Electrospun nanofibers as a new solid phase microextraction coating for determination of volatile organic impurities in biological products.

In this study, preparation of a solid-phase microextraction fiber of polydimethylsiloxane (PDMS) polymer in its blends with polystyrene (PS) via electrospinning process, on a stainless steel wire has been reported. The electrospining enhancement of PDMS solutions in the presence of PS is found due to the increase in the viscosity of the mixed solutions. Characteristics of the fibers were inspected by energy dispersive X-ray spectroscopy, scanning electron microscopy and field emission scanning electron microscopy. The applicability of the coating was assessed for headspace solid phase microextraction (SPME) of some residual solvents (Diethyl Ether, Toluene and Chloroform) from biological products (Anti venin and Foot-and-mouth disease vaccine) followed by gas chromatography-mass spectrometry. The effects of extraction time and temperature, desorption time and temperature, agitation rate and ionic strength on the extraction efficiency were investigated. Under optimized conditions, limits of detection in the range of 2-10 µg L-1 and limits of quantification in the range of 10-50 µg L-1 were obtained. The method showed linearity in a wide range with a correlation coefficient greater than 0.99. In addition, the obtained inter-day and intra-day precisions were in the range of 1.57-8.28% and 4.87-11.72%, respectively. The thermal stability of the fiber was also investigated and it was found to be durable at 230 °C for 450 min. Furthermore, the proposed method was successfully applied for quantification of chloroform in Foot-and-mouth disease vaccine and diethyl ether and toluene in Anti-venin with recoveries in the range of 78.84-123.01%.

Zinc oxide-gold nanocomposite as a proper platform for label-free DNA biosensor.

In this study, a simple and cost-effective electrochemical DNA biosensor was developed for sensitive detection of mycobacterium tuberculosis (TB). Nanocomposite of zinc oxide (ZnO) and gold nanoparticles (AuNPs) was used as a platform for immobilizing thiolated TB DNA (probe DNA). ZnO was electrodeposited on a glassy carbon electrode by potentiostat electrolysis of Zn (NO3)2 solution at -1.0 V (vs. Ag/AgCl), then AuNPs were loaded as the second layer at -0.4 V from HAuCl4 solution. Thiolated probe DNA was then covalently attached to AuNPs. Anodic peak current of Fe (CN)63-/4- was followed in hybridization experiments and a linear calibration curve was obtained in concentration range of 2.5-250 pM and limit of detection (LOD) of 1.8 pM for target DNA. The label-free TB biosensor exhibited high selectivity, suitable stability, and reproducibility.

Application of metal-organic framework as redox probe in an electrochemical aptasensor for sensitive detection of MUC1.

In this work, an electrochemical aptasensor was developed for sensitive detection of MUC1 based on metal-organic framework-reduced graphene oxide nanocomposite (Cu-MOF-RGO). Cu- MOF-RGO appeared to be suitable as a platform for immobilization of MUC1 aptamer, and also as an electrochemical probe, which exhibited well-defined peaks with good stability and reproducibility. Cu-MOF-graphene oxide (Cu-MOF-GO) nanocomposite was prepared and cast on the electrode surface, then in order to increase the conductivity of the electrode, GO was electrochemically reduced to RGO. In the presence of MUC1, the peak current of Cu in the nanocomposite decreased, which could be explained based on the formation of MUC1-aptamer complexes on the electrode, and consequence blocking the electron transfer of Cu at the electrode surface. Under optimum experimental conditions, a linear calibration curve was obtained by differential pulse voltammetry in the concentration range of 0.1 pM-10 nM (25 pg mL-1 - 2500 ng mL-1) with a limit of detection (LOD) of 0.033 pM (7.5 pg mL-1) of MUC1. The proposed aptasensor offers acceptable selectivity, stability, and reproducibility in the determination of MUC1 spiked to human blood serum samples.

Simultaneous determination of l­DOPA, l­tyrosine and uric acid by cysteic acid - modified glassy carbon electrode.

