Magnetic micro‐solid‐phase extraction based on magnetite‐MCM‐41 with gas chromatography–mass spectrometry for the determination of antidepressant drugs in biological fluids

A new facile magnetic micro‐solid‐phase extraction coupled to gas chromatography and mass spectrometry detection was developed for the extraction and determination of selected antidepressant drugs in biological fluids using magnetite‐MCM‐41 as adsorbent. The synthesized sorbent was characterized by several spectroscopic techniques. The maximum extraction efficiency for extraction of 500 μg/L antidepressant drugs from aqueous solution was obtained with 15 mg of magnetite‐MCM‐41 at pH 12. The analyte was desorbed using 100 μL of acetonitrile prior to gas chromatography determination. This method was rapid in which the adsorption procedure was completed in 60 s. Under the optimized conditions using 15 mL of antidepressant drugs sample, the calibration curve showed good linearity in the range of 0.05–500 μg/L (r ² = 0.996–0.999). Good limits of detection (0.008–0.010 μg/L) were obtained for the analytes with good relative standard deviations of <8.0% (n  = 5) for the determination of 0.1, 5.0, and 500.0 μg/L of antidepressant drugs. This method was successfully applied to the determination of amitriptyline and chlorpromazine in plasma and urine samples. The recoveries of spiked plasma and urine samples were in the range of 86.1–115.4%. Results indicate that magnetite micro‐solid‐phase extraction with gas chromatography and mass spectrometry is a convenient, fast, and economical method for the extraction and determination of amitriptyline and chlorpromazine in biological samples.     

Determination of four bisphenols in water and urine samples by magnetic dispersive solid‐phase extraction using a modified zeolite/iron oxide composite prior to liquid chromatography diode array detection

A novel approach is presented to determine four bisphenols in water and urine samples, employing magnetic dispersive solid‐phase extraction combined with liquid chromatography and diode array detection. A modified zeolite‐based magnetic composite was used as an efficient sorbent, combining the advantages of magnetic materials with the remarkable properties of zeolites. A multivariate optimization design was employed to optimize some experimental factors affecting magnetic dispersive solid‐phase extraction. The method was evaluated under optimized conditions (i.e., amount of sorbent, 50 mg; sample pH, unadjusted; NaCl concentration, 1.25%; extraction and elution time, 2 min; eluent solvent, ethanol; eluent solvent volume, 400 µL), obtaining good linearity with correlation coefficients ranging between 0.995 and 0.999 (N = 5) (from 2 to 250 µg/L for bisphenol A, bisphenol AP, and bisphenol P and from 5 to 250 µg/L for bisphenol AF). Method repeatability was assessed obtaining coefficients of variation between 3 and 11% (n = 6). Finally, the method was applied to spiked real samples, obtaining for water samples relative recoveries between 83 and 105%, and for urine samples between 81 and 108% for bisphenol A, bisphenol AP, and bisphenol AF, and between 47 and 59% for bisphenol P.     

Core-shell magnetic porous organic polymer for magnetic solid-phase extraction of fluoroquinolone antibiotics in honey samples followed by high-performance liquid chromatography with fluorescence detection

By monomer-mediated in-situ growth synthesis strategy, with hydroquinone and 1,3,5-tris(4-aminophenyl)benzene as monomers, a core-shell magnetic porous organic polymer was synthesized through a simple azo reaction. Based on this, a magnetic solid-phase extraction–high-performance liquid chromatography–fluorescence detection method was proposed for the analysis of fluoroquinolones in a honey sample. With ofloxacin, ciprofloxacin, enrofloxacin, lomefloxacin, and difloxacin as target analytes, factors affecting the extraction efficiency had been optimized. The LODs were 1.5–5.4 ng/L (corresponding to 0.23–0.81 μg/kg in honey). The linear range was 0.005–20 μg/L for difloxacin, 0.01–20 μg/L for ofloxacin, ciprofloxacin and lomefloxacin, and 0.02–20 μg/L for enrofloxacin. The enrichment factor was 84.4–91.7-fold with a high extraction efficiency of 84.4–91.7%. The method was assessed by the analysis of target fluoroquinolones in honey samples, and the recoveries for the spiked samples were 79.3–95.8%. The results indicated that the established magnetic solid-phase extraction–high-performance liquid chromatography–fluorescence detection method is efficient for the analysis of trace fluoroquinolones in honey.     

