On-line preconcentration of perfluorooctanoic acid and perfluorooctanesulfonic acid by nonaqueous capillary electrophoresis

Separation of major environmental pollutants as perfluorooctanoic acid (PFOA) and perfluorooctanesulfonic acid (PFOS) by capillary electrophoresis is reported for the first time. It is not possible to resolve the solutes in an aqueous media. However, the use of methanol and acetonitrile as the background electrolyte (BGE) solvents allowed their rapid separation in an uncoated capillary. A major effort was put into BGE optimization in respect to both separation efficiency and detection for further on‐line preconcentration. 5 mmol.L^?1 naphthalene‐1‐sulfonic acid and 10 mmol.L^?1 triethylamine dissolved in ACN/MeOH (50:50 v/v) provided best separation and detection conditions. Next, the large‐volume sample stacking and the field‐amplified sample injection were applied and compared. Large‐volume sample stacking improved limits of detection (LODs) with regard to the standard injection by 69 times for PFOA and 143 times for PFOS with LODs of 280 and 230 nmol.L^?1, respectively. Field‐amplified sample injection improved LODs 624 times for PFOAand 806 times for PFOS with LODs 31 and 40 nmol.L^?1, respectively. Both preconcentration methods showed repeatabilities of migration times less than 1.2% RSD intraday and 6.6% RSD interday. The method was applied on PFOA and PFOS analysis in a sample of river water treated with solid‐phase extraction, which further improved LOD toward 5.6 × 10^?10 mol.L^?1 for PFOS and 6.4 × 10^?10 mol.L^?1 for PFOA and allows the method to be used for river water contamination screening or decomposition studies.     

Determination of tetracyclines in human urine samples by capillary electrophoresis in combination with field amplified sample injection

A sensitive method using CZE‐UV detection has been developed for the determination of five tetracycline antibiotics in human urine samples. To improve the sensitivity of the method, an on‐line preconcentration strategy, named field‐amplified sample injection, has been developed, based on the electrokinetic injection of the sample, which requires only a 1:100 dilution with sample solvent before injection. Under optimum conditions, sensitivity enhancement factors ranged from 450 to 800 for the studied compounds. The applicability of the proposed method was demonstrated by the determination of these antibiotics in spiked urine samples. The limits of quantification were lower than 0.8 mg/L and the precision (intra‐ and inter‐day), expressed as %RSD was below 14%. Recoveries ranged from 92.1 to 96.7%. Thus, the proposed procedure is a simple, fast and efficient strategy which could be used as therapeutic drug monitoring in human urine samples.     

Development and validation of a capillary electrophoresis method for the determination of phenothiazines in human urine in the low nanogram per milliliter concentration range using field-amplified sample injection

A capillary zone electrophoresis (CZE) method with ultraviolet-visible detection has been established and validated for the determination of five phenothiazines: thiazinamium methylsulfate, promazine hydrochloride, chlorpromazine hydrochloride, thioridazine hydrochloride, and promethazine hydrochloride in human urine. Optimum separation was obtained on a 64.5 cm x 75 microm bubble cell capillary using a buffer containing 150 mM tris(hydroxymethyl)aminomethane and 25% acetonitrile at pH 8.2, with temperature and voltage of 25 degrees C and 20 kV, respectively. Naphazoline hydrochloride was used as an internal standard. Field-amplified sample injection (FASI) has been applied to improve the sensitivity of the detection. Considering the influence of parameters affecting the on-line preconcentration (nature of preinjection plug, sample solvent composition, injection times, and injection voltage) and due to the significant interactions among them, in this paper we propose for the first time the application of a multivariate approach to carry out the study. The optimized conditions were as follows: preinjection plug of water for 7 s at 50 mbar, electrokinetic injection for 40 s at 6.2 kV, and 32 microm of H3PO4 in the sample solvent. Also, a solid-phase extraction (SPE) procedure is developed to obtain low detection limits and an adequate selectivity for urine samples. The combination of SPE and FASI-CZE-UV allows adequate linearities and recoveries, low detection limits (from 2 to 5 ng/mL), and satisfactory precisions (3.0-7.2% for an intermediate RSD %).     

