On-line Preconcentration Method Research Articles

Sensitive analysis of selenium speciation in natural seawater by isotope-dilution and large volume injection using PTV-GC-ICP-MS

Although oceans play a key role in the global selenium (Se) cycle, there is currently very little quantitative information available on the distribution of Se concentrations and Se speciation in marine environments. In general, determining Se concentration and speciation in seawater is highly challenging due to very low Se levels ((sub)ng⋅L−1), whereas matrix elements interfering Se pre-concentration and detection are up to the g⋅L−1 levels. In this study, we established a sensitive method for the determination of the various Se chemical fractions present in natural seawater, i.e. selenite (SeIV), selenate (SeVI), organic Se-II + Se0 and total Se, using species-specific isotope dilution gas chromatography coupled to inductively coupled plasma mass spectrometry (ID-GC-ICP-MS). We compared different derivatization reagents and optimized specific pre-treatment protocols, including a microwave assisted oxidation protocol for the determination of total Se and organic Se-II + Se0 using H2O2. To increase sensitivity, we developed an online pre-concentration method based on large volume injection (LVI) using a programmed temperature vaporization (PTV) inlet. Eventually, the developed method achieved low absolute and methodological detection limits, i.e., respectively, 0.1–0.3 pg and 0.9–3.1 ng.L-1 for the different fractions. The accuracy of our method was of 2% for a certified reference material (CRM) diluted in artificial seawater while the precision was better than 4% for a freshwater CRM in artificial seawater matrix as well as two common seawater CRMs certified for trace elements excluding Se. As a proof-of-concept, we quantified the various Se fractions in a large number of natural water samples from the Baltic and North Seas, encompassing a wide range of salinity (7–35 psu), which shows that its detection limits are sufficient to determine total Se, SeIV, SeVI and organic Se-II + Se0 concentrations in brackish and marine systems.

On-line preconcentration of fluorescein derivatives of essential amino acids using sweeping in a nonionic micellar system

A rapid and effective method of nonionic micellar electrokinetic chromatography with laser-induced fluorescence detection and sweeping as an online preconcentration method was developed for the simultaneous determination of free essential amino acids in supplement formulations. The analytes were derivatized using fluorescein isothiocyanate isomer I dissolved in dimethyl sulfoxide. Optimal separation conditions were achieved using an uncoated fused silica capillary (40.0 cm effective length, 50.2 cm total length, 50 µm internal diameter), 20 mM Na2B4O7, (pH 9.3), 50 mM Brij-35, and 15% (v v-1) methanol as running buffer. Complete separation of all analytes was achieved within 15 min. The optimized method exhibited good linearity (r2 > 0.997), acceptable precision (1.3 – 12.4%), and accuracy (88 – 112%) in concentration range 20 – 1000 ng mL−1. The limit of detection was 6 ng mL−1. The method successfully determined the quantity of free essential amino acids in a dietary supplement containing spirulina.

A pH-mediated field amplification sample stacking technique based on portable microchip electrophoresis heavy metal ion detection system.

We built a portable microchip electrophoresis heavy metal ion detection system and proposed a pH-mediated field amplified sample stacking (pH-mediated FASS) online preconcentration method. The pH-mediated FASS focuses and stacks heavy metal cations by controlling electrophoretic mobilities with a pH change between the analyte and the background electrolyte (BGE) in solution to improve the detection sensitivity of the system. We optimized and adjusted sample matrix solution (SMS) ratios and pH to create concentration and pH gradients for SMS and BGE. Furthermore, we optimize the microchannel width to improve the preconcentration effect further. The system and method analyzed soil leachates polluted with heavy metals and separated Pb2+ and Cd2+ within 90s, obtaining their levels at 58.01mg/L and 4.91mg/L with sensitivity enhancement factors (SEF) of 26.40 and 43.73. Compared with inductively coupled plasma atomic emission spectrometry (ICP-AES), the detection error of the system was less than 8.80%.

