Urinary Hydrocarbons Research Articles

Determination of Hydroxylated Polyaromatic Hydrocarbons in Urine by Gas Chromatography–Mass Spectrometry Using Dispersive Liquid-Liquid Microextraction and Injector-Port Derivatization

Determination of Hydroxylated Polyaromatic Hydrocarbons in Urine by Gas Chromatography–Mass Spectrometry Using Dispersive Liquid-Liquid Microextraction and Injector-Port Derivatization

Oxidative derivatives of polycyclic aromatic hydrocarbons in urine of smokers during transition to e-cigarettes

Oxidative derivatives of polycyclic aromatic hydrocarbons in urine of smokers during transition to e-cigarettes

Determination of hydroxypolycyclic aromatic hydrocarbons in urine by electromembrane extraction coupled with liquid chromatography

Polycyclic aromatic hydrocarbons (PAHs) have been classified as the earliest discovered class of environmental carcinogens, seriously endangering human health and ecological safety, and the hydroxylpolycyclic aromatic hydrocarbons (OH-PAHs) in human urine are considered as the main biomarkers to evaluate the exposure levels of PAHs in human body. Therefore, it is necessary to develop an accurate and sensitive method for the determination of urinary OH-PAHs. Herein, we proposed electromembrane extraction (EME) as a simple, effective and ecofriendly sample pretreatment technique for selective extraction, purification and enrichment of four typical OH-PAHs (2-naphthol, 2- and 3-phenanthrol, 2-hydroxyfluorene) in human urine for the first time. Under the optimum conditions, an analytical method of EME coupled with HPLC was established, which provided wide linear ranges for four OH-PAHs from 1 to 500 ng mL-1 with low LODs of 0.05–0.3 ng mL-1. The average recoveries of four OH-PAHs at three spiked levels in human urine were 81.6–102.5% with RSDs all below 9.4%. The present method has been successfully applied for the sensitive determination of trace four OH-PAHs in urine samples from non-smokers and smokers with a maximum concentration of 2.24 and 3.56 ng mL-1, respectively, which showed a great potential for the analysis of trace OH-PAHs in biological samples.

Online SPE-LC-MS-MS method for eight hydroxylated metabolites of polycyclic aromatic hydrocarbons in urine and determination of optimal sampling time after firefighter training

Polycyclic aromatic compounds (PAHs) are formed during incomplete combustion and firefighters are inadvertently at risk of being exposed to these and other hazardous compounds. Exposure to PAHs is often estimated by measuring their hydroxylated metabolites (OH-PAH) in urine. Here, an online-SPE LC-MS-MS method was set up for eight OH-PAHs thus increasing sample throughput and minimizing manual handling. The method was validated over a 5-month period and showed good reproducibility with intra- and inter-day variation of 2.4–8.1 % and 1.6–6.5 %, respectively, of low-level samples and accuracy (91.6–104.8 %) for a standard reference material. The method was applied to urine samples from conscripts training to become firefighters to determine the optimal sampling time for this training activity before a large intervention study. In total, six conscripts sampled urine 6–8 times over a 40-hr period during a 3-day training course. All eight metabolites were detected in ≥ 97 % of the samples and showed peak excretion 4–6 hrs after the training corresponding to 8–10 hrs after first exposure. Samples taken the morning after the exercise contained low levels of most metabolites. Consequently, 4–6 hrs post exposure is recommended as the optimal sampling time for quantification of PAH exposure and monitoring of potential differences in exposure.

Magnetic solid phase extraction coupled to HPLC-UV for highly sensitive analysis of mono-hydroxy polycyclic aromatic hydrocarbons in urine

