Quantification of polycyclic aromatic hydrocarbons (PAHs) in human hair by HPLC with fluorescence detection: a biological monitoring method to evaluate the exposure to PAHs

A high-performance liquid chromatographic (HPLC) method with fluorescence detection was developed for the quantification of polycyclic aromatic hydrocarbons (PAHs) in human hair. Fifteen kinds of PAHs classified as priority pollutants by the US EPA were quantified with four perdeuterated PAHs as internal standards. After 50 mg hair samples were washed with n-hexane to remove external contamination of PAHs, the samples were digested in 2.5 M sodium hydroxide. The digests were extracted with n-hexane and then analyzed by HPLC. Eleven kinds of PAHs were identified in hair samples of 20 subjects, and 10 kinds of PAHs were eventually quantified using the internal standards. For anthracene, chrysene and benzo[k]fluoranthene, significant differences were observed between smokers and non-smokers. Although benzo[b]fluoranthene, dibenz[a,h]anthracene, benzo[ghi]perylene and indeno[1,2,3-cd]pyrene were observed in the particulates of indoor and outdoor air, they were not detected in all hair samples. The analysis of PAHs in human hair should be useful as a new biomarker to evaluate the exposure to PAHs.     

Results of a European inter-laboratory comparison study on the determination of EU priority polycyclic aromatic hydrocarbons (PAHs) in edible vegetable oils

A collaborative study on the analysis for 15 + 1 EU priority PAHs in edible oils was organised to investigate the state-of-the-art of respective analytical methods. Three spiked vegetable oils, one contaminated native sunflower oil, and one standard solution were investigated in this study. The results of 52 laboratories using either high-performance liquid chromatography with fluorescence detection or gas chromatography with mass-selective detectors were evaluated by application of robust statistics. About 95% of the laboratories were able to quantify benzo[a]pyrene together with five other PAHs included in the commonly known list of 16 US-EPA PAHs. About 80% of the participants also quantified seven additional PAHs in most samples, two of which were benzo[b]fluoranthene and benzo[k]fluoranthene, which were also known from the EPA list. Only about 50% of the participants quantified cyclopenta[cd]pyrene, benzo[j]fluoranthene, and benzo[c]fluorene. The robust relative standard deviations of the submitted results without discrimination between the methods applied ranged between 100% for 5-methylchrysene in spiked olive oil and 11% for the same analyte in spiked sunflower oil. The results clearly showed that for these analytes the methods of analysis are not yet well established in European laboratories, and more collaborative trials are needed to promote further development and to improve the performances of the respective methods.     

Solid vapor pressure for five heavy PAHs via the Knudsen effusion method

Polycyclic aromatic hydrocarbons (PAHs) are compounds resulting from incomplete combustion and many fuel processing operations, and they are commonly found as subsurface environmental contaminants at sites of former manufactured gas plants. Knowledge of their vapor pressures is the key to predict their fate and transport in the environment. The present study involves five heavy PAHs, i.e. benzo[b]fluoranthene, benzo[k]fluoranthene, benzo[ghi]perylene, indeno[1,2,3-cd]pyrene, and dibenz[a,h]anthracene, which are all as priority pollutants classified by the US EPA. The vapor pressures of these heavy PAHs were measured by using Knudsen effusion method over the temperature range of 364 K to 454 K. The corresponding values of the enthalpy of sublimation were calculated from the Clausius-Clapeyron equation. The enthalpy of fusion for the 5 PAHs was also measured by using differential scanning calorimetry and used to convert earlier published sub-cooled liquid vapor pressure data to solid vapor pressure in order to compare with the present results. These adjusted values do not agree with the present measured actual solid vapor pressure values for these PAHs, but there is good agreement between present results and other earlier published sublimation data.     

高效液相色谱法测定黄浦江表层沉积物中的痕量多环芳烃

建立了高效液相色谱-二极管阵列检测器(PDA)测定上海市黄浦江表层沉积物中16种多环芳烃(PAHs)的方法。在保留时间定性分析的基础上,利用PDA获取的紫外扫描光谱图对目标组分进行了准确的定性,并通过异构体紫外光谱图中特征峰的差异有效地识别了样品中的4种异构体,即苯并［b］荧蒽、 苯并［k］荧蒽、 苯并［a］芘和苯并［e］芘。通过检测波长的优化,减少了干扰物的影响,提高了检测灵敏度;针对分离度较差的两种目标组分(苯并［b］荧蒽和苯并［k］荧蒽)的定量进行了分析讨论。该方法对16种PAHs的检出限(以干基计)介于1.1～18.3 ng/g之间,具有较高的方法灵敏度。黄浦江表层沉积物测定结果表明,除二氢苊外的15种PAHs都被检出,含量为10.1～253.0 ng/g。     

Resolution of 13 polycyclic aromatic hydrocarbons by constant-wavelength synchronous spectrofluorometry.

