Ultra-preconcentration and determination of thirteen organophosphorus pesticides in water samples using solid-phase extraction followed by dispersive liquid–liquid microextraction and gas chromatography with flame photometric detection

An ultra-preconcentration technique composed of solid-phase extraction (SPE) and dispersive liquid–liquid microextraction (DLLME) coupled with gas chromatography–flame photometric detection (GC–FPD) was used for determination of thirteen organophosphorus pesticides (OPPs) including phorate, diazinon, disolfotane, methyl parathion, sumithion, chlorpyrifos, malathion, fenthion, profenphose, ethion, phosalone, azinphose-methyl and co-ral in aqueous samples. The analytes were collected from large volumes of aqueous solutions (100 mL) into 100 mg of a SPE C₁₈ sorbent. The effective variables of SPE including type and volume of elution solvent, volume and flow rate of sample solution, and salt concentration were investigated and optimized. Acetone was selected as eluent in SPE and disperser solvent in DLLME and chlorobenzene was used as extraction solvent. Under the optimal conditions, the enrichment factors were between 15,160 and 21,000 and extraction recoveries were 75.8–105.0%. The linear range was 1–10,000 ng L^?1 and limits of detection (LODs) were between 0.2 and 1.5 ng L^?1. The relative standard deviations (RSDs) for 50 ng L^?1 of OPPs in water with and without an internal standard, were in the range of 1.4–7.9% (n = 5) and 4.0–11.6%, respectively. The relative recoveries of OPPs from well and farm water sat spiking levels of 25 and 250 ng L^?1 were 88–109%.     

Polyaniline-nylon-6 electrospun nanofibers for headspace adsorptive microextraction

A headspace adsorptive microextraction technique was developed using a novel polyaniline-nylon-6 (PANI-N6) nanofiber sheet, fabricated by electrospinning. The homogeneity and the porosity of the prepared PANI-N6 sheet were studied using the scanning electron microscopy (SEM) and nanofibers diameters were found to be around 200 nm. The novel nanofiber sheet was examined as an extracting medium to isolate some selected chlorobenzenes (CBs), as model compounds, from aquatic media. The extracted analytes were desorbed using μL-amounts of solvent and eventually an aliquot of extractant was injected into gas chromatography–mass spectrometry (GC–MS). Various parameters affecting the extraction and desorption processes were optimized. The developed method proved to be convenient and offers sufficient sensitivity and a good reproducibility. Limits of detection achieved for CBs with the developed analytical procedure ranged from 19 to 33 ng L^?1, while limits of quantification were from 50 to 60 ng L^?1. The relative standard deviations (RSD) at a concentration level of 0.1 ng mL^?1 and 1 ng mL^?1 were in the range of 8–14% and 5–11% (n = 3), respectively. The calibration curves of analytes were investigated in the range of 50–1000 ng L^?1 and R² between 0.9739 and 0.9932 were obtained. The developed method was successfully applied to the extraction of selected CBs from tap and river water samples. The relative recovery (RR) percentage obtained for the spiked real water samples at 0.1 ng mL^?1 and 1 ng mL^?1 level were 93–103% and 95–104%, respectively. The whole procedure showed to be conveniently applicable and quite easy to handle.     

Ultrasensitive determination of serum albumin using resonance light scattering based on ZnS-polyacrylic acid nanoparticles

ZnS-polyacrylic acid (ZnS-PAA) was prepared by an in situ polymerization method using nano-ZnS as core in the presence of acrylic acid (AA), and ZnS-PAA nanoparticles was characterized by ultraviolet spectrometry (UV) and transmission electron microscopy (TEM). Based on the significant increase of the resonance light scattering (RLS) intensity with the interaction between nanoparticles and serum albumin, RLS method was developed for the sensitive determination of serum albumin (BSA and HSA). Under optimum conditions, the change of the intensity (ΔI) of the RLS spectra at λ = 392 nm was linearly proportional to the concentration of BSA and HSA. The linear range was 1–100 ng mL^?1 for HSA and 1–120 ng mL^?1 for BSA, and the limit of detection (LOD) was 0.4 ng mL^?1 for HSA and 0.5 ng mL^?1 for BSA. This method proved to be very sensitive, rapid, simple and tolerant of most interfering substances.     