On-line preconcentration and determination of sulfadiazine in food samples using surface molecularly imprinted polymer coating by capillary electrophoresis

In this study, on-line preconcentration and selective determination of the trace sulfadiazine (SDZ) existing in milk and hen egg white samples were realized by the capillary electrophoresis using molecularly imprinted polymer (MIP) coated capillary. The capillary coated with MIP was firstly prepared through the surface imprinted techniques, using SDZ as template molecule and dopamine as function monomer and crosslinker, and then amine-terminated poly(2-methyl-2-oxazoline) (PMOXA-NH2) was introduced onto polydopamine layer to reduce the non-specific adsorption. Successful preparation of SDZ-MIP-PMOXA coating was verified by zeta potential, as well as water contact angle. The SDZ-MIP-PMOXA coated capillary performed well on-line preconcentration of SDZ and the obtained peak area of SDZ was 46 times higher than that one obtained in bare capillary using the same procedure. Then the proposed on-line preconcentration method was fully validated and displayed good linear behavior in the concentration from 5.0 to 100.0 ng/mL, with the limit of detection was low to 1.5 ng/mL; and this method presented excellent accuracy and robustness. The prepared SDZ-MIP-PMOXA coated capillary also showed high selectivity with the imprinting factor of 5.85 and good repeatability during five consecutive runs with the relative standard deviation value of peak area was 1.6%. At last, the application of the prepared SDZ-MIP-PMOXA coated capillary in the detection of SDZ in spiked food samples was investigated, and good recoveries of 98.7–109.3% were obtained.

Determination of pepsin by capillary electrophoresis using mixed polymer coated capillary with switchable properties toward protein adsorption/desorption.

In this work, a simple online preconcentration method for quantitative detection of pepsin was realized by using the binary mixed polymer brushes coated capillary with switchable properties toward protein adsorption. Firstly, the binary mixed polymer brushes were prepared by grafting poly(2-methyl-2-oxazoline) and poly(4-vinylpyridine) onto the inner wall of the capillary through a polydopamine anchor. Then the coatings were characterized by X-ray photoelectron spectrometer and electroosmotic flow measurement. The results indicated that the composition of coating could be controlled by varying the feed ratio of poly(2-methyl-2-oxazoline) to poly(4-vinylpyridine) and the inner surface charge could be tuned toward the change of pH and ionic strength. The results showed that when the poly(2-methyl-2-oxazoline)/poly(4-vinylpyridine) mass ratio was 80/20, the highest online preconcentration effect was obtained and the sensitivity enhancement factor was 6.3. Moreover, satisfactory sensitivity (limit of detection: 7.5ng/mL) and good repeatability were obtained with the online preconcentration method. The polymer-coated capillary was still stable for online preconcentration and detection of pepsin after 50 consecutive runs. Last, the proposed method was used successfully to online preconcentrate pepsin in the saliva matrix.

Analysis of Seventeen Certified Water Reference Materials for Trace and Technology‐Critical Elements

Concentrations of elements in the aquatic environment are a key parameter for various scientific fields such as biogeochemistry, biology and environmental science. Within this context, the scientific community asks for new analytical protocols to be able to quantify more and more elements of the periodic table. Therefore, the requirements for aqueous reference materials have increased drastically. Even though a wide variety of CRMs of different water matrices are available, certified values of many elements (e.g., rare earth elements (REE), technology‐critical elements, such as Ga and In, and generally those elements which are not part of current monitoring regulations) do not yet exist. Therefore, the scientific community relies on published elemental concentrations of many CRMs provided by other researchers. Some elements of interest, such as the REE, are well studied and plenty of literature values exist. However, less studied elements, such as Ga and In, are rarely studied. In this study, an ‘externally’ calibrated quantification method based on an optimised online preconcentration method coupled with ICP‐MS/MS was used for the quantification of thirty‐four elements. The method was applied to seventeen water CRMs covering freshwater, brackish water and seawater. The measured data are combined with a comprehensive literature review on non‐certified values in selected water CRMs, and new consensus values are suggested for various non‐certified elements.

