On-line Sample Preconcentration Research Articles

Application of on-line sample preconcentration by large-volume dual preconcentration by isotachophoresis and stacking (LDIS) on straight-channel microchips.

In this study, large-volume dual preconcentration by isotachophoresis and stacking (LDIS) which is an on-line sample preconcentration technique coupling large-volume sample stacking with an electroosmotic flow pump (LVSEP) with transient isotachophoresis (tITP) was applied to microchip electrophoresis (MCE) for improving both detection sensitivities and peak shapes. To realize LDIS in MCE, we investigated experimental procedures for injecting a short plug of a leading electrolyte (LE) solution into a straight microchannel without any sophisticated injector apparatus. We found that a short LE plug could be injected into a sample-filled straight-channel only by making the liquid level of the LE solution in an outlet reservoir higher than that in an inlet one. By applying a reversed-polarity voltage to the microchip, anionic analytes injected throughout the microchannel were first enriched by LVSEP, followed by tITP. Through the second preconcentration effect by tITP in LDIS, sensitivity enhancement factor (SEF) and asymmetry factor for a standard dye were improved from 878 and 0.62 to 1330 and 1.14, respectively, relative to those in conventional LVSEP. It should be noted that more viscous running buffer containing sieving polymers could be employed to the LDIS analysis, which was effective for improving the SEF and the separation efficiencies, especially for bio-polymeric compounds. Finally, LDIS was applied to the oligosaccharide and protein analyses in MCE, resulting in the SEFs of 1410 and ca. 50 for maltotriose and bovine milk casein, respectively.

Innovative analysis of diazepam, zolpidem and their main metabolites in human urine by micelle-to-solvent stacking in capillary electrophoresis

Diazepam and zolpidem are the most widely used medications for managing insomnia. However, significant concerns regarding the potential risks of misuse and abuse problems arose in many literatures. While urine analysis is a valuable diagnostic tool, a challenge arises from the fact that some parent drugs may remain undetectable in urine. This necessitates concurrent monitoring of their metabolites. Here, we described an innovative on-line sample preconcentration technique known as micelle to solvent stacking (MSS) for the analysis of diazepam, zolpidem, and their main metabolites in urine. Several key parameters warrant further discussion to optimize the MSS model, enhancing its performance in terms of sensitivity and resolution. After optimizing the conditions, we conducted a validation test, achieving high correlation coefficients (greater than 0.9977) for intra-day and inter-day regression lines. Additionally, both the relative standard deviation (RSD) and relative error (RE) remained below 6.10% and 12.55%, respectively. The limits of detection (LODs, S/N = 3) for all five analytes ranged from 2.0 to 56 ng/mL. Compared to the conventional capillary zone electrophoresis method, this new approach exhibited remarkable sensitivity enhancements, ranging from 123 to 235-fold. Upon applying this method to actual urine samples from patients, we successfully detected nordiazepam, zolpidem, and its metabolites. This simple and sensitive approach has promising applications in supporting patient medication safety and bolstering forensic investigations.

On-line sample preconcentration by LVSEP-FASI with simple voltage control on Y-channel chips.

To eliminate complicated voltage controls for highly sensitive microchip electrophoresis (MCE) analyses on the basis of combining two online sample preconcentration techniques, large-volume sample stacking with an electroosmotic flow (EOF) pump (LVSEP) and field-amplified sample injection (FASI), cross-channel microchips and a multichannel high-voltage power supply were replaced to Y-channel chips and a conventional power supply designed for capillary electrophoresis, respectively. By simple switching of the electric circuit after the LVSEP-FASI sample enrichments, the focused analytes could be separated during anodic migration in a separation channel. In the LVSEP-FASI analysis of fluorescein using the Y-channel microchip, the maximum sensitivity enhancement factor (SEF) of 7400 was achieved, resulting in a 30-fold detectability increase compared to the conventional LVSEP. The developed method was applied to the oligosaccharide analysis in MCE. As a result, the SEF for maltotriose was improved from 450 to 2300 and the baseline separation of the oligosaccharides was achieved without any complicated voltage control in LVSEP-FASI on the Y-channel chips.