Determination of 1-hydroxypyrene in human urine by HPLC with electrochemical detection at a boron-doped diamond film electrode

A high-performance liquid chromatographic method with electrochemical detection (HPLC-ED) at a boron-doped diamond film electrode with preliminary separation and preconcentration by solid-phase extraction (SPE) has been developed for the determination of 1-hydroxypyrene (1-HP) in human urine. 1-HP is among the most widely used biomarkers of exposure to polycyclic aromatic hydrocarbons. Optimal HPLC-ED conditions have been found: mobile phase methanol-0.05 mol L(-1) phosphate buffer pH 5.0 (80:20, v/v), detection potential +1,000 mV versus Ag/AgCl (3 mol L(-1) KCl), and flow rate 0.8 mL min(-1). For SPE, LiChrolut(?) RP-18 E cartridges were used. The extraction yield was (87.0 ± 5.8)% (n = 5). The concentration dependence of 1-HP was measured in the concentration range from 0.01 to 10 μmol L(-1) (2.18-2,180 μg L(-1)) using methanolic solutions resulting from the SPE pretreatment of spiked human urine samples. The limit of detection (signal-to-noise ratio 3) and the limit of quantification (signal-to-noise ratio 10) of the biomarker were 0.013 μmol L(-1) (2.84 μg L(-1)) and 0.043 μmol L(-1) (9.39 μg L(-1)), respectively, which is sufficient for its determination in the urine of persons exposed to polycyclic aromatic hydrocarbons.     

A natural and renewable biosorbent phase as a low‐cost approach in disposable pipette extraction technique for the determination of emerging contaminants in lake water samples

This study proposes an efficient analytical methodology using a biosorbent (cork) as an extraction phase in disposable pipette extraction technique for the rapid determination of the emerging contaminants methyl paraben, ethyl paraben, benzophenone, 3‐(4‐methylbenzylidene) camphor and 2‐(ethylhexyl)‐4‐(dimethylamino) benzoate in lake water samples using high‐performance liquid chromatography with diode array detection. The optimized conditions were comprised of 800 μL of sample, three cycles of 30 s each for the extraction, pH 6, addition of 30% w/v of NaCl. For the desorption step, the optimized desorption conditions were achieved with 100 μL of a mixture comprised of 50% methanol and 50% acetonitrile v/v, using one cycle of 30 s. Excellent analytical performance was achieved with limits of detection of 0.6 μg/L for methyl paraben to 1.4 μg/L for 3‐(4‐methylbenzylidene) camphor, and the limit of quantitation varied from 2 μg/L for methyl paraben to 4.3 μg/L 3‐(4‐methylbenzylidene) camphor, respectively. The correlation coefficients ranged from 0.9962 for ethyl paraben to 0.9980 for methyl paraben. The method accuracy varied from 71–132%, and the intraday precision ranged from 3 to 23% (n = 3) and interday from 9 to 23% (n = 9). The robustness was evaluated through Youden and Lenth's methods and indicated no significant variations in the results.     

Application of a magnetic molecularly imprinted polymer for the selective extraction and trace detection of lamotrigine in urine and plasma samples

Magnetic molecularly imprinted polymers have been synthesized for the selective preconcentration and trace determination of lamotrigine (LTG) in urine and plasma samples. The magnetic nanoparticles were modified by tetraethyl orthosilicate and 3‐methacryloxypropyl trimethoxysilane before imprinting. The magnetic molecularly imprinted polymers were prepared via surface molecular imprinting technique, using Fe₃O₄ as a magnetic component, LTG as template molecule, methacrylic acid as a functional monomer, ethylene glycol dimethacrylate as a cross‐linker, and 2,2′‐azobisisobutyronitrile as a radical initiator in methanol/acetonitrile (50:50, v/v) as the porogen. The obtained sorbent was characterized using scanning electron microscopy, Fourier‐transform infrared spectroscopy, X‐ray diffraction, and thermal analysis. Separation of the sorbent from the sample solution was simply achieved by applying an external magnetic field. Determination of the extracted drug was performed by high‐performance liquid chromatography with UV detection. Under the optimum extraction conditions, the method detection limits were 0.7 μg/L (based on S/N of 3) for urine samples and 0.5 μg/L for plasma samples. A linear dynamic range of 1–1000 μg/L was obtained for LTF in plasma and urine samples. Finally, the applicability of the proposed method was evaluated by extraction and determination of LTG in urine and plasma samples.     