A rapid, automated system for the separation, preconcentration and measurement of 90Sr, and U, Am and Pu isotopes

An online separation and preconcentration method, using an automated flow injection setup and solid phase extraction followed by ICP-MS/MS, was developed for the analysis of 90Sr, and U, Am and Pu isotopes in various liquid sample matrices. The radionuclide analytes were separated from interferences and complex matrices using DGA-branched resin and Sr resin, then specific gases were used in the reaction/collision cell in the ICP-MS/MS to measure the different analytes. The system requires smaller sample volumes (10 mL), less sample preparation and shorter processing time (46 min per sample) compared to traditional radiometric and other MS techniques. Based on a 10 mL sample, the limits of detection were 1.48 pg L−1 (8257 mBq L−1) for 90Sr, 1.75 pg L−1 (0.40 mBq L−1) for 234U, 0.65 pg L−1 (77.65 mBq L−1) for 241Am, and 0.56 pg L−1 (1.25 mBq L−1) for 239Pu when all target analytes were measured in one analysis. The analytical figures of merit were evaluated for a range of sample matrices including lake water, seawater and urine and were comparable to those reported in the literature. This online system thus provides a novel, fully automated analytical tool with faster analysis time, smaller sample requirements, minimum sample preparation, low detection limits and the flexibility to handle single and multiple measurements of various radionuclides.

Optimisation of preconcentration for determination of dicarboxylic acids using ion chromatography

ABSTRACT An automated on-line preconcentration and detection method for determination of dicarboxylic acids (DCAs) in aqueous samples is proposed. Parameters of two different off-line preconcentration methods (solid phase extraction – SPE, and concentrator columns) were tested and optimised. Compared with SPE columns, the concentrator column AC15 was chosen as the best choice for the next optimization followed by separation and detection by ion chromatography (IC). The sensitive method for determination of 7 DCAs (oxalic, malonic, succinic, maleic, glutaric, pimelic and azelaic acid) was devised with detection limits of 0.12–0.40 nM. The presented system describes an alternative on-line method usable for automated monitoring of selected analytes.

An automatic on-line sorptive extraction platform for palladium determination in automobile exhaust catalysts based on a PTFE-turnings packed column, flow injection analysis and flame atomic absorption spectrometry

ABSTRACT A novel simple sensitive and effective on-line flow injection column preconcentration platform using poly-tetrafluoroethylene turnings as sorbent was developed prior to flame atomic absorption spectrometric (FAAS) determination of palladium in vehicle exhaust catalysts, hydrogenation catalysts and environmental water samples. The method is based on the sorption of palladium(II)-diethyldithiophosphate, which was found to be more effective than dithiocarbamate ones. Methyl isobutyl ketone (MIBK) was used as eluent prior atomisation and quantification. All parameters like pH, DDPA concentration, flow rates, preconcentration time that affect the sensitivity of the method as well as interferences of other metals were investigated in detail. The enhancement factor calculated as 114 for 60 s preconcentration time while the sampling frequency as 40 h−1. The detection limit was 0.9 μg L−1 and the relative standard deviation (RSD) was 2.8% at 20.0 μg L−1 concentration level. The accuracy of the proposed method was estimated by analysing certified reference material and spiked environmental samples. The analytical features of the method were compared with other on-line column preconcentration methods reported in the literature.