Sensitive liquid chromatography–tandem mass spectrometry method for the determination of pantoprazole sodium in human urine

A sensitive and selective liquid chromatographic–tandem mass spectrometric (LC–MS–MS) method was developed to determine pantoprazole sodium (PNT) in human urine. After solid-phase extraction with SPE cartridge, the urine sample was analysed on a C₁₈ column (symmetry 3.5 μm; 75 mm × 4.6 mm i.d) interfaced with a triple quadrupole tandem mass spectrometer. Positive electrospray ionization was employed as the ionization source. The mobile phase consisted of acetonitrile–water (90:10, v/v). The method was linear over a concentration range of 1–100 ng mL^?1. The lower limit of quantitation was 1 ng mL^?1. The intra-day and inter-day relative standard deviation across three validation runs over the entire concentration range was <10.5%. The accuracy determined at three concentrations (8.0, 50.0 and 85.0 ng mL^?1 PNT) was within ±1.25% in terms of relative errors.     

A novel approach for determining total titanium from titanium dioxide nanoparticles suspended in water and biosolids by digestion with ammonium persulfate

Titanium dioxide (i.e. TiO₂) in nano-form is a constituent of many nanomaterials that are used in sunscreens, cosmetics, industrial products and in biomedical applications. Quantification of TiO₂ nanoparticles in various matrixes is a topic of great interest for researchers studying the potential health and environmental impacts of nanoparticles. However, analysis of TiO₂ as Ti⁴⁺ is difficult because current digestion techniques require use of strong acids that may be a health and safety risk in the laboratory. To overcome this problem, we developed a new method to digest TiO₂ nanoparticles using ammonium persulfate as a fusing reagent. The digestion technique requires short times to completion and optimally requires only 1 g of fusing reagent. The fusion method showed >95% recovery of Ti⁴⁺ from 6 μg mL^?1 aqueous suspensions prepared from 10 μg mL^?1 suspension of different forms of TiO_2, including anatase, rutile and mixed nanosized crystals, and amorphous particles. These recoveries were greater than open hot-plate digestion with a tri-acid solution and comparable to microwave digestion with a tri-acid solution. Cations and anions commonly found in natural waters showed no significant interferences when added to samples in amounts of 10 ng to 110 mg, which is a much broader range of these ions than expected in environmental samples. Using ICP-MS for analysis, the method detection limit (MDL) was determined to be 0.06 ng mL^?1, and the limit of quantification (LOQ) was 0.20 ng mL^?1. Analysis of samples of untreated and treated wastewater and biosolids collected from wastewater treatment plants yielded concentrations of TiO₂ of 1.8 and 1.6 ng mL^?1 for the wastewater samples, respectively, and 317.4 ng mg^?1 dry weights for the biosolids. The reactions between persulfate ions and TiO₂ were evaluated using stoichiometric methods and FTIR and XRD analysis. A formula for the fusing reaction is proposed that involves the formation of sulfate radicals.     

Determination of sulfonamides in food samples by membrane-protected micro-solid phase extraction coupled with high performance liquid chromatography

In the present study, a simple and sensitive extraction method based on polypropylene membrane-protected micro-solid phase extraction (MP-μ-SPE) has been developed for analysis of sulfonamides in food samples. Poly (methacrylic acid-ethylene glycol dimethacrylate) (p-MAA-EDMA) was synthesized using orthogonal array experimental design, optimized with three factors at four levels and evaluated on yield, hydrophobic and cation-exchange properties. The optimized p-MAA-EDMA was then employed as the sorbent in the MP-μ-SPE for extraction of sulfonamides from milk and chicken muscle samples, followed by high performance liquid chromatographic analysis with ultraviolet detection. Under optimized extraction conditions, good linearities (0.010–1.0 μg mL^?1 with r² > 0.9900), low limits of detection (0.38–0.62 ng mL^?1), and acceptable intra-day (2.7–13.7%) and inter-day (6.7–15.2%) relative standard deviations were obtained. It was demonstrated to be an effective approach to handle semi-solid/solid samples with good resistance to interference from “dirty” samples.     

