Development of a Global Metabo-Lipid-Prote-omics Workflow to Compare Healthy Proximal and Distal Colonic Epithelium in Mice.

A multimetabo-lipid-prote-omics workflow was developed to characterize the molecular interplay within proximal (PC) and distal (DC) colonic epithelium of healthy mice. This multiomics data set lays the foundation to better understand the two tissue types and can be used to study, for example, colon-related diseases like colorectal cancer or inflammatory bowel disease. First, the methyl tert-butyl ether extraction method was optimized, so that from a single tissue biopsy >350 reference-matched metabolites, >1850 reference-matched lipids, and >4500 proteins were detected by using targeted and untargeted metabolomics, untargeted lipidomics, and proteomics. Next, each omics-data set was analyzed individually and then merged with the additional omics disciplines to generate a deep understanding of the underlying complex regulatory network within the colon. Our data demonstrates, for example, differences in mucin formation, detected on substrate level as well as on enzyme level, and altered lipid metabolism by the detection of phospholipases hydrolyzing sphingomyelins to ceramides. In conclusion, the combination of the three mass spectrometry-based omics techniques can better entangle the functional and regional differences between PC and DC tissue compared to each single omics technique.

Importance of gut microbiota metabolites in the development of cardiovascular diseases (CVD).

Cardiovascular disease (CVD) remains the most common cause of death worldwide and has become a public health concern. The proven notable risk factors for CVD are atherosclerosis, hypertension, diabetes, dyslipidemia, inflammation, and some genetic defects. However, research has shown a correlation between metabolic health, gut microbiota, and dietary risk factors. The gut microbiota makes an important contribution to human functional metabolic pathways by contributing enzymes that are not encoded by the human genome, for instance, the breakdown of polysaccharides, polyphenols and vitamins synthesis. TMAO and SCFAs, human gut microbiota compounds, have respective immunomodulatory and pro-inflammatory effects. Choline and l-carnitine are abundant in high-fat diets and are transformed into TMA by gut bacteria. The liver's phase of metabolism then changes TMA into TMAO. In turn, TMAO promotes the activation of macrophages, damages vascular endothelium, and results in CVD-however, dysbiosis decreases SCFAs and bile acids, which raises intestinal permeability. Congestion in the portal vein, a drop in cardiac output, a reduction in intestinal perfusion, and intestinal leakage are all caused by heart failure. These factors induce systemic inflammation by increasing intestinal leakage. By raising CRP and pro-inflammatory reactions, human gut dysbiosis and elevated TMAO levels promote the development of arterial plaque, hasten the beginning of atherosclerosis, and raise the risk of CAD. A healthy symbiosis between the gut microbiota and host is a key factor in shaping the biochemical profile of the diet, therefore which are crucial for maintaining the intestinal epithelial barrier, growing mucosa, reducing inflammation, and controlling blood pressure.

Adaptive Real-Time Object Detection for Autonomous Driving Systems.

Accurate and reliable detection is one of the main tasks of Autonomous Driving Systems (ADS). While detecting the obstacles on the road during various environmental circumstances add to the reliability of ADS, it results in more intensive computations and more complicated systems. The stringent real-time requirements of ADS, resource constraints, and energy efficiency considerations add to the design complications. This work presents an adaptive system that detects pedestrians and vehicles in different lighting conditions on the road. We take a hardware-software co-design approach on Zynq UltraScale+ MPSoC and develop a dynamically reconfigurable ADS that employs hardware accelerators for pedestrian and vehicle detection and adapts its detection method to the environment lighting conditions. The results show that the system maintains real-time performance and achieves adaptability with minimal resource overhead.

Blood Microsampling to Monitor Metabolic Profiles During Physical Exercise.