A method capable of determining 13 PAHs (acenaphthene, anthracene, benzo[a]anthracene, benzo[a]pyrene, benzo[b]fluoranthene, benzo[k]fluoranthene, chrysene, dibenzo[ah]anthracene, fluoranthene, fluorene, indene[1,2,3-cd]pyrene, phenanthrene and pyrene) in a mixture of 16 EPA PAHs by second derivative synchronous spectrofluorometry in the constant wavelength mode was developed. It has not been possible to determine the following PAHs in the mixture: acenaphthylene, benzo[ghi]perylene and naphthalene. The approach studied allows the sensitive, rapid and inexpensive identification and quantitation of 13 PAHs in a solution of hexane. The detection limits are <1 microg L(-1) (except for chrysene and phenanthrene).     

Optimization of the matrix solid-phase dispersion sample preparation procedure for analysis of polycyclic aromatic hydrocarbons in soils: comparison with microwave-assisted extraction

A fast and simple preparation procedure based on the matrix solid-phase dispersion (MSPD) technique is proposed for the first time for the isolation of 16 polycyclic aromatic hydrocarbons (PAHs) from soil samples. Naphthalene, acenaphthene, fluorene, phenanthrene, anthracene, fluoranthene, pyrene, benz[a]anthracene, chrysene, benzo[e]pyrene, benzo[b]fluoranthene, benzo[k]fluoranthene, benzo[a]pyrene, dibenzo[a,h]anthracene, benzo[g,h,i]perylene, and indeno[1,2,3-c,d]pyrene were considered in the study. Extraction and clean-up of samples were carried out in a single step. The main parameters that affect extraction yield, such as dispersant, type and amount of additives, clean-up co-sorbent and extractive solvent were evaluated and optimized. The addition of an alkali solution in MSPD was required to provide quantitative recoveries. Analytical determinations were carried out by high performance liquid chromatography (HPLC) with fluorescence detection. Quantification limits (between 0.01 and 0.6 ng g(-1) dry mass) were well below the regulatory limits for all the compounds considered. The extraction yields for the different compounds obtained by MSPD were compared with the yields obtained by microwave-assisted extraction (MAE). To test the accuracy of the MSPD technique, the optimized methodology was applied to the analysis of standard reference material BCR-524 (contaminated industrial soil), with excellent results.     

Aromatic hydrocarbon metabolites in fish: automated extraction and high-performance liquid chromatographic separation into conjugate and non-conjugate fractions

An automated extractor-concentrator was used to extract metabolites of naphthalene, 2,6-dimethylnaphthalene, and benzo[a]pyrene from serum, bile and liver homogenate of rainbow trout (Salmo gairdneri). The extracts were analyzed by reversed-phase high-performance liquid chromatography (HPLC) with fluorescence detection. Recoveries of naphthalene and 2,6-dimethylnaphthalene metabolites from all matrices were generally greater than 90%; however, the recoveries of benzo[a]pyrene metabolites from serum ranged from 37-99%. In addition, conjugated metabolites of polycyclic aromatic hydrocarbons (PAHs) were separated from non-conjugated metabolites and parent PAHs by using two diol columns with normal-phase HPLC. The extraction and separation techniques were also applied to isolate metabolites in samples from fish fed 2,6-dimethylnaphthalene.     

Supercritical fluid extraction of polycyclic aromatic hydrocarbons from liver samples and determination by HPLC-FL

An extraction/clean-up procedure by SFE was developed for isolating PAHs from liver samples for subsequent HPLC-FL determination of ten PAHs in the enriched extract. Recoveries (90-115%) and RSD % (< or =7.7) were satisfactory. When applied to 11 samples of bird of prey (Tyto alba) protected species and classified of special interest, from the Galicia (Northwest to Spain), benzo[ghi]perylene and indeno[1,2,3-cd]pyrene were undetectable; chrysene and benzo[a]pyrene are only detected in one sample; benzo[a]anthracene and benzo[k]fluoranthene are only quantified in one sample and benzo[b]fluoranthene in two samples. The other PAHs, anthracene, fluoranthene and pyrene are present in almost all the samples.     