BackgroundAs a common pollutant, the carcinogenic properties of polycyclic aromatic hydrocarbons have garnered considerable attention. Trace metabolites of polycyclic aromatic hydrocarbons can be detected in urine as a non-invasively approach to monitor the exposure level. Nonetheless, the urine samples have the disadvantages of being large in volume and containing numerous impurities. Given the growing demand to study metabolites with low abundance and potential biomarkers, there is a pressing need for a preconcentration and high-throughput technique for effectively handling complex liquid samples. ResultsPolystyrene-coated magnetic nanoparticles were used to establish a novel magnetic extraction method for monohydroxy polycyclic aromatic hydrocarbons in urine samples. Polystyrene magnetic nanoparticles are an ideal absorbent for solid-phase extraction. After the material was mixed with the sample and adsorbed the target analyte, the analytes on the material were eluted and quantified using high-performance liquid chromatography. Influencing factors were optimized, and the proposed method achieved desirable sensitivity in analyzing low-abundance metabolites in large volumes of complex urine samples. The recoveries of intra-day and inter-day were 78.0–118.0 % and 81.0 %–115.0 %, respectively. The intra-day and inter-day reproducibility were less than 4.5 % and 8.6 %, respectively. The limits of detection were in the range of 0.009–0.041 ng mL−1, and the limits of quantification were in the range of 0.030–0.135 ng mL−1. Significance and noveltyThe application of reusable polystyrene-coated magnetic solid-phase nanoparticles as adsorbents makes the extraction of monohydroxy polycyclic aromatic hydrocarbons from urine samples economical and environmentally benign. The proposed method is simple, sensitive, and efficient compared to existing techniques. The nanoparticles are easy to prepare, showing potential for rapid screening of complex bulk bio-samples in batches with high efficiency and low budget.

Simultaneous determination of multiple isomeric hydroxylated polycyclic aromatic hydrocarbons in urine by using ultra-high performance liquid chromatography tandem mass spectrometry

Monitoring the level of hydroxylated polycyclic aromatic hydrocarbons (OH-PAHs) in urine is the key to exploring human metabolic changes and comprehensive potential toxicity of PAHs. The OH-PAHs with isomeric structure have different biological functions, indicating that their quantification is indispensable. However, the quantitation method is still dissatisfactory due to the poor separation of these isomeric OH-PAHs. The current study established a ultra-high performance liquid chromatography (UHPLC) tandem mass spectrometry (MS) method to complete the simultaneous determination of 17 OH-PAHs, including two naphthalene metabolites (1-hydroxynaphthalene, 2-hydroxynaphthalene), two fluorene metabolites (2-hydroxyfluorene, 3-hydroxyfluorene), five phenanthrene metabolites (1-hydroxyphenanthrene, 2-hydroxyphenanthrene, 3-hydroxyphenanthrene, 4-hydroxyphenanthrene, 9-hydroxyphenanthrene), a pyrene metabolite (1-hydroxypyrene), five chrysene metabolites (1-hydroxychrysene, 2-hydroxychrysene, 3-hydroxychrysene, 4-hydroxychrysene, 6-hydroxychrysene) and two benzo[a]pyrene metabolites (3-hydroxybenzo[a]pyrene, 9-hydroxybenzo[a]pyrene). The method validation results showed good selectivity, linearity (r2 > 0.999), inter-day and intra-day precision (relative standard deviation (RSD) < 5.5% and RSD < 6.3%), stability (RSD < 19.3%), matrix effect (-8.3%-11.5%) and recovery (65.9%-116.2%). This method is convenient, sensitive and efficient, saving expensive materials and complicated derivatization procedures. The practical applicability of developed approach was also tested in urine samples to identify potential biomarkers of PAHs exposure in humans, and a great compromise was obtained between recoveries and extract convenience. The developed approach may be widely utilized for specific determination of OH-PAHs with isomer structure in urine samples. It is expected that the application of this method may provide powerful references for PAHs exposure assessment.

Contributions of dietary, demographic, disease, lifestyle and other factors in explaining variabilities in concentrations of selected monohydroxylated polycyclic aromatic hydrocarbons in urine: Data for US children, adolescents, and adults