Determination of tobramycin in human serum by capillary electrophoresis with contactless conductivity detection

A study on the determination of the antibiotic tobramycin by CE with capacitively coupled contactless conductivity detection is presented. This method enabled the direct quantification of the non-UV-absorbing species without incurring the disadvantages of the indirect approaches which would be needed for optical detection. The separation of tobramycin from inorganic cations present in serum samples was achieved by optimizing the composition of the acetic acid buffer. Field-amplified sample stacking was employed to enhance the sensitivity of the method and a detection limit of 50 microg/L (S/N = 3) was reached. The RSDs obtained for migration time and peak area using kanamycin B as internal standard were typically 0.12 and 4%, respectively. The newly developed method was validated by measuring the concentration of tobramycin in serum standards containing typical therapeutic concentrations of 2 and 10 mg/L. The recoveries were 96 and 97% for the two concentrations, respectively.     

Field-amplified online sample stacking capillary electrophoresis UV detection for plasma malondialdehyde measurement

Malondialdehyde (MDA) determination is the most widely used method for monitoring lipid peroxidation. Here, we describe an easy field-amplified sample injection (FASI) CE method with UV detection for the detection of free plasma MDA. MDA was detected within 8 min by using 200 mmol/L Tris phosphate pH 5.0 as running buffer. Plasma samples treated with ACN for protein elimination were directly injected on capillary without complex cleanup and/or sample derivatization procedures. Using electrokinetic injection, the detection limit in real sample was 3 nmol/L, thus improving of about 100-fold the LOD of the previous described methods based on CE. Precision tests indicate a good repeatability of our method both for migration times (CV = 1.11%) and for areas (CV = 2.05%). Moreover, a good reproducibility of intra- and inter-assay tests was obtained (CV = 2.55% and CV = 5.14%, respectively). Suitability of the method was tested by measuring MDA levels in 44 healthy volunteers.     

Determination of quinocide as impurity in primaquine tablets by capillary zone electrophoresis

A capillary zone electrophoretic method has been developed and validated for the determination of the impurity quinocide (QC) in the antimalarial drug primaquine (PQ). Different buffer additives such as native cyclodextrins and crown ethers were evaluated. Promising results were obtained when either β‐cyclodextrin (β‐CD) or 18‐crown‐6 ether (18C6) were used. Their separation conditions such as type of buffer and its pH, buffer additive concentration, applied voltage capillary temperature and injection time were optimized. The use of 18C6 offers slight advantages over β‐CD such as faster elution times and improved resolution. Nevertheless, migration times of less than 5 min and resolution factors (R_s) in the range of 2–4 were obtained when both additives were used. The method was validated with respect to selectivity, linearity, limits of detection and quantitation, analytical precision (intra‐ and inter‐day variability) and repeatability. Concentrations of 2.12 and 2.71% (w/w) of QC were found in pharmaceutical preparations of PQ from two different manufacturers. A possible mechanism for the successful separation of the isomers is also discussed. Copyright © 2008 John Wiley & Sons, Ltd.     

Determination of lipoic acid and dihydrolipoic acid in human plasma and urine by high-performance liquid chromatography with fluorimetric detection

A highly sensitive method for the determination of -lipoic acid (LA) and dihydrolipoic acid (DHLA) in human plasma and urine has been developed. Samples were acidified and extracted with organic solvent, and the free sulfhydryls of DHLA protected as the dicarboxyethylate by treatment with ethylchloroformate. The free carboxylic function of LA and the SH-protected DHLA were converted into their amide derivatives with the strong fluorophore 2-(4-aminophenyl)-6-methylbenzothiazole in the presence of a coupling agent and a base catalyst. The resulting fluorescent amides of both LA and DHLA were separated on a reversed-phase column (Ultrasphere C₈) using simple isocratic elution with acetonitrile–water (80:20) and detected fluorimetrically (excitation 343, emission 423 nm). The method is highly sensitive, reproducible, and is easily applied for the simultaneous determination of LA and DHLA in biological samples.     

Identification and determination of inorganic anions in real extracts from pre- and post-blast residues by capillary electrophoresis

Fast, selective, and sensitive analysis of inorganic anions is compulsory for the identification of explosives in post-blast or environmental samples. For the last twenty years, capillary electrophoresis (CE) has become a valuable alternative to ion chromatography (IC) for the analysis of inorganic-based explosives because of its low running costs and its simplicity of use. This article focuses on the development and validation of a CE method for the simultaneous analysis of 10 anions (chloride, nitrite, nitrate, thiosulphate, perchlorate, chlorate, thiocyanate, carbonate, sulphate, and phosphate) which can be found in post-blast residues, plus for the first time azide anion, possibly present in the composition of detonators, and the internal standard (formate) in 20 min total runtime. Intermediate precisions were 2.11% for normalized areas and 0.72% for normalized migration times. Limits of detection close to 0.5 ppm for all anions were obtained with the use of preconcentration techniques, thanks to a fast and simple sample preparation allowing the analysis of a large variety of matrices with the developed generic CE method. The matrix effects were statistically studied for the first time in the explosive field for different matrices, containing interfering anions and cations, sometimes at high levels. In fact, no significant matrix effect occurred (tests with blank matrix extracts of soil, cloth, glass, plastic, paper, cotton, and metal). Finally, analyses of real post-blast residues and real detonator extracts were performed. The CE results were compared with those obtained with the IC method used routinely and showed excellent correlation.     