Electrochemical immunosensors for interleukin-6. Comparison of carbon nanotube forest and gold nanoparticle platforms

Electrochemical immunosensors based on single wall nanotube (SWNT) forests and 5 nm glutathione-protected gold nanoparticles (GSH-AuNP) were developed and compared for the measurement of human cancer biomarker interleukin-6 (IL-6) in serum. Detection was based on sandwich immunoassays using multiple (14–16) horseradish peroxidase labels conjugated to a secondary antibody. Performance was optimized by effective blocking of non-specific binding (NSB) of the labels using bovine serum albumin. The GSH-AuNP immunosensor gave a detection limit (DL) of 10 pg mL^?1 IL-6 (500 amol mL^?1) in 10 μL calf serum, which was 3-fold better than 30 pg mL^?1 found for the SWNT forest immunosensor for the same assay protocol. The GSH-AuNPs platform also gave a much larger linear dynamic range (20–4000 pg mL^?1) than the SWNT system (40–150 pg mL^?1), but the SWNTs had 2-fold better sensitivity in the low pg mL^?1 range.     

Determination of ofloxacin and gatifloxacin by mixed micelle-mediated cloud point extraction-fluorimetry combined methodology

A cloud point extraction process using mixed micelle of the anionic surfactant sodium dodecyl sulfate (SDS) and the non-ionic surfactant polyoxyethylene(7.5)nonylphenylether (PONPE 7.5) to extract two fluoroquinolone antimicrobial agents, ofloxacin and gatifloxacin, from aqueous media was investigated. The method is based on the mixed micelle-mediated extraction of fluoroquinolones in the presence of NaCl as an inducing agent in phase separation, followed by spectrofluorimetric determination. The effect of different variables such as pH, PONPE7.5 concentration, SDS concentration, NaCl concentration, cloud point temperature, and time was investigated, and optimum conditions were established. At optimum conditions, the rectilinear calibration graphs were obtained in the concentration range of 0.1–150 and 0.1–250 ng mL^?1 for ofloxacin and gatifloxacin, and the limits of detection were 0.04 and 0.06 ng mL^?1, respectively. The proposed procedure was applied successfully for the detection of the investigated drugs in their pharmaceutical dosage forms, in spiked plasma, spiked urine, and urine samples, with good precision and accuracy.     

Extraction and preconcentration of trace levels of cobalt using functionalized magnetic nanoparticles in a sequential injection lab-on-valve system with detection by electrothermal atomic absorption spectrometry

A new approach to performing extraction and preconcentration employing functionalized magnetic nanoparticles for the determination of trace metals is presented. Alumina-coated iron oxide nanoparticles were synthesized and used as the solid support. The nanoparticles were functionalized with sodium dodecyl sulfate and used as adsorbents for solid phase extraction of the analyte. Extraction, elution, and detection procedures were performed sequentially in the sequential injection lab-on-valve (SI-LOV) system followed by electrothermal atomic absorption spectrometry (ETAAS). Mixtures of hydrophobic analytes were successfully extracted from solution using the synthesized magnetic adsorbents. The potential use of the established scheme was demonstrated by taking cobalt as a model analyte. Under the optimal conditions, the calibration curve showed an excellent linearity in the concentration range of 0.01–5 μg L^?1, and the relative standard deviation was 2.8% at the 0.5 μg L^?1 level (n = 11). The limit of detection was 6 ng L^?1 with a sampling frequency of 18 h^?1. The present method has been successfully applied to cobalt determination in water samples and two certified reference materials.     

Application of modified multiwalled carbon nanotubes as solid sorbent for separation and preconcentration of trace amounts of manganese ions

In this work, the potential of modified multiwalled carbon nanotubes for separation and preconcentration of trace amounts of manganese ion is studied. Multiwalled carbon nanotubes were oxidized with concentrated HNO₃ and then modified with loading 1-(2-pyridylazo)-2-naphtol. Mn(II) ions could be quantitatively retained by modified multiwalled carbon nanotubes in the pH range of 8–9.5. Elution of the adsorbed manganese was carried out with 5.0 mL of 0.1 mol L^?1 HNO₃. Detection limit is 0.058 ng mL^?1 and analytical curve is linear in the range of 0.1 ng mL^?1–5.0 μg mL^?1 in the initial solution with a correlation coefficient 0.9977 and the preconcentration factor is 100. Relative standard deviation for eight replicate determination of 0.5 μg mL^?1 of manganese in the final solution is 0.41%. The effects of the experimental parameters, including the sample pH, flow rates of sample and eluent solution, eluent type, breakthrough volume and interference ions, were studied for preconcentration of Mn(II) ions in detail to optimize the conditions. The method was successfully applied for separation, preconcentration and determination of manganese in different samples.     