Monitoring approaches and technical improvements are key factors to improve a sportsman's health, training, and recovery after an injury. In this study, a targeted metabolomics approach using microsampling with hemaPEN® was developed to measure changes in blood concentrations of nine amino acids and four organic acids before, during, and after exercise. The aim of this research project was to investigate if a reliable monitoring of metabolite levels during sports activity can be achieved by collecting one drop of whole blood at different time points. A hemaPEN device is an easy-to-use and noninvasive microsampling technique designed to collect four accurate and precise blood volumes simultaneously (10.96 µl). Twenty healthy volunteers between 19 and 30 years of age were included in this study. Physical activity consisted in running as fast as possible 1,600 m after 400 m warm-up. One drop of blood was collected at five time points: before exercise, after 800-m running, after 1,600 m, and 30 min and 60 min after finishing the exercise. The influence of physical activity on metabolite levels was evaluated using two ultrahigh-performance liquid chromatography coupled to tandem mass spectrometry (UHPLC-MS/MS) methods. Analytical performance criteria such as metabolite stability, method precision, trueness, and accuracy were found to be satisfactory. Expected significant metabolic changes were identified for lactic acid, main TCA cycle intermediates, and some amino acids (e.g., creatinine, choline, and taurine). This preliminary study performed on a small cohort demonstrated a high interest of using microsampling for fluxomics analysis, not only to collect quickly and easily biological samples during sports events but also because it is much easier to store and to process the samples than classical plasma/serum samples obtained by venipuncture. The present results open new avenue for fluxomics analysis in the context of health care.

CO2-effervescence assisted dispersive micro solid-phase extraction based on a magnetic layered double hydroxide modified with polyaniline and a surfactant for efficient pre-concentration of heavy metals in cosmetic samples.

In this paper, a CO2-effervescence assisted dispersive micro solid-phase extraction procedure (CO2-EA-DµSPE) using a magnetic layered double hydroxide modified with polyaniline and a surfactant (Zn-Al-LDH-PA-DBSNa-Fe3O4) was applied for the pre-concentration of heavy metals (Ni2+, Pb2+, Co2+, and Cd2+). The final analysis of the analytes was carried out by atomic absorption spectroscopy. XRD, FTIR, and SEM studies were used for the characterization of the synthesized nanoadsorbent. For the maximum extraction efficiency, effective factors (including pH, nanoadsorbent dosage, and volume of the eluent) were investigated using the central composite design (CCD) method. Under the optimum conditions, the preconcentration factor was more than 20. The linear ranges for Ni2+, Pb2+, Co2+, and Cd2+ were obtained as (5-550), (7-750), (5-500), and (3-100) ng mL-1, respectively. The proposed method provided low detection limits (1.4, 2.1, 1.5, and 0.9 ng mL-1 for Ni2+, Pb2+, Co2+, and Cd2+, respectively) and suitable repeatability (relative standard deviation values (RSDs) below 6.1%, n = 6). Finally, the current method was successfully used for the extraction of heavy metals from cosmetic samples.

Epitope Prediction by Novel Immunoinformatics Approach: A State-of-the-art Review.

Immunoinformatics is a science that helps to create significant immunological information using bioinformatics softwares and applications. One of the most important applications of immunoinformatics is the prediction of a variety of specific epitopes for B cell recognition and T cell through MHC class I and II molecules. This method reduces costs and time compared to laboratory tests. In this state-of-the-art review, we review about 50 papers to find the latest and most used immunoinformatic tools as well as their applications for predicting the viral, bacterial and tumoral structural and linear epitopes of B and T cells. In the clinic, the main application of prediction of epitopes is for designing peptide-based vaccines. Peptide-based vaccines are a considerably potential alternative to low-cost vaccines that may reduce the risks related to the production of common vaccines.

Preparation of Diphtheria and Pseudomonas Exotoxin A Immunotoxins and Evaluation of Their Cytotoxicity Effect on SK-BR-3, BT-474, and MDA-MB-231 Breast Cancer Cell Lines.

Immunotoxin targeted therapy is a promising way of cancer therapy that is made from a toxin attached to an antibody which target a specific protein presented on cancer cells. In this study, we introduce immunotoxins comprising of truncated pseudomonas exotoxin A (PEA) and diphtheria toxin (DT) conjugated to trastuzumab. The effectiveness of 20 and 30 µg/ml immunotoxins and trastuzumab were studied on SK-BR-3 and BT-474 HER2/neu positive breast cancer cell lines by a cell death assay test. The produced immunotoxins have the potential to reduce the therapeutic dose of the trastuzumab and in the same time achieve higher efficiency.

Monitoring of cyanotoxins in water from hypersaline microalgae colonies by ultra high performance liquid chromatography with diode array and tandem mass spectrometry detection following salting-out liquid-liquid extraction.