Data from National Health and Nutrition Examination Survey for 2003–2014 for US children aged 6–11 years (N = 2097), adolescents aged 12–19 ears (N = 2642), and adults aged ≥ 20 years (N = 9170) were analyzed to investigate the effects of dietary, demographic, disease, lifestyle, and other factors on concentrations of nine metabolites of polycyclic aromatic hydrocarbons (PAH) in urine. PAHs analyzed were: 1-hydroxynaphthalene, 2-hydroxynaphthalene, 2-hydroxyfluorene, 3-hydroxyfluorene, 9-hydroxyfluorene, 1-hydroxyphenanthrene, 2-hydroxyphenanthrene, 3-hydroxyphenanthrene, and 1-hydroxypyrene. Adults with diabetes were found to have higher adjusted levels of 1-hydroxynaphthalene (4139 vs. 3622 ng/L, p < 0.01) than nondiabetics. Adults with albuminuria had higher adjusted levels of 1-hydroxynaphthalene (4140 vs.3621 ng/L, p < 0.01) and 2-hydroxynaphthalene (6039 vs. 5468 ng/L, p < 0.01) than those without albuminuria. Children with albuminuria had lower adjusted levels of 9-hydroxyfluorene (162 vs. 187 ng/L, p = 0.04), 1-hydroxyphenanthrene (92 vs. 108 ng/L, p < 0.01), and 1-hydroxypyrene (118 vs. 138 ng/L, p < 0.01) than those without albuminuria. The ratios of smoker to nonsmoker adjusted levels for adults varied from a low of 1.4 for 2-hydroxyphenanthrene to a high of 5.6 for 3-hydroxyfluorene. Exposure to environmental tobacco smoke at home was associated with higher levels of most OH-PAHs among children, adolescents, and adults. Consumption of red meat not processed at high temperatures was associated with increased levels of 1-hydroxypyrene (β = 0.00040, p = 0.01), 1-, 2-, and 3-hydroxyphenanthrene, 3-, and 9-hydroxyfluorene. Consumption of red meat processed at high temperatures was associated with increased levels of 2-hydroxynaphthalene (β = 0.00046, p = 0.02) among adults. Consumption of fish processed at high temperatures was associated with decreased levels of 1-hydroxynaphtahlene (β = − 0.00088, p < 0.01), 2-, 3-, and 9-hydroxyfluorene, 1-, 2-, and 3-hydroxyphenanthrene. Among adults, alcohol consumption and caffeine may be associated with increased levels of certain OH-PAHs. Oxidative stress and inflammation associated with exposure to PAHs are associated with albuminuria and have the potential to lead to the development of diabetes.

Preparation of ferrofluid from toner powder and deep eutectic solvent used in air-assisted liquid-liquid microextraction: Application in analysis of sixteen polycyclic aromatic hydrocarbons in urine and saliva samples of tobacco smokers

A new ferrofluid prepared from toner powder and deep eutectic solvent was used in air-assisted liquid-liquid microextraction procedure for extraction of sixteen polycyclic aromatic hydrocarbons from urine and saliva samples of tobacco smokers. In this work, initially, a water-immiscible deep eutectic solvent is synthesized by mixing choline chloride and stearic acid under mild conditions and used as a support solvent in the preparation of ferrofluid from toner powder. Then, a few microliters of the prepared ferrofluid is mixed with sample solution and dispersed in all parts of the solution by performing aspirating/dispersing cycles. To collect the ferrofluid, a magnet is deposited at the outside of the tube and the solution is discarded. The analytes in the collected phase are back-extracted into n-heptane and determined by gas chromatography–mass spectrometry. The validation parameters including limit of detection and quantification, stability, accuracy, linearity, selectivity, precision, enrichment factor, and extraction recovery were studied and the data showed that the method have low limits of detection (in the range of 6–21 ng L−1 in deionized water and 18–63 ng L−1 in saliva and urine samples, respectively) and quantification (in the range of 20–72 ng L−1 in deionized water and 60–216 ng L−1 in saliva and urine samples). Extraction recoveries and enrichment factors ranged from 61 to 84% and 305 to 420, respectively. Relative standard deviations of ≤9% for the extraction of 200 ng L−1of each analyte were obtained for intra– (n = 6) and inter–day (n = 6) precisions. Finally, urine and saliva samples of tobacco smokers were successfully analyzed using the proposed method.

Determination of 1-Hydroxypyrene as a Biomarker for the Effects of Polycyclic Aromatic Hydrocarbons in Urine by Chromatography–Mass Spectrometry

A simple and sensitive procedure has been developed for the determination of 1-hydroxypyrene in urine as a biological marker of the effects of polycyclic aromatic hydrocarbons. After enzymatic hydrolysis for conjugate cleavage, the analyte was extracted from the matrix by liquid–liquid extraction with n-hexane, the extract was evaporated to dryness, and the analyte was derivatized with the silylating reagent N,O-bis(trimethylsilyl)trifluoroacetamide into trimethylsilyl ether at room temperature. The trimethylsilyl extract was analyzed by capillary gas chromatography with mass-selective detection. Quantitative high-precision determination was achieved with the use of the isotope-labeled 1-hydroxypyrene-d9 reference standard. No matrix effects were observed. The detection limit was 0.02 ng/mL, and the linearity range was 0.1–100 ng/mL. Relative standard deviations were 0.044 and 0.064 under the conditions of repeatability and intralaboratory precision, respectively. The percentage recovery was 96–102%.

Determination of trace hydroxyl polycyclic aromatic hydrocarbons in urine using graphene oxide incorporated monolith solid-phase extraction coupled with LC-MS/MS.