Electrochemical immunoassay of cotinine in serum based on nanoparticle probe and immunochromatographic strip

A disposable sensor for the determination of cotinine in human serum was developed based on immunochromatographic test strip and quantum dot label. In this assay, cotinine linked with quantum dot competes with cotinine in sample to bind to anti-cotinine antibody in the test strip and the quantum dots serve as signal vehicles for electrochemical readout. Some parameters governing the performance of the sensor were optimized. The sensor shows a wide linear range from 1 ng mL^?1 to 100 ng mL^?1 cotinine with a detection limit of 1.0 ng mL^?1. The sensor was validated with spiked human serum samples and it was found that this method was reliable in measuring cotinine in human serum. The results demonstrate that this sensor is rapid, accurate, and less expensive and has the potential for point of care (POC) detection of cotinine and fast screening of tobacco smoke exposure.     

Determination of triclosan,triclocarban and methyl-triclosan in aqueous samples by dispersive liquid–liquid microextraction combined with rapid liquid chromatography

In this study, dispersive liquid–liquid microextraction (DLLME) combined with ultra-high-pressure liquid chromatography (UHPLC)–tunable ultraviolet detection (TUV), has been developed for pre-concentration and determination of triclosan (TCS), triclocarban (TCC) and methyl-triclosan (M-TCS) in aqueous samples. The key factors, including the kind and volume of extraction solvent and dispersive solvent, extraction time, salt effect and pH, which probably affect the extraction efficiencies were examined and optimized. Under the optimum conditions, linearity of the method was observed in the range of 0.0500–100 μg L^?1 for TCS, 0.0250–50.0 μg L^?1 for TCC, and 0.500–100 μg L^?1 for M-TCS, respectively, with correlation coefficients (r²) > 0.9945. The limits of detection (LODs) ranged from 45.1 to 236 ng L^?1. TCS in domestic waters was detected with the concentration of 2.08 μg L^?1. The spiked recoveries of three target compounds in river water, irrigating water, reclaimed water and domestic water samples were achieved in the range of 96.4–121%, 64.3–84.9%, 77.2–115% and 75.5–106%, respectively. As a result, this method can be successfully applied for the rapid and convenient determination of TCS, TCC and M-TCS in real water samples.     

Capabilities of mixed-mode liquid chromatography coupled to inductively coupled plasma mass spectrometry for the simultaneous speciation analysis of inorganic and organically-bound selenium

This work investigates for the first time the potential of mixed-mode (anion-exchange with reversed-phase) high performance liquid chromatography coupled to inductively coupled plasma mass spectrometry (ICP-MS) for the simultaneous retention and selective separation of a range of inorganic and organically-bound selenium (Se) species. Baseline separation and detection of selenocystine (SeCys₂), Se-methyl-selenocysteine (SeMC), selenomethionine (SeMet), methylseleninic acid (MSA), selenite, γ-glutamyl-methyl-selenocysteine (γ-glutamyl-SeMC), and selenate in a Se standard mixture by mixed-mode HPLC-ICP-MS was achieved by switching between two citrate mobile phases of different pH and ionic strength within a single chromatographic run of 20 min. Limits of detection obtained for these Se species ranged from 80 ng kg^?1 (for SeMC) to 123 ng kg^?1 (for selenate). Using this approach as developed for selenium speciation, an adequate separation of inorganic and organic As compounds was also achieved. These include arsenite, arsenate, arsenobetaine (AsB) and dimethylarsenic acid (DMA), which may coexist with Se species in biological samples. Application of the newly proposed methodology to the investigation of the elemental species distribution in watercress (used as the model sample) after enzymatic hydrolysis or leaching in water by accelerated solvent extraction (ASE) was addressed. Only SeMet, SeMC and selenate could be tentatively identified in watercress extracts by mixed-mode HPLC-ICP-MS and retention time matching with standards. Recoveries (n = 3) of these Se species from samples spiked with standards averaged 102% (for SeMC), 94.9% (for SeMet) and 98.3% (for selenate). Verification of the presence of SeMet and SeMC in an enzymatic watercress extract was achieved by on-line HPLC-ESI MS/MS in selected reaction monitoring (SRM) mode.     