Sensitivity enhancement of capillary electrophoresis-frontal analysis-based method for characterization of drug-protein interactions using on-line sample preconcentration.

Capillary electrophoresis-frontal analysis is one of the most frequently used approaches for the study of plasma protein-drug interactions as a substantial part of new drug development. However, the capillary electrophoresis-frontal analysis typically combined with ultraviolet-visible detection suffers from insufficient concentration sensitivity, particularly for substances with limited solubility and low molar absorption coefficient. The sensitivity problem has been solved in this work by its combination with an on-line sample preconcentration. According to the knowledge of the authors this combination has never been used to characterize plasma protein-drug binding. It resulted in a fully automated and versatile methodology for the characterization of binding interactions. Further, the validated method minimalizes the experimental errors due to a reduction in the manipulation of samples. Moreover, employing an on-line preconcentration strategy with capillary electrophoresis-frontal analysis using human serum albumin-salicylic acid as a model system improves the drug concentration sensitivity 17-fold compared to the conventional method. The value of binding constant (1.51±0.63) ·104 L/mol obtained by this new capillary electrophoresis-frontal analysis modification is in agreement with the value (1.13±0.28) ·104 L/mol estimated by a conventional variant of capillary electrophoresis-frontal analysis without the preconcentration step, as well as with literature data obtained using different techniques.

Recent Applications of Dynamic On-Line Sample Preconcentration Techniques in Capillary Electrophoresis

This review highlights recent applications of on-line sample preconcentration techniques in capillary electrophoresis (CE) from 2019 to 2021. Various preconcentration techniques based on the analyte velocity change in two or three discontinuous solutions system including field-amplified sample stacking, transient isotachophoresis, pH-mediated stacking, sweeping, and their modified and combined techniques have been employed to enrich and separate biological, environmental, food samples, and so on, in CE. More than 60 published research articles collected from Scopus databases described the dynamic on-line sample preconcentration techniques. This review provides comprehensive tables listing the applications of the on-line sample preconcentration techniques.

Capillary Electrophoresis-Mass Spectrometry for the Direct Analysis of Metabolites in Highly Saline Samples Using In-Capillary Preconcentration.

Capillary electrophoresis-mass spectrometry (CE-MS) is gaining interest for metabolomics studies because of its high separation efficiency, selectivity, and versatility. The ability to inject nanoliters from only a few microliters of sample in the injection vial makes this approach very suited for volume-limited applications. However, the low injection volumes could compromise the detection sensitivity of CE-MS, thereby potentially limiting its scope in metabolomics. To overcome this issue, online sample preconcentration methods have been developed to increase sample-loading volumes without hampering the intrinsic high separation efficiency of CE. In this protocol, online preconcentration with sample stacking based on pH junction was assessed for the direct profiling of endogenous metabolites in rat brain microdialysates. Sample stacking was realized by a pre-injection of ammonium hydroxide, followed by a large sample injection (i.e., about 17% of the total capillary volume). It is shown that this relatively simple and fast preconcentration procedure is fully compatible with the high-salt concentration in microdialysates and significantly improves the detection sensitivity of the CE-MS method.

Development of transient trapping micellar electrokinetic chromatography coupled with mass spectrometry for steroids analysis.

