High Resolution Gas Chromatography Coupled To High Resolution Mass Spectrometry Research Articles

Diterpenes content of the brown alga Dictyota ciliolata (Dictyotales, Phaeophyceae) and recognition of a Brazilian haplotype based on psbA sequences

ABSTRACTMacroalgae of the genus Dictyota produce large amount of diterpenes, which have many ecological and biological functions. We analysed the crude extracts of three Brazilian populations of the brown alga Dictyota ciliolata by hydrogen nuclear magnetic resonance (1H-NMR) spectroscopy and high-resolution gas chromatography coupled to mass spectrometry (HRGC/MS). All of the HRGC/MS profiles showed that D. ciliolata contained prenylated guaiane (Group I), xeniane, and crenulidane diterpenes (Group III) as major constituents. Eleven diterpenes were detected, dictyol B acetate, dictyol B, dictyol C, dictyoxide, isopachydictyol A, pachydictyol A, 4β-acetoxydictyodial A, and four crenulidane diterpenes. Dictyol B acetate was the major product in all of algal populations in Brazil. The diterpene profiles of the Brazilian populations were compared to other ones obtained in populations from the Atlantic and Pacific Oceans. The genomic DNA of a separate Brazilian population was extracted and amplified for psbA, the sequences of which were used to build a haplotype network with other sequences available in Genbank. Ten psbA haplotypes were found, five in the Indian Ocean, four in the Western Pacific Ocean and three in the Atlantic Ocean. The eastern (Western Pacific Ocean) and the western (Indian Ocean) coasts of Australia were the most diverse areas and they contained most of known haplotypes. One haplotype identified via the molecular network was found to be common in all of the oceans and thus is proposed as the ancestral form. The chemical and molecular data are consistent at the recent proposal in which D. ciliolata was classified as an essentially pantropical species, which may prove to be highly beneficial to bioprospection studies as this species is a source of potential bioactive products.

Meeting the European Commission performance criteria for the use of triple quadrupole GC-MS/MS as a confirmatory method for PCDD/Fs and dl-PCBs in food and feed samples.

Until recently, European Union (EU) legislation required the use of high-resolution gas chromatography coupled to high-resolution mass spectrometry (HRGC-HRMS) based on magnetic sector analyzers as a standard approach for confirmatory analysis of dioxins (PCDDs) and furans (PCDFs) in feed and food. However, recent technological advances in MS instruments enabled other alternative analytical techniques to meet the same analytical criteria as those requested for HRGC-HRMS. In this sense, triple quadrupoles (GC-MS/MS) can be a realistic alternative for the analysis of dioxins. In this work, the performance of GC-MS/MS technology was evaluated against the criteria demanded by the EU for confirmatory analysis of dioxins and PCBs in food and feed. Thus, the study comprises a number of parameters including chromatographic separation, limit of quantification, linearity, repeatability, and ion ratio precision. Analyses of solvent standards as well as sample extracts (inter-calibration extracts and certified reference materials) were also considered within the scope of this study. Additionally, direct comparisons of the results obtained by GC-MS/MS with those from GC-HRMS were made. The results of this work suggested that GC-MS/MS was highly sensitive and selective for confirmatory analysis of PCDD/Fs and related compounds in food and feed samples and meets all the criteria requested by the European Commission.

Congener-specific analysis of polychlorinated naphthalenes (PCNs) in the major Chinese technical PCB formulation from a stored Chinese electrical capacitor.

Impurity of polychlorinated naphthalenes (PCNs) in commercial polychlorinated biphenyl (PCB) formulations has been recognized as a relevant source of PCNs in the environment. Congener-specific analysis of most main PCB formulations has been accomplished previously, excluding the Chinese product. The insulating oil in a stored Chinese electric capacitor containing the major Chinese technical formulation "PCB3" was sampled and tested by isotope dilution technology using high-resolution gas chromatography coupled to high-resolution mass spectrometry (HRGC/HRMS). The detected concentration of PCNs in the Chinese PCB oil sample was 1,307.5 μg/g and therefore significantly higher than that reported in PCB formulations from other countries, as well as that in the transformer oil (ASKAREL Nr 1740) additionally tested in the present study for comparison. Based on the measurement, the total amount of PCNs in Chinese PCB3 oil is estimated to be 7.8 t, which would mean only 0.005 % of global production of PCNs of 150,000 t. The homolog profile is similar to those of PCN in Aroclor 1262 and Clophen A40, where the contributions from hexa-CNs and hepta-CNs are predominant and accounted for similar proportions. The Toxic Equivalent Quantity (TEQ) concentration of dioxin-like PCN congeners is 0.47 μg TEQ/g, with the dominant contributors of CN-73 and CN-66/67. This TEQ content from PCN is higher than that in most other PCB formulations with the exemption of the Russian Sovol formulation. The total TEQ in the historic 6,000 t of the Chinese PCB3 formulation is estimated to be 2.8 kg TEQ.