Automated separation, preconcentration and measurement of 90Sr in liquid samples with complex matrices by online ion exchange chromatography coupled with inductively coupled plasma mass spectrometry (ICP-MS)

An online separation and preconcentration method, employing a lab-on-valve system using solid phase extraction, followed by inductively coupled plasma tandem mass spectrometry (ICP-MS/MS), was developed for the analysis of 90Sr. The 90Sr was separated from 90Zr, an isobaric interference present at high concentrations in many samples, and other matrix components using a dual-column setup (Eichrom DGA-B and Sr resins). Any remaining 90Zr was then chemically resolved from the 90Sr in the ICP-MS/MS using O2 and H2 reaction gases. The system requires small sample volumes (10 mL), minimal sample preparation compared to traditional radiometric and other MS techniques and has a processing time of 22 min per sample. Based on a 10 mL sample size, the system limit of detection, limit of quantification and method detection limit (MDL) were 0.47 Bq L−1 (0.09 pg L−1), 1.57 Bq L−1 (0.32 pg L−1) and 1.79 Bq L−1 (0.34 pg L−1), respectively. The robustness of the system and suitability for use in various sample matrices was demonstrated using spiked lake water, spiked groundwater, spiked seawater and radioactive water samples. Recovery of the IAEA 2018 Proficiency Test Exercise water sample (n = 5) was 99% with an RSD of 11.9%. The method thus provides a powerful tool for the rapid analysis of low levels of 90Sr in various water/wastewater samples.

Simultaneous and highly sensitive determination of selenium and tellurium species in environmental samples by on-line ionic liquid based in-situ solvent formation microextraction with hydride generation atomic fluorescence spectrometry detection

A novel on-line preconcentration and speciation analysis method for the simultaneous determination of inorganic Se and Te species is presented in this work. The methodology is based on the on-line formation of a hydrophobic ionic liquid (IL) directly in the liquid sample stream of a flow injection system, thus achieving an efficient and rapid extraction of the analytes complexed with ammonium pyrrolidine dithiocarbamate into the finely dispersed extractant droplets, that were then retained in a column filled with cotton. A full study of the chemical and hydrodynamical parameters was developed, including the right selection of the IL used as extractant and its concentration, pH, complexing reagent, sample and ion-exchange reagent volumes and column design. Additionally, a miniaturized external hydride generator was adapted to the spectrometer in order to increase the sensitivity of the atomic fluorescence measurements using only 250 μL of 5 mol L−1 HNO3 in methanol as eluent. The analytical figures of merit obtained for 15 mL of sample included sensitivity enhancement factors of 71, 70, 49 and 40 for Te(IV), Te(VI), Se(IV) and Se(VI), respectively, and limits of detection of 1.8 ng L−1 for both Te species, 2.6 ng L−1 for Se(IV) and 3.2 ng L−1 for Se(VI). After optimization, the method was successfully applied for the analysis of environmental samples: soils and sediments, as well as sea, river, underground and tap water.

Restricted access material-ion imprinted polymer-based method for on-line flow preconcentration of Cd2+ prior to flame atomic absorption spectrometry determination

The present paper describes a new restricted access material-ion imprinted polymer for on-line preconcentration of Cd2+ from natural water samples, and simultaneous clean-up of humic acid. Cadmium-imprinted poly(allylthiourea) was synthesized and modified with 2-hydroxyethyl methacrylate (IIP-HEMA) to obtain the restricted access ability. The on-line flow injection system coupled to FAAS (FIA-FAAS) was based on percolation of 20.0 mL of the sample (pH 8.70 and in presence of organic matter) through a mini-column (100.0 mg of IIP-HEMA) at 7.5 mL min−1 flow rate. The elution step was performed using 2.0 mol L−1 HNO3 at 5.0 mL min−1 flow rate. The method provided linearity in the range 0.56–50.00 μg L−1 and limit of detection of 0.17 μg L−1. The preconcentration factor was found to be 37-fold, consumption index of 0.54 mL, preconcentration efficiency of 13.10 min−1 and sample throughput of 21 h−1. The precision of the method was evaluated in terms of intra-day and inter-day repeatability and the relative standard deviation percentage (%RSD) obtained by two-level concentration (2.50 and 40.00 μg L−1) in the presence and absence of humic acid were <5.0%. Low interferences of ions commonly found in water samples as well as the humic acid were observed. The accuracy was attested by addition and recovery tests at two-level of Cd2+ (2.00 and 20.00 μg L−1, both prepared in presence of humic acid) in a real water sample, and the recovery percentages ranged from 93.8 to 108.7%. Cadmium was also analyzed in a certified reference material TORT-2 (lobster hepatopancreas). The method was applied for Cd2+ determination on water after spiking procedure with known concentration, food (linseed, 301.8 ± 15.3 μg kg−1), herbal medicine (Ginkgo biloba L., 221.2 ± 5.8 μg kg−1) and cigarette (77.9 ± 6.3 μg kg−1).

Determination of dicarboxylic acids in atmospheric aerosols using continuous aerosol sampler with on-line connected ion chromatography system

The aim of the presented study was to use a recently developed CGU-ACTJU-IC system for on-line determination of dicarboxylic acids (DCAs) in aerosols over short time periods involving aerosol sampling into deionized water using a Condensation Growth Unit – Aerosol Counterflow Two-Jets Unit (CGU-ACTJU) sampler, quantitative preconcentration of analytes and determination by ion chromatography (IC). The parameters of automated on-line preconcentration method, using two parallel preconcentration columns and piston pump, were optimized. Limits of detection of studied DCAs, i.e., oxalic, malonic, succinic, glutaric/adipic, pimelic, azelaic and maleic acid, were determined to 0.27, 0.92, 1.74, 0.82, 1.17, 1.41 and 0.48 ng m−3, respectively.The optimized method was used for the analysis of DCAs in urban aerosol in Brno in two campaigns: from 5th to 9th December 2016 and from 12th to 21st July 2017. The results obtained by the developed method were compared with two reference methods including sampling of aerosols on filters and subsequent analysis of DCAs by gas chromatography with mass spectrometry (GC/MS) or IC.The presented study describes an alternative on-line method, compared to filter sampling, for monitoring of DCAs in atmospheric aerosols, allowing measurement with 1-h time resolution.

Large‐volume sample stacking with polarity switching for analysis of azo dyes in water samples by capillary electrophoresis

ABSTRACTA simple, fast and efficient on-line pre-concentration method (large-volume sample stacking) by capillary electrophoresis was proposed for determination of azo dyes residues (allura red [AR], sunset yellow [SY] and tartrazine [TAR]) in water samples. Pre-concentration variables involved in the system were optimised using of a Box–Bhenken design. Under the optimal conditions: injection time 150.0 s, pre-concentration time 120.0 s and reverse potential −8.0 kV, the proposed methodology improved the analytical sensibility achieving limits of detection of 21.0–41.4 µg L−1 with enrichment factors of 82.1–210.8 fold. The large-volume sample stacking-capillary electrophoresis method was validated and applied to determine azo dyes residues in 20 water samples (bottled, spring and tap water). Two samples were positive for sunset yellow and tartrazine with a concentration of 25.3 and 30.2 µg L−1 and % RSD less than 10.0% in all cases.

Performance of restricted access copper-imprinted poly(allylthiourea) in an on-line preconcentration and sample clean-up FIA-FAAS system for copper determination in milk samples

A new on-line solid phase preconcentration method using an ion-imprinted polymer with restrict access material based on copper-imprinted poly(allylthiourea) modified with 2-hydroxyethyl methacrylate and bovine serum (IIP-HEMA-BSA) to preconcentrate Cu2+ ions and exclude protein from milk samples via FIA-FAAS was developed. The samples were on-line preconcentrated at pH 4.5 through a mini-column packed with 50.0 mg of IIP-HEMA-BSA at a flow rate of 7.6 mL min−1 followed by a counter-current elution with 1.00 mol L−1 HCl towards the FAAS detector. The interference of ions commonly found in milk samples and the presence of BSA were investigated in the Cu2+ ions preconcentration. The method provided an analytical curve ranging from 3.6 to 100.0 μg L−1, preconcentration factor of 24-fold, limit of detection of 1.1 μg L−1 and sample throughput of 20 h−1. The accuracy of the method was attested by Cu2+ ions addition/recovery tests as well as by using GFAAS of samples microwave-assisted digested. The proposed method was successfully applied to the copper determination in milk samples, requiring only pH adjustment followed by preconcentration step as sample pretreatment. Copper content in bovine milk sample was 0.635 ± 0.042 mg kg−1, while to soybean milk samples were between 0.048 ± 0.008 and 0.094 ± 0.005 mg kg−1. To the best of our knowledge, the potential of IIP-HEMA-BSA for Cu2+ ions extraction from biological samples and simultaneous removal of macromolecules employing minimal sample pretreatment was demonstrated for the first time. The proposed extraction method stands out by being simple, fast and a low-cost analytical strategy when compared to the conventional microwave-assisted acid digestion.

