control methods in crops. Many of these pesticides are degraded through oxidative mechanisms, and their persistence in juices can be enhanced by antioxidants. This study covers the degradation of four pesticides, aldicarb, demeton-S-methyl, fenamiphos, and methiocarb, to their respective sulfoxide and sulfone in grape juices, water (pH 3.5) and water (pH 3.5) with quercetin (one of the most important flavonoids of grape) added in an attempt to establish whether the presence of antioxidants can affect the degradation rate of pesticides. For this purpose, a multiresidue method based on solid-phase extraction (SPE) was developed for the simultaneous determination of these pesticides and their metabolites in commercial juices. The extraction procedure was carried out in C(18) columns. The subsequent elution of pesticides was performed with dichloromethane prior to the determination by liquid chromatography-tandem mass spectrometry (LC-MS/MS), using two precursor-product ion transitions. Average recoveries for all the pesticides studied were higher than 80%, with relative standard deviations lower than 15% in the concentration range 0.005-0.05 microg/mL, and the quantification limits achieved ranged from 0.1 to 4.6 microg/L. The results demonstrated that degradation was slower in fruit juices and aqueous solutions with quercetin than in water. Several commercial grape juices were also analyzed to establish the levels of these pesticides. Methiocarb, fenamiphos, and demeton-S-methyl were found at low levels in some samples.

Seeds of nutmeg are used as spice, but they are also abused because of psychotropic effects described after ingestion of large doses. It was postulated that these effects could be attributable to metabolic formation of amphetamine derivatives from the main nutmeg ingredients elemicin (EL), myristicin (MY), and safrole (SA). In a case of a suspected nutmeg abuse, neither such amphetamine derivatives nor the main nutmeg ingredients could be detected in urine. The metabolites of EL, MY, and SA were identified using gas chromatography-mass spectrometry in rat urine and their presence in human urine of the nutmeg abuser was confirmed. The identified metabolites indicated that EL, MY, and SA were once and twice hydroxylated at the side chain. In addition, EL was O-demethylated at 2 positions followed by side chain hydroxylation. MY and SA were demethylenated and subsequently methylated. In the human urine sample, the following metabolites could be identified: O-demethyl elemicin, O-demethyl dihydroxy elemicin, demethylenyl myristicin, dihydroxy myristicin, and demethylenyl safrole. As in the human urine sample, neither amphetamine derivatives nor the main nutmeg ingredients could be detected in the rat urine samples. Finally, toxicologic detection of nutmeg abuse was possible by identification of the described metabolites of the EL, MY, and SA in urine applying the authors' systematic toxicologic analysis procedure using full-scan gas chromatography-mass spectrometry after acid hydrolysis, liquid-liquid extraction of analytes, and microwave-assisted acetylation of extracted analytes.

A peak considered to be due to disulfoton sulfoxide as a metabolite of disulfoton was detected in the analysis of the chingentsuai extraction solution of vegetables by GC-FPD. In the analysis by GC/MS for identification, though the retention time and the mass spectrum were the same as those of the standard, the spectrum was different from MS library data. It appeared that decomposition of disulfoton sulfoxide occurred at the inlet. Therefore, we used a programmed temperature vaporization (PTV) inlet in the pulsed splitless mode to achieve a low inlet temperature and high injection pressure. As a result, the expected mass spectrum of disulfoton sulfoxide was obtained. Under this condition, the peak obtained from chingentsuai was identified as disulfoton sulfoxide. Disulfoton sulfoxide was detected from 2 of 25 chingentsuai samples, at concentrations of 0.66 microg/g and 0.14 microg/g.

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.

Cinnamomum carolinense, locally known as madeu, is a tree endemic to the volcanic mountains of the Island of Pohnpei in the Eastern Carolines of the South Pacific. The bark is harvested from trees and brewed to make a medicinal tea and hot beverage that is regularly consumed. Many species of Cinnamomum contain the known hepatocarcinogen safrole, sparking concern regarding habitual consumption of this beverage. HPLC-PDA analysis confirmed the presence of the carcinogen in alcoholic extracts of Cinnamomum carolinense bark shavings (0.435%, w/w), but safrole was not detected in the tea. The limit of detection and limit of quantitation of safrole were determined to be 1.25 and 3.75 microg/mL, respectively. The traditional preparation method, which boils the bark shavings, degrades the safrole.

