An international round-robin study for the analysis of particulate semi-volatile organics by thermal desorption gas chromatography mass spectrometry

Thermal desorption gas chromatography mass spectrometry (TD-GC/MS) is becoming more commonly used for the quantification and identification of organic compounds in particulate matter (PM), including ambient and source PM such as diesel particulate matter (DPM). It has been proven as an alternative to the traditional solvent extraction (SE) method and liquid injection gas chromatograph mass spectrometry (LI-GC/MS). However, little information is available on how different types of TD-GC/MS systems compare to each other for analysis of real-world PM samples or to direct LI-GC/MS for analysis of PM components in a test solution. To address this, CanmetENERGY Characterization Laboratory initiated a round robin with the participation of 10 laboratories worldwide. Three sample types were analysed: (i) a test solution with a suite of pure compounds commonly found in PM, analysed by TD-GC/MS and LI-GC/MS; (ii) a DPM sample, analysed by TD-GC/MS and SE; and (iii) an ambient PM sample, analysed by TD-GC/MS. The first part of the study showed good overall performance and comparability between the different TD-GC/MS systems and LI-GC/MS method for the analysis of PM components in a test solution, with some variability of results due to system types and parameters used, concentration of calibration standards, and whether or not an internal standards was used. The analysis of the DPM sample showed greater variability between laboratories and methods as many PM components were present near the detection limit and matrix effects particularly affected the TD-GC/MS analysis of heavier n-alkanes. In the last part of the study, for the analysis of an ambient PM sample by TD-GC/MS, the analysis of variance showed good comparison between labs for polycyclic aromatic hydrocarbons (94% non-significant), but slightly lower for n-alkanes (68%) and biomarkers (57%).     

Determination of nitrated polycyclic aromatic hydrocarbons in diesel particulate-related standard reference materials by using gas chromatography/mass spectrometry with negative ion chemical ionization

Gas chromatography/mass spectrometry (GC/MS) with negative ion chemical ionization (NICI) detection was utilized for quantitative determination of nitrated polycyclic aromatic hydrocarbons (nitro-PAHs) in diesel particulate-related standard reference materials (SRMs). Prior to GC/MS analysis, isolation of the nitro-PAHs from the complex diesel particulate extract was accomplished using solid phase extraction (SPE) and normal-phase liquid chromatographic (LC) fractionation using an amino/cyano stationary phase. Concentrations of eight to ten mononitro-PAHs and three dinitropyrenes were determined in three diesel particulate-related SRMs: SRM 1650a Diesel Particulate Matter, SRM 1975 Diesel Particulate Extract, and SRM 2975 Diesel Particulate Matter (Industrial Forklift). The results from GC/MS NICI using two different columns (5% phenyl methylpolysiloxane and 50% phenyl methylpolysiloxane) were compared to each other and to results from two other laboratories for selected nitro-PAHs. 1-Nitropyrene was the most abundant nitro-PAHs in each of the diesel particulate SRMs (19.8+/-1.1 micro g g(-1) particle in SRM 1650a and 33.1+/-0.6 micro g g(-1) particle in SRM 2975). Three dinitropyrene isomers were measured in SRM 1975 at 0.5-1.4 micro g g(-1) extract and in SRM 2975 at 1-3 micro g g(-1) particle.     

Simple and sensitive method for determination of nitrated polycyclic aromatic hydrocarbons in diesel exhaust particles by gas chromatography-negative ion chemical ionisation tandem mass spectrometry

An extremely simple and sensitive method was developed for determination of nitrated polycyclic aromatic hydrocarbons (nitro-PAHs; mono-nitro-PAHs and dinitropyrenes) in diesel exhaust particles (DEPs) by gas chromatography-negative ion chemical ionisation tandem mass spectrometry (GC/NCI/MS/MS). We used two types of column in GC/NCI/MS/MS analysis. A polar column was used for determination of mono-nitro-PAHs, and a non-polar column was used for determination of dinitropyrenes and mono-nitro-PAHs except nitrofluoranthenes. The proposed method requires no clean-up procedure. The limits of detection ranged from 0.01 to 0.09 pg for all compounds tested. The applicability of the method to DEP samples was validated using diesel particulate standard reference materials (SRMs). Although DEPs contain complex matrices, all compounds could be detected easily in SRM2975 (diesel particulate matter) and SRM1975 (diesel particulate extract) without a clean-up procedure. The RSDs were less than 5% for all compounds examined. The quantitative results for SRMs exhibited good agreement with the available data in the literature. These results indicate that the proposed GC/NCI/MS/MS method is useful for determination of nitro-PAHs in DEP samples.     