Electrochemical oxidation of l­cysteine resulted in the formation of a film on glassy carbon electrode. In a solution of levodopa (l­DOPA), l­tyrosine (Tyr) and uric acid (UA), three separated intense anodic peaks were appeared in differential pulse voltammetry regime. Experimental conditions were optimized for simultaneous determination of the three compounds. All experiments were carried out in phosphate buffer solution (0.1 M, pH 8). Calibration curves were obtained in the presence of various concentrations of l­DOPA, Tyr, and UA. Linear concentration ranges were 0.65-22 µM for l­DOPA, 3.5-96 µM for Tyr, and 1.0-19 µM for UA. The limits of detection (LODs) were calculated as 0.2, 1.1 and 0.36 µM for l­DOPA, Tyr, and UA, respectively. The electrochemical sensor was used successfully for the simultaneous determination of l­DOPA, Tyr, and UA species in human blood serum samples.

Electrochemical fabrication of a novel ZnO/cysteic acid nanocomposite modified electrode and its application to simultaneous determination of sunset yellow and tartrazine.

A new nanocomposite (ZnO/Cysteic acid) was deposited on glassy carbon electrode by cyclic voltammetry. Uniform deposition of the nanocomposite was observed by scanning electron microscopy. The electron transfer characteristics of two food additives, sunset yellow and tartrazine, were greatly improved on the modified electrode. The prepared electrode was used in the sensitive simultaneous determination of sunset yellow and tartrazine by differential pulse voltammetry. Linear calibration curves were obtained in the concentration ranges of 0.1-3.0, and 0.07-1.86µM, and detection limits of 0.03 and 0.01µM for sunset yellow and tartrazine, respectively. The proposed method was evaluated by determination of the dyes in processed soft drinks with satisfactory results (recovery>95% and RSD%<5%).

Application of carbon nanotubes-ionic liquid hybrid in a sensitive atorvastatin ion-selective electrode.

Atorvastatin (ATR) was determined by a potentiometric method. The ion-pair of ATR and cetyltrimethylammonium bromide (CTAB) was used as a suitable ionophore. A graphite paste electrode was modified with ATR-CTAB ion-pair, multiwalled carbon nanotubes (MWCNTs), and an ionic liquid, 1-butyl-3-mtehyl-imidazolium hexafluorophosphate (BMIMPF6). The amounts of electrode ingredients were optimized (graphite powder: paraffin oil: ATR-CTAB: MWCNTs: BMIMPF6 (58:26:5:8:3 w/w%). Surface characterization was done by using scanning electron microscopy. The potential measurements were recorded at optimized pH by using acetate buffer solution (0.1molL(-1), pH5.5). At the above experimental conditions, calibration curve (E vs. log [ATR]) was linear (R(2)=0.9977) in the concentration range of 1.0×10(-9)-1.0×10(-3)molL(-1) (0.0012-1209mgL(-1)) of ATR with a Nernstian slope of 58.14±0.2mV decade(-1), and detection limit of 1.0×10(-9)molL(-1) (0.0013mgL(-1)). After each injection of ATR to the buffer solution, the potential was stabilized in a very short time (average response time~6s) at 25°C. The modified graphite paste electrode had a long lifetime (>4months). Recovery of the spiked drug to blood serum samples (95.3-98.2%) revealed the reliability of electrode response to ATR. Blood serum samples from consumers were analyzed by the proposed method; the results were comparable with those from HPLC standard method. The potentiometric analysis of ATR tablets by the proposed electrode resulted in a relative error of 0.8% and 1.5% for 20 and 40mg per tablets, respectively. Finally, the electrode was used in potentiometric titration of ATR (1.0×10(-3)molL(-1)) by CTAB (1.0×10(-3)molL(-1)). Excellent accuracy (≈100%) was obtained from the volume of the titrant at the endpoint.

New nanostructure of polydimethylsiloxane coating as a solid-phase microextraction fiber: Application to analysis of BTEX in aquatic environmental samples.