축산물 사료중의 Polybrominated Diphenyl Ethers(PBDEs) 평가

Polybrominated diphenyl ethers (PBDEs) are a group of brominated flame retardants (BFRs), which are used in a variety of consumer products. Several of those are produced in large quantities. Their chemical structure similarities to polychlorinated dibenzo-p-dioxins and polychlorinated biphenyls (PCBs), as well as their toxicity, has been studied. PBDEs are persistent and lipophilic, which results in their bioaccumulation in the fatty tissues of organisms and enrichment throughout food chains. In addition, a number of studies also reported high levels of PBDEs in animals and food resulting from the use of contaminated animal feed Public concern about PBDEs levels in animals and food has been raised. Feed contamination by toxic chemicals has been the cause of the contamination of poultry products. The purpose of this study was to evaluate PBDEs in pig feed to search the origin chase of POPs in pigs. Feed samples were obtained wheat from East Europe, corn from South America and America, soybean meal from Korea, America, South America and India and tallow from Korea. The preparation of samples was based on the EPA method 1614. Instrumental analysis was based on the use of high resolution gas chromatography coupled to high resolution mass spectrometry (HRGC/HRMS). Quantification was carried out by the isotopic dilution method. The analysis of <TEX>${\Sigma}PBDEs$</TEX> involved 22 PBDE congeners, including BDE-17, 28, 47, 49, 66, 71, 77, 85, 99, 100, 119, 126, 138, 153, 154, 156, 183, 184, 190, 191, 196 and 197.

돼지사료 중의 다이옥신 잔류 평가

ABSTRACTDioxins are chemical substances that are not properly decomposed degradation under natural photochemical, biological and chemical conditions in the environment, and bio-accumulate through the food chain, so pose a risk of causing adverse effects to human health and the environment. Ninety seven percent of human exposure to dioxins comes through meat, fish, dairy consumption and so on. Pork is the largest proportion meat aspect of daily intake in the Korean food. Pigs are three times more than Cattle in terms of breeding population and production amount in the world, and the amount is increasing every year. Therefore, it is necessary to monitor the exposure level of dioxins in pig’s body. Also, it is required to evaluate the presence of these chemical substances in the pig’s feed grain such as corn, soybeans, and tallow. The purpose of this study was to evaluate PCDD/Fs in pig's feed to search the origin of PCDD/Fs. Feed samples obtained were wheat from East Europe, corn from South America and America, soybean meal from Korea, America, South America and India and tallow from Korea. The preparation of samples was based on the EPA method 1613. Instrumental analysis was based on the use of high resolution gas chromatography coupled to high resolution mass spectrometry (HRGC/HRMS). The concentrations of 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) and 1,2,3,7,8-pentachlorodibenzo-p-dioxin (PCDD), which are the most toxic potent dioxins, were not observed in any of the samples. However OCDD was detected in most of the samples. The TEQs in the feed samples were ranged from 0 pg TEQ/g to 0.09841 pg TEQ/g. Especially, the high level of TEQs was observed in the samples of wheat from East Europe and soybean meal from Korea.Key words: PCDD/Fs, HRGC/HRMS, feed, EPA method 1613

A rapid method for screening of the Stockholm Convention POPs in small amounts of human plasma using SPE and HRGC/HRMS

A rapid analytical screening method allowing simultaneous analysis of 23 persistent organic pollutants (POPs) in human plasma was developed. Sample preparation based on solid-phase extraction (SPE) with additional clean-up using small multilayer silica gel columns. SPE was performed using a custom made polystyrene–divinylbenzene sorbent for the extraction of chlorinated and brominated POPs. Special efforts to reduce sample volume and improve speed and efficiency of the analytical procedure were made. Determination of 16 polychlorinated biphenyls (PCBs), 5 organochlorine (OC) pesticides, octachlorinated dibenzo-p-dioxin (OCDD) and polybrominated diphenyl ether (BDE #47) in 0.5mL human plasma was performed by using high resolution gas chromatography coupled to high resolution mass spectrometry (HRGC/HRMS). Recovery of POPs ranged between 46% and 110%, and reproducibility was below 25% relative standard deviation (RSD) for all target compounds, except for trans-nonachlor and OCDD, which were present only at low levels. Limits of detection (LOD) were for the PCBs between 0.8 and 117.7pgmL−1 plasma and for the OC pesticides between 5.9 and 89.1pgmL−1 plasma. The LOD for OCDD and BDE #47 were 1.4pgmL−1 plasma, and 9.2pgmL−1 plasma, respectively. The presented method was successfully applied to 1016 human plasma samples from an epidemiological study on cardiovascular disease.