Amplification of lysozyme signal detected in capillary electrophoresis using mixed polymer brushes coating with switchable properties

In this work, a mixed polymer brushes based on poly(2-methyl-2-oxazoline) (PMOXA) and poly(acrylic acid) (PAA) coated capillary with switchable protein adsorption/desorption properties was developed and applied for on-line extraction and preconcentration of lysozyme. The study of electroosmotic flow (EOF) and fluorescence microscope showed that the inner surface charge of PMOXA/PAA mixed brush coated capillary displayed the switchable behavior toward the change of pH value and ionic strength (I), and PMOXA/PAA mixed brushes coated capillary could adsorb high amounts of lysozyme at pH 7 (I = 10−5 M), and the most of adsorbed lysozyme could then be desorbed at pH 3 (I = 10−1 M). Subsequently, this coated capillary with switchable lysozyme adsorption/desorption ability was applied for on-line extraction and preconcentration of lysozyme during capillary electrophoresis (CE) performance. Under the process of on-line preconcentration, the detection signal (peak area) of lysozyme obtained in PMOXA/PAA coated capillary was 26 times that obtained in bare capillary under normal CE while the contour chain length of PAA was 1.56 times that of PMOXA. Moreover, the value of low detection limit (LOD) of lysozyme using above coated capillary under on-line preconcentration method reached to 4.5 × 10−9 mg/mL, and 1 × 105-fold sensitivity enhancement was realized for lysozyme as compared with the bare capillary under normal CE.

On-line preconcentration of sodium dodecyl sulfate-protein complexes using electrokinetic supercharging method with a prefilled water plug in capillary sieving electrophoresis.

An electrokinetic supercharging (EKS) method with a prefilled water plug at the head column of capillary was developed for on-line preconcentration of sodium dodecyl sulfate (SDS)-protein complexes in capillary sieving electrophoresis (CSE). Conventional EKS is a combination of electrokinetic injection with transient isotachophoresis (tr-ITP). The capillary is first filled with background electrolyte, then an appropriate amount of a leading electrolyte is filled and electro-injection is carried out for certain duration. After that, terminating electrolyte is filled, and tr-ITP is subsequently initiated, followed by capillary electrophoresis (CE) separation. In this work, the performance of EKS was evaluated by integrating multiple sub-methods step by step, and a water plug containing polymer was introduced before electrokinetic injection in order to further improve the concentration effect. The positive effects of the sub-methods were verified, including molecular sieving effect of polymer, field enhanced sample injection (FESI) with and without a water plug, and transient isotachophoretic electrophoresis-based FESI. It was observed that analyte discrimination usually encountered in conventional electrokinetic injection was eliminated due to the similar charge to mass ratios of SDS-protein complexes. Based on these results, a hybrid on-line preconcentration method, EKS with injecting a water plug containing polymer before sample electrokinetic injection, was proposed and used to indiscriminately preconcentrate SDS-protein complexes, which provided a sensitivity enhancement factor of more than 1000. It was very suitable for the analysis of low-abundance proteins, providing the information of their molecular mass.