Electrospinning technique was used to convert polydimethyl siloxane (PDMS) sol-gel solution to a new nanostructure on a stainless steel wire. The surface morphology of the fiber was observed by scanning electron microscopy (SEM). It showed a diameter range of 30-60nm for PDMS nanoparticles with a homogeneous and porous surface structure. The applicability of this coating was assessed for the headspace SPME (HS-SPME) of benzene, toluene, ethylbenzene and xylenes (BTEX) from water samples followed by gas chromatography-mass spectrometry. The important parameters affecting extraction efficiency such as extraction time and temperature, desorption conditions, agitation rate and ionic strength were investigated and optimized. Under the optimized conditions, LODs and LOQs of 0.3-5µgL(-1) and 1-10µgL(-1) were obtained, respectively. The method showed linearity in the broad range of 1-5000µgL(-1) with correlation coefficient of >0.99. Inter-day and intra-day precisions of the developed method ranged from 2.43% to 6.54% and from 5.24% to 13.73%, respectively. The thermal stability of the fiber was investigated on stainless steel wire. It was found to be durable at 260°C for more than 360min. Furthermore, the proposed method was successfully applied for quantification of BTEX in real water samples.

Highly selective electrode for potentiometric analysis of methadone in biological fluids and pharmaceutical formulations.

In order to develop a fast and simple procedure for methadone analysis in biological fluids, a graphite paste electrode (GPE) was modified with the ion-pair of methadone-phosphotungstic acid, and multiwalled carbon nanotubes (MWCNTs). Optimized composition of the electrode with respect to graphite powder:paraffin oil:MWCNTs:ion pair, was 58:30:8:4 (w/w%). The electrode showed a near-Nernstian slope of 58.9 ± 0.3 mV/decade for methadone in a wide linear range of 1.0 × 10(-8)-4.6 × 10(-3)M, with a detection limit of 1.0 × 10(-8)M. The electrode response was independent of pH in the range of 5-11, with a fast response time (~4s) at 25 °C. The sensor showed high selectivity and was successfully applied to the determination of sub-micromolar concentrations of methadone in human blood serum and urine samples, with recoveries in the range of 95-99.8%. The average recovery of methadone from tablets (5 mg/tablet) by using the proposed method was 98%. The life time of the modified electrode was more than 5 months, due to the characteristic of GPE which can be cut off and fresh electrode surface be available. A titration procedure was performed for methadone analysis by using phosphotungstic acid, as titrating agent, which showed an accurate end point and 1:1 stoichiometry for the ion-pair formed (methadone:phosphotungstic acid). The simple and rapid procedure as well as excellent detection limit and selectivity are some of the advantages of the proposed sensor for methadone.

Differential pulse voltammetric determination of nanomolar concentrations of antiviral drug acyclovir at polymer film modified glassy carbon electrode.

An electrochemical sensor for the sensitive detection of acyclovir was developed by the electropolymerization of Eriochrome black T at a pretreated glassy carbon electrode. The surface morphology of the modified electrode was characterized by field emission scanning electron microscopy. Under the optimized conditions, a significant electrochemical improvement was observed toward the electrooxidation of acyclovir on the modified electrode surface relative to the unmodified electrode. The detection limit of 12 nM and two linear calibration ranges of 0.03-0.3 µM and 0.3-1.5 µM were obtained for acyclovir determination using a differential pulse voltammetric method in acetate buffer (0.1 M, pH 4.0). Real sample studies were carried out in human blood serum and pharmaceutical formulations, which offered good recovery (98-102%). The electrode showed excellent reproducibility, selectivity and antifouling effects.

Fast Electrocatalytic Determination of Methimazole at an Activated Glassy Carbon Electrode.

A fast and simple voltammetric method for the determination of methimazole in pharmaceutical products was reported. A glassy carbon electrode was pretreated by anodization at +1.75 V (vs. SCE) for 5 min, followed by potential cycling in the range of 0.3-1.3 V (20 cycles). The pretreated electrode showed an excellent electrocatalytic effect on the oxidation of methimazole. Compared with untreated electrode, a large decrease (~300 mV) in the oxidation peak of methimazole was observed. The oxidation peak current at the new potential (0.4 V vs. SCE) was linearly dependent on the concentration of methimazole in the range of 7.0 - 130 µM with a detection limit of 3.7 µM (S/N = 3). The method was successfully used in the determination of methimazole in thyramozol tablets. Due to the simple and fast electrode preparation, there is no need for electrode cleaning or storage.