Polyaniline-silica doped with oxalic acid as a novel extractor phase in thin film solid-phase microextraction for determination of hormones in urine

In this study, different polyanilines were synthesized and evaluated for the determination of three hormones, including 17-β-estradiol, 17-α-ethinylestradiol, and estrone, in urine using a novel methodology based on thin film solid-phase microextraction technique, employing the sampling well plate system. The extractor phases, designated as polyaniline doped with hydrochloric acid, polyaniline doped with oxalic acid, polyaniline-silica doped with hydrochloric acid, and polyaniline-silica doped with oxalic acid, were characterized by electrical conductivity measurements, scanning electron microscopy, and Fourier transform infrared spectroscopy. The optimized extraction conditions were composed of 1.5 mL of urine and pH adjusted to 10, with no need to dilute sample and the desorption step, 300 μL of acetonitrile was used. The calibration curves were performed in the sample matrix, with detection and quantification limits ranged from 0.30 to 3.03 μg L⁻¹ and from 1.0 to 10.0 μg L⁻¹, respectively, with r ≥ 0.9969. The relative recoveries ranged from 71% to 115%, and intraday precision showed values ≤12% and interday ≤20%. The applicability of the method was successfully evaluated, and six urine samples from female volunteers were analyzed. The analytes were not detected or were below the limits of quantification in these samples.     

Inside-tube solid-phase microextraction as an interlink between solid-phase microextraction and needle device for n-hexane evaluation in air and urine headspace

Monitoring the trace amount of chemicals in various samples remains a challenge. This study was conducted to develop a new solid-phase microextraction (SPME) system (inside-tube SPME) for trace analysis of n-hexane in air and urine matrix. The inside-tube SPME system was prepared based on the phase separation technique. A mixture of carbon aerogel and polystyrene was loaded inside the needle using methanol as the anti-solvent. The air matrix of n-hexane was prepared in a Tedlar bag, and n-hexane vapor was sampled at a flow rate of 0.1 L/min. Urine samples spiked with n-hexane were used to simulate the sampling method. The limit of detection using the inside-tube SPME was 0.0003 μg/sample with 2.5 mg of adsorbent, whereas that using the packed needle was 0.004 μg/sample with 5 mg of carbon aerogel. For n-hexane analysis, the day-to-day and within-day coefficient variation were lower than 1.37%, with recoveries over 98.41% achieved. The inside-tube SPME is an inter-link device between two sample preparation methods, namely, a needle trap device and an SPME system. The result of this study suggested the use of the inside-tube SPME containing carbon aerogel (adsorbent) as a simple and fast method with low cost for n-hexane evaluation.     

Carbonized silk fibers for in‐tube solid‐phase microextraction to detect polycyclic aromatic hydrocarbons in water samples

Silk fibers were carbonized to develop a biomass carbon material as an adsorbent for solid‐phase microextraction. The surface structure of the carbonized silk fibers was characterized by scanning electron microscopy, and the graphitization degree was determined by Raman spectrometry. After carbonization under high temperature, the orderliness and structural regularity of carbon atoms on silk fibers were promoted. Extraction tube packed with carbonized silk fibers was prepared for in‐tube solid‐phase microextraction. Coupled with high performance liquid chromatography, it exhibited good extraction performance for hydrophobic polycyclic aromatic hydrocarbons. Main parameters including sampling volume, sampling rate, methanol content in sample, and desorption time were systematically investigated. Under the optimum conditions, the analysis method was established and it exhibited wide linear range (0.016–20 μg/L) with good linearity (correlation coefficient ≥ 0.9947), low limits of detection (0.005–0.050 μg/L), and high enrichment factors (1189–2775). Relative standard deviations (n = 3) for intraday (≤3.3%) and interday (≤9.6%) tests indicated that the extraction material had satisfactory repeatability. Finally, the analytical method was successfully applied to detect trace polycyclic aromatic hydrocarbons in real water samples, demonstrating its satisfactory practicability.     

Fabrication of boronate‐decorated polyhedral oligomeric silsesquioxanes grafted cotton fiber for the selective enrichment of nucleosides in urine

Various cotton fiber based boronate‐affinity adsorbents are recently developed for the sample pretreatment of cis‐diol‐containing biomolecules, but most do not have efficient capacity due to limited binding sites on the surface of cotton fibers. To increase the density of boronate groups on the surface of cotton fiber, polyhedral oligomeric silsesquioxanes were used to modify cotton fiber to provide plentiful reactive sites for subsequent functionalization with 4‐formylphenylboronic acid. The new adsorbent showed special recognition ability towards cis‐diols and high adsorption capacity (175 μg/g for catechol, 250 μg/g for dopamine, 400 μg/g for adenosine). The in‐pipette‐tip solid‐phase extraction was investigated under different conditions, including pH and ionic strength of solution, adsorbent amount, pipette times, washing solvent, and elution solvent. The in‐pipette‐tip solid‐phase extraction coupled with high‐performance liquid chromatography was used to analyze four nucleosides in urine samples. Under the optimal extraction conditions, the detection limits were determined to be between 5.1 and 6.1 ng/mL (S/N  =  3), and the linearity ranged from 20 to 500 ng/mL for these analytes. The accuracy of the analytical method was examined by studying the relative recoveries of analytes in real urine samples with recoveries varying from 83 to 104% (RSD = 3.9–10.2%, n = 3).     