Data trend shifts induced by method of concentration for trace metals in seawater: Automated online preconcentration vs. borohydride reductive coprecipitation of nearshore seawater samples for analysis of Ni, Cu, Zn, Cd, and Pb via ICP‐MS

AbstractThis research compares performance, reproducibility, and detection limits of ambient seawater analysis for trace metals using both borohydride reductive coprecipitation and an automated chelation column (seaFAST™ 2) preconcentration for matrix interferent elimination on total and dissolved grab samples in nearshore to marine waters, over a broad concentration range, prior to inductively coupled plasma mass spectrometry (ICP‐MS) injection. A move to an online preconcentration method both minimizes sample preparation, and eliminates correction errors when accounting for trace impurities in precipitated samples, induced via reagents. The reproducability of the online preconcentration method described, coupled with low blanks and method detection limits (MDLs), demonstrates the effectiveness of the automated procedure using ethylenediaminetriacetic and iminodiacetate acid chelation exchange resin and multianalyte determination by ICP‐MS for total and dissolved Ni, Cu, Zn, Cd, and Pb in marine water samples. Average CASS‐5 recoveries using the online preconcentration method (n = 9) were 109% ± 7%, 104% ± 5%, 103% ± 7%, 101% ± 3%, and 86% ± 8%, respectively. The MDLs obtained from the automated method for Ni, Cu, Zn, Cd, and Pb were 3.3, 1.8, 13.5, 4, and 10 times lower, respectively, than for the Borohydride method. There were statistically significant differences between the methods for CASS‐5 recoveries of Ni, Cu, Zn (p &lt; 0.0001), and Pb (p = 0.0024). Comparison of methods gave high concordance (rC ≥ 0.90) between methods for total and dissolved Ni, Cu, Zn, and Pb, and total Cd.

Sample Preconcentration Protocols in Microfluidic Electrophoresis.

Electrophoretic on-line sample preconcentration techniques in microfluidic channels improve the sensitivity prior to the separation. Among various techniques, the most important field-amplified sample stacking and sweeping on cross-channel microchips are demonstrated. As a novel microfluidic preconcentration approach, a large-volume sample stacking with electroosmotic flow pump (LVSEP) on straight-channel chips is also presented, which can omit a complicated voltage program for sample injection processes. In this chapter, we describe how to prepare and how to run these on-line sample preconcentration methods in microchip electrophoresis.

Enantioselective analysis of pheniramine in rat using large volume sample stacking or cation-selective exhaustive injection and sweeping coupled with cyclodextrin modified electrokinetic chromatography

For the aim of simultaneously performing the enantioseparation and determination of the trace enantiomers in plasma samples, enantioseparation by HPLC using five kinds of chiral stationary phases were initially investigated. But unfortunately, enantioseparation could not be detected in reversed mobile phase mode with all the five columns. For this reason, two simple, economical and highly efficient online preconcentration methods, large volume sample stacking and sweeping (LVSS-sweeping) and cation-selective exhaustive injection and sweeping (CSEI-sweeping) both followed by the cyclodextrin modified electrokinetic chromatography (CDEKC) were examined in the present work. Parameters affecting the enantioseparation and enhancement efficiency of these two injection modes were monitored in detail, and migration order of the two enantiomers was identified by circular dichroism (CD) and HPLC. Upon optimization, two enantiomers were best separated with the improvement of sensitivity reaching 160-fold and 4000-fold respectively for LVSS-sweeping and CSEI-sweeping comparing with the normal CDEKC separation. Then the optimal condition of CSEI-sweeping-CDEKC was validated and showed high sensitivity (10 ng/mL for lower limit of quantification, LLOQ), satisfactory accuracy (96.8–111.6%) and precision (relative standard deviation, RSD within 9.4%). This demonstrated it to be a suitable strategy for the rapid enantioselective determination and quantitative analysis of pheniramine enantiomers in plasma samples. Therefore, the method was further applied in the enantiomeric analysis of pheniramine in rat pharmacokinetics and plasma protein binding investigations. Stereoselectivity in pharmacokinetics as well as plasma protein binding were observed, suggesting that the stereoselective protein binding might be responsible for the stereoselectivity in pharmacokinetics.