An on-line sample preconcentration technique based on transient trapping (tr-trapping) in micellar electrokinetic chromatography (MEKC) was applied for steroid detection with UV (tr-trapping-UV) and electrospray ionization mass spectrometry detection (tr-trapping-ESI-MS). ESI-MS was used to improve the sensitivity in MEKC. The MEKC separation was carried out using volatile ammonium formate as a background solution to facilitate the coupling with ESI-MS. The partial introduction of a sodium dodecyl sulfate (SDS) micellar solution before the introduction of a sample solution to the capillary provided the effective preconcentration of analytes. At the same time, the SDS micelle would not enter the ESI-MS system, so its interference in ESI-MS detection was suppressed under the optimal condition, then five steroids can be separated by the developed method. In tr-trapping-ESI-MS, an acidic condition of pH 3.5 was employed to suppress the electroosmotic flow, which can avoid micellar solution migrating to the MS instrument. The developed method showed that the micellar solution requires a twofold slower time than the sample to migrate along the column, which can prohibit the cause of the problem with the MS instrument and interference signal of SDS in the steroid's detection. The tr-trapping-ESI-MS protocol showed up to 540-fold enhancements of the peak intensity and 50-fold improvement of the limit of detection compared with capillary zone electrophoresis using androsterone as a model sample.

Capillary electrophoresis-mass spectrometry as a tool for the noninvasive target metabolomic analysis of underivatized amino acids for evaluating embryo viability in assisted reproduction.

Monitoring metabolite uptake and excretion in the culture medium is a noninvasive technique that is used for the metabolic study of cleaving embryos after in vitro fertilization. Low sample consumption, the versatility of the detection, and optimal sensitivity and selectivity are essential elements for extracellular metabolome analyses, and can be conveniently achieved by combining CE with mass spectrometric detection. This paper reports a method for amino acid determination in a limited volume sample (8 μL)ofspent culture media collected after the cultivation of in vitro fertilized embryos. Special attention was focused on the sample preparation procedure. The sample was processed with acetonitrile, which facilitates online sample preconcentration via field-amplified sample stacking, and undesired sample evaporation was significantly reduced by the simultaneous addition of dimethyl sulfoxide. Key parameters that affected electrophoretic separation and mass spectrometric detection were investigated, including the type of buffers and organic solvent, optimization of their concentrations, and finally the settings for their ionization. The separation and quantification of 19 amino acids were achieved using 15% acetic acid as the background electrolyte with a sheath liquid consisting of an equimolar mixture of methanol and water. The applicability of the optimized system was demonstrated by determining the amino acid profile in 40 samples of spent cultivation medium in this pilot study. This developed method also has great potential for amino acid analyses in minute sample volumes of other biological matrices.

High-throughput and high-sensitivity capillary electrophoresis\u2013mass spectrometry method for sulfur-containing amino acids

Biological thiol amino acids have been suggested as biomarkers for pathological changes because they are reactive chemicals that participate in various physiological processes. In this study, multisegmented injection capillary electrophoresis–mass spectrometry with online sample preconcentration was used for analysis of thiol amino acids and intermediates of sulfur metabolism in human glioma cell line U-251 with high accuracy, throughput, and sensitivity. This was achieved using multiple, large-volume injections for online sample preconcentration. The 16 intermediates of sulfur metabolism had a good linear correlation coefficient range of 0.984–1 and the limit of detection range was 1.4–203.9 ng/mL. The recovery ranges of most amino acids were 88.1–114.5%, 89.0–104.3%, and 76.9–104.5% at 0.3, 0.75, and 1.5 μg/mL, respectively. The relative standard deviation ranges for the inter- and intra-day precision were 1.8–10.7% and 4.3–18.8%, respectively. Compared with the traditional injection method, the analytical time for compounds in sulfur metabolism was reduced to 4 min/sample, the method throughput was enhanced five times, and the sensitivity was increased 14.4–33.1 times. Customized injection sequences were applied in experimental optimization. The developed method simplified the experimental optimization to one injection and is suitable for the analysis of sulfur metabolites in biological samples and has high sensitivity, throughput, speed, and accuracy.

Online sample preconcentration techniques in nonaqueous capillary and microchip electrophoresis

One of the major drawbacks of electrophoresis in both capillary and microchip is the unsatisfactory sensitivity. Online sample preconcentration techniques can be regarded as the most common and powerful approaches commonly applied to enhance overall detection sensitