CE-UV Method Research Articles

Determination of trace bisphenol A in complex samples using selective molecularly imprinted solid-phase extraction coupled with capillary electrophoresis

The highly selective, fast and effective sample pretreatment technique molecularly imprinted solid-phase extraction (MISPE) can overcome the low sensitivity of the highly efficient capillary electrophoresis–UV method (CE–UV). In this work, narrowly dispersible bisphenol A (BPA)-imprinted polymeric microspheres with a high capacity factor of k′=6.8 and an imprinted factor of I=6.53 were investigated as selective solid-phase extraction (SPE) sorbents for use in extraction of BPA from different sample matrices (tap water, wastewater, Yangtze River water, soil from the Yangtze River, shrimp and human urine). Washing and eluting protocols of MISPE were optimized. Under optimal conditions, recoveries of MISPE were investigated. Recoveries were basically constant and the relative standard deviation (RSD) was lower than 5.8% when loading volumes changed from 1 to 50mL. Recoveries ranged from 71.20% to 86.23% for different sample matrices. Compared with C18 SPE, MISPE had higher selectivity and recovery for BPA. BPA was determined with good accuracy and precision in different complex samples using CE–UV coupled with MISPE. Spiked recoveries ranged from 95.20% to 105.40%, and the RSD was less than 7.2%. Because a large loading volume was achieved, the enrichment efficiency of pretreatment and the sensitivity of this method were improved. The limits of detection of this MISPE–CE–UV method for BPA in tap water, wastewater, Yangtze River water, soil from the Yangtze River, shrimp and human urine were 3.0μgL−1, 5.4μgL−1, 6.9μgL−1, 2.1μgL−1, 1.8μgL−1 and 84μgL−1, respectively.

Comparative Study of Captopril Derivatization Reaction by LC-UV, LC-MS and CE-UV Methods

The LC-UV, LC-MS and CE-UV study of chemical reaction between captopril and p-bromo- phenacyl bromide as derivatizing reagent is reported. During transformation of captopril, its thiol group is involved and the reaction is irreversible. Neutral or alkaline environments favor derivatization. The yield of reaction increases linear with the concentration of the reagent, while changes in temperature do not influence it significantly. Kinetic studies show that derivatization of captopril flows quickly, reaching a plateau after 30 minutes. Based on the study of these effects some important parameters such as deri- va- tizing agent concentration, reaction time, temperature and pH were confined. The results were used for development of captopril determination methods both in pharmaceutical formulations and biological samples. (doi: 10.5562/cca1787)

Noninvasive detection of counterfeited ampoules of dexamethasone using NIR with confirmation by HPLC-DAD-MS and CE-UV methods

Application of near-infrared (NIR) measurements together with chemometric data processing is widely used for counterfeit drug detection. The most difficult counterfeits to detect are the "high quality fakes", which have the proper composition but are produced in violation of technological regulations by underground manufacturers. This study uses such forgeries and addresses important issues. The first is the possibility of applying the NIR/chemometric approach to the detection of injectable formulations of drugs (in this case dexamethasone), which are aqueous solutions with low concentration of active ingredients, directly in the closed ampoules. The second issue is the comparison of NIR/chemometric conclusions with detailed chemical analysis.

Analysis of aristolochic acids by CE‐MS with carboxymethyl chitosan‐coated capillary

A CE-MS method for rapid determination of aristolochic acid-I and aristolochic acid-II (AA-II) in traditional Chinese medicines and biological samples was described in the present paper. AA-I and AA-II can be baseline separated within 6 min by CE-MS with carboxymethyl-chitosan-coated capillary. CZE conditions including pH, concentration of buffer, applied voltage, and capillary temperature were systematically investigated, and the composition and flow rate of sheath liquid were also optimized for CE-MS. Furthermore, the CE-UV method without any additives in BGE solution was established and compared with the CE-MS method. The results showed that the two methods could achieve satisfactory separation efficiency, repeatability, and linearity, while the LOD was 0.6 microg/mL for CE-UV and 0.05 microg/mL for CE-MS. Compared with the CE-UV method, the sensitivity of CE-MS was significantly improved, in addition to the structure information provided by MS detection at the same time. As an application example, a spiked sample in human serum was analyzed by the CE-MS method, indicating that the new CE-MS method can be applied to analyze AAs in biological samples.

Simultaneous analysis by capillary electrophoresis of five amyloid peptides as potential biomarkers of Alzheimer's disease

We report here a CE method for the separation and quantitation of five amyloid peptides (Aβ1-42, 1-40, 1-39, 1-38, and 1-37) considered as potential biomarkers of Alzheimer's disease. These amyloid peptides have very similar structures. Sample preparation and storage conditions are critical parameters to ensure their solubility and to avoid the aggregation process in particular for Aβ1-42. Their solubility was found fully dependent on the NH 4OH concentration that was employed initially to dissolve the lyophilized amyloid peptides. Conditions to achieve a full separation of these peptides were found using a dynamic coating with 1,4-diaminobutane (DAB). The linear decrease of their electrophoretic mobility highlighted an ion-pairing phenomenon between the peptides and DAB. The optimal background electrolyte was a 40 mM borate buffer, pH 9 containing 3 mM of DAB. Under these conditions, resolutions ranged from 1.3 to 2.4 with theoretical plates reaching 300,000. Under the retained conditions, we showed that adsorption of peptides to silica was negligible (recovery over 94.5%) and depletion effect of the background electrolyte was overcome. The method was finally validated in terms of linearity and repeatability and the limits of detection for the five Aβ peptides were estimated. The inter-day repeatability of the migration times was very satisfactory with RSDs less than 1.55%. The RSDs of the peak areas were below 5%. With this CE–UV method, limits of detection of the peptides ranged from 300 to 500 nM. We finally demonstrated that this method can be applied to real biological samples such as CSF.

Sample stacking for the analysis of penicillins by microemulsion electrokinetic chromatography

In this study, on-line sample concentration methods, which coupled field-amplified sample injection and sweeping technology with MEEKC, were used to detect and analyze eight common penicillin antibiotics (nafcillin, dicloxacillin, ampicillin, oxacillin, penicillin V, cloxacillin, penicillin G, and amoxicillin). During the optimization of field-amplified sample injection-sweeping MEEKC, the composition of sample matrix and the length of acidic plug were found to be the predominant influences for penicillin stacking. Both zwitterionic ampicillin and amoxicillin could only be stacked through cation-selective-exhaustive-injection sweeping, whereas the other six penicillin compounds were found to be concentrated by anion-selective-exhaustive-injection sweeping. Hence, in order to simultaneously concentrate the eight penicillins in a single-run sweeping step, a combination of successive anion- and cation-selective injections was used. When compared with previous CE-UV methods, the proposed on-line concentration MEEKC method provided better detection sensitivity and faster separation for these penicillins either in single ion-selective injection or in successive anion-/cation-selective injection where the LODs were in the range of 0.2-2.8 microg/L and 0.5-5.8 microg/L, respectively.

Enantiomeric Separation of Omeperazole Enantiomers by Aqueous CE Using UV and MS Detection

This paper details the analysis of the enantiomers of omeprazole, using aqueous CE coupled with MS detection. Following our previously published work: where a non-aqueous CE–UV method was developed for omeprazole and 5-hydroxy-omeprazole; coupling to electro-spray ionization (ESI) MS detection has now been investigated, using a sheath-flow interface for introduction. An aqueous CE method was developed and designed to afford increased compatibility with ESI–MS detection, employing an ammonium acetate buffer system (pH 5.8). Common partial filling methods could not be utilized to avoid the entrance of cyclodextrin into the MS, and therefore a modified method of non-continuous-flow CE–MS was applied, with the CE separation carried out without applied ESI voltage, before reapplying and allowing flow into the MS for data collection. A chiral CE separation of omeprazole and 5-hydroxyomeprazole was achieved, and chiral CE resolution of omeprazole has been demonstrated using MS detection.

A sensitivity enhanced high-performance liquid chromatography fluorescence method for the detection of nephrotoxic and carcinogenic aristolochic acid in herbal medicines

A new, sensitive and selective HPLC method with fluorescence detector (HPLC–FLD) for the determination of nephrotoxic and carcinogenic aristolochic acid (AA) in herbal medicines by using pre-column derivatization with zinc powder in acetic acid is presented. Variables governing the derivatization reaction, such as the amount of zinc powder and acetic acid, as well as the derivatization time were studied and optimized. An extended linear dynamic range over three orders of magnitude was observed for AA-I and AA-II ( R 2 > 0.9998). Method accuracy at low, medium and high spiked AA levels determined by the percentage mean deviation was below 4.4% and 7.2% for AA-I and AA-II, respectively. The detection limits of 0.39 ng/mL (AA-I) and 0.52 ng/mL (AA-II) were 2 orders of magnitude lower than those obtained from HPLC-MS or CE-ECD analyses, 3–4 orders of magnitude lower than those from HPLC-UV or CE-UV methods. The developed method has been applied for the determination of AA in herbal medicines. Among the tested samples, Guanmutong had the highest AA concentration (2607.0 μg/g AA-I, 711.2 μg/g AA-II). Comparison studies between HPLC–FLD and HPLC–MS/MS demonstrated that the two methods gave similar quantitative results for the selected herb samples.

Characterization of transferrin glycoforms in human serum by CE‐UV and CE‐ESI‐MS

Human transferrin (Tf) is a model glycoprotein for congenital disorders of glycosylation (CDG) diagnosis. In the last few years, new CE-UV methods for intact Tf glycoforms analysis have been developed using nonvolatile BGEs and organic modifiers. However, the use of these BGEs does not allow the coupling of these procedures with electrospray MS (ESI-MS). In this study, a new CE-UV separation method of Tf glycoforms is developed, using a double-layer stable coating and a volatile BGE based on ammonium acetate. The separation method is optimized using standard Tf and their potential is demonstrated applying the method to the analysis of sera Tf from healthy individuals and CDG patients. The CE-UV separation method has been coupled to ESI-MS detection. Main parameters such as sheath liquid composition are optimized in order to obtain a good sensitivity. The CE-ESI-MS method has also been used in serum samples obtaining the separation of the different proteins present in serum and partial separation of Tf glycoforms. Different mass spectra and deconvoluted molecular masses were obtained for each sialoform, allowing unequivocal glycoform identification.

Capillary electrophoresis-mass spectrometry for impurity profiling of basic pharmaceuticals using non-volatile background electrolytes

A generic approach has been developed for coupling capillary electrophoresis (CE) using non-volatile background electrolytes (BGEs) with mass spectrometry (MS) using a sheath liquid interface. CE-MS has been applied for basic and bi-functional compounds using a BGE consisting of 100 mM of TRIS adjusted to pH 2.5 using phosphoric acid. A liquid sheath effect is observed which may influence the CZE separation and hence may complicate the correlation between CE-UV and CE-MS methods. The influence of the liquid sheath effect on the migration behavior of basic pharmaceuticals has been studied by simulation experiments, in which the BGE outlet vial is replaced by sheath liquid in a CE-UV experiment. As a consequence of the liquid sheath effect, phosphate based BGEs can be used without significant loss of MS sensitivity compared to volatile BGEs. The use of buffer constituents such as TRIS can lead to lower detection limits as loss of MS sensitivity can be compensated by better CE performance. TRIS based BGEs permit relatively high injection amounts of about 100 pmol while maintaining high resolution. The ESI-MS parameters were optimized for a generic method with maximum sensitivity and stable operation, in which the composition of the sheath liquid and the position of the capillary were found to be important. Furthermore, the nebulizing pressure strongly influenced the separation efficiency. The system showed stable performance for several days and a reproducibility of about 15% RSD in peak area has been obtained. Nearly all test compounds used in this study could be analyzed with an MS detection limit of 0.05% measured in scan mode using extracted ion chromatograms. As a result, CE-MS was found to be a valuable analytical tool for pharmaceutical impurity profiling.

Simultaneous determination of allantoin, hypoxanthine, xanthine, and uric acid in serum/plasma by CE

Allantoin (All) is an oxidative end product of purines in mammals. The small amount of All present in human plasma or serum results from free radical action on urate and may provide a stable marker of in vivo free radical activity. Because free radicals have been implicated in the development and progression of atherosclerosis, this study focused on the metabolic compounds of the All pathway. We propose a new fast CE (CE/UV) method for the simultaneous determination of All, uric acid (UA), hypoxanthine (HX), and xanthine (X) in human plasma. These products were quantified in the plasma of patients with chronic renal failure before hemodialysis (n = 6), patients with chronic heart failure (n = 6) and controls (n = 6). The filtered plasma were diluted ten-fold before the direct injection in CE/UV (195 nm), which allows separating the four compounds in less than 13 min. The metabolites were detectable at concentrations of 0.3-0.6 micromol/L. The method was linear over the range 0.5-150 micromol/L for All, HX, and X and 10-1500 micromol/L for UA (r > 0.99). The analytical performance of this method is satisfactory with intra-assay CV < 3.4%, inter-assay CV < 5% (HX and X < 7%), and recovery (93-101%). The proposed CE-UV method appears to be a useful tool for studying physiological and pathological changes of HX, UA, and All levels in plasma samples, the latter being a possible indicator of free radical damage in vivo.

Evaluation of free hydroxyl radical scavenging activities of some Chinese herbs by capillary zone electrophoresis with amperometric detection.

Due to the severe damage caused by free hydroxyl radicals (OH.) to cells and tissues, there is much interest in finding and studying effective and non-toxic OH. scavengers, including traditional Chinese herbs. In this paper, the simple and highly-sensitive technique of capillary zone electrophoresis with amperometric detection (CZE-AD) was used to study the OH. scavenging activities of aqueous extracts from some traditional Chinese herbs. Salicylic acid (SAL) was used as an OH. trap, and the content of OH. could be determined by assaying their products, 2,3-dihydroxybenzoic acid (2,3-DHBA) and 2,5-dihydroxybenzoic acid (2,5-DHBA). The optimum conditions for CZE-AD for the determination of 2,3-DHBA and 2,5-DHBA were explored. The linearity ranges of 2,3-DHBA and 2,5-DHBA were 1.0 x10(-7) approximately 1.0 x10(-4) mol L(-1), and their detection limits were as low as 2 x 10(-8) mol L(-1), which were much better than the CE-UV method often used. The traditional Chinese herbs studied included Radix angelicae sinensis, Rhizoma coptidis, Ligustrum lucidum, Ligusticum wallichii, Radices glycyrrhizae and Semen plantaginis. The experiments showed that the aqueous extracts from all of the above traditional Chinese herds had free OH. scavenging activities, although to different degrees.

Determination of hydroxyl radical by capillary zone electrophoresis with amperometric detection

Hydroxyl radical (OH) can cause severe damage to cells and tissues. However, its analysis is very difficult for its high reactivity and very short half-life. In this paper, a simple and highly sensitive method, capillary zone electrophoresis with amperometric detection (CZE-AD) was introduced indirectly to determine OH by determining its reaction products with salicylic acid (SAL), 2,3-dihydroxybenzoic acid (2,3-DHBA) and 2,5-dihydroxybenzoic acid (2,5-DHBA). The optimum conditions of CZE-AD for the determination of 2,3-DHBA and 2,5-DHBA were explored. Under the optimum conditions, SAL, 2,3-DHBA and 2,5-DHBA could be perfectly separated within 15min, and the linearity ranges of 2,3-DHBA and 2,5-DHBA were between 1.0×10−7 and 1.0×10−4moll−1. Their detection limits were as low as 2×10−8moll−1, which were much lower than that in CE-UV method. The method was also applied to study the free OH scavenging activity of angelica polysaccharide. The experimental results showed that this CZE-AD method was very sensitive and practical in both the determination of free OH and the evaluation of free OH scavenging activities of antioxidants.

Determination of triamterene and its main metabolite hydroxytriamterene sulfate in human urine by capillary electrophoresis using ultraviolet absorbance and laser-induced fluorescence detection

Two capillary electrophoresis methods have been developed for the direct determination of triamterene and its main metabolite hydroxytriamterene sulfate in human urine. Analytes were detected using conventional UV detection as well as laser-induced fluorescence (LIF) detection with an HeCd-laser operating at a wavelength of 325 nm. The results of both detection techniques were compared. Indeed, the limit of quantification was eightfold lower using LIF detection (50 ng/ml) in comparison to UV detection (400 ng/ml). As no interference due to endogenous urine compounds was observed, direct urine analysis was feasible. Analysis was very simple and fast—one run could be performed within less than 10 min (CE–UV method) and 2.5 min (CE–LIF method), respectively. Both assays were fully validated and applied to urine samples from a human volunteer. The results of the application of the CE–LIF method to human urine samples are presented in this publication.

Determination of linear alkylbenzenesulfonates in wastewater treatment plants and coastal waters by automated solid-phase extraction followed by capillary electrophoresis–UV detection and confirmation by capillary electrophoresis–mass spectrometry

Linear alkylbenzenesulfonates (LASs) were determined in wastewaters and coastal waters by solid-phase extraction, using two different sample preparation protocols depending on the sample treated, followed by capillary electrophoresis and ultraviolet detection (CE–UV). The linear range of the proposed method varied from 3 to 53 and from 25 to 495 μg/l, depending on the compound, with a limit of detection of 1 μg/l when 250 ml of coastal water was preconcentrated. [M−H] − ions were used for CE–MS confirmation after quantification by CE–UV. CE–MS diagnostic ions were the same ones used in LC–electrospray (ESI) MS and corresponded to m/ z 297, 311, 325 and 339 for C 10, C 11, C 12 and C 13 LASs, respectively. LASs were determined in wastewater samples of the influent and effluent of three wastewater treatment plants (WWTPs), two of them using biological treatment with secondary settlement and receiving mainly domestic wastewaters whereas one of the plants was operated with physicochemical treatment and received mainly industrial wastewaters. LASs were also analyzed in two samples from coastal waters of the bay of Cadiz (Spain) receiving untreated domestic effluents. All samples were also analyzed by LC–ESI-MS and the results are compared with the CE–UV method developed in this work. The concentration levels of total LASs varied from 988 to 1309 μg/l in the influents of WWTPs, whereas in the effluents the concentrations varied from 136 to 197 μg/l. The levels of LASs in coastal wastewaters of the bay of Cadiz varied from 739 to 911 μg/l, indicating that the wastewaters discharged into the bay did not undergo any treatment at all.