Sensitive profiling of biogenic amines in human urine by capillary electrophoresis with field amplified sample injection

In order to monitor biogenic amines in human urine, a method based on field‐amplified sample injection combined with capillary electrophoresis and direct UV absorption detection was developed. Dopamine, tyramine, tryptamine, serotonin and epinephrine were effectively separated and identified in human urine samples, and detection limits were 0.072, 0.010, 0.027, 0.010 and 0.120 µmol/L, respectively. Detection limits comparable to laser‐induced fluorescence detection or solid phase extraction combined with capillary electrophoresis were achieved. Parameters affecting electrophoretic system detection sensitivity were investigated. Optimal separation conditions were obtained using as background electrolyte a pH 6.5 mixture of 2‐(morpholino)ethanesulfonic acid 20 mmol/L and 30 mmol/L phosphate buffer, containing 0.05% hydroxypropylcellulose and 10% v/v methanol. Injections of the sample solution were performed by applying a voltage of 12 kV for 50 s. Recovery and accuracy ranged between 89.4 and 94.9%, and 89 and 112%, respectively. The method was successfully applied on actual urine samples (from a healthy volunteer): target bioamine content was consistent with endogenous levels reported in the literature. The proposed method is simple, fast and inexpensive and can be conveniently employed in work‐related stress studies. The affordability and noninvasive sampling of the method allow epidemiological studies on large number of exposed persons to be performed. Copyright © 2013 John Wiley & Sons, Ltd.     

Ultrasound-assisted emulsification microextraction for the determination of ephedrines in human urine by capillary electrophoresis with direct injection. Comparison with dispersive liquid-liquid microextraction

Ultrasound-assisted emulsification microextraction and dispersive liquid-liquid microextraction were compared for extraction of ephedrine, norephedrine, and pseudoephedrine from human urine samples prior to their determination by capillary electrophoresis. Formation of a microemulsion of the organic extract with an aqueous solution (at pH 3.2) containing 10% methanol facilitated the direct injection of the final extract into the capillary. Influential parameters affecting extraction efficiency were systematically studied and optimized. In order to enhance the sensitivity further, field-amplified sample injection was applied. Under optimum extraction and stacking conditions, enrichment factors of up to 140 and 1750 as compared to conventional capillary zone electrophoresis were obtained resulting in limits of detection of 12-33 μg/L and 1.0-2.8 μg/L with dispersive liquid-liquid microextraction and ultrasound-assisted emulsification microextraction when combined with field-amplified sample injection. Calibration graphs showed good linearity for urine samples by both methods with coefficients of determination higher than 0.9973 and percent relative standard deviations of the analyses in the range of 3.4-8.2% for (n = 5). The results showed that the use of ultrasound to assist microextraction provided higher extraction efficiencies than disperser solvents, regarding the hydrophilic nature of the investigated analytes.     

Different strategies for the preconcentration and separation of parabens by capillary electrophoresis

Several strategies, namely, large volume sample stacking (LVSS), field‐amplified sample injection (FASI), sweeping, and in‐line SPE‐CE, were investigated for the simultaneous separation and preconcentration of a group of parabens. A BGE consisting of 20 mM sodium dihydrogenphosphate (pH 2.28) and 150 mM SDS with 15% ACN was used for the separation and preconcentration of the compounds by sweeping, and a BGE consisting of 30 mM sodium borate (pH 9.5) was used for the separation and preconcentration of the compounds by LVSS, FASI, and in‐line SPE‐CE. Several factors affecting the preconcentration process were investigated in order to obtain the maximum enhancement of sensitivity. The LODs obtained for parabens were in the range of 18–27, 3–4, 2, and 0.01–0.02 ng/mL, and the sensitivity evaluated in terms of LODs was improved up to 29‐, 77‐, 120‐, and 18 400‐fold for sweeping, LVSS, FASI, and in‐line SPE‐CE, respectively. These preconcentration techniques showed potential as good strategies for focusing parabens. The four methods were validated with standard samples to show the potential of these techniques for future applications in real samples, such as biological and environmental samples.     