Uncertainty assessment of polychlorinated dibenzo- p-dioxins and dibenzofuran and dioxin-like polychlorinated biphenyl analysis in stationary source sample emissions in accordance with the impending European standard EN-1948 using fly ashes

The analysis of polychlorinated dibenzo -p-dioxins and dibenzofurans (PCDD/Fs) and dioxin-like polychlorinated biphenyls (dl-PCBs) present in stack gas emissions and solid residues from incinerators will be mandatory in the foreseeable future. European standard EN-1948 is in the process of being updated through the addition of a new Part 4 related to the analysis of the 12 dl-PCBs. Therefore, either a comprehensive and reliable method capable of analyzing all of these 29 compounds (12 dl-PCBs and 17 2,3,7,8-PCDD/Fs) needs to be developed, or the existing PCDD/F analytical procedure must be adapted to include the dl-PCBs. This study has taken the latter approach of modifying PCDD/F methodology and in particular the fractionation step, by isolating dioxins and dl-PCBs into separate fractions ready for high resolution gas chromatography coupled to high resolution mass spectrometry (HRGC/HRMS) analysis. Results obtained from the analysis of Certified Reference Materials (CRM-490 and CRM-615) and fly ashes from the European Committee for Standardization (CEN) intercalibration study demonstrated that the proposed methodology is appropriate to determine the dl-PCBs in accordance with the impending European standard EN-1948. Uncertainty values obtained during the validation of the analytical methodology were 13% total I-TEQ (International Toxic Equivalent) for PCDD/Fs and 31% total WHO-TEQ (World Health Organization Toxic Equivalent) in the case of dl-PCBs. In addition, ‘real’ samples such as emissions and fly ashes were successfully analyzed following the proposed analytical method.

Baseline levels of dioxin and furan emissions from waste thermal treatment in Colombia

Background data of polychlorinated dibenzodioxin and dibenzofuran (PCDD/Fs) emissions from the incineration sector in Colombia are presented. Monitoring was carried out during a two-year period, 2003–2005. Twelve plants were sampled for stack gas emissions of dioxins and total solid particulate (TSP). Additionally, PCDD/Fs in several fly ash samples were analyzed. Most incinerators burned industrial refuse materials and medical residues. A wide range of PCDD/Fs emission levels were found. In particular, levels ranging from 6.9 to 343.8 ng I-TEQ/N m 3 were determined in plants without any air pollution control system (APCS). In contrast, 0.5–39.2 ng I-TEQ/N m 3 levels were found in plants with APCS while 8.5–67.5 ng I-TEQ/g were measured in fly ash samples. TSP values ranged from 14 to 448 mg/N m 3. This study also evaluated the impact of implementing different control systems in an incinerator. Finally, for comparison purposes several samples were analyzed by both high resolution gas chromatography coupled to high resolution mass spectrometry (HRGC-HRMS) and high resolution gas chromatography coupled to ion-trap low resolution mass spectrometry–mass spectrometry (HRGC-IT LRMS/MS). Overall, I-TEQ values deviated about 20–30% between both techniques.

Performances and limitations of the HRMS method for dioxins, furans and dioxin-like PCBs analysis in animal feedingstuffs: Part I: Results of an inter-laboratory study

The European strategy for dioxin monitoring of the food chain has defined high-resolution gas chromatography coupled to high-resolution mass spectrometry (HRGC/HRMS) method as the confirmatory method that can provide reliable and comparable results at sub-parts per trillion (ppt) level. This paper describes the first inter-laboratory study on dioxins, furans and dioxin-like PCBs by HRGC/HRMS method in animal feedingstuffs. Two different statistical approaches (ISO 5725 and Cofino’s statistics) were used for the statistical evaluation. For this particular study, the performances of the HRGC/HRMS method seem to be congener-independent in repeatability and reproducibility conditions over a concentration range covering more than four orders of magnitude. Results clearly show the effect of precision loss below 0.1 ppt level per congener in repeatability conditions and below 0.2 ppt level per congener in reproducibility conditions. LODs reported by the laboratories give median values of 0.02 ng/kg for most of the toxic congeners. Relative standard deviation between the laboratories’ mean values using upper-bound approach for TEQ calculation is 6.2%, more than twice the maximum level set at 0.75 ng TEQ/kg of product.

도심 지역의 다이옥신류 폴리염화비페닐류의 침적 플럭스

Atmospheric bulk (wet and dry) samples were monthly collected in an urban environment (Daeyeon-dong) of Busan over a year, to assess the deposition flux and seasonality of dioxin-like polychlorinated biphenyls (DLPCBs) using stainless steel pots. Deposition fluxes of DLPCBs in bulk samples were determined using high resolution gas chromatography coupled to high resolution mass spectrometry (HRGC/HRMS). Particle deposition fluxes in the urban environment varied from 23 to 98 <TEX>$mg^2$</TEX>/year (mean 41 <TEX>$gm^2$</TEX>/year). DLPCB deposition fluxes in atmospheric bulk samples ranged from 0.09 to 0.77 ng-<TEX>$TEQ/m^2$</TEX>/year (mean 0.35 ng-<TEX>$TEQ/m^2$</TEX>/year). Seasonal atmospheric deposition fluxes of DLPCBs were high in winter and low in summer. Atmospheric deposition fluxes of particles and DLPCBs in this study were comparable to or slightly lower values than those of different locations in the world. Monthly DLPCB profiles in deposition bulk samples were similar over a year. Non-ortho PCBs were higher contributions to the total DLPCBs fluxes than mono-ortho PCBs. In particular, PCB 126 had the highest concentrartion (>75%) in all deposition samples, followed by PCB 169 and PCB 156. A highly positive correlation was found among the deposition fluxes of DLPCB species, suggesting the possibility of that the DLPCB contamination originated from one source. The deposition fluxes of DLPCBs were not significantly correlated with temperature and the amount of precipitation even though the summer season with the highest temperature and the largest amount of precipitation showed the lowest DLPCB deposition flux.

Time and space patterns of volatile organic compounds in a sewage treatment plant

47 regulated and non-regulated volatile organic compounds (VOCs) were characterised by closed-loop stripping analysis (CLSA) and high resolution gas chromatography coupled to mass spectrometry (HRGC/MS) in 28 aqueous samples from 4 sampling points along a sewage treatment plant in Manresa, Catalonia, Spain. A 4×2 2 factorial design (16 samples) was first prepared for the sampling, and reinforced with 12 additional samples at the plant influent. The total analyte weighted mean concentration was 232 μg l −1 at the plant influent, with a mass flow of 2231 kg yr −1. Petroleum solvents and terpenic compounds accounted for 79% of the influent analyte concentration. VOC concentration in influent was clearly higher for most VOCs from 12 to 22 h (high organic load hours), and lower from 24 to 10 h (lower organic load). Differences between time bands were confirmed through t tests. Differences between weekdays and the weekend were not so clear, and could not be confirmed through t tests. VOC concentrations along the plant are discussed. Overall analyte removal in the plant was 89%.

Two xanthones from Polygala paniculata and confirmation of the 1-hydroxy-2,3,5-trimethoxy-xanthone at trace level by HRGC-MS.

Polygala paniculata L. yielded the xanthones 1-hydroxy-5-methoxy-2,3-methylenedioxyxanthone (1) and 1,5-dihydroxy-2,3-dimethoxyxanthone (2), together with coumarin murragatin and flavonol rutin. Their structures were established by chemical and spectroscopic methods (EIMS, IR, 1H and 13C NMR, NOE difference spectroscopy). By posterior analysis of an apolar crude extract using high resolution gas chromatography coupled to mass spectrometry (HRGC-MS) it was possible to characterize two sterol (spinasterol and delta25-spinasterol) and the minor 1-hydroxy-2,3,5-trimethoxyxanthone (3). Thus, the xanthone 3 was confirmed through of co-injection HRGC-MS of the respective extract with a certified standard obtained by methylation of 2 with diazomethane.

Flavonoids and trypanocidal activity of Bulgarian propolis

Acetone and ethanol extracts of two Bulgarian propolis samples ( Bur and Lov) were investigated by high temperature high resolution gas chromatography coupled to mass spectrometry (HT-HRGC-MS), and their activity against Trypanosoma cruzi was evaluated. The ethanol extracts— Et- Bur and Et- Lov—showed similar composition, with a high content of flavonoids, and strong inhibitory activity against T. cruzi proliferative epimastigotes, which were more susceptible than trypomastigotes. In the presence of blood, the activity of Et- Bur or Et- Lov against trypomastigotes was similar to that of the standard drug, crystal violet. Both extracts also showed similar and significant activity against Staphylococcus aureus and Candida albicans, while being inactive against Escherichia coli. The acetone extract, Ket- Bur, was more active than Et- Bur against both forms of T. cruzi.

HRGC-MS analysis of terpenoids from Maytenus ilicifolia and Maytenus aquifolium ("espinheira santa")

This work describes the identification of some of the minor chemical constituents of espinheira santa (Maytenus ilicifolia and Maytenus aquifolium, Celastraceae), a medicinal plant widely utilized in Brazil. By using high resolution gas chromatography coupled to mass spectrometry (HRGC-MS), it was possible to identify mainly triterpenoids and steroids in espinheira santa, most of them reported for the first time in both Maytenus species.