Electrospun nanostructured polystyrene as a new coating material for solid-phase microextraction: Application to separation of multipesticides from honey samples.

For the first time, electrospun polystyrene nanostructure was used as coating material on a stainless steel wire for solid-phase microextraction. Surface morphology of the coating was studied by scanning electron microscopy which showed the formation of nanofibers on the wire. The coating was stable after conditioning at 250°C for 2h. The efficiency of the polystyrene coating was approved by extracting a mixture of seven pesticides (polar and apolar) from head space of honey samples followed by gas chromatography-mass spectrometry. The important parameters affecting extraction efficiency such as, extraction time and temperature, desorption conditions, agitation rate and ionic strength were investigated. Under optimized experimental conditions, detection limits for the investigated pesticides ranged from 0.1-2µgL(-1). The intra- and inter-day precisions of the developed method were 3.5-17.6% and 10.0-25.0%, respectively. Finally, all the investigated pesticides were spiked to honey samples and extracted by the proposed method. The accuracies of determination of all the species were found to be in the range of 81-125%.

Electrochemical, spectroscopic, and theoretical studies on the interaction between azathioprine and DNA.

Possible interaction between immunosuppressive drug, azathioprine, and calf thymus DNA was explored by cyclic voltammetry, spectrophotometry, competitive spectrofluorimetry, circular dichroism spectroscopy (CD), and viscosity measurements. Cyclic voltammetry showed negative shift in the reduction peak of azathioprine in the presence of DNA, and large decrease in peak current, referring to the predominance of electrostatic forces. The binding constant was calculated to be 1.22×10(3)M(-1). Absorption hyperchromism without shift in wavelength was observed when DNA was added to azathioprine solution. Competitive fluorescence experiments were conducted by using Hoechst 33258 and methylene blue as probes for minor groove and intercalation binding modes, respectively. The studies showed that azathioprine could release Hoechst 33258, while negligible effect was detected in the case of methylene blue. Stern-Volmer quenching constant (KSV) and complex formation constant (Kf) were obtained from the fluorescence measurements to be 7.6×10(3)M(-1) and 7.76×10(4)M(-1), respectively, at 298K. Enthalpy and entropy changes during the interaction between azathioprine and DNA were calculated from Van't Hoff plot (ΔH=-20.2kJmol(-1); ΔS=26.11Jmol(-1)K(-1) at 298K) which showed an exothermic spontaneous reaction, and involvement of electrostatic forces in the complex formation with DNA. Moreover, circular dichroism studies revealed that azathioprine induced detectable changes in the negative band of DNA spectrum. Viscosity of DNA solution decreased in the presence of azathioprine, showed a non-intercalative mode of interaction. Finally, molecular docking calculations showed that in the lowest energy level of drug-DNA complex, azathioprine approaches the minor grooves of DNA.

Sensitive determination of methadone in human serum and urine by dispersive liquid-liquid microextraction based on the solidification of a floating organic droplet followed by HPLC-UV.

Dispersive liquid-liquid microextraction based on solidification of floating organic droplet was developed for the extraction of methadone and determination by high-performance liquid chromatography with UV detection. In this method, no microsyringe or fiber is required to support the organic microdrop due to the usage of an organic solvent with a low density and appropriate melting point. Furthermore, the extractant droplet can be collected easily by solidifying it at low temperature. 1-Undecanol and methanol were chosen as extraction and disperser solvents, respectively. Parameters that influence extraction efficiency, i.e. volumes of extracting and dispersing solvents, pH, and salt effect, were optimized by using response surface methodology. Under optimal conditions, enrichment factor for methadone was 134 and 160 in serum and urine samples, respectively. The limit of detection was 3.34 ng/mmL in serum and 1.67 ng/mL in urine samples. Compared with the traditional dispersive liquid-liquid microextraction, the proposed method obtained lower limit of detection. Moreover, the solidification of floating organic solvent facilitated the phase transfer. And most importantly, it avoided using high-density and toxic solvents of traditional dispersive liquid-liquid microextraction method. The proposed method was successfully applied to the determination of methadone in serum and urine samples of an addicted individual under methadone therapy.