In this study two different analytical approaches have been developed to determine the presence of several cyanotoxins in saline water samples from a continental salt marsh. A salting-out assisted liquid-liquid extraction (SALLE) has been used in combination with ultra-high performance liquid chromatography-tandem mass spectrometry and UV-diode array detection (UHPLC-MS/MS and UHPLC-DAD). The target analytes are eight microcystins named MC-RR, MC-YR, MC-LR, MC-WR, MC-LA, MC-LY, MC-LW, MC-LF and nodularin (NOD), covering a wide range of polarities. The separation was achieved using a Zorbax Eclipse Plus RRHD C18 column (50 × 2.1 mm, 1.8 µm) in less than 7.5 and 5.5 min for UV and MS/MS detection, respectively. The mobile phase used consisted of water (solvent A) and acetonitrile (MeCN) (solvent B), both containing 0.01% of formic acid for DAD and 0.4% of formic acid for MS/MS detection, at a flow rate of 0.4 mL min-1. The temperature of the column was set at 25 °C and 20 µL of sample were injected. The main parameters affecting the SALLE procedure were studied and the following optimum values were obtained: neutral pH, 2 mL of acetonitrile as extraction solvent and 1.2 g of ammonium sulfate as salting-out agent for 4 mL of water sample. The validation protocols for both methods were accomplished with real water samples obtaining LODs ranging from 1.0 to 3.4 µg L-1 and 0.02 to 0.11 µg L-1 for DAD and MS/MS respectively. Although the SALLE-UHPLC-DAD methodology is easier and cheaper than UHPLC-MS/MS significantly better detection limits were achieved with tandem mass spectrometry as well as allowing for unambiguous identification. Extraction recoveries were higher than 77.0% (except for MC-RR and NOD which were 53.2% and 54.3, respectively) with satisfactory inter-day and intra-day precisions (RSD below 13.3%).

Switchable fatty acid based CO2-effervescence ameliorated emulsification microextraction prior to high performance liquid chromatography for efficient analyses of toxic azo dyes in foodstuffs.

An in-situ chemical reaction based dispersive procedure termed as CO2-effervescence assisted emulsification microextraction was first utilized for pre-concentration of Sudan dyes in spices. It is simple, quick, and efficient; based on a switchable fatty acid (hexanoic acid, pKa ∼ 4.8). The extraction agent is able to be switched into hydrophobic/hydrophilic forms in pH-changeable media, leading to facile and effective dispersion and phase's separation. The method was proved by the analysis of spiked samples. Optimization was statistically performed, resulted in maximum extraction efficiencies. Under optimal conditions, method validation was assayed in the aqueous solutions. Wide linearity with high R2 values and low detection limits (1-5 ng mL-1) were obtainable. Also, enrichment factors were 62-66 with proper repeatability (%RSDs < 6.8, n = 3). Good validations in terms of recovery (86.6-104.5%) and repeatability (%RSDs < 7.8, n = 3, C = 15 and 30 µg g-1) in foodstuffs confirmed a great effectiveness of the method in complex matrices.

Highly effective and safe intermediate based on deep eutectic medium for carrier less-three phase hollow fiber microextraction of antiarrhythmic agents in complex matrices.

For the first time, three phase hollow fiber liquid phase microextraction using an influential, and green middle phase comprised a new relatively-hydrophobic deep eutectic solvent (three-phase HF-LPME-DES) was developed for trace analyses of antiarrhythmic drugs in biological and environmental samples. The extraction solvent was easily synthesized by mixing the green and cheap raw materials, namely choline chloride and 1-phenylethanol (ChCl: Ph-ETOH), in the ambient temperature. Good compatibility to pores of hollow-fiber, high ability for extraction of ionizable organic compounds with no need to any carrier agents, and easy availability in the laboratory environment turned this new proposed deep eutectic intermediate to a worthy generation of the supported liquid membrane (SLM). Final determination was accurately done by high performance liquid chromatography-ultraviolet detection (HPLC-UV). After effective statistical optimization of main parameters, the valid analytical features were found to be: wide linear dynamic ranges (LDRs) of 0.8 to 500 ng mL-1 with the determination coefficients (R2s) higher than 0.98, low detection limits (LODs) of 0.3-0.8 ng mL-1, and logical precision (relative standard deviations (%RSDs, n = 3) of 5.2-6.5%). Also, enrichment factors and extraction recoveries were 110-135 and 44-54%, respectively. These satisfactory results confirmed the potent effectiveness of the proposed microextraction procedure for achievement to clean and proper enrichment of the aforesaid compounds in highly complex real samples.

Emulsification microextraction of amphetamine and methamphetamine in complex matrices using an up-to-date generation of eco-friendly and relatively hydrophobic deep eutectic solvent.

In this study, a new extraction medium based on a quite bio-compatible and bio-degradable deep eutectic solvent comprising choline chloride and phenylethanol (ChCl: Ph-ETOH) was simply and cheaply synthesized at room temperature. At the next step, it was effectively utilized at the service of air agitated-emulsification microextraction (AA-EME) of two major amphetamine-type stimulants (ATSs) in human plasma and pharmaceutical wastewater pursued by high performance liquid chromatography-ultraviolet detection (HPLC-UV). This safe, effective, and rapid enrichment process based on the new low-density DES was easily practicable via a homemade extraction cell possessing a narrow neck and with no extra demand the emulsifier intermediates. Statistical study of main parameters effects using central composite design (CCD) combined with desirability function (DF) demonstrated that pH 12, 250 µL of extraction solvent, 8 air agitation cycles, and 5% of salt amount resulted in maximum extraction efficiencies (63-66%) with DF value close to 0.98. Under optimal conditions, wide linear dynamic ranges (LDRs) of 15.0-2000 and 8.0-3000 ng mL-1 with the determination coefficients (R2s) close to 0.99 were obtainable for amphetamine and methamphetamine, respectively. Low limits of detection (LODs) as well as relative standard deviations (%RSDs, n = 3) were found to be 2.0-5.0 ng mL-1 and 5.7-7.8%, respectively. Also, enrichment factors (EFs) were quantitative in the span of 47-50. On the other hand, satisfactory and accurate assessment at low levels close to therapeutic and toxic domains in human plasma sample and pharmaceutical wastewater was successfully obtained.

Magnetic nanoparticle based solid-phase extraction of heavy metal ions: A review on recent advances.

This review (with 151 refs) focuses on recent progress that has been made in magnetic nanoparticle-based solid phase extraction (SPE), pre-concentration and speciation of heavy metal ions. In addition, it discusses applications to complex real samples such as environmental, food, and biological matrices. The introduction addresses current obstacles and limitations associated with established SPE approaches and discusses the present state of the art in different formats of off-line and on-line SPE. The next section covers magnetized inorganic nanomaterials for use in SPE, with subsections on magnetic silica, magnetic alumina and titania, and on magnetic layered double oxides. A further section treats magnetized carbonaceous nanomaterials for use in SPE, with subsections on magnetic graphene and/or graphene oxides, magnetic carbon nanotubes and magnetic carbon nitrides. We then discuss the progress made in SPE based on the use of magnetized organic polymers (mainly non-imprinted and ion-imprinted polymer). This is followed by shorter sections on the use of magnetized metal organic frameworks, magnetized ionic liquids and magnetized biosorbents. All sections include discussions of the nanomaterials in terms of selectivity, sorption capacity, mechanisms of sorption and common routes for material synthesis. A concluding section addresses actual challenges and discusses perspective routes towards further improvements. Graphical abstract An overview on booster nanomaterials (ionic liquids, inorganic, organic and biological materials, and metal-organic frameworks) for use in magnetic nanoparticle-based solid-phase extraction of heavy metal ions.

A twin purification/enrichment procedure based on two versatile solid/liquid extracting agents for efficient uptake of ultra-trace levels of lorazepam and clonazepam from complex bio-matrices.

In this research work, two consecutive dispersive solid/liquid phase microextractions based on efficient extraction media were developed for the influential and clean pre-concentration of clonazepam and lorazepam from complicated bio-samples. The magnetism nature of the proposed nanoadsorbent proceeded the clean-up step conveniently and swiftly (∼5min), pursued by a further enrichment via a highly effective and rapid emulsification microextraction process (∼4min) based on a deep eutectic solvent (DES). Finally, the instrumental analysis step was practicable via high performance liquid chromatography-ultraviolet detection. The solid phase used was an adequate magnetic nanocomposite termed as polythiophene-sodium dodecyl benzene sulfonate/iron oxide (PTh-DBSNa/Fe3O4), easily and cost-effectively prepared by the impressive co-precipitation method followed by the efficient in situ sonochemical oxidative polymerization approach. The identification techniques viz. FESEM, XRD, and EDX certified the supreme physico-chemical properties of this effective nanosorbent. Also the powerful liquid extraction agent, DES, based on bio-degradable choline chloride, possessed a high efficiency, tolerable safety, low cost, and facile and mild synthesis route. The parameters involved in this versatile hyphenated procedure, efficiently evaluated via the central composite design (CCD), showed that the best extraction conditions consisted of an initial pH value of 7.2, 17mg of the PTh-DBSNa/Fe3O4 nanocomposite, 20 air-agitation cycles (first step), 245µL of methanol, 250µL of DES, 440µL of THF, and 8 air-agitation cycles (second step). Under the optimal conditions, the understudied drugs could be accurately determined in the wide linear dynamic ranges (LDRs) of 4.0-3000ngmL-1 and 2.0-2000ngmL-1 for clonazepam and lorazepam, respectively, with low limits of detection (LODs) ranged from 0.7 to 1.0ngmL-1. The enrichment factor (EF) and percentage extraction recovery (%ER) values were found to be 75 and 57% for clonazepam and 56 and 42% for lorazepam at the spiked level of 75.0ngmL-1, possessing proper repeatabilities (relative standard deviation values (RSDs) below 5.9%, n=3). These valid analytical features provided quite accurate drug analyses at therapeutically low spans and levels below potentially toxic domains, implying a proper purification/enrichment of the proposed microextraction procedure.

Ultrasound-promoted dispersive micro solid-phase extraction of trace anti-hypertensive drugs from biological matrices using a sonochemically synthesized conductive polymer nanocomposite.

In this work, a rapid and efficient procedure named ultrasound meliorated dispersive micro solid-phase extraction followed by high performance liquid chromatography-ultra violet detection (US-D-µSPE-HPLC-UV) was developed for the pre-concentration of the main trace anti-hypertensive drugs in complex matrices. The basis of this procedure was a polypyrrole-sodium dodecylbenzenesulfonate/zinc oxide (PPy-DBSNa/ZnO) nanocomposite. It was readily synthesized by the impressive way of in situ sonochemical oxidative polymerization in the presence of some additives such as FeCl3 and DBSNa, ultimately leading to the effective coating of PPy on the ZnO nanoparticle cores. Characterization of the proposed nanosorbent was performed by different techniques such as FESEM, XRD,EDX, and TGA, confirming the high quality and proper physico-chemical properties of the proposed sorbent. In order to better investigate the input variables, the central composite design (CCD) combined with the desirability function (DF) was utilized. The enriched optimum conditions consisted of the initial pH value of 11.8, 15mg of the PPy-DBSNa/ZnO nanocomposite, a sonication time of 4.6min, and 100µL of methanol, resulting in maximum responses at a relatively low extraction time with a logical DF. Under the optimum conditions, good linearity (5-5000, 2.5-3500, and 2.5-3000ngmL-1 for metoprolol, propranolol, and carvedilol, respectively, with the correlation of determinations (R2s) higher than 0.99), low limits of detection (LODs) (0.8-1.5ngmL-1), proper repeatabilities (relative standard deviation values (RSDs) below 6.3%, n=3), reasonable enrichment factors (EFs) (60-72), and good extraction recoveries (ERs) (higher than %75) were obtainable. These appropriate validations corroborated a good effectiveness of ultrasonic waves in the achievement of a supreme solid phase as well as a facile and efficient microextraction of the low therapeutic concentrations in human plasma and urine samples.

Efficient and clean pre-concentration of ultra-trace calcium channel blockers from biological matrices via a hyphenated procedure of two sequential dispersive solid/liquid phase microextractions.