The biomonitoring of hydroxy polycyclic aromatic hydrocarbons in urine, as a direct way to access multiple exposures to polycyclic aromatic hydrocarbons, has raised great concerns due to their increasing hazardous health effects on humans. Solid-phase extraction is an effective and useful technique to preconcentrate trace analytes from biological samples. Here, we report a novel solid-phase extraction method using a graphene oxide incorporated monolithic syringe for the determination of six hydroxy polycyclic aromatic hydrocarbons in urine coupled with liquid chromatography-tandem mass spectrometry. The effect of graphene oxide amount, washing solvent, eluting solvent, and its volume on the extraction performance were investigated. The fabricated monoliths gave higher adsorption efficiency and capacity than the neat polymer monolith and commercial C18 sorbent. Under the optimum conditions, the developed method provided the detection limits (S/N = 3) of 0.02-0.1ng/mL and the linear ranges of 0.1-1500ng/mL for six analytes in urine sample. The recoveries at three spiked levels ranged from 77.5 to 97.1%. Besides, the intra column-to-column (n = 3) and inter batch-to-batch (n = 3) precisions were ≤ 9.8%. The developed method was successfully applied for the determination of hydroxy polycyclic aromatic hydrocarbons in urine samples of coke oven workers.

Determination of three hydroxyl polycyclic aromatic hydrocarbons in urine using magnetic solid-phase extraction with magnetic carbon nitride composites coupled with high performance liquid chromatography

A method based on the magnetic solid-phase extraction using magnetic carbon nitride composites coupled with high performance liquid chromatography-fluorescence detector (HPLC-FLD) was developed for the simultaneous determination of three hydroxyl polycyclic aromatic hydrocarbons (OH-PAHs) in urine. The synthesized sorbent was characterized by scanning electron microscope, X-ray diffractometer, vibrating sample magnetometer and surface area analyzer. The extraction parameters such as amount of sorbent, extraction time, eluting solvent and volume (single eluting volume×eluting times) were investigated in detail. Under the optimized conditions, the method showed a linear range of 0.25-250 μg/L (correlation coefficient=0.999). The limit of detection was 0.08 μg/L and the limit of quantification was 0.25 μg/L for each analyte. The proposed method gave a recovery of 90.1%-102%. The relative standard deviations for the intra-day and inter-day were 1.5%-7.7% and 2.2%-8.7%, respectively. The feasibility of the developed method was further demonstrated during the analysis of real samples. The results indicate that the magnetic carbon nitride composites can be used for the effective purification and enrichment of three OH-PAHs in urine. In addition, the method can be applied for analysis of urinary OH-PAHs in a simple, fast, and effective manner.

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.

Inorganic-organic hybrid coating material for the online in-tube solid-phase microextraction of monohydroxy polycyclic aromatic hydrocarbons in urine.

An inorganic-organic hybrid nanocomposite (zinc oxide/polypyrrole) that represents a novel kind of coating for in-tube solid-phase microextraction is reported. The composite coating was prepared by a facile electrochemical polymerization strategy on the inner surface of a stainless-steel tube. Based on the coated tube, a novel online in-tube solid-phase microextraction with liquid chromatography and mass spectrometry method was developed and applied for the extraction of three monohydroxy polycyclic aromatic hydrocarbons in human urine. The coating displayed good extraction ability toward monohydroxy polycyclic aromatic hydrocarbons. In addition, long lifespan, excellent stability, and good compression resistance were also obtained for the coating. The experimental conditions affecting the extraction were optimized systematically. Under the optimal conditions, the limits of detection and quantification were in the range of 0.039-0.050 and 0.130-0.167 ng/mL, respectively. Good linearity (0.2-100 ng/mL) was obtained with correlation coefficients larger than 0.9967. The repeatability, expressed as relative standard deviation, ranged between 2.5% and 9.4%. The method offered the advantage of process simplicity, rapidity, automation, and sensitivity in the analysis of human urinary monohydroxy polycyclic aromatic hydrocarbons in two different cities of Hubei province. An acceptable recovery of monohydroxy polycyclic aromatic hydrocarbons (64-122%) represented the additional attractive features of the method in real urine analysis.