Simple and sensitive determination of malondialdehyde in human urine and saliva using UHPLC–MS/MS after derivatization with 3,4‐diaminobenzophenone

Malondialdehyde has been used as a biomarker for lipid peroxidation in biological samples. An ultra‐high performance liquid chromatography with tandem mass spectrometry method was developed to determine the levels of malondialdehyde in human urine and saliva samples. To select the optimum derivatization reagent from four diamino compounds, the reactivity and sensitivity of their derivatives were compared, and 3,4‐diaminobenzophenone was selected. The optimum reaction conditions for malondialdehyde with 3,4‐diaminobenzophenone were as follows: a reagent dosage of 50 mg/L, pH of 4, and reaction for 30 min at 50°C. The formed derivative product was analyzed using ultra‐high performance liquid chromatography with tandem mass spectrometry without additional extraction or concentration steps. In the optimal conditions, the method was used to determine malondialdehyde concentration in human urine and saliva samples. The limits of quantification for malondialdehyde in biological samples were over a concentration range of 0.1–0.3 μg/L. Additionally, the calibration curve showed a linearity greater than r ² = 0.997. The method was used to analyze 14 human urine and saliva samples from healthy volunteers. Malondialdehyde was detected in the concentration range of 1.7–33.6 μg/g creatinine in all human urine samples and 0.1–1.3 μg/L in all human saliva samples.     

Determination of fipronil and its metabolites in environmental water samples by meltblown nonwoven fabric-based solid-phase extraction combined with gas chromatography-electron capture detection

In this study, a new method for the determination of fipronil and its three metabolites in environmental water samples was developed based on meltblown nonwoven fabric solid-phase extraction combined with gas chromatography-electron capture detection. As the core material of medical masks, meltblown nonwoven fabric is made of polypropylene superfine fibers which are randomly distributed and bonded together with a relatively large specific surface area and good permeability. Polypropylene as a high molecular hydrocarbon-based polymer has the characteristics of good hydrophobicity and lipophilicity, which can be applied for the separation and enrichment of hydrophobic substances in food, environment, and biological samples. The meltblown nonwoven fabric is soft and can fill the solid-phase extraction cartridge tightly. This aspect also makes it suitable to be used as an ideal solid-phase extraction sorbent. A series of parameters influencing the extraction efficiency were investigated, and under the optimized conditions, fipronil and its three metabolites had a good linear relationship in the range of 0.2–100 μg/L with a correlation coefficient R^{2 of} more than 0.999. The recoveries at three spiked concentrations were in the range of 99.2–107.3% with the relative standard deviations less than 9.8% (intra-day) and 8.1% (inter-day). The limit of detection for the four target analytes was in the range of 0.02–0.06 μg/L. Finally, this method was successfully applied in the analysis of fipronil and its three metabolites in various types of environmental water samples.     

Cellulose membrane modified with polypyrrole as an extraction device for the determination of emerging contaminants in river water with gas chromatography–mass spectrometry

In this study, a simple, efficient, and reusable device based on cellulose membranes modified with polypyrrole was developed to extract 14 emerging contaminants from aqueous matrices. For chemical polymerization, a low‐cost cellulose membrane was immersed in 0.1 mol/L pyrrole and 0.5 mol/L ammonium persulfate for 40 min in an ice/water bath. The cellulose membranes modified with polypyrrole were accommodated in a polycarbonate holder suitable for solid‐phase extraction disks. Solid‐phase extraction parameters that affect extraction efficiency, such as sample volume, pH, flow rate, and desorption were optimized. Subsequently, determination of target compounds was performed by gas chromatography with mass spectrometry. The linear range for analytes ranged from 0.05 to 500 μg/L, with coefficients of determination above 0.990. The limits of quantification varied between 0.05 and 10 μg/L, with relative standard deviations lower than 17%. The performance of the proposed cellulose membranes modified with polypyrrole device for real samples was evaluated after extraction of emerging contaminants from a river water sample from the city of Curitiba, Brazil. Bisphenol A (6.39 μg/L), caffeine (17.83 μg/L), and paracetamol (19.28 μg/L) were found in these samples.     

