Determination of triclosan as its pentafluorobenzoyl ester in human plasma and milk using electron capture negative ionization mass spectrometry

A sensitive method for the determination of triclosan in plasma and milk is presented. Following hydrolysis of possible conjugates, triclosan is extracted with n-hexane/acetone, partitioned into alcoholic potassium hydroxide, and converted into its pentafluorobenzoyl ester. After sulfuric acid cleanup, sample extracts are analyzed by gas chromatography/electron capture negative ionization mass spectrometry. The limit of quantification was 0.009 ng/g for a 5-g plasma sample and 0.018 ng/g for a 3-g milk sample. The coefficient of variation for the method was 6%. The method was tested on more than 70 human plasma and milk samples, of which all plasma samples and more than half of the milk samples were above the limit of quantification. The presented method has lowered the limit of quantification for triclosan in human matrixes significantly as compared to previous methods and makes possible the analysis of triclosan in humans under normal exposure conditions.     

Determination of Pentachlorophenol and Its Oil Solvent in Wood Pole Samples by SFE and GC with Postcolumn Flow Splitting for Simultaneous Detection of the Species

An alternative approach is described for the measurement of pentachlorophenol (PCP) and its oil solvent in wood samples by supercritical fluid extraction (SFE) and gas chromatography (GC). The determination is achieved over a single chromatographic run using postcolumn flow splitting for simultaneous ECD/FID detection of the SFE extracted species. First, PCP and oil components are quantitatively extracted from a 0.3-g wood sample using 10% MeOH/CO(2) supercritical fluid at 0.65 g/mL and 120 °C. An aliquot of the SFE solution is then mixed with 10 mL of a buffered aqueous phase at pH 9.4. After PCP is acetylated by the addition of 500 μL of acetic anhydride, it is followed by its extraction with 2.00 mL of hexane along with oil. Then, 0.5 μL of supernatant organic phase is injected into the GC for a selective and simultaneous determination of the species. The method has a linear response over 3 orders of magnitude for both species with a linear regression correlation coefficient higher than 0.98 (95% confidence limit) and an absolute detection limit of 60 ng of PCP and 80 μg of oil per 0.1-g wood sample. The precision (relative standard deviation) is 4% for PCP and 1% for oil as established for a typical average concentration sample. The accuracy of the SFE GC-ECD/FID combined technique for PCP and oil was assessed by analyzing wood samples collected from newly and in-service PCP/oil-impregnated red pine poles.     

Use of solid-phase microextraction for the quantitative determination of herbicides in soil and water samples

An in-depth study of SPME optimization and application has been made, considering not only aqueous (surface water and groundwater samples) but also the more complex soil samples. Seven herbicides widely used in the area of study have been selected including five triazine herbicides (atrazine, simazine, terbumeton, terbuthylazine, terbutryn), molinate, and bromacil. linearity range was between 0.1 and 10 ng/mL and the repeatability below 10% when applying the optimized SPME procedure to water samples. Reproducibility was found to be lower than 20% at the 1 ng/mL level, and the limits of determination in environmental water samples using GC/MS (SIM mode) were well below 0.1 ng/mL (values ranging from 10 to 60 ng/L). Extraction of selected herbicides from soil was carried out by microwave-assisted solvent extraction using methanol in screw-capped vials, leading to recoveries over 80% in spiked soil samples at the 5-200 ng/g level. SPME application over methanolic soil extracts required a 10-fold dilution with distilled water. The recommended procedure was found to be fully applicable for quantitative determination of selected herbicides in soils containing low organic matter content with coefficients of variation below or around 10% and limits of determination ranging from 1 to 10 ng/g. Both procedures were applied to real-world surface water and soil samples where several pesticides were detected including atrazine, simazine, terbuthylazine, and molinate.     

Evaluation of a four-channel multiplexed electrospray triple quadrupole mass spectrometer for the simultaneous validation of LC/MS/MS methods in four different preclinical matrixes

A four-channel multiplexed electrospray interface on a triple quadrupole mass spectrometer was evaluated for the simultaneous validation of LC/MS/MS methods for the quantitation of loratadine and its metabolite, descarboethoxyloratadine, in four different biological matrixes. The assays were performed in rat, rabbit, mouse, and dog plasma from 1 to 1000 ng/mL using 96-well solid-phase extraction for sample preparation. The limit of quantitation of 1 ng/mL corresponded to 5.56 pg of each analyte injected on-column. For the drug, quality control samples (n = 6 at four concentrations) had precision ranging from 0.967 to 16.0% and accuracy ranging from -8.44 to 10.5% across all four species. For the metabolite, the precision ranged from 0.684 to 11.0% and the accuracy was between 6.36 and -9.06%. Intersprayer cross talk for the multiplexed electrospray ion source was evaluated as a function of analyte concentration and was less than 0.08% at concentrations as high as 1000 ng/mL. These results demonstrate the feasibility of using parallel analysis to reduce the time required for method validation and to increase sample throughput in drug development studies.     

Two trace analytical methods for determination of hydroxylated PCBs and other halogenated phenolic compounds in eggs from Norwegian birds of prey

Two new trace analytical methods are presented for identification and quantification of phenolic compounds in complex biological matrixes such as bird of prey eggs. One method is based on derivatization with methyl chloroformate prior to GC/high-resolution MS (HRMS) analysis in electron impact ionization mode. Alternatively, the underivatized phenolic analytes were separated and detected by HPLC coupled to time-of-flight MS (TOF-MS) in the negative ion electrospray ionization mode. For both methods, the egg samples were homogenized and dried with acidified sodium sulfate, cold column-extracted, and cleaned up by gel permeation chromatography and subsequently a Florisil column. Recovery rates for pentachlorophenol (PCP), tetrabromobisphenol A (TBBPA), and selected hydroxylated PCBs (HO-PCBs) from spiked hen's eggs (spiking level 1 ng/g of wet weight (ww)) were in the range of 56-98% for the HPLC/MS method and 57-108% for GC/MS including derivatization. Typical detection limits of the HPLC/TOF-MS method were 5 pg/g ww (1-2 pg injected) for HO-PCBs and PCP and 20 pg/g ww (3 pg injected) for TBBPA. The GC/HRMS method achieved detection limits of approximately 1 pg/g ww in predatory bird eggs for all analytes (0.2 pg injected for derivatized TBBPA and 0.05 pg injected for derivatized HO-PCBs and PCP). Eight eggs from four different Norwegian predatory bird species were analyzed. The concentrations determined with the two different quantification methods corresponded well with each other. PCP and TBBPA were found in all samples at concentrations up to 1350 and 13 pg/g ww, respectively (GC/HRMS values). A total of 55 penta- to nonachloro-HO-PCB congeners were detected in the eight eggs, 10 of those could be structurally identified. The maximum HO-PCB congener concentration was found for 4-HO-CB 187 in a peregrine falcon egg with estimated 388 pg/g ww. Another peregrine falcon egg was highest contaminated with sum HO-PCBs (estimated 2.1 ng/g ww). This level was 1.2 per thousand of the sum PCBs value for the same egg. Furthermore, indications were found that the HO-PCB congener distribution pattern could be species specific for predatory birds.     

Quantifying phthalate metabolites in human meconium and semen using automated off-line solid-phase extraction coupled with on-line SPE and isotope-dilution high-performance liquid chromatography--tandem mass spectrometry

We developed an analytical method using off-line solid-phase extraction (SPE) coupled with on-line SPE and isotope-dilution high-performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS) to determine the concentrations of phthalate metabolites in human meconium and in semen. First, we used off-line SPE to remove interfering proteins and other biomolecules from the samples. Then, we preconcentrated the phthalate metabolites in the extract using on-line SPE before measuring them by HPLC-MS/MS. For most of the analytes, the limits of detection ranged between 0.2 and 0.7 ng/g for meconium and between 0.3 and 0.7 ng/mL for semen. The recovery after off-line SPE varied for most analytes between 65 and 99% at concentrations ranging from 3.0 to 30.0 ng/mL in semen and between 67 and 103% at concentrations ranging from 2.0 to 10.0 ng/mL in meconium. Precision measured by the relative standard deviation ranged from 3.2 to 19.1% for intraday and from 3.9 to 18.6% for interday. We validated this novel approach--which is applicable to other biological matrixes, including serum and breast milk--on spiked samples and on five meconium samples and one pooled semen sample from people with no known occupational exposure to phthalates.     

Speciation of inorganic lead and ionic alkyllead compounds by GC/MS in prescreened rainwaters

A simple and novel screening method for lead compounds in environmental waters is proposed. The analytes, in an acetic medium, are sorbed on a C60 fullerene column as diethyldithiocarbamate complexes and subsequently eluted with isobutyl methyl ketone (IBMK), the lead being determined by flame atomic absorption spectrometry. The screening method acts as filter and indicates whether the target analytes are present above or below the detection limit of the method (0.5 ng/mL), giving no false positives. Positive samples were speciated by GC/MS, using a flow system similar to that of the screening method. Lead(II) and ionic di- and trialkyllead are derivatized with a Grignard reagent. Both methods use similar chemical and flow variables except for the eluent, IBMK in the screening method and n-hexane in the speciation method. The GC/MS speciation method is very sensitive; it exhibits a linear range of 0.02-5 ng/mL, and detection limits of 1-4 ng/L are achieved. Also, its repeatibility, as RSD, is less than 6%. The method was applied to the speciation of Pb2+ and di- and trialkyllead in positive prescreened rainwater samples from three different urban areas.     

Determination of low molecular weight silicones in plasma and blood of women after exposure to silicone breast implants by GC/MS

A sensitive, one-step sample preparation method for detection of volatile, low molecular weight (LMW) cyclic silicones hexamethylcyclotrisiloxane (D3), octamethyl-cyclotetrasiloxane (D4), decamethylcyclopentasiloxane (D5), and dodecamethylcyclohexasiloxane (D6) in plasma and blood using gas chromatography coupled with mass spectrometry (GC/MS, SIM mode) is presented. In spiked experiments, extraction efficiencies for these siloxanes (100-20 000 ng/mL) were approximately 90% for plasma and approximately 80% for blood; only in the case of D3 was the recovery very low. Plasma and blood of women who are or were exposed to silicone gel-filled implants and of control subjects were analyzed for low molecular weight silicones. D3-D6 were not detectable in control plasma or blood. Although the investigated numbers of patients samples are very limited, and thus, no statistical analysis is possible, our data clearly show a general increase in the amount of LMW cyclic siloxanes in the bodies of women with silicone implants. In particular, several years after ruptured silicone implants were removed, siloxanes could still be found in blood samples from several women. Siloxane compound D3 varied between 6 and 12 ng/mL (plasma) and between 20 and 28 ng/mL (blood), whereas the concentration range of D4 was 14-50 ng/mL (plasma) and 79-92 ng/mL (blood). D5 and D6, with one exception, could not be detected.     

Applications of Hadamard transform to gas chromatography/mass spectrometry and liquid chromatography/mass spectrometry

Successful application of the Hadamard transform (HT) technique to gas chromatography/mass spectrometry (GC/MS) and liquid chromatography/mass spectrometry (LC/MS) is described. Novel sample injection devices were developed to achieve multiple sample injections in both GC and LC instruments. Air pressure was controlled by an electromagnetic valve in GC, while a syringe pump and Tee connector were employed for the injection device in LC. Two well-known, abused drugs, 3,4-methylenedioxy-N-methylamphetamine (MDMA) and N, N-dimethyltryptamine (DMT), were employed as model samples. Both of the injection devices permitted precise successive injections, resulting in clearly modulated chromatograms encoded by Hadamard matrices. After inverse Hadamard transformation of the encoded chromatogram, the signal-to-noise (S/N) ratios of the signals were substantially improved compared with those expected from theoretical values. The S/N ratios were enhanced approximately 10-fold in HT-GC/MS and 6.8 in HT-LC/MS, using the matrices of 1023 and 511, respectively. The HT-GC/MS was successfully applied to the determination of MDMA in the urine sample of a suspect.     

Comparison of MALDI-TOF-MS and HPLC-ESI-MS/MS for Endopeptidase Activity-Based Quantification of Anthrax Lethal Factor in Serum

Diagnosing and treating anthrax at the earliest stage of disease is critical. We developed a method to diagnose anthrax at early stages of infection by detecting anthrax lethal factor (LF) at the attomol/mL level in plasma or serum. This method uses antibody capture and quantification of LF endoproteinase activity by isotope dilution matrix-assisted laser-desorption ionization (MALDI) time-of-flight (TOF) mass spectrometry (MS). Many public health laboratories do not use MALDI-TOF-MS; thus, we have adapted the LF method for detection by electrospray ionization (ESI) tandem MS (MS/MS), which allowed comparison of both MS platforms for LF quantification. Calibration curves were linear from 0.05-2.5 ng/mL when measured after 2 h and from 0.005-1.0 ng/mL after 18 h incubation time. The limit of detection was 0.005 ng/mL using a 200 μL sample. The coefficient of variation for quality control samples was 6-12% for both MS platforms. Samples used to perform cross-validation included 158 serum samples from a study in rabbits exposed to anthrax spores by inhalation. Some were treated with anthrax immune globulin before exposure. Concentrations measured by ESI-MS/MS matched those by MALDI-TOF-MS with p = 0.99 (r(2) = 0.997) and -0.25% mean relative difference (±9% standard deviation). This study shows that isotope dilution MALDI-TOF-MS is a robust and precise quantitative MS platform.     

A device for automated direct sampling and quantitation from solid-phase sorbent extraction cards by electrospray tandem mass spectrometry

A new solid-phase extraction (SPE) device in the 96-well format (SPE Card) has been employed for automated off-line sample preparation of low-volume urine samples. On-line automated analyte elution via SPE and direct quantitation by micro ion spray mass spectrometry is reported. This sample preparation device has the format of a microtiter plate and is molded in a plastic frame which houses 96 separate sandwiched 3M Empore sorbents (0.5-mm-thickness, 8-microm particles) covered on both sides by a microfiber support material. Ninety-six discrete SPE zones, each 7 mm in diameter, are imbedded into the sheet in the conventional 9-mm pitch (spacing) of a 96-well microtiter plate. In this study one-quarter of an SPE Card (24 individual zones) was used merely as a convenience. After automated off-line interference elution of applied human urine from 24 samples, a section of SPE Card is mounted vertically on a computer-controlled X, Y, Z positioner in front of a micro ion spray direct sampling tube equipped with a beveled tip. The beveled tip of this needle robotically penetrates each SPE elution zone (sorbent disk) or stationary phase in a serial fashion. The eluted analytes are sequentially transferred directly to a microelectrosprayer to obtain tandem mass spectrometric (MS/MS) analysis. This strategy precludes any HPLC separation and the associated method development. The quantitative determination of Ritalin (methylphenidate) from fortified human urine samples is demonstrated. A trideuterated internal standard of methylphenidate was used to obtain ion current response ratios between the parent drug and the internal standard. Human control urine samples fortified from 6.6 to 3300 ng/mL (normal therapeutic levels have been determined in other studies to be between 50 and 100 ng/mL urine) were analyzed and a linear calibration curve was obtained with a correlation coefficient of 0.9999, where the precision of the quality control (QC) samples ranged from 9.6% at the 24 ng/mL QC level to 1.2% at the 3000 ng/mL QC level, and the accuracy for the four levels of QC samples ranged from 98.1% to 100.3%. The QC samples were prepared at four concentrations which included 24, 240, 1200, and 3000 ng/mL, respectively. The run time per sample in this work was 1.5 min not including the sample preparation time.     

顶空气相色谱/质谱法分析大豆油中正己醛含量

建立大豆油中正己醛含量的顶空气质分析方法。将大豆油试样放入密封的气化瓶中,在55℃温度下,使正己醛气化达到平衡时,取液上气体注入GC/MS,经非极性毛细管柱HP-5ms分离,质谱选择离子监测对正己醛进行定量测定。该方法在0.05~1.00μg/mL范围内线性关系良好,相关系数0.9997;最小检测限为3.15ng/mL,方法回收率在90.0~107%之间,相对标准偏差3.4%。该方法前处理简单、快速,定量准确,灵敏度高,可用于实验室检测大豆油中正己醛含量。     

厦门港和筼筜湖沉积物中多溴联苯醚类防火剂(PBDEs)的色谱-质谱联机同位素稀释法测定

利用^13C标记的多溴联苯醚标准作内标对厦门沿岸海域和筼筜湖沉积物进行色谱-质谱联机测定并采用同位索稀释法进行数据收集和计算,分别测定PBDE3、15、28、47、60、85、99、100、138、153、154和183共12个同类化合物,其回收率范围为75．8—100．5％,最低检测限0．02ng／g（d）,沉积物中总PBDE最高值为厦门第一码头2．06ng／g（d）,最低是厦门大学海水浴场为0．10ng／g（d）。     

Species-specific GC/ICP-IDMS for trimethyllead determinations in biological and environmental samples

An accurate and sensitive species-specific isotope dilution GC/ICPMS method was developed for the determination of trimethyllead (Me3Pb+) in biological and environmental samples. A trimethyllead spike was synthesized from 206Pb-enriched metallic lead by reaction of lead halide with methyllithium and subsequent formation of trimethyllead iodide. The isotopic composition of the spike solution was determined by GC/ICPMS after derivatization with tetraethylborate, and its concentration was determined by reverse isotope dilution analysis. The species-specific GC/ICP-IDMS method was validated by reference material CRM 605 (urban dust) certified for Me3Pb+. The method was also applied to determine the Me3Pb+ content in six biological reference materials (DORM 2, CRM 278, CRM 422, CRM 463, CRM 477, MURST-ISS-A2) and one sediment reference material (CRM 580) for which no certified values of this species exist. The Me3Pb+ concentrations in the biological reference materials vary in the range of 0.3-17 ng g(-1) (as Pb) except for the Antarctic Krill (MURST-ISS-A2), where the concentration was less than the detection limit of 0.09 ng g(-1), which was also found for the sediment. Up to 20% of total lead was methylated in the biological reference materials, whereas much higher methylation fractions were found for mercury. The method was also applied to seafood samples purchased from a supermarket with Me3Pb+ concentrations in the limited range of 0.3-0.7 ng g(-1). On the contrary, the portion of methylated lead in these samples varied over more than 2 orders of magnitude from 0.02 to 7.5%.     

厦门港和筼筜湖沉积物中多溴联苯醚类防火剂(PBDEs)的色谱一质谱联机同位素稀释法测定

利用13C标记的多溴联苯醚标准作内标对厦门沿岸海域和筼筜湖沉积物进行色谱—质谱联机测定并采用同位素稀释法进行数据收集和计算,分别测定PBDE3、15、28、47、60、85、99、100、138、153、154和183共12个同类化合物,其回收率范围为75.8-100.5%,最低检测限0.02ng/g(d),沉积物中总PBDE最高值为厦门第一码头2.06ng/g(d),最低是厦门大学海水浴场为0.10ng/g(d)。     

An analytical method for trifluoroacetic Acid in water and air samples using headspace gas chromatographic determination of the methyl ester

An analytical method has been developed for the determination of trace levels of trifluoroacetic acid (TFA), an atmospheric breakdown product of several of the hydrofluorocarbon (HFC) and hydrochlorofluorocarbon (HCFC) replacements for the chlorofluorocarbon (CFC) refrigerants, in water and air. TFA is derivatized to the volatile methyl trifluoroacetate (MTFA) and determined by automated headspace gas chromatography (HSGC) with electron-capture detection or manual HSGC using GC/MS in the selected ion monitoring (SIM) mode. The method is based on the reaction of an aqueous sample containing TFA with dimethyl sulfate (DMS) in concentrated sulfuric acid in a sealed headspace vial under conditions favoring distribution of MTFA to the vapor phase. Water samples are prepared by evaporative concentration, during which TFA is retained as the anion, followed by extraction with diethyl ether of the acidified sample and then back-extraction of TFA (as the anion) in aqueous bicarbonate solution. The extraction step is required for samples with a relatively high background of other salts and organic materials. Air samples are collected in sodium bicarbonate-glycerin-coated glass denuder tubes and prepared by rinsing the denuder contents with water to form an aqueous sample for derivatization and analysis. Recoveries of TFA from spiked water, with and without evaporative concentration, and from spiked air were quantitative, with estimated detection limits of 10 ng/mL (unconcentrated) and 25 pg/mL (concentrated 250 mL:1 mL) for water and 1 ng/m(3) (72 h at 5 L/min) for air. Several environmental air, fogwater, rainwater, and surface water samples were successfully analyzed; many showed the presence of TFA.     

Comparison of three coupled gas chromatographic detectors (MS, MIP-AES, ICP-TOFMS) for organolead speciation analysis

An automatic unit for the screening of rainwater is used for the determination of organolead compounds using different detectors coupled to a gas chromatograph. A systematic overview is given of the advantages and disadvantages of several detectors (electron ionization mass spectrometry, EI-MS; microwave induced plasma atomic emission spectrometry, MIP-AES; and inductively coupled plasma time-of-flight mass spectrometry, ICP-TOFMS, for the speciation of organolead compounds on the basis of sensitivity, selectivity and reliability. C60 fullerene and RP-C18 were used as sorbent materials for these compounds. The primary assets of the fullerene sorbent, as compared to C18 sorbent, are high sensitivity and selectivity resulting from efficient adsorption due to large surface area and interstitial volume. Among the detection systems, GC/ ICP-TOFMS is the most sensitive, with absolute detection limits of approximately 15 fg of organolead compounds (as lead) using 5-mL sample volumes. Except for diethyllead, similar sensitivities were obtained by MIP-AES. GC/MS is intrinsically the most specific option, because the species are detected directly from molecular information. The precision is similar for all detectors. The screening of rainwater from different locations showed that samples collected in countries in which leaded gasolines are now banned contain organolead species at concentrations below 2 pg/ mL, levels that can be detected only for sample volumes of 25 mL and using MIP-AES or ICP-TOFMS as detectors, their determination being impossible by GC/MS.     

液液萃取-GC/MS法分析地表水中的多氯联苯

建立了液液萃取-GC/MS联用分析地表水中的多氯联苯的方法,并对实际水样进行了测定。各目标物的线性回归R值均大于0.999,MDL为0.08-3.9ng/L,RSD为0.25%-6.2%,回收率为87.6%-113%。     

A 384-well solid-phase extraction for LC/MS/MS determination of methotrexate and its 7-hydroxy metabolite in human urine and plasma

A solid-phase extraction procedure, in a 384-well format, has been developed for methotrexate and its primary metabolite, 7-hydroxymethotrexate, in human urine and plasma. This format has not been utilized previously for solid-phase extraction of drugs from biological fluids. The 384-well plates contained a C-18 stationary phase bonded to silica particles which are incorporated into a glass-fiber membrane. Methotrexate and 7-hydroxymethotrexate have been quantified across the curve range of 1 to 50 microg/mL and 50 to 1000 ng/mL, respectively, in urine and from 5 to 250 ng/mL and 5 to 100 ng/mL, respectively, in plasma. Both analytes are quantified by linear regression using 20-microL sample aliquots. Experiments to evaluate the influence of particle size, elution volume, and injection volume on signal intensity were conducted and are reported, along with the results of experiments examining cross contamination between wells. Recovery was determined to be > or = 95% from urine. Results from a run of 384 samples analyzed over a 14-h period indicate that 384-well SPE can be successfully utilized to increase analytical run sizes and sample throughput for LC/MS/MS determination of small drug molecules in biological samples.     

Simultaneous measurement of urinary bisphenol A and alkylphenols by automated solid-phase extractive derivatization gas chromatography/mass spectrometry

Bisphenol A (BPA) and alkylphenols (APs) are widely used industrial chemicals. BPA is used to manufacture polycarbonate plastic and epoxy resins; APs are used to make alkylphenol ethoxylates, common nonionic surfactants. BPA and APs can leach into the environment during industrial production and after degradation of the polycarbonate plastics and nonionic surfactants. Environmental exposure to these phenolic compounds has been associated with adverse reproductive and developmental effects in wildlife. We developed a sensitive and robust method for measuring BPA and six APs; 3-tert-butylphenol, 4-tert-butylphenol, 4-n-octylphenol, 4-tert-octylphenol, 4-n-nonylphenol, and technical-grade nonylphenol in urine. The method is based on the use of automated solid-phase extraction (SPE) coupled to isotope dilution-gas chromatography/mass spectrometry (GC/MS). During the automated SPE process, the phenols are both extracted from the urine matrix and derivatized, using pentafluorobenzyl bromide, on commercially available styrene-divinylbenzene copolymer-based SPE cartridges. After elution from the SPE column, the derivatized phenols in the SPE eluate are analyzed by GC/MS. The method, validated on spiked pooled urine samples and on urine samples from exposed persons, has limits of detection of approximately 0.1 ng in 1 mL of urine.