Simultaneous Determination of Seven Metabolites of Polycyclic Aromatic Hydrocarbons in Human Urine by Online Solid Phase Extraction-High Performance Liquid Chromatography

Abstract A high performance liquid chromatographic method for simultaneous determination of seven metabolites of polycyclic aromatic hydrocarbons (PAHs), including 1-hydroxynaphthalene, 2-hydroxynaphthalene, 2-hydroxyfluorene, 2-hydroxyphenanthrene, 4-hydroxyphenanthrene, 1-hydroxypyrene and 6-hydroxychrysene in human urine, was developed coupled with online solid phase extraction by use of a double ternary liquid chromatography system with a fluorescence detector. The target compounds were online concentrated on the Turboflow Cyclone solid phase extraction column, and then transferred by a six-way valve to the Hypersil Green PAH column for separation with acetonitrile and water as mobile phase at a flow rate of 1.0 mL min−1 and 35 °C. The analysis was completed in 20 min. Under the optimized chromatographic conditions, the method showed good linearity (r ≥ 0.999) in the range of 5−2000 ng L−1 or 50−20000 ng L−1 with the LODs of 0.5−15 ng L−1 and the recoveries of 80.7%−110.7%. The proposed method was successfully applied to the detection of metabolites of polycyclic aromatic hydrocarbons in urine from several smokers and non-smokers. The concentrations of 2-hydroxynaphthalene, 1-hydroxynaphthalene, 2-hydroxyfluorene, 2-hydroxyphenanthrene, 4-hydroxyphenanthrene and 6-hydroxychrysene in the smoker urine were much higher than those in non-smoker.

Magnetic graphene oxide as adsorbent for the determination of polycyclic aromatic hydrocarbon metabolites in human urine.

Detection of monohydroxy polycyclic aromatic hydrocarbons metabolites in urine is an advisable and valid method to assess human environmental exposure to polycyclic aromatic hydrocarbons. In this work, novel Fe3O4/graphene oxide composites were prepared and their application in the magnetic solid-phase extraction of monohydroxy polycyclic aromatic hydrocarbons in urine was investigated by coupling with liquid chromatography and mass spectrometry. In the hybrid material, superparamagnetic Fe3O4 nanoparticles provide fast separation to simplify the analytical process and graphene oxide provides a large functional surface for the adsorption. The prepared magnetic nanocomposites were characterized by X-ray diffraction, scanning electron microscopy, transmission electron microscopy, and vibrating sample magnetometry. The experimental conditions were optimized systematically. Under the optimal conditions, the recoveries of these compounds were in the range of 98.3-125.2%, the relative standard deviations ranged between 6.8 and 15.5%, and the limits of detection were in the range of 0.01-0.15 ng/mL. The simple, quick, and affordable method was successfully used in the analysis of human urinary monohydroxy polycyclic aromatic hydrocarbons in two different cities. The results indicated that the monohydroxy polycyclic aromatic hydrocarbons level in human urine can provide useful information for environmental exposure to polycyclic aromatic hydrocarbons.

Determination of four hydroxyl polycyclic aromatic hydrocarbons in urine by solid-phase extraction with monolithic column coupled with high performance liquid chromatography

A method based on monolithic column solid-phase extraction (SPE) coupled with high performance liquid chromatography (HPLC) was developed for the simultaneous determination of four hydroxyl polycyclic aromatic hydrocarbons (OH-PAHs) in urine.A poly (butyl methacrylate-co-ethylene dimethacrylate) monolithic column was prepared in a syringe and was used as the sorbent.The parameters influencing SPE, such as loading volume, washing solvent, eluent, and elution volume, were investigated in detail.Under the optimized conditions, the linearities were obtained in range of 1.2-200.0 ng/mL, and the LODs and LOQs of the analytes were 0.06-0.09 ng/mL and 0.20-0.30 ng/mL, respectively.The intra-day and inter-day relative standard deviations were 1.4%-5.3% and 2.6%-7.3%, respectively.The recoveries evaluated using the spiked (3 ng/mL) urine samples of coke oven workers ranged from 78.2% to 117.0%.The feasibility of the developed method was further demonstrated for the analysis of the real samples.The results indicated that the reusable monolithic column enabled effective purification and enrichment of the four OH-PAHs in urine, and can be applied to the analysis of OH-PAHs in urine due to its simplicity and accuracy.

