Development of the thin film solid phase microextraction (TF-SPME) method for metabolomics profiling of steroidal hormones from urine samples using LC-QTOF/MS.

In the present study, the development and optimization of a thin film solid phase microextraction method (TF-SPME) was conducted for metabolomics profiling of eight steroid compounds (androsterone, dihydrotestosterone, dihydroepiandrosterone, estradiol, hydroxyprogesterone, pregnenolone, progesterone and testosterone) from urine samples. For optimization of extraction method, two extraction sorbents (PAN-C18 and PS-DVB) were used as they are known to be effective for isolation of low-polarity analytes. The stages of sample extraction and analyte desorption were considered as the most crucial steps in the process. Regarding the selection of the most suitable desorption solution, six different mixtures were analyzed. As a result, the mixture of ACN: MeOH (1:1, v/v) was chosen in terms of the highest analytes' abundances that were achieved using the chosen solvent. Besides other factors were examined such as the volume of desorption solvent and the time of both extraction and desorption processes. The analytical determination was carried out using the ultra-high performance liquid chromatography coupled with high resolution tandem mass spectrometry detection in electrospray ionization and positive polarity in a scan mode (UHPLC-ESI-QTOF/MS). The developed and optimized TF-SPME method was validated in terms of such parameters as extraction efficiency, recovery as well as matrix effect. As a result, the extraction efficiency and recovery were in a range from 79.3% to 99.2% and from 88.8% to 111.8%, respectively. Matrix effect, calculated as coefficient of variation was less than 15% and was in a range from 1.4% to 11.1%. The values of both validation parameters (recovery and matrix effect) were acceptable in terms of EMA criteria. The proposed TF-SPME method was used successfully for isolation of steroids hormones from pooled urine samples before and after enzymatic hydrolysis of analytes.

Silicone Wristbands in Exposure Assessment: Analytical Considerations and Comparison with Other Approaches.

Humans are exposed to numerous potentially harmful chemicals throughout their lifetime. Although many studies have addressed this issue, the data on chronic exposure is still lacking. Hence, there is a growing interest in methods and tools allowing to longitudinally track personal exposure to multiple chemicals via different routes. Since the seminal work, silicone wristbands (WBs) have been increasingly used to facilitate human exposure assessment, as using WBs as a wearable sampler offers new insights into measuring chemical risks involved in many ambient and occupational scenarios. However, the literature lacks a detailed overview regarding methodologies being used; a comprehensive comparison with other approaches of personal exposure assessment is needed as well. Therefore, the aim of this review is fourfold. First, we summarize hitherto conducted research that employed silicone WBs as personal passive samplers. Second, all pre-analytical and analytical steps used to obtain exposure data are discussed. Third, we compare main characteristics of WBs with key features of selected matrices used in exposure assessment, namely urine, blood, hand wipes, active air sampling, and settled dust. Finally, we discuss future needs of research employing silicone WBs. Our work shows a variety of possibilities, advantages, and caveats associated with employment of silicone WBs as personal passive samplers. Although further research is necessary, silicone WBs have already been proven valuable as a tool for longitudinal assessment of personal exposure.

Concentrations of urinary biomarkers and predictors of exposure to pyrethroid insecticides in young, Polish, urban-dwelling men.

Pyrethroid insecticides are a class of pesticides with multiple agricultural and residential applications. However, widespread use of these chemicals may pose a threat to human health. Biomarkers of pyrethroid exposure are frequently detected in populations around the world, but some groups may be underrepresented. Moreover, there is an ongoing debate on factors contributing to pyrethroid burden in humans. To address these problems, we measured urinary biomarkers of pyrethroid exposure in urine samples from 306 young men living in urban area of Lódz, Poland, and gathered questionnaire data to identify predictors of exposure. Limit of detection (LOD) of gas chromatography-mass spectrometry (GC-MS) method was 0.1 ng/mL for all quantified pyrethroid metabolites, namely cis-3-(2,2-dichlorovinyl)-2,2-dimethylcyclopropane carboxylic acid (cis-DCCA), trans-3-(2,2-dichlorovinyl)-2,2-dimethylcyclopropane carboxylic acid (trans-DCCA), cis-3-(2,2-dibromovinyl)-2,2-dimethylcyclopropane carboxylic acid (cis-DBCA), and 3-phenoxybenzoic acid (3-PBA). Detection rate ranged from 32% (cis-DBCA) to 76% (trans-DCCA). Concentrations of urinary biomarkers in studied sample were in lower range of these observed in similar studies, with unadjusted geometric means (GMs) of most prevalent biomarkers, trans-DCCA and 3-PBA, equal to 0.268 and 0.228 ng/mL, respectively. As for questionnaire data, the statistical analysis revealed that non-dietary factors, especially dog ownership and pesticide use on household pets, contribute significantly to urinary trans-DCCA and 3-PBA concentrations (p ≤ 0.009). Moreover, a few dietary sources of exposure were identified, such as seeds and nuts consumption for 3-PBA (p < 0.001) and vegetable juice intake for trans-DCCA (p = 0.015). Multivariate analyses further highlighted the importance of non-dietary factors in pyrethroid exposure. Compared to other works, our results confirm widespread exposure to pyrethroids observed in other studies and stress the role of residential pyrethroid use in pyrethroid burden in humans.