In the present work, we propose a safe, simple, and relatively rapid procedure for the efficient clean-up and pre-concentration of ultra-trace calcium channel blockers (CCBs) from the human plasma and urine samples followed by high performance liquid chromatography-ultraviolet detection. The proposed sample preparation method is a combination of two microextraction methods termed as ultrasound-assisted dispersive micro solid-phase extraction coupled with air-agitated liquid-liquid microextraction based on solidification of a floating organic droplet (UA-dµSPE-AA-LLME-SFO). A superior clean-up and a higher pre-concentration factor are two valuable outcomes of the mentioned procedure, leading to an accurate measurement of the therapeutically low concentrations in the biological samples. The basis of the first dispersive micro solid-phase extraction is an effective nano-adsorbent named the PANI-DBSNa/TiO2 nano-composite. It was easily synthesized sonochemically by the in situ chemical oxidative polymerization method as core-sell structures, and subsequently, characterized by different techniques including FESEM, XRD, and TGA. The optimum conditions enriched via the response surface methodology (RSM) consisted of pH 12.1, 23 mg of the PANI-DBSNa/TiO2 nano-composite, a sonication time of 4.3 min, 225 µL of methanol, and 78 µL of 1-undecanol. Under the optimum experimental conditions, the good linear ranges of 4.0-4000, 8.0-10000, and 7.0-8000 ng mL-1 were obtainable for diltiazem, amlodipine, and verapamil, respectively, with the correlation of determinations (R2s) higher than 0.99 and the low limits of detection (LODs) of 1.5-3.0 ng mL-1. The relative standard deviations (%RSDs) were in the span of 5.5-6.5% (n = 3); implying on the satisfactory repeatability (or reproducibility). The enrichment factor (EF) and extraction recovery percentage (%ER) values were found to be 68 and 65% for diltiazem, 80 and 75% for amlodipine, and 46 and 45% for verapamil, respectively.

Rapid determination of some beta-blockers in complicated matrices by tandem dispersive liquid-liquid microextraction followed by high performance liquid chromatography.

In this research work, an efficient tandem dispersive liquid-liquid microextraction (TDLLME) procedure coupled with high performance liquid chromatography-ultraviolet detection (HPLC-UV) was successfully applied for the determination of beta-blockers in human plasma and pharmaceutical wastewater samples. High clean-up and preconcentration factor are easily and rapidly feasible via this novel, cheap, and safe microextraction method, leading to high quality experimental data. It consists of two sequential dispersive liquid-liquid microextraction methods, accomplished via air/ultrasonic agitation and air agitation, respectively. In order to enrich the optimal values for the mentioned procedures, the Box-Behnken design (BBD) combined with the desirability function (DF) was used. The optimum values were found to be 11.0 % (w/v) of the salt amount, an initial pH value of 12.0, 103 µL of organic extractant phase, and 45 µL of aqueous extractant phase with pH value of 2.0, resulted in reasonable recovery percentages with a logical desirability. Under optimal experimental conditions, good linear ranges (3-2000 ng mL-1 for metoprolol and 2.5-2500 ng mL-1 for propranolol with the correlation of determinations (R 2s) higher than 0.99) and low limits of detection (0.8 and 1.0 ng mL-1 for propranolol and metoprolol, respectively) were obtainable. Also, TDLLME-HPLC-UV provided good proper repeatabilities (relative standard deviations (RSDs) below 5.7 %, n = 3) and high enrichment factors (EFs) of 75-100. Graphical abstract TDLLME of beta-blockers from complicated matrices.

Tandem air-agitated liquid-liquid microextraction as an efficient method for determination of acidic drugs in complicated matrices.

A rapid and simple microextraction method with a high sample clean-up, termed as tandem air-agitated liquid-liquid microextraction (TAALLME), is described. This method is based upon the tandem implementation of the air-agitated liquid-liquid microextraction (AALLME), and this approach improves the applicability of the dispersive liquid-liquid microextraction (DLLME) methods in complicated matrices. With very simple tools, the three non-steroidal anti-inflammatory drugs diclofenac, ibuprofen, and mefenamic acid were efficiently extracted, with an overall extraction time of 7 min. By performing the first AALLME, these acidic analytes, contained in an aqueous sample solution (donor phase, 8.0 mL), were extracted into the organic solvent (1,2-dichloroethane, 37 µL), and their simple back-extraction into the aqueous acceptor solution (pH, 10.01, 51 µL) was obtained in 2 min by a second implementation of AALLME. Response surface methodology (RSM) was used for optimization of the experimental parameters. The pH values 2.94 and 10.01 were obtained for the donor and acceptor phases, respectively, and the volumes 99.5 and 51 µL were obtained for the organic solvent and the acceptor phase, respectively, as the optimal extraction conditions. Under the optimized conditions, tandem AALLME-HPLC-UV provided a good linearity in the range of 0.5-4000 ng mL(-1), limits of detection (0.1-0.3 ng mL(-1)), extraction repeatabilities (relative standard deviations (RSDs) below 7.7%, n = 5), and the enrichment factors (EFs) of 80-104. Finally, the applicability of the proposed method was evaluated by the extraction and determination of the drugs under study in the wastewater and human plasma samples.