To defend themselves against herbivory, plants use a variety of direct and indirect strategies involving induced increases in secondary substances. Species of the Allium genus (Alliaceae), such as the leek Allium porrum (L.), produce nonprotein sulfur amino acids derived from cysteine, i.e., alk(en)yl-cysteine sulfoxides that are precursors of volatile thiosulfinates and disulfides. These defend most species including the specialist leek moth, Acrolepiopsis assectella. We determined by measuring the increase in the sulfur precursor propyl-cysteine sulfoxide (PCSO) if production of this precursor is induced in response to moth attack and mechanical wounding. The concentration of PCSO was determined by HPLC in 2- or 6-mo-old leeks after attacks of various intensity either by the specialist leek moth or by a generalist moth, Agrotis ipsilon. Injury-induced release of sulfur volatiles was measured by GC/MS after the attacks. Results showed an increase in the production of sulfur compounds in both the precursor and volatile form, occurring only in association with intensive attacks by leek moths. The increase in sulfur precursors also led to an increase in the release of sulfur volatiles. This induced response may provide an effective defense strategy against the plant's main natural enemy, both directly and indirectly by attracting entomophagous insects.

Objective: To establish a method for GC fingerprint determination of the chemical constituents in Herba Asari., Method: GC and GC-MS were used to optimize the fingerprint determination method, and identify the main peaks in the GC fingerprint., Result: A preferable method for GC fingerprint determination of the chemical constituents in Herba Asari was established., Conclusion: A general acquaintance of the chemical constituents in Herba Asari can be obtained by using the preferable GC fingerprint determination method, which is useful for quality evaluation of the crude drug of Herba Asari.

A rapid and sensitive biological monitoring (BM) method for assessing exposure to the environmental carcinogen safrole has been developed. The method is an isocratic high-performance liquid chromatographic (HPLC) analysis of urinary dihydroxychavicol (DHAB) and eugenol, the urinary metabolites of safrole. Good linearity, precision, and accuracy were demonstrated. A recovery of 98.8 +/- 5.4% (SD, n = 3) was found for DHAB and 84.1 +/- 3.4% (n = 3) for eugenol. The quantitation limits of the method were 8 ng for DHAB and 10 ng for eugenol. The validity of the method was demonstrated by a linear dose-response relationship observed in rats given oral doses of safrole at 30, 75, and 150 mg/kg body weight. The method was also used to monitor the environmental exposure to the Taiwanese betel quid (TBQ) chewing, because TBQ used in Taiwan not only contains areca (betel) nut, slaked lime, and catechu but also Piper betle inflorescence or its leaves. Both of the latter have a high content of safrole. The feasibility of the method to monitor TBQ chewing was demonstrated by an analysis of 153 spot human urine samples. The results showed that the p value of the nonparametric group comparison was < 0.001 for DHAB and 0.832 for eugenol. The TBQ chewers also exhibited a significantly higher rate of urinary DHAB (but not eugenol) than the nonchewers with an odd ratio of 3.47 (95% CI, 1.61-7.51). However, when only the eugenol-positive subjects were taken into analysis, the ratio rose to 24.38 (95% CI, 3.00-197.90).

Little is known about the possible health effects associated with inhaling alkenylbenzenes through cigarette smoking, even though these flavor-related compounds have known toxic effects in animals. We developed a rapid and sensitive solid-phase extraction (SPE) method to quantify seven alkenylbenzenes and piperonal in mainstream cigarette smoke particulate. The smoke particulate fraction of a single cigarette was collected on Cambridge filter pads, solvent extracted, concentrated, purified with SPE, and analyzed by selected ion monitoring gas chromatography-mass spectrometry. We positively identified and quantified five alkenylbenzenes compounds (eugenol, isoeugenol, methyleugenol myristicin, and elemicin) and piperonal in the smoke particulate from eight U.S. brands with mean levels (measured in triplicate) ranging from 6.6 to 4210 ng per cigarette. Additionally, complete blocking of nearly invisible ventilation holes in the cigarette filter increased 2- to 7-fold the percent transfer of alkenylbenzenes from tobacco to the particulate fraction of mainstream smoke.