Determination of 1-nitropyrene in low volume ambient air samples by high-performance liquid chromatography with fluorescence detection

To measure the actual exposure of a person to 1-nitropyrene (1-NP) in airborne particulate matter, it is considered more accurate to collect air samples with a portable air sampler than to sample at a fixed location. However, because the portable samplers can sample only small volumes, a sensitive method is needed to analyze the compounds that are collected on a filter. Here we describe a high-performance liquid chromatographic (HPLC) method with fluorescence detection that is sensitive and precise enough for use with portable air samplers. The developed column-switching system successfully removed the interfering substances in the samples with only a simple pretreatment. To improve the precision of the measurement, deuterated 1-NP was used as an internal standard, and it eluted immediately prior to 1-NP with sufficient resolution (R_s, 1.50). The detection limit was 0.32 fmol/injection, and the calibration range was from 2 to 100 fmol. The proposed method was applied to determining 1-NP in fine airborne particulate matter (PM_2.5) at two sites with low pollution levels. 1-NP was detected in all samples at concentrations in the low fmol/m³ range. The proposed method has enough sensitivity and precision to determine 1-NP in the limited air volume of the portable sampler.     

An automatic method for the determination of carcinogenic 1-nitropyrene in extracts from automobile exhaust particulate matter

An automatic method for the determination of carcinogenic 1-nitropyrene in extracts from automobile exhaust particulate matter has been developed. Organic matter was extracted from particulate matter and the extract was concentrated. Then the extract was injected into a two-dimensional (2D) high-performance liquid chromatograph (HPLC) with a fluorescence detection system. To improve the sensitivity a large sample volume (300 microl) was applied to the HPLC system using the double on-column focusing technique. To achieve full automation and high selectivity a reduction column packed with platinum black was introduced after the 2D HPLC system using a C18 column as the primary column and a pentabromobenzyl column as the secondary column. This HPLC system could determine 0.01-3,000 ng ml(-1) of 1-nitropyrene in an extract from diesel and gasoline exhaust particulate matter at 40 min intervals.     

Analytical approaches for arsenic determination in air: A critical review

This review describes the different steps involved in the determination of arsenic in air, considering the particulate matter (PM) and the gaseous phase. The review focuses on sampling, sample preparation and instrumental analytical techniques for both total arsenic determination and speciation analysis. The origin, concentration and legislation concerning arsenic in ambient air are also considered. The review intends to describe the procedures for sample collection of total suspended particles (TSP) or particles with a certain diameter expressed in microns (e.g. PM10 and PM2.5), or the collection of the gaseous phase containing gaseous arsenic species. Sample digestion of the collecting media for PM is described, indicating proposed and established procedures that use acids or mixtures of acids aided with different heating procedures. The detection techniques are summarized and compared (ICP-MS, ICP-OES and ET-AAS), as well those techniques capable of direct analysis of the solid sample (PIXE, INAA and XRF). The studies about speciation in PM are also discussed, considering the initial works that employed a cold trap in combination with atomic spectroscopy detectors, or the more recent studies based on chromatography (GC or HPLC) combined with atomic or mass detectors (AFS, ICP-MS and MS). Further trends and challenges about determination of As in air are also addressed.     

Determination of paraquat and diquat in human body fluids by high-performance liquid chromatography/tandem mass spectrometry

Paraquat (PQ) and diquat (DQ) in human whole blood and urine were analyzed by high-performance liquid chromatography/tandem mass spectrometry (HPLC/MS/MS) with positive ion electrospray ionization (ESI). The compounds were extracted with Sep-Pak C18 cartridges from whole blood and urine samples containing ethyl paraquat as an internal standard. The separation of PQ and DQ was carried out using ion-pair chromatography with heptafluorobutyric acid in 20 mM ammonium acetate and acetonitrile gradient elution for successful coupling with MS. Both compounds formed base peaks due to [M-H]+ ions by HPLC/ESI-MS and the product ions produced from each [M-H]+ ion by HPLC/MS/MS. Selective reaction monitoring (SRM) showed much higher sensitivity for both body fluids. Therefore, a detailed procedure for the detection of compounds by SRM with HPLC/MS/MS was established and carefully validated. The recoveries of PQ and DQ were 80.8-95.4% for whole blood and 84.2-96.7% for urine. The calibration curves for PQ and DQ showed excellent linearity in the range of 25-400 ng ml(-1) of whole blood and urine. The detection limits were 10 ng ml(-1) for PQ and 5 ng ml(-1) for DQ in both body fluids. The intra- and inter-day precision for both compounds in whole blood and urine samples were not greater than 13.0%. The data obtained from the determination of PQ and DQ in rat blood after oral administration of the compounds are also presented.     

Development of a high performance liquid chromatography method and a liquid chromatography-tandem mass spectrometry method with pressurized liquid extraction for simultaneous quantification and confirmation of cyromazine, melamine and its metabolites in foods of animal origin

Simple and sensitive methods have been developed for simultaneous detection of cyromazine, melamine and their metabolites (ammeline, ammelide and cyanuric acid) in samples of animal origins. These include a high performance liquid chromatography (HPLC) method and a liquid chromatography-tandem mass spectrometry (LC-MS/MS) method and are useful in regular monitoring and in toxicity studies of these molecules. Representative samples used in this study include muscles and livers of swine, bovine, sheep and chicken, kidneys of swine, bovine and sheep, and milk powder. A new sample preparation procedure with pressurized liquid extraction (PLE) at 1400psi and 70°C was investigated. Quantification of these five compounds by HPLC was achieved using an APS-2 column with UV detection at 230 nm. Limit of detection (LOD) was at 10 μgkg(-1), and limit of quantification (LOQ) was at 40 μgkg(-1). Recoveries of the five analytes in spiked samples ranged from 72.2% to 115.4% with RSD less than 12%. Confirmatory analysis of the analytes was performed using LC-MS/MS in selected reaction monitoring (SRM) mode. The LOD and LOQ were 5 μgkg(-1) and 15 μgkg(-1), respectively. This is the first simultaneous analysis of cyromazine, melamine, ammeline, ammelide and cyanuric acid residues in complex tissue samples using PLE and HPLC. It is expected that these methods will find many practical applications in evaluating the safety of cyromazine, melamine and their metabolites.     

The rapid identification of drug metabolites using capillary liquid chromatography coupled to an ion trap mass spectrometer

Capillary liquid chromatography (LC) using a 320 microns column and a flow rate of 10 microL/min has been coupled to an ion trap mass spectrometer using electrospray ionisation (ESI) to enable the rapid and effective identification of metabolites in urine, following oral administration of a novel human neutrophil elastase inhibitor, GW311616. Metabolites were identified from their mass (MS) spectra and tandem (MS/MS) mass spectra using minimal sample (1 microL of urine) and no sample pretreatment. Sensitivity assessment has shown that both molecular weight and structural information is obtainable on as little as 5 pg of compound, making the capillary LC/ion trap system as described an ideal analytical tool for the detection and characterisation of low level metabolites in biofluids (particularly when sample volume is limited). This level of detection was unattainable using a triple quadrupole mass spectrometer operating in full-scan mode, although 200 fg on column was detected using selected reaction monitoring target analysis.     

Determination of ultra-trace levels of gold in size-segregated atmospheric particulate samples by laser induced fluorescence: towards an aerosol tracer

A method is described for measuring the gold content of integral and size-segregated samples of atmospheric particulate matter. After acid digestion and a liquid/ liquid extraction, the sample are analysed by Two-Colour Laser Induced Fluorescence in a graphite furnace. An absolute instrumental detection limit of 1 fg is achieved. Assuming a sampled volume of 1 m(3), this corresponds to an atmospheric concentration of 50 fg m(-3). Due to blank limited noise, the above limits increase to 20 fg and 1 pg m(-3), respectively. Results of the analysis of filter samples as well as of size-segregated impactor samples are presented.     

Determination of polycyclic aromatic hydrocarbons from ambient air particulate matter using a cold fiber solid phase microextraction gas chromatography-mass spectrometry method

Polycyclic aromatic hydrocarbons (PAH) from ambient air particulate matter (PM) were analyzed by a new method that utilized direct immersion (DI) and cold fiber (CF) SPME-GC/MS. Experimental design was used to optimize the conditions of extraction by DI-CF-SPME with a 100μm polydimethylsiloxane (PDMS) fiber. The optimal conditions included a 5min equilibration at 70°C time in an ultrasonic bath with an extraction time of 60min. The optimized method was validated by the analysis of a NIST standard reference material (SRM), 1649b urban dust. The results obtained were in good agreement with certified values. PAH recoveries for reference materials were between 88 and 98%, with a relative standard deviation ranging from 5 to 17%. Detection limits (LOD) varied from 0.02 to 1.16ng and the quantification limits (LOQ) varied from 0.05 to 3.86ng. The optimized and validated method was applied to the determination of PAH from real particulate matter (PM10) and total suspended particulate (TPS) samples collected on quartz fiber filters with high volume samplers.     

Determination of bromethalin in commercial rodenticides found in consumer product samples by HPLC-UV-vis spectrophotometry and HPLC-negative-ion APCI-MS

A small amount of green particulate material is encountered in a consumer complaint sample. The green particulates in the sample are identified as a bromethalin-containing rodenticide using high-performance liquid chromatographic (HPLC)-UV-vis spectrophotometric and HPLC-negative-ion atmospheric pressure chemical ionization (APCI)-mass spectrometric (MS) approaches, which are commonly used for the detection and confirmation of bromethalin in grain-based rodenticides. The selective and sensitive nature of the MS detector makes it possible to determine bromethalin without extensive sample cleanup and preconcentration. The estimated detection limit with the UV-vis detector is 500 pg of bromethalin injected into the column. The extensive fragmentation of the bromethalin molecule under APCI conditions provides sufficient structural information for positive identification.     

Effect of ionization suppression by trace impurities in mobile phase water on the accuracy of quantification by high‐performance liquid chromatography/mass spectrometry

The high‐performance liquid chromatography (HPLC) column is capable of enrichment/pre‐concentration of trace impurities in the mobile phase during the column equilibration, prior to sample injection and elution. These impurities elute during gradient elution and result in significant chromatographic peaks. Three types of purified water were tested for their impurity levels, and hence their performances as mobile phase, in HPLC followed by total ion current (TIC) mode of MS. Two types of HPLC‐grade water produced 3–4 significant peaks in solvent blanks while LC/MS‐grade water produced no peaks (although peaks were produced by LC/MS‐grade water also after a few days of standing). None of the three waters produced peaks in HPLC followed by UV‐Vis detection. These peaks, if co‐eluted with analyte, are capable of suppressing or enhancing the analyte signal in a MS detector. As it is not common practice to run solvent blanks in TIC mode, when quantification is commonly carried out using single ion monitoring (SIM) or single or multiple reaction monitoring (SRM or MRM), the effect of co‐eluting impurities on the analyte signal and hence on the accuracy of the results is often unknown to the analyst. Running solvent blanks in TIC mode, regardless of the MS mode used for quantification, is essential in order to detect this problem and to take subsequent precautions. Copyright © 2010 John Wiley & Sons, Ltd.     

Evaluation of an in-injection port thermal desorption-gas chromatography/mass spectrometry method for analysis of non-polar organic compounds in ambient aerosol samples

Thermal desorption coupled with gas chromatography/mass spectrometry (TD-GC/MS) is an alternative to solvent extraction (SE)-based GC/MS (SE-GC/MS) for the analysis of non-polar organic compounds in filter or impactor-collected aerosols. TD-GC/MS has no sample pretreatment and requires a small filter aliquot for detecting individual organic compounds. The performance of an in-injection port TD-GC/MS is evaluated for polycyclic aromatic hydrocarbons (PAHs), n-alkanes, iso-/anteiso-alkanes, hopanes, steranes, branched alkanes, cyclohexanes, alkenes, and phthalates in standards and ambient air samples. Replicate analysis for 132 organic compounds showed relative standard deviations <10%, with the majority <5%. Accuracy for 15 PAHs, determined with NIST standard reference material (SRM) 1649a urban dust, was within +/-5% of the certified values. TD-GC/MS and SE-GC/MS method comparisons for 14 Hong Kong ambient samples agreed within 11% for 106 non-polar compounds. For 19 Tong Liang, China samples, agreement was within 13% for 23 PAHs.     

Ternary-column system for high-throughput direct-injection bioanalysis by liquid chromatography/tandem mass spectrometry

As a continuation of our efforts to improve our high-flow on-line bioanalytical approach for high-throughput quantitation of drugs and metabolites in biological matrices by high-performance liquid chromatography (LC) and tandem mass spectrometry (MS/MS), we have developed a ternary-column on-line LC/MS/MS system with dual extraction columns used in parallel for purification and an analytical column for analysis. The advantage of the dual extraction column system is that sample analysis can take place in one of the extraction columns while the other column is being equilibrated. Thus, the equilibration time does not add to the run time, hence shortening the injection cycle time and increasing the sample throughput. Moreover, the use of two extraction columns in parallel increases the number of samples that can be injected before the system fails due to an overused extraction column. Such a system has successfully been used to develop and validate a positive ion electrospray LC/MS/MS bioanalytical method for the quantitative determination of a guanidine-containing drug candidate in rat plasma. The system used for this work utilized two Oasis HLB extraction columns (1 x 50 mm, 30 microm), one C18 analytical column (3.9 x 50 mm, 5 microm), a ten-port switching value and a tandem mass spectrometer. The on-line analysis was accomplished by the direct injection of 10 microL of the sample, obtained by mixing a rat plasma sample 1:1 with an aqueous internal standard solution. Selected reaction monitoring (SRM) was utilized for the detection of the analyte and internal standard. The standard curve range was 1.00-200 ng/mL. The intra- and inter-day precision and accuracy were within 6.6%. The on-line purification step lasted for only 0.3 min and total run time was only 1.6 min.     

Detection of monohydroxylated polycyclic aromatic hydrocarbons in urine and particulate matter using LC separations coupled with integrated SPE and fluorescence detection or coupled with high‐resolution time‐of‐flight mass spectrometry

A high‐performance liquid chromatographic (HPLC) method with integrated solid‐phase extraction for the determination of 1‐hydroxypyrene and 1‐, 2‐, 3‐, 4‐ and 9‐hydroxyphenanthrene in urine was developed and validated. After enzymatic treatment and centrifugation of 500 μL urine, 100 μL of the sample was directly injected into the HPLC system. Integrated solid‐phase extraction was performed on a selective, copper phthalocyanine modified packing material. Subsequent chromatographic separation was achieved on a pentafluorophenyl core–shell column using a methanol gradient. For quantification, time‐programmed fluorescence detection was used. Matrix‐dependent recoveries were between 94.8 and 102.4%, repeatability and reproducibility ranged from 2.2 to 17.9% and detection limits lay between 2.6 and 13.6 ng/L urine. A set of 16 samples from normally exposed adults was analyzed using this HPLC‐fluorescence detection method. Results were comparable with those reported in other studies. The chromatographic separation of the method was transferred to an ultra‐high‐performance liquid chromatography pentafluorophenyl core–shell column and coupled to a high‐resolution time‐of‐flight mass spectrometer (HR‐TOF‐MS). The resulting method was used to demonstrate the applicability of LC‐HR‐TOF‐MS for simultaneous target and suspect screening of monohydroxylated polycyclic aromatic hydrocarbons in extracts of urine and particulate matter.     

Determination of phenothiazines in human body fluids by solid-phase microextraction and liquid chromatography/tandem mass spectrometry

Eleven phenothiazine derivatives with heavy side-chains were found to be extractable from human whole blood and urine samples by solid-phase microextraction (SPME) with a polyacrylate-coated fiber. The fiber was then injected into the desorption chamber of an SPME-liquid chromatography (LC) interface for LC/tandem mass spectrometry (MS/MS) with positive ion electrospray (ES) ionization. All compounds formed base peaks due to [M + 1](+) ions by LC/ES-MS/MS. By use of LC/ES-MS/MS, the product ions produced from each [M + 1](+) ion showed base peaks due to side-chain liberation. Selected reaction monitoring (SRM) and selected ion monitoring (SIM) were compared for the detection of the 11 phenothiazine derivatives from human whole blood and urine. SRM showed much higher sensitivity than SIM for both types of sample. Therefore, a detailed procedure for the detection of drugs by SRM with SPME-LC/MS/MS was established and carefully validated. The extraction efficiencies of the 11 phenothiazine derivatives spiked into whole blood and urine were 0. 0002-0.12 and 2.6-39.8%, respectively. The regression equations for the 11 phenothiazine derivatives showed excellent linearity with detection limits of 0.2-200 ng ml(-1) for whole blood and 4-22 pg ml(-1) for urine. The intra- and inter-day precisions for whole blood and urine samples were not greater than 15.1%. The data obtained after oral administration of perazine or flupentixol to a male subject are presented.     

A new high-performance liquid chromatographic/electrospray ionization tandem mass spectrometric method for the simultaneous determination of cyclophosphamide, methotrexate and 5-fluorouracil as markers of surface contamination for occupational exposure monitoring

A new high-performance liquid chromatographic/electrospray ionization tandem mass spectrometric (HPLC/ESI-MS/MS) method was developed for the simultaneous quantification of 5-fluorouracil (5FU), methotrexate (MTX) and cyclophosphamide (CP) in environmental samples. These compounds, commonly used in the treatment of cancer, are recognized as genotoxic. In order to estimate the occupational exposure of hospital personnel handling these drugs, wipe samples were taken from the working surfaces and directly analyzed (with trophosphamide as internal standard) using a reversed-phase capillary column and MS/MS detection. This is the first HPLC/MS/MS method for the simultaneous determination of 5FU, MTX and CP. The present method offers high sensitivity, with detection limits of 1.1 microg l(-1) for MTX and CP and 33.3 microg l(-1) for 5FU, avoiding any sample preconcentration procedure. Rapidity, specificity, high accuracy (mean values between 92.4 and 99.9%) and precision (mean RSD values between 3.4 and 12.1%) make the method suitable for the routine determination of these three antineoplastic drugs.     

Determination of (R)- and (S)-phenprocoumon in human plasma by enantioselective liquid chromatography/electrospray ionisation tandem mass spectrometry

Phenprocoumon is a commonly used oral anticoagulant of the coumarin type, and has found extensive clinical use in the treatment of thrombophlebitis, pulmonary embolism and atrial fibrillation. In the course of a clinical study to investigate the influence of genetic polymorphisms of the CYP2C9 enzyme on phenprocoumon metabolism, we developed a new enantioselective liquid chromatography/electrospray ionisation tandem mass spectrometry (LC/MS/MS) method to quantify (R)- and (S)-phenprocoumon in human plasma. HPLC separation of the enantiomers was achieved on a Chira-Grom-2 column under isocratic conditions using a water/acetonitrile/formic acid eluent. For detection and quantification a triple-quadrupole MS system was used in the selected reaction monitoring (SRM) mode. As an internal standard the structurally homologous compound warfarin was chosen. The detector response was linear with a correlation coefficient of 0.988-0.999 for (R)-phenprocoumon and 0.989-0.999 for (S)-phenprocoumon in the investigated concentration range between 62.5 and 1000 ng/mL (per enantiomer). The limit of detection (LOD) was 12.5 ng/mL.     

Quantitative analysis of cimetidine in human plasma using LC/APCI/SRM/MS.

A quantitative method was developed and validated for rapid and sensitive analysis of cimetidine in human plasma. The method involved the use of liquid chromatography (LC) coupled with atmospheric pressure chemical ionization (APCI) and selected reaction monitoring (SRM) mass spectrometry (MS). A cimetidine analog, SKF92374, was used as the internal standard. Separation of cimetidine and the internal standard was accomplished using a reverse-phase HPLC column (C18). The eluted components were ionized by the APCI source and subsequently detected by a highly selective triple quadrupole mass spectrometer in the SRM mode. Linear standard curves were obtained from 5 ng/mL (lower limit of quantitation) to 10,000 ng/mL. The results demonstrated excellent precision (%RSD 1. 1-8.9%) and accuracy (94.7-108.0%) over this range. In addition, the amount of plasma sample needed for analysis was small (50 muL), and the plasma pretreatment (analyte recovery >94%) was simple and time saving. This assay was used to evaluate cimetidine levels in premature infants following intravenous infusion of cimetidine.