Expanding the applicability of magnetic ionic liquids for multiclass determination in biological matrices based on dispersive liquid–liquid microextraction and HPLC with diode array detector analysis

Monitoring biological samples at trace levels of chemicals from anthropogenic actions such as pesticides, pharmaceuticals, and hormones has become a very important subject. This work describes a method for the determination of eight compounds of different chemical classes in human urine samples. Dispersive liquid–liquid microextraction based on magnetic ionic liquids was used as the sample preparation procedure. The main parameters of the method, such as sample dilution, type, and volume of disperser solvent, amount of magnetic ionic liquids, extraction time, and pH were optimized by univariate and multivariate procedures. Validation was performed using a urine sample of a male volunteer in order to obtain a calibration curve and the main analytical parameters of merit such as limits of detection and quantification. Values varied from 3.0 to 7.5 µg/L and from 10 to 25 µg/L, respectively. Satisfactory precisions of 21% for intraday (n = 3) and 16% for interday (n = 9) were achieved. Accuracy was evaluated by relative recovery assays using different urine samples and ranged from 75 to 130%. Robustness was assured by the Lenth method. The validated procedure was applied to five urine samples from different volunteers and the hormone estrone was found in one sample.     

Immunochemical analysis of 3-phenoxybenzoic acid,a biomarker of forestry worker exposure to pyrethroid insecticides

Pyrethroid insecticides widely used in forestry, agricultural, industrial, and residential applications have potential for human exposure. Short sample preparation time and sensitive, economical high-throughput assays are needed for biomonitoring studies that analyze a large number of samples. An enzyme-linked immunosorbent assay (ELISA) was used for determining 3-phenoxybenzoic acid (3-PBA), a general urinary biomarker of exposure to some pyrethroid insecticides. A mixed-mode solid-phase extraction reduced interferences from acid hydrolyzed urine and gave 110 ± 6% recoveries from spiked samples. The method limit of quantification was 2 μg/L. Urine samples were collected from forestry workers that harvest pine cone seeds where pyrethroid insecticides were applied at ten different orchards. At least four samples for each worker were collected in a 1-week period. The 3-PBA in workers classified as high, low, or no exposure based on job analysis over all sampling days was 6.40 ± 9.60 (n = 200), 5.27 ± 5.39 (n = 52), and 3.56 ± 2.64 ng/mL (n = 34), respectively. Pair-wise comparison of the differences in least squares means of 3-PBA concentrations among groups only showed a significant difference between high and no exposure. Although this difference was not significant when 3-PBA excretion was normalized by creatinine excretion, the general trend was still apparent. No significant differences were observed among days or orchards. This ELISA method using a 96-well plate was performed as a high-throughput tool for analyzing around 300 urine samples measured in triplicate to provide data for workers exposure assessment.     

Evaluation of measurement uncertainty in the determination of jasmonic acid in <Emphasis Type="Italic">Lemna minor</Emphasis> L. by liquid chromatography with fluorescence detection

The uncertainty was estimated for the determination of jasmonic acid (JA) content in Lemna minor L. plant extracts using a reverse-phase liquid chromatography with fluorescence detection. JA was extracted from plant material, followed by solid-phase extraction procedures, derivatisation and quantification. In the estimation of uncertainty, the sampling, sample processing and chromatographic determination that may significantly influence the uncertainty of analytical data were considered. The results show that the method recovery and sample homogeneity are the two main contributors to uncertainty. The method has a relative expanded uncertainty (coverage factor k = 2) of about 17% at the JA content of approximately 100 ng/g.     

Application of magnetized MOF‐74 to phthalate esters extraction from Chinese liquor