Effects of smoking on the concentrations of urinary 10 metabolites of polycyclic aromatic hydrocarbons in coke oven workers

To explore the effects of smoking on urinary 10 metabolites of polycyclic aromatic hydrocarbons (PAHs) in the coke oven workers. Occupational health examination was performed on 1401 coke oven workers in one coking plant, their urine were collected respectively. The concentrations of the ten monohydroxy polycyclic aromatic hydrocarbons in urine were detected by gas chromatography/mass spectrometry. The 1401 workers were divided into four groups, namely control, adjunct workplaces, bottom and side, top group according to their workplaces and the different concentrations of PAHs in the environment. The concentrations of the ten monohydroxy polycyclic aromatic hydrocarbons between smokers and nonsmokers in each workplace group were compared using analysis of covariance, respectively. The levels of concentrations of the sixteen polycyclic aromatic hydrocarbons we detected at control were significantly higher than those at other areas (P < 0.05). Comparing the ten monohydroxy polycyclic aromatic hydrocarbons levels between smokers and nonsmokers, the levels of 1-hydroxynaphthalene and 2-hydroxynaphthalene among smokers were higher than nonsmokers with statistically significance in control, adjunct workplaces, bottom and side and top groups (P < 0.05). However, the levels of 1-hydroxypyrene had no statistically significant differences between the four areas. Urinary 1-hydroxynaphthalene and 2-hydroxynaphthalene may be used as biomarkers for the impact of smoking on monohydroxy polycyclic aromatic hydrocarbons in the coke oven workers.

Development of Simultaneous Determination Method of Hydroxylated Polycyclic Aromatic Hydrocarbons in Urine by LC/MS/MS and Its Application to Assessment of Polycyclic Aromatic Hydrocarbons Exposure

Development of Simultaneous Determination Method of Hydroxylated Polycyclic Aromatic Hydrocarbons in Urine by LC/MS/MS and Its Application to Assessment of Polycyclic Aromatic Hydrocarbons Exposure

Using liquid chromatography–tandem mass spectrometry to quantify monohydroxylated metabolites of polycyclic aromatic hydrocarbons in urine

We present an assay which employs enzyme digestion and solid phase extraction followed by liquid chromatography–tandem mass spectrometry to simultaneously quantify 16 hydroxylated polycyclic aromatic hydrocarbons (OHPAHs) in 3-ml samples of urine. The analytes consisted of 2-, 3-, and 4-ring OHPAHs, namely, 1- and 2-hydroxynaphthalene (1- and 2-OHNAP), 2-hydroxyfluorine (2-OHFLU), 1-, 2-, 3-, 4-, and 9-hydroxyphenanthrene (1-, 2-, 3-, 4-, and 9-OHPHE), 1-hydroxypyrene (1-OHPYR), 1- and 2-hydroxybenzo( a)anthracene (1- and 2-OHBAA), 3- and 6-hydroxychrysene (3- and 6-OHCHR) and 3-, 7-, and 9-hydroxybenzo( a)pyrene (3-, 7-, and 9-OHBAP). The method was validated using urine samples from steel workers and control subjects. The coefficients of variation of the method for the particular analytes were between 7% and 27% and the limits of quantitation were between 0.002 and 0.010 μg/l urine. The 2- and 3-ring OHPAHs were easily quantified in all subjects. However, 1-OHPYR was the only representative of the 4- and 5-ring metabolites that could be quantified. Pairwise correlations showed that all OHPAHs were highly correlated with each other (0.553 ≤ r ≤ 0.910) and with 1-OHPYR (0.614 ≤ r ≤ 0.910), the metabolite most widely accepted as a short-term biomarker of exposure to PAHs. The analyte, 2-OHNAP exhibited the lowest pairwise correlations with the other OHPAHs (0.542 ≤ r ≤ 0.628), presumably due to confounding by smoking. Metabolites of phenanthrene, an abundant PAH and the smallest to possess a bay region, are promising OHPAHs for characterizing both exposures to PAHs and the various metabolic pathways.

Analysis of hydroxylated polycyclic aromatic hydrocarbons in urine using comprehensive two-dimensional gas chromatography with a flame ionization detector

The determination of polycyclic aromatic hydrocarbon (PAH) metabolites in human urine is the method of choice for assessing exposure to carcinogenic compounds. The objective of this study was the development of a comprehensive two-dimensional gas chromatography (GC × GC) method using a flame ionisation detector (FID) to simultaneously determine 10 hydroxylated PAH. The method was based on enzymatic deconjugation, liquid–liquid extraction, and trimethylsilyl (TMS) derivatization of the analytes by microwave heating. Satisfactory separation was achieved. The coefficient of variance was 3.8–12.8%. LOD was 0.03–0.18 μg/L, and LOQ was 0.1–0.5 μg/L. The mean recovery was 76%. The method was applied to the analysis of urine from smokers and non-smokers.