Determination of polycyclic aromatic hydrocarbons from buzzards (Buteo buteo) and tawny owl (Strix aluco) by liquid chromatography with fluorescence detection

Supercritical fluid extraction was applied to the determination of naturally contaminated polycyclic aromatic hydrocarbons (PAHs) in bird tissue by liquid chromatography with fluorescence detection (LC-FL). Recoveries (> 90%) and relative standard deviations (< or = 7.7%) were satisfactory. The levels of 10 PAHs were analyzed in 6 classes of tissues (heart, liver, intestine, muscle, lung, and kidney) of 10 buzzards and 2 tawny owls, predatory birds from the Galicia (northwest Spain). The PAHs found most abundantly were pyrene, fluoranthene, benzo[a]anthracene, and anthracene. Chrysene, benzo[b]fluoranthene, benzo[k]fluoranthene, benzo[ghi]perylene, and indeno[1,2,3-cd]pyrene were not detected. Intestine, kidney, and lung were more polluted than other tissues.     

Rapid SPE-HPLC determination of the 16 European priority polycyclic aromatic hydrocarbons in olive oils

Polycyclic aromatic hydrocarbons (PAHs) are a large class of organic compounds. It has been established that the main source of exposure to these compounds for human beings is through food, particularly fats and oils, due to the lipophilic nature of these polycyclic compounds. The aim of this work was to optimise and validate a method involving SPE and HPLC for rapid determination of the 16 European Union (EU) priority PAHs (required by the Recommendation 2005/108/EC) in vegetable oils. Two spectrofluorometric detectors and a UV-Visible detector in series were used to identify and quantify the target compounds. Linearity, recoveries, LOD, and LOQ were found to be in agreement with the performance criteria for benzo[a]pyrene (BaP) analysis as required by the Commission Directive 2005/10/EC, and satisfactory for all the compounds of interest, except for cyclopenta[c,d]pyrene, which presented a very low signal in the UV. Optimised chromatographic conditions for the separation of 25 PAHs, comprising both EPA and EU priority PAHs plus benzo[e]pyrene and benzo[b]chrysene, have been also proposed.     

The development of solid-surface fluorescence characterization of polycyclic aromatic hydrocarbons for potential screening tests in environmental samples

This paper presents the characterization of polycyclic aromatic hydrocarbons (PAHs) in solid-surface fluorescence as the first step for obtaining new optical sensors for PAHs screening. The fluorescence properties of the EPA-PAHs (naphthalene, acenaphthene, acenaphthylene, fluorene, phenanthrene, anthracene, fluoranthene, pyrene, chrysene, benzo[a]anthracene, benzo[k]fluoranthene, benzo[b]fluoranthene, benzo[a]pyrene, indeno [1,2,3-cd]pyrene, benzo[g,h,i]perylene and dibenzo[a,h]anthracene) on five types of solid-surfaces were evaluated. The experimental variables (pH and percentage of organic solvent in samples) were studied, obtaining different possibilities for making individual sensors for some of these PAHs and the best conditions for developing sensors for PAH screening were also studied.     

New approach for screening polycyclic aromatic hydrocarbons in water samples

For the first time, solid-phase extraction (SPE) has been combined to room-temperature phosphorimetry (RTP) to determine the 16 polycylic aromatic hydrocarbons related as major pollutants by the US Environmental Protection Agency (EPA). These include naphthalene, anthracene, acenaphthylene, acenaphthene, fluorene, fluoranthene, benzo(a)anthracene, benzo(k)fluoranthene, benzo(b)fluoranthene, benzo(a)pyrene, indeno(1,2,3-cd)pyrene, pyrene, chrysene, phenanthrene, benzo(g,h,i)perylene and dibenzo(a,h)anthracene. The pre-concentration factor obtained by SPE, combined with the sensitivity of RTP, resulted in calibration curves with linear dynamic ranges at the parts-per-billion level (ng ml(-1)). The limits of detection were estimated at the parts-per-trillion level (pg ml(-1)). Several pollutants usually encountered in water samples were tested for interference. These included polychlorinated biphenyls, pesticides, and volatile organic compounds. As a result of the appropriate combination of excitation wavelength (330 nm) and phosphorescence enhancers (0.1 M TlNO(3) and 0.05 M sodium dodecyl sulfate, SDS), no interference was observed. The results demonstrate the potential of SPE-RTP for screening polycyclic aromatic hydrocarbons (PAHs) in environmental waters.     