Sensitive Analysis of Idarubicin in Human Urine and Plasma by Liquid Chromatography with Fluorescence Detection: An Application in Drug Monitoring.

A new approach for the sensitive, robust and rapid determination of idarubicin (IDA) in human plasma and urine samples based on liquid chromatography with fluorescence detection (LC-FL) was developed. Satisfactory chromatographic separation of the analyte after solid-phase extraction (SPE) was performed on a Discovery HS C18 analytical column using a mixture of acetonitrile and 0.1% formic acid in water as the mobile phase in isocratic mode. IDA and daunorubicin hydrochloride used as an internal standard (I.S.) were monitored at the excitation and emission wavelengths of 487 and 547 nm, respectively. The method was validated according to the FDA and ICH guidelines. The linearity was confirmed in the range of 0.1-50 ng/mL and 0.25-200 ng/mL, while the limit of detection (LOD) was 0.05 and 0.125 ng/mL in plasma and urine samples, respectively. The developed LC-FL method was successfully applied for drug determinations in human plasma and urine after oral administration of IDA at a dose of 10 mg to a patient with highly advanced alveolar rhabdomyosarcoma (RMA). Moreover, the potential exposure to IDA present in both fluids for healthcare workers and the caregivers of patients has been evaluated. The present LC-FL method can be a useful tool in pharmacokinetic and clinical investigations, in the monitoring of chemotherapy containing IDA, as well as for sensitive and reliable IDA quantitation in biological fluids.

Evaluation of 1-year urinary excretion of eight metabolites of synthetic pyrethroids, chlorpyrifos, and neonicotinoids.

Synthetic pyrethroids, chlorpyrifos, and neonicotinoids are representatives of non-persistent insecticides ubiquitously used against insects all over the world. Their widespread use causes prevalent exposure to these compounds, which may be hazardous to human health. The insecticides have short biological half-lives and are mostly excreted in urine within 24 h after entering the human body; thus, the urinary concentration of their metabolites is highly dependent on the time elapsed between exposure and sample collection. Considering the within-day fluctuations in urinary concentration, one randomly collected sample may cause misclassification of long-term exposure. We evaluated the variability of excretion of eight insecticide metabolites in 24-h urine samples collected from 14 volunteers once or twice per month over 12 consecutive months. High detection frequency above 70% for non-specific metabolites of pyrethroid, chlorpyrifos, and neonicotinoids confirmed widespread exposure to these insecticides in the studied population. A long-term variability of exposure was assessed based on intraclass correlation coefficient (ICC). We found relatively low variability of excretion for non-specific pyrethroid metabolites and 3,5,6-trichloro-2-pyridinol (ICC > 0.75), but poor repeatability for 6-chloronicotinic acid. Constantly higher ICCs were observed for daily excretion than for unadjusted concentrations. Seasonal differences were observed for 3,5,6-trichloro-2-pyridinol and 6-chloronicotinic acid, with the highest and the lowest median concentration, respectively, in the summer. Due to high ICC values and lack of seasonal variations, one 24-h urine sample was considered sufficient to characterize long-term excretion of non-specific pyrethroid metabolites in non-occupationally exposed population. In addition, we calculated the daily intake (DI) for cypermethrin, permethrin, deltamethrin, and chlorpyrifos. The estimated DI values were mostly below the acceptable daily intake, which indicates that the evaluated exposure is non-hazardous to the population.