Sensitive amperometric determination of methimazole based on the electrocatalytic effect of rutin/multi-walled carbon nanotube film.

Electrochemical deposition was used to prepare a glassy carbon electrode modified with multi-walled carbon nanotubes and the glycosidic compound, rutin (R/MWCNTs/GCE). Cyclic voltammetry of the modified electrode in aqueous solution (pH8) showed a pair of well-defined, stable and reversible redox peaks with surface confined characteristics. The catechol moiety of rutin produced the voltammetric peaks via a 2 electron, 2 proton mechanism in the range of 0.0-0.4V (vs. Ag/AgCl). The transfer coefficient (α), heterogeneous electron transfer rate constant (ks), and surface concentration (Γ) for R/MWCNTs/GCE were calculated by using the cyclic voltammetric data. The modified electrode showed excellent catalytic activity toward oxidation of methimazole. Fixed-potential amperometry was used for sub-micromolar determination of methimazole at pH8. Linear dependence of anodic current to methimazole concentration was obtained in the range of 0.1-26µM of the drug with a limit of detection at 18nM. The modified electrode retained its initial response for at least 2weeks if stored in dry ambient conditions. The electrode was used for the amperometric determination of methimazole in formulations and spiked blood serum samples, successfully.

Sensitive determination of atorvastatin in human plasma by dispersive liquid-liquid microextraction and solidification of floating organic drop followed by high-performance liquid chromatography.

A novel and sensitive dispersive liquid-liquid microextraction method based on the solidification of the floating organic drop combined with high-performance liquid chromatography and ultraviolet detection was used for the determination of atorvastatine in blood serum samples. The chromatographic separation of atorvastatin was carried out using methanol as the mobile phase organic modifier. Various parameters affecting the extraction efficiency were optimized, such as the kind and volume of extraction solvent (1-undecanol) and disperser solvent (acetonitrile), pH, and the extraction time. The calibration curve was linear in the range of 0.2-6000 µg/L of atorvastatin (r(2) = 0.995) with a limit of detection of 0.07 µg/L. The relative standard deviation for 100 µg/L of atorvastatin in human plasma was 8.4% (n = 4). The recoveries of plasma samples spiked with atorvastatin were in the range of 98.8-113.8%. The obtained results showed that the proposed method is fast, simple, and reliable for the determination of very low concentrations of atorvastatin in human plasma samples.

Determination of free formaldehyde in vaccines and biological samples using solid-phase microextraction coupled to GC-MS.

A headspace solid-phase microextraction method coupled to GC-MS was successfully developed for the trace determination of formaldehyde in veterinary bacterial and human vaccines, and diphtheria-tetanus antigen. The formaldehyde was derivatized by means of the Hantzsch reaction prior to extraction and subsequent determination. Three different types of solid-phase microextraction fibers, polar, and nonpolar poly(dimethylsiloxane) and polyethylene glycol were prepared by using a sol-gel technique. The effects of different parameters such as type of fiber coating, extraction time and temperature, desorption conditions, agitation rate, and salt effect were investigated. Under the optimized conditions, the detection limit of the method was 979 ng/L using the selected ion-monitoring mode. The interday and intraday precisions of the developed method under the optimized conditions were below 13%, and the method shows linearity in the range of 1.75-800 µg/L with a correlation coefficient of 0.9963. The optimized method was applied to the determination of formaldehyde from some biological products. The results were satisfactory compared to the standard method.

Electrochemical and spectroscopic studies of the interaction between the neuroleptic drug, gabapentin, and DNA.