Highly selective solid-phase extraction of trace Pd(II) by murexide functionalized halloysite nanotubes

The originality on the high efficiency of murexide modified halloysite nanotubes as a new adsorbent of solid phase extraction has been reported to preconcentrate and separate Pd(II) in solution samples. The new adsorbent was confirmed by Fourier transformed infrared spectra, X-ray diffraction, scanning electron microscope, transmission electron microscope and N₂ adsorption–desorption isotherms. Effective preconcentration conditions of analyte were examined using column procedures prior to detection by inductively coupled plasma-optical emission spectrometry (ICP-OES). The effects of pH, the amount of adsorbent, the sample flow rate and volume, the elution condition and the interfering ions were optimized in detail. Under the optimized conditions, Pd(II) could be retained on the column at pH 1.0 and quantitatively eluted by 2.5 mL of 0.01 mol L^?1 HCl–3% thiourea solution at a flow rate of 2.0 mL min^?1. The analysis time was 5 min. An enrichment factor of 120 was accomplished. Common interfering ions did not interfere in both separation and determination. The maximum adsorption capacity of the adsorbent at optimum conditions was found to be 42.86 mg g^?1 for Pd(II).The detection limit (3σ) of the method was 0.29 ng mL^?1, and the relative standard deviation (RSD) was 3.1% (n = 11). The method was validated using certified reference material, and has been applied for the determination of trace Pd(II) in actual samples with satisfactory results.     

Determination of atorvastatin in human serum by salting out assisted solvent extraction and reversed-phase high-performance liquid chromatography–UV detection

A simple and rapid technique based on salting out assisted solvent extraction was developed for extraction of atorvastatin from serum sample and high performance liquid chromatography–UV was used for its detection. In the present study, 1.0 mL serum was extracted by 0.5 mL of acetonitrile and some parameters that can affect extraction such as type and volume of extraction solvent, type of salt, and pH were optimized. Under optimized experimental conditions, the calibration curve was found to be linear in the range of 0.001–10 ng mL⁻¹ in human serum and the correlation coefficient (R²) and the limits of detection were >0.99 and 0.0005 ng mL⁻¹, respectively. The accuracy of the method in terms of average recovery of the compound in spiked serum and water samples was better than 90%.     

Preconcentration of atrazine and simazine with multiwalled carbon nanotubes as solid-phase extraction disk

A sensitive and selective preconcentration method using solid-phase extraction (SPE) disk, namely multiwalled carbon nanotubes (MWCNTs) disk, is proposed for the determination of atrazine and simazine in water samples. Atrazine and simazine were extracted on MWCNTs disk and then determined by gas chromatography–mass spectrometry (GC/MS). Several parameters on the enrichment factor of the analytes were investigated. The experimental results showed that it was possible to obtain quantitative analysis when the solution pH was 5 using 200 mL of validation solution containing 0.1 μg of triazines and 5 mL of acetone as an eluent. The maximum enrichment factors for atrazine and simazine were 3900 ± 250 and 4000 ± 110, respectively when 200 mL of sample solution volume was used. Relative standard deviations for seven determinations were 6.9% (atrazine) and 3.0% (simazine) under optimum conditions. The linear range of calibration curves were 0.1 to 1 ng mL^− 1 for each analyte with good correlation coefficients. The detection limits (3S/N) were 2.5 and 5.0 pg mL^− 1 for atrazine and simazine, respectively. The proposed method was successfully applied to the determination of atrazine and simazine in environmental water samples with high precision and accuracy.     

Analysis of hydrocarbon contamination with membrane-assisted solvent extraction: Comparison of agitation and sonication methods

Membrane-assisted solvent extraction (MASE) coupled to large volume injection was applied to the determination of (gasoline-type) hydrocarbon contamination in water samples. Hexane was used as acceptor phase. 50 μL extract was injected in the programmed temperature vaporizer injector using combined split–splitless evaporation. The extraction conditions were optimized both for MASE with agitation and for MASE with sonication. In the course of optimization the effect of extraction time, extraction temperature, agitation speed, solvent volume, pH, ionic strength and the addition of methanol were tested. Over 75% recovery was accomplished in the range of diesel oil hydrocarbons (n-C₉–n-C₂₄). The developed method was validated. Linearity, accuracy and precision were tested. The method showed excellent linearity between 1 and 1000 μg L^?1 for n-alkanes and between 0.05 and 50 mg L^?1 for gasoline. The method was tested with comprehensive GC × GC as well and found to be non-discriminative to all major compounds of diesel oil.     