In this study, magnetized MOF‐74 (Ni) was prepared using an ultrasound‐assisted synthesis method. This novel functional magnetic adsorbent was characterized using various techniques. Using the prepared material as adsorbents, a magnetic solid‐phase extraction method coupled with high‐performance liquid chromatography was proposed for determining four phthalate esters in Chinese liquor samples. The extraction parameters, including solution pH, adsorbent amount, extraction time, and eluent type and volume, were optimized. Under the optimized conditions, proposed method showed good linearity within the range of 1.53–200 μg/L for diphenyl phthalate, 2.03–200 μg/L for butyl benzyl phthalate, 7.02–200 μg/L for diamyl phthalate, and 6.03–200 μg/L for dicyclohexyl phthalate, with correlation coefficients > 0.9944, low limits of detection (0.46–2.10 μg/L, S/N = 3), and good extraction repeatability (relative standard deviations of 3.7%, n = 6). This method was successfully used to analyze phthalate esters in Chinese liquor samples with recoveries of 74.4–104.8%. Two phthalate esters were detected in two samples, both at concentrations that satisfied the Chinese national standard, indicating this method has practical application prospects. The extraction efficiency of this method was also compared with conventional solid‐phase extraction using commercial C₁₈ cartridges. The results demonstrated that the proposed magnetic solid‐phase extraction is a simple, time‐saving, efficient, and low‐cost method.     

Development of microextraction techniques in combination with GC–MS/MS for the determination of mycotoxins and metabolites in human urine

Simple and highly efficient sample preparation procedures, namely, dispersive liquid–liquid microextraction and salting‐out liquid–liquid extraction for the analysis of ten Fusarium mycotoxins and metabolites in human urine were compared. Various parameters affecting extraction efficiency were carefully evaluated. Under optimal extraction conditions, salting‐out liquid–liquid extraction showed a better accuracy (84–96%) and precision (<14%) than dispersive liquid–liquid microextraction. Hence, a multibiomarker method based on salting‐out liquid–liquid extraction followed by gas chromatography with tandem mass spectrometry was proposed. Satisfactory results in terms of validation were achieved. The method resulted in low limits of detection and quantitation within the range of 0.12–4 and 0.25–8 μg/L, respectively. The method accuracy and precision were evaluated at three spiking levels (8, 25 and 100 μg/L) and the recoveries were in a range from 70 to 120% with relative standard deviations lower than 15%. Matrix effect was evaluated and matrix‐matched calibrations were used for quantitation purpose. The developed method was applied in 12 human urine samples as a pilot study before and after sample treatment with β‐glucuronidase before the analysis to quantify the mycotoxin conjugates. Total deoxynivalenol (free + conjugated) was found in 83% of samples at an average concentration in positive samples of 31.6 μg/L.     

Preparation and utilization of a molecularly imprinted polymer for solid phase extraction of tramadol

In this paper, a highly selective molecularly imprinted polymer (MIP) for tramadol hydrochloride, a drug used to treat moderate to severe pain, was prepared and its use as solid-phase extraction (SPE) sorbent was demonstrated. The molecularly imprinted solid-phase extraction procedure followed by high performance liquid chromatography with ultraviolet detector (MISPE-HPLC) was developed for selective extraction and determination of tramadol in human plasma and urine. The optimal conditions for molecularly imprinted solid-phase extraction (MISPE) consisted of conditioning with 1 mL methanol and 1 mL of deionized water at neutral pH, loading of tramadol sample (50 μg L⁻¹) at pH 7.5, washing using 1 mL acetone and elution with 3 × 1 mL of 10% (v/v) acetic acid in methanol. The MIP selectivity was evaluated by checking several substances with similar molecular structures to that of tramadol. Results from the HPLC analyses showed that the calibration curve of tramadol (using MIP from human plasma and urine) is linear in the ranges of 6–100 and 3–120 μg L⁻¹ with good precisions (1.9% and 2.9% for 5.0 μg L⁻¹), respectively. The recoveries for plasma and urine samples were higher than 81%.      

Modified dispersive liquid‐phase microextraction based on sequential injection solidified floating organic drop combined with HPLC for the determination of phenobarbital and phenytoin

A modified dispersive liquid phase microextraction based on sequential injection solidified floating organic drop was developed for simultaneous separation/preconcentration of trace amounts of phenobarbital and phenytoin. The important factors affecting on the extraction recovery including pH, the volume of extraction solvent, ionic strength, and the number of injections were investigated and optimized by Box–Behnken design and desirability function. Under the optimum experimental conditions, the calibration graph was linear in the concentration range of 1.0–300.0 μg/L (r² = 0.997) for phenobarbital and 2.0–400.0 μg/L (r² = 0.996) for phenytoin. The limit of detection and limit of quantification were 0.35 and 1.2 μg/L for phenobarbital and 0.65 and 2.2 μg/L for phenytoin, respectively. The relative standard deviation for six replicate determinations at 10 μg/L was 3.3 and 4.1% for phenobarbital and phenytoin, respectively. The developed method was successfully applied to the determination of phenobarbital and phenytoin in urine and plasma samples.