Simultaneous determination of oleanolic acid,ursolic acid,quercetin and apigenin in Swertia mussotii Franch by capillary zone electrophoresis with running buffer modifier

The method of capillary zone electrophoresis (CZE) with direct UV detection was developed for the determination of oleanolic acid, ursolic acid, quercetin and apigenin. and then for the first time successfully applied to the analysis of four analytes in Swertia mussotii Franch and its preparations. Various factors affecting the CZE procedure were investigated and optimized, and the optimal conditions were: 50 × 10^?3 mol/L borate‐phosphate buffer (pH 9.5) with 5.0 × 10^?3 mol/L β‐cyclodextrin, 15 kV separation voltage, 20 °C column temperature, 250 nm detection wavelength and 5 s electrokinetic injection time (voltage 20 psi). Under the conditions, oleanolic acid, ursolic acid, quercetin and apigenin could be determined within the test ranges with a good correlation coefficient (r² > 0.9991). The limits of detection for conditions, oleanolic acid, ursolic acid, quercetin and apigenin were 0.3415, 0.2003, 0.0062 and 0.2538 µg/mL, respectively, and the intra‐ and inter‐day relative standard deviations were no more than 4.72%. This procedure provided a convenient, sensitive and accurate method for simultaneous determination of oleanolic acid, ursolic acid, quercetin and apigenin in S. mussotii Franch. Copyright © 2014 John Wiley & Sons, Ltd.     

Determination of quinolone antibiotics in urine by capillary electrophoresis with chemiluminescence detection

A novel and simple method is presented for the determination of norfloxacin, ciprofloxacin, and ofloxacin by capillary electrophoresis with chemiluminescence detection. This method is based on the enhancing effect of quinolones on the chemiluminescence reaction of the Ce(SO₄)₂–Ru(bpy)₃²⁺–HNO₃ system. Three quinolones were successfully separated and detected under optimum conditions. The obtained detection limits were 2.3×10^–7 mol/L, 5.2×10^–8 mol/L, and 7.8×10^–8 mol/L for ciprofloxacin, norfloxacin, and ofloxacin, respectively. The RSD of migration time and peak area were less than 1.8 and 3.8% (n = 5), respectively. The applicability of the proposed method was illustrated in the determination of ofloxacin in eye drops and of norfloxacin in human urine samples, and the monitoring of pharmacokinetics for norfloxacin.     

Determination of eight illegal drugs in human urine by combination of magnetic solid‐phase extraction with capillary zone electrophoresis

Using magnetite/silica/poly(methacrylic acid‐co‐ethylene glycol dimethacrylate) (Fe₃O₄/SiO₂/poly(MAA‐co‐EDMA)) magnetic microspheres, a rapid and high‐throughput magnetic solid‐phase extraction coupled with capillary zone electrophoresis (MSPE‐CZE) method was developed for the determination of illegal drugs (ketamine, amphetamines, opiates, and metabolites). The MSPE of target analytes could be completed within 2 min, and the eight target analytes could be baseline separated within 15 min by CZE with 30 mM phosphate buffer solution (PBS, pH 2.0) containing 15% v/v ACN as background electrolyte. Furthermore, hydrodynamic injection with field‐amplified sample stacking (FASS) was employed to enhance the sensitivity of this MSPE‐CZE method. Under such optimal conditions, the limits of detection for the eight target analytes ranged from 0.015 to 0.105 μg/mL. The application feasibility of MSPE‐CZE in illegal drugs monitoring was demonstrated by analyzing urine samples, and the recoveries of target drugs for the spiked sample ranging from 85.4 to 110.1%. The method reproducibility was tested by evaluating the intra‐ and interday precisions, and relative standard deviations of <10.3 and 12.4%, respectively, were obtained. To increase throughput of the analysis, a home‐made MSPE array that has potential application to the treatment of 96 samples simultaneously was used.     

Application of simultaneous separation and derivatization for the determination of α‐lipoic acid in urine samples by high‐performance liquid chromatography with spectrofluorimetric detection

To help to clarify therapeutic functions of lipoic acid (LA) in biochemical and clinical practice we have elaborated a fast, simple and accurate HPLC method enabling determination of LA in human urine. The proposed analytical approach includes reduction of LA with tris(2‐carboxyethyl)phosphine and simultaneous separation and derivatization of the analyte with butylamine and o‐phthaldialdehyde followed by spectrofluorimetric detection at λ_ex = 340 nm and λ_em = 440 nm. The assay was performed using gradient elution and the mobile phase containing 0.0025 mol L^?1 o‐phthaldialdehyde in 0.0025 mol L^?1 NaOH and acetonitrile. Linearity of the detector response for LA was observed in the range of 0.3–8 μmol L^?1. Limits of detection and quantification for LA in urine samples were 0.02 and 0.03 μmol L^?1, respectively. The total analysis time, including sample work‐up, was <20 min. The analytical procedure was successfully applied to analysis of real urine samples delivered from six healthy volunteers who received a single 100 mg dose of LA.     