Simultaneous preconcentration of cadmium,cobalt and nickel in water samples by cationic micellar precipitation and their determination by inductively coupled plasma-optical emission spectrometry

In the present study an easy micellar precipitation process at ambient temperature using cationic surfactant cetyl-trimethyl ammonium bromide (CTAB) was applied to the simultaneous extraction of cadmium, cobalt and nickel from aqueous samples. The analytical procedure involved the complex formation of these cations with 1,8-dihydroxyanthrone as a chelating agent in buffer media of pH 7.0. After the phase separation, the precipitated complexes were first dissolved in 0.25 mL of 80:20 propanol–water mixture containing 0.03 mL HNO₃ and then subjected to the inductively coupled plasma-optical emission spectrometry (ICP-OES) analysis. The concentration of 1,8-dihydroxyanthrone, pH, amount of CTAB and centrifuge time was optimized. Under the optimum conditions, the preconcentration factor was 40 and the improvement factors of 11.6, 9.5 and 14.4 with detection limits of 0.008, 0.009 and 0.004 (ng mL^? 1) were obtained for Cd, Co and Ni respectively. Under the presence of foreign ions no significant interference was observed. Finally, the proposed extraction method was successfully applied to the determination of these elements in various water and geological CRM samples with recovery percentages of 97–104% and RSD values of 1.87–2.36%.     

Selective determination of inorganic cobalt in nutritional supplements by ultrasound-assisted temperature-controlled ionic liquid dispersive liquid phase microextraction and electrothermal atomic absorption spectrometry

In the present work, a simple and rapid analytical method based on application of ionic liquids (ILs) for inorganic Co(II) species (iCo) microextraction in a variety of nutrient supplements was developed. Inorganic Co was initially chelated with 1-nitroso-2-naphtol (1N2N) reagent followed by a modern technique named ultrasound-assisted temperature-controlled ionic liquid dispersive liquid phase microextraction (USA-TILDLME). The extraction was performed with 1-hexyl-3-methylimidazolium hexafluorophosphate [C₆mim][PF₆] with the aid of ultrasound to improve iCo recovery. Finally, the iCo-enriched IL phase was solubilized in methanol and directly injected into an electrothermal atomic absorption spectrometer (ETAAS). Several parameters that could influence iCo microextraction and detection were carefully studied. Since the main difficulty in these samples is caused by high concentrations of potential interfering ions, different approaches were evaluated to eliminate interferences. The limit of detection (LOD) was 5.4 ng L^?1, while the relative standard deviation (RSD) was 4.7% (at 0.5 μg L^?1 Co level and n = 10), calculated from the peak height of absorbance signals. Selective microextraction of iCo species was achieved only by controlling the pH value during the procedure. The method was thus successfully applied for determination of iCo species in nutritional supplements.     

Simultaneous quantification of nimesulide,phenylpropanolamine, caffeine and chlorpheniramine in rat plasma by RP–HPLC/PDA method and application to pharmacokinetic studies in healthy rat subjects

Simultaneous determination of nimesulide, phenylpropanolamine, chlorpheniramine and caffeine in rat plasma by reversed-phase high performance liquid-chromatography (RP–HPLC) with photodiode array (PDA) detection method was developed and validated. Sample preparation based on a simple extraction procedure consisting of deproteination and extraction with methanol solution followed by volume make up with the aqueous component of the mobile phase obtained best recoveries of the analytes. The chromatographic conditions were optimized and the analytes were separated on XBridge™ C₁₈ (3.5 μm, 4.6 × 150 mm) column in isocratic elution with the mobile phase composition of acetonitrile and 10 mM ammonium acetate buffer (pH 4.0, 0.1% formic acid) (18:82 v/v%) at the flow rate of 1 mL min⁻¹ and the effluents were monitored in the wavelength range of 220–275 nm. The method was linear for all analytes over the following concentration (ng mL⁻¹) ranges: nimesulide 250–4000; phenylpropanolamine 100–1500; chlorpheniramine 20–500; and caffeine 10–100. Acceptable precision, accuracy and recoveries were obtained for quality control (QC) samples at three concentrations (low QC, middle QC and high QC). The percentage of relative standard deviation (% RSD) of Inter and intra-run precision of all molecules was <15% and the percentage of accuracy was 100 ± 10. The analytes were more stable in rat plasma at different storage conditions. Finally the method was efficiently applied to pharmacokinetics study in rat plasma.