Validation of a method for extraction of polycyclic aromatic hydrocarbons from sewage sludge and analysis by GC-MS

In the present study, the solid–liquid extraction with low temperature purification was validated for the determination of 16 polycyclic aromatic hydrocarbons from sewage sludge by gas chromatography-mass spectrometry. Recoveries ranged 70–114% for naphthalene, acenaphthylene, acenaphthene, fluorene, phenanthrene, anthracene, fluoranthene, pyrene, benzo[a]anthracene and chrysene, while the compounds benzo[b]fluoranthene, benzo[k]fluoranthene, benzo[a]pyrene, indeno[1,2,3-cd]pyrene, dibenzo[a,h]anthracene and benzo[g,h,i]perylene showed recoveries of between 40 and 70%. The relative standard deviation was less than 13% for all of the compounds. Negative matrix effect was observed on the 10 compounds with less retention time in the chromatographic analysis and positive matrix effect noticed on the others. The limits of quantification were from 2 to 20 μg kg^?1, about 30 times less than the maximum residue limit allowed in sludge by the European Union. The validated method produced quantification of 11 PAHs in one sludge sample at concentrations ranging 20–2000 μg kg^?1.     

Gas chromatographic-mass spectrometric determination of polycyclic aromatic hydrocarbons formed during the pyrolysis of phenylalanine

The formation of polycyclic aromatic hydrocarbons (PAHs) during pyrolysis process of phenylalanine had been studied. Ten PAHs, including fluorene, phenanthrene, anthracene, fluoranthene, pyrene, benzo[a]anthracene, chrysene, benzo[k]fluoranthene, benzo[e]pyrene, and benzo[a]pyrene were analyzed by gas chromatography-mass spectrometry using selective ion monitoring mode. This technique offers the capability to analyze trace amounts of PAHs in phenylalanine pyrolyzates. The pyrolysis was carried out in a micro-furnace with quartz furnace liner. The injection was conducted with glass pelletizer syringe to avoid metal contamination. Qualitative results were obtained at 900 degrees C and quantitative analysis of 10 PAHs was done for 700 and 900 degrees C.     

Ozonation of pyrene and benzo[a]pyrene in silica and soil –14C-mass balances and chemical analysis of oxidation products as a first step to ecotoxicological evaluation

Ozonation of polycyclic aromatic hydrocarbons (PAHs) in soil is a process that can be used for in-situ soil remediation or in combination with bioremediation techniques. First steps to a comprehensive ecotoxicological evaluation of this method is done by ozonation of radioactively labeled (¹⁴C) pyrene and benzo[a]pyrene in a silty soil (LUFA 2.2) under mass-balancing conditions and GC-MS analysis of aromatic ozonation products. ¹⁴C-Mass-balances for pyrene and benzo[a]pyrene (b[a]p) in soil showed that, apart from ¹⁴CO₂ formation, considerable percentages of both PAHs are oxidized to water soluble substances (20–30%) or to non-extractable or bound residues (10% for pyrene, 30% for b[a]p). TLC and GC-MS analysis of ozonation products extractable from artificially contaminated silica and soil by organic solvents revealed a large number of aromatic substances. PAH-quinones and ten ring fission products with formyl- and carboxy-groups of both pyrene and b[a]p could be identified.     

Collection and analytical characterisation of the atmospheric particulate in the city of Bari

In this paper an application of new procedures for atmospheric particulate analysis is illustrated. PM10, PAHs (benzo[a]anthracene (BaA), benzo[b]fluoranthene (BbF), benzo[j]fluoranthene (BjF), benzo[k]fluoranthene (BkF), benzo[a]pyrene (BaP), indeno[1, 2, 3-cd]pyrene (Ip), dibenzo[a, h]anthracene (DbA)) and heavy metals (Cu, Ni, Zn, Co, Mn, Cd, Fe and Pb) were investigated. PM10 determination was performed by gravimetric method, PAHs were measured by GC-MS, and heavy metals by HPIC. An air quality monitoring campaign on the territory of Bari municipality has been organised, and its results are shown.     

Error propagation as a factor in selection of measurement intervals for the determination of polycyclic aromatic hydrocarbons by second-derivative spectrofluorimetry

The most suitable wavelength intervals were selected for the determination of 4 polycyclic aromatic hydrocarbons (PAHs; benzo[g,h,i]perylene, dibenzo[a,h]anthracene, pyrene, and triphenylene) in very complex mixtures of 11 PAHs: anthracene, benz[a]anthracene, benzo[a]pyrene, benzo[b]fluoranthene, benzo[g,h,i]perylene, benzo[k]fluoranthene, chrysene, dibenz[a,h]anthracene, phenanthrene, pyrene, and triphenylene. The multiple linear regression algorithm was applied to measurements made in several wavelength intervals previously selected on the basis of sensitivity and minimum number of interfering compounds. Of the different models obtained, those displaying minimum error propagation in the analytical result were selected. By applying the models proposed in this study, we precisely and accurately determined benzo[g,h,i]perylene, dibenz[a,h]anthracene, pyrene, and triphenylene in complex mixtures--a feat that could not be achieved by the use of constant-wavelength spectrofluorimetry in combination with second-derivative techniques.     

Ozonation of pyrene and benzo[a]pyrene in silica and soil –14C-mass balances and chemical analysis of oxidation products as a first step to ecotoxicological evaluation

Ozonation of polycyclic aromatic hydrocarbons (PAHs) in soil is a process that can be used for in-situ soil remediation or in combination with bioremediation techniques. First steps to a comprehensive ecotoxicological evaluation of this method is done by ozonation of radioactively labeled (¹⁴C) pyrene and benzo[a]pyrene in a silty soil (LUFA 2.2) under mass-balancing conditions and GC-MS analysis of aromatic ozonation products. ¹⁴C-Mass-balances for pyrene and benzo[a]pyrene (b[a]p) in soil showed that, apart from ¹⁴CO₂ formation, considerable percentages of both PAHs are oxidized to water soluble substances (20–30%) or to non-extractable or bound residues (10% for pyrene, 30% for b[a]p). TLC and GC-MS analysis of ozonation products extractable from artificially contaminated silica and soil by organic solvents revealed a large number of aromatic substances. PAH-quinones and ten ring fission products with formyl- and carboxy-groups of both pyrene and b[a]p could be identified. Received: 11 March 1997 / Accepted: 25 April 1997     

Determination of polycyclic aromatic hydrocarbons (PAHs) in particulate matter collected with low volume samplers

The determination of polycyclic aromatic hydrocarbons (PAHs) contained in samples of particulate matter (PM), collected with low volume pumps, were carried out with an high sensitivity method that comes from several revisions of a previous method. The present work describes how, by using programmable temperature vaporization (PTV) and a mass selective detector with inert ionic source for the GC-MS analysis and the modifications of microwave-assisted extraction (MAE), the sensitivity of the method can be increased.The PAHs chosen for testing the method are: benzo[a]anthracene (BaA), benzo[b]fluoranthene (BbF), benzo[k]fluoranthene (BkF), benzo[a]pyrene (BaP), indeno[1,2,3-cd]pyrene (Ip) and dibenzo[a,h]anthracene (DbA). They, in fact, belong to that group of substances that are the most harmful for human health for their carcinogenicity.PAHs recoveries for spiked standard solutions at different concentrations were between 95 and 100% with relative standard deviation ranging from 1 to 3%. The revised method was validated using a 1649a urban dust standard reference material (SRM). The results obtained were in good agreement with certified values. The high sensitivity of the method allows to carry out analyses using only a half of the sampled filter (usually 47 mm diameter membranes). In this way, the other half can be used for the characterization of the other components of PM (heavy metals, organic carbon, ions, etc). The last step has been constituted by application of the optimized method on real samples collected in two cities located in Southern Italy (Bari and Taranto).     

Comparison of ultrasonic and pressurized liquid extraction for the analysis of polycyclic aromatic compounds in soil samples by gas chromatography coupled to tandem mass spectrometry

A pressurized liquid extraction (PLE) method has been optimized for the determination of polycyclic aromatic hydrocarbons (PAHs) in soil samples and it was compared with ultrasonic extraction. The extraction step was followed by gas chromatography-triple quadrupole mass spectrometry (GC-QqQ-MS/MS) analysis. Parameters such as type of solvent, extraction time, extraction temperature and number of extractions were optimized. There were no significant differences among the two extraction methods although better extraction efficiencies were obtained when PLE was used, minimizing extraction time and solvent consumption. PLE procedure was validated, obtaining limits of detection (LODs) ranging from 0.02 to 0.75 μg kg⁻¹ and limits of quantification (LOQs) ranging from 0.07 to 2.50 μg kg⁻¹ for the selected PAHs. Recoveries were in the range of 59-110%, except for naphthalene, which was the most volatile PAH. Finally, the method was applied to real soil samples from Southeast of Spain. PAHs concentrations were low, and phenanthrene, pyrene, fluorene, benzo[a]pyrene and chrysene were the most frequently detected analytes in the samples.