The interaction between gabapentin (GP), a neuroleptic drug, and calf-thymus DNA was investigated. Cyclic voltammetry and different spectroscopic methods (UV/vis spectrophotometry, spectroflourimetry and circular dichroism) were used to study the process. The oxidation current of GP was considerably decreased in the presence of calf-thymus DNA. The negative shift of the peak potential in cyclic voltammetry suggested an electrostatic interaction, while spectroscopic methods show minor groove binding as the predominant mode. The values of binding constants obtained from UV absorption, spectrofluorimetry and voltammetric measurements were in close agreement. Standard Gibbs free energy change, ΔG(0), of the interaction was calculated to be -21.99 kJ mol(-1) which indicated the spontaneity of the binding interaction. A hyperchromic effect in the maximum absorption band of DNA as well as quenching in fluorescence experiments were observed in the presence of GP.

Preparation of a novel iodide-selective electrode based on iodide-miconazole ion-pair and its application to pharmaceutical analysis.

An iodide-miconazole ion-paired complex was used as a suitable ion-exchanger for the preparation of a plasticized-PVC membrane electrode. Among different solvent mediators tested, dioctylsebacate exhibited the proper response characteristics, including Nernstian slope of the calibration curve, fast response time and good reproducibility of the emf values. The electrode exhibits a Nernstian slope of -59.8 +/- 0.5 mV decade(-1) for I- ion over a concentration range of 1.0 x 10(-5) - 1.0 x 10(-2) M with a limit of detection of 7.0 x 10(-6) M. The electrode displays a good selectivity for I- with respect to a number of inorganic and organic species. It canbe used over a pH range of 2.5 - 8.5. The membrane sensor was successfully applied to the determination of iodide in water samples and blood serum, as well as in pharmaceutical products such as iodoquinol and thyroxin.

Preparation of a diclofenac potentiometric sensor and its application to pharmaceutical analysis and to drug recovery from biological fluids.

A novel diclofenac ion-selective electrode is prepared, characterized and used in pharmaceutical analysis. The diclofenac complex with hexadecylpyridinium bromide is obtained in situ by soaking the PVC-membranes in a 1x10(-2) M diclofenac solution. Among four different solvent mediators tested, dibutyl phthalate (DBP) exhibited a proper behavior including Nernstian slope of the calibration curve, fast response time and good reproducibility of the emf values. The electrode exhibits a Nernstian slope of -59+/-1 mV decade(-1) for diclofenac in the concentration range 1.0x10(-5) to 1.0x10(-2) M with a limit of detection of 4.0x10(-6) M. The electrode displays a good selectivity for diclofenac with respect to a number of common inorganic and organic species. It can be used in a pH range of 6.0-9.0. The membrane sensor was successfully applied to the determination of diclofenac in its tablets as well as for its recovery from blood serum and urine samples.

Fluorometric chemosensors. Interaction of toxic heavy metal ions Pb(II), Cd(II), and Hg(II) with novel mixed-donor phenanthroline-containing macrocycles: spectrofluorometric, conductometric, and crystallographic studies.

The macrocycles L(1)-L(3) incorporating N(2)S(3)-, N(2)S(2)O-, and N(2)S(2)-donor sets, respectively, and containing the 1,10-phenanthroline unit interact in acetonitrile solution with heavy metal ions such as Pb(II), Cd(II), and Hg(II) to give 1:1 ML, 1:2 ML(2), and 2:1 M(2)L complex species, which specifically modulate the photochemical properties of the ligands. The stoichiometry of the complex species formed during spectrofluorometric titrations and their formation constants in MeCN at 25 degrees C were determined from fluorescence vs M(II)/L molar ratio data. The complexes [Pb(L(1))][ClO(4)](2).(1)/(2)H(2)O (1), [Pb(L(2))][ClO(4)](2).MeNO(2) (1a), [Pb(L(3))(2)][ClO(4)](2).2MeCN (1b), and [Cd(L(3))][NO(3)](2) (2b) were also characterized by X-ray diffraction studies. The conformation adopted by L(1)-L(3) in these species reveals the aliphatic portion of the rings folded over the plane containing the heteroaromatic moiety with the ligands trying to encapsulate the metal center within their cavity. In 1, 1a, and 2b the metal ion completes the coordination sphere by interacting with counteranion units and solvent molecules. On the contrary, the 1:2 complex 1b shows Pb(II) sandwiched between two symmetry-related molecules of L(3) reaching an overall [4N + 4S] eight-coordination.