The in vitro metabolism of RWJ-34130, an antiarrhythmic agent, was conducted using rat hepatic 9000 x g supernatant (S9) and microsomes in an NADPH-generating system, and the rat liver perfusion. The 100 and 20 microg ml(-1) concentrations of RWJ-34130 aqueous solution were used for microsomal incubation and liver perfusion, respectively. Unchanged RWJ-34130 (approximately 77-78% of the sample in both S9 and microsomes) plus a major metabolite, RWJ-34130 sulfoxide (20% of the sample in both S9 and microsomes) were profiled, isolated and identified from both hepatic S9 and microsomal incubates (60 min) using HPLC and mass spectrometry (MS), and by comparison to a synthetic RWJ-34130 sulfoxide, which was synthesized by reacting RWJ-34130 with MCPBA (meta-chloroperoxy benzoic acid). No unchanged RWJ-34130 was detected in the 3 h liver perfusate, however, 1-phenyl-2-oxo-pyrrolidine was profiled, isolated and identified as a major hydrolyzed metabolite of liver perfusate. RWJ-34130 is not extensively metabolized in vitro in rat hepatic S9 and microsomes. All HPLC metabolic profiles of hepatic S9 and microsomal samples (30 min, 60 min) were qualitatively and nearly quantitatively identical.

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.

A liquid chromatographic (LC) method was developed for determining safrole in herbal products derived from sassafras (Sassafras albidum), as well as related compounds such as isosafrole and dihydrosafrole. The procedure involves solvent extraction and isolation of analyte by reversed-phase LC with UV detection at 235 nm. Safrole is resolved from related compounds and other sample constituents including thymol, a component of thyme. A linear concentration range of 0.003-0.200 mg/mL was obtained for safrole, isosafrole, and dihydrosafrole. Limits of detection (LOD) and quantitation (LOQ) were e0.0015 and 0.0051 micrograms/mL for safrole, 0.0018 and 0.0061 micrograms/mL for isosafrole, and 0.0038 and 0.0125 micrograms/mL for dihydrosafrole, respectively. Intraday relative standard deviations (RSDs) for safrole (n = 5) from various samples ranged from 1.30 to 5.39% at analyte levels of 0.01-1.5%. Safrole contents of 26 samples including root bark powder, leaves, oils, tea concentrate, herbal extract tinctures, and herbal powder capsules ranged from < LOD for most leaf samples to 92.4% for an oil. Recoveries of safrole from fortified samples ranged from 83.6% for an oil to 117.2% for a tincture preparation. Safrole contents of 0.09-4.66 mg/cup were found for brewed teas prepared from sassafras root bark powders and tinctures.

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.

We established a method for quantitative analysis of methyleugenol and safrole in Asarum heterotropoides by GC to study their season variation. The results showed that the content of methyleugenol in the essential oil of the subterranean part of different seasons was 21%-39%, and that for safrole was 17%-33%. In the upterranean part, methyleugenol was 1.4%-9.6%, and safrole was 0.36%-2.1%. It is clear that methyleugenol and safrole are provided mainly by the subterranean part. The quantity of methyleugenol is the richest in the period of sprouting and after-fruiting, and for the safrole content the richest periods are that of sprouting, blooming and after-fruiting. In the case of upterranean part, the richest time is blooming period both for methyleugenol and safrole, and after blooming they decrease gradually.

Safrole (4-allyl-1,2-methylenedioxybenzene), a natural plant component of the aromatic oil of sassafras root bark, possesses carcinogenic and mutagenic activity. Legal restrictions have been placed on safrole as a food additive. However, sassafras teas continue to be accessible from health food establishments in the United States. Supercritical fluid extraction (SFE) with gas chromatographic-mass spectrometric (GC-MS) determination is utilized in the formulation of a rapid, accurate, and specific method for the determination of safrole and related allylbenzenes in unbrewed sassafras teas. Samples are extracted in a static-dynamic mode with CO2 at 690 bar and 80 degrees C with methanol as an extractor-added modifier. Levels of safrole exceeding 10,000 mg/kg (1.0%) are commonly encountered. Lesser amounts of other allylbenzenes, including eugenol and 4-allyl-1,2-dimethoxybenzene, are also reported. Recoveries of safrole and related compounds from previously extracted tea samples fortified at 100 and 1000 mg/kg ranged from 96 to 101%.

An HPLC method for the simultaneous determination of safrole (S), dihydrosafrole (DHS), and chloromethyldihydrosafrole (CM-DHS) in piperonyl butoxide, with fluorimetric detection and elution gradient, is described. Samples with internal standard (piperonyl isobutyrate) are adsorbed on silica cartridges, then eluted with an appropriate solvent. Internal standard, S, DHS, and CMDHS are eluted together and injected into a reversed-phase chromatography column. The concentrations of the three products are calculated from calibration graphs, and linearity and reproducibility are verified. The limit of detection is 2 mg.kg-1. This analytical method allows for control of the synthesis process of piperonyl butoxide and determination of some carcinogenic substances, such as S and DHS.

The suitability of C18 reversed-phase thin-layer chromatography (TLC) for enrichment of adducts in the 32P-postlabeling assay was investigated for structurally diverse classes of DNA adducts derived from benzo[a]pyrene, 2-acetylaminofluorene, benzoquinone, safrole, and mitomycin C. The TLC enrichment involved retention of adducts to the C18 phase followed by elution with organic solvent-water. Adduct patterns obtained by the C18 purification were qualitatively similar to those obtained by the nuclease P1 and butanol procedures, the two commonly used enrichment methods. Adduct recoveries by the C18 method varied for different adducts and were significantly lower than those obtained by the other two techniques.

This article reports the detecting result of trace elements in processed products of Pinellia ternata and of the contents of As, Hg, Pb in the products processed with sulfur. The result shows that all the processing procedures are necessary and safe.

A direct second-derivative spectrofluorimetric procedure for determining sulphoxide impurity in phenothiazines and their formulations has been developed. The method, which has been applied to the analysis of chlorpromazine hydrochloride and prochlorperazine mesylate, as examples of typical phenothiazine substances, and to their formulations, is based on the measurement of the amplitude taken from the minimum at ca. 270 nm to the longer wavelength maximum in the second-derivative excitation spectrum of the sulphoxide in pH 8 buffer solution. The method is rapid, accurate and precise, and can be used to measure the concentration of sulphoxide in phenothiazines and their formulations at concentrations down to 0.1% m/m of that of the parent phenothiazine.

The technique of selected ion monitoring (SIM) allows the mass spectrometer to be used as a selective detector when coupled to a gas chromatograph. The addition of a data acquisition system enables the selective quantitative determination of compounds to be made when present at low levels in a complex mixture. The detection limit of a compound may be lowered by 2 or 3 orders of magnitude, since the instrument is set to one or more ions which are specific for the compound(s) to be measured, for the maximum amount of time. This lowering of the detection limit and increase in selectivity makes it possible not only to determine compounds at ppb levels, but also to carry out rapid and selective determinations of compounds at the ppm level, using a smaller sample and a minimum of preliminary sample concentration. A fuller description of the principles of SIM is given. This technique has already found wide application in medical and environmental fields, and some examples of its application to alcoholic beverages are given: detection and determination of possible contaminants, such as phthalate, esters residual monomers from plastic materials (styrene, vinyl chloride), and traces of pesticides; determination of certain non-volatile compounds in wines; and the selective determination of certain compounds for which levels in finished products are limited such as the thujones, safrole, beta-asarone, and coumarine.

A liquid chromatographic method is described for determination of perphenazine and perphenazine sulfoxide in representative dosage forms. Sulfoxide levels were nondetectable or less than 1% in tablets and in an injectable product. Sulfoxide levels increase with time in some syrup formulations and may be as high as 11% in syrup formulations before their expiration date.

Interest in the quantitative analysis of flavouring agents in alcoholic beverages arises from the possible toxicological consequences deriving from their use. The method reported here, based on gas chromatography with selected-ion monitoring mass spectrometry (SIM), allows the simultaneous measurement of alpha, beta-thujone, pulegone, safrole, coumarin and beta-asarone at the ppb level in alcoholic beverages.