Hyphenation of field‐amplified sample injection and transient isotachophoresis in CE for the determination of sotalol and metoprolol in human urine samples

A sensitive approach of capillary electrophoresis coupled with field‐amplified sample injection and transient isotachophoresis was developed for the simultaneous determination of two β‐blockers: sotalol and metoprolol. In this dual focusing technique, the samples were prepared via only dissolution in ultrapure water and then injected electrokinetically. Phosphate acted as both the background electrolyte and the leading electrolyte. Its optimized concentration was 80 mM. A total of 25 mM of glycine was used as the terminating electrolyte. Under optimum conditions, good separation of sotalol and metoprolol was achieved within 10 min. In comparison with the conventional method, the sensitivity enhancement factors were up to 1031 and 919 for sotalol and metoprolol, respectively. The proposed method was employed in the determination of sotalol and metoprolol in spiked human urine samples. The limits of detection and limits of quantitation obtained via ultraviolet detection were 5 and 12 ng/mL, respectively, for sotalol, and 10 and 25 ng/mL, respectively, for metoprolol. The intraday repeatability values were lower than 2.7 and 1.7% for peak area and migration time, respectively. The assay is a simple and efficient strategy with potential for application in clinical and biochemical laboratories for monitoring sotalol and metoprolol.     

乙醛酸和草酸的毛细管区带电泳分析

建立了同时测定乙醛酸和草酸的毛细管区带电泳法。考察了缓冲溶液的种类、浓度和pH以及分离电压等因素对分离结果的影响。在缓冲溶液为20 mmol/L硼砂-5.5 mmol/L邻苯二甲酸氢钾(pH 9.0)、分离电压20 kV、检测波长212 nm的优化条件下,11 min内即可实现对目标物的分离。乙醛酸和草酸分别在0.8～20 g/L和1.2～20 g/L范围内线性关系良好,相关系数分别为0.9993和0.9975;方法的检出限分别为0.2和0.4 g/L(信噪比为3);样品的加标回收率为98.3%～102.5%,相对标准偏差为0.35%～0.61%。该方法操作简便、快速、成本低廉,已应用于实际样品的分析,并获得了令人满意的结果。     

Determination of norfloxacin in human urine by capillary electrophoresis with electrochemiluminescence detection

A fast and sensitive approach that can be used to detect norfloxacin in human urine using capillary electrophoresis with end-column electrochemiluminescence (ECL) detection of is described. The separation column was a 75-μm i.d. capillary. The running buffer was 15 mmol L⁻¹ sodium phosphate (pH 8.2). The solution in the detection cell was 50 mmol L⁻¹ sodium phosphate (pH 8.0) and 5 mmol L⁻¹ The ECL intensity varied linearly with norfloxacin concentration from 0.05 to 10 μmol L⁻¹. The detection limit (S/N=3) was 0.0048 μmol L⁻¹, and the relative standard deviations of the ECL intensity and the migration time for eleven consecutive injections of 1.0 μmol L⁻¹ norfloxacin (n=11) were 2.6% and 0.8%, respectively. The method was successfully applied to the determination of norfloxacin spiked in human urine without sample pretreatment. The recoveries were 92.7–97.9%.      

Combination of cationic surfactant-assisted solid-phase extraction with field-amplified sample stacking for highly sensitive analysis of chlorinated acid herbicides by capillary zone electrophoresis

This report describes a novel online field-amplified sample stacking (FASS) procedure to analyze 16 chlorinated acid herbicides. By using a poly(vinyl alcohol) (PVA)-coated capillary to reduce electroosmotic flow and introducing a methanol-water plug before sample loading, the sample injection time could be very long without loss of sample and separation efficiency. Under the optimized condition, the FASS procedure could provide great sensitivity enhancement (5000-10 000-fold) and satisfactory reproducibility (relative standard deviations of migration times less than 2.4%, relative standard deviations of peak areas less than 8.0%). Combined with cationic surfactant-assisted solid-phase extraction (CSA-SPE), the limit of detection of the herbicides ranged from 0.269 to 20.3 ppt, which are two orders lower than those of the US Environmental Protection Agency standard method 515.1. The CSA-SPE-FASS-CE method was successfully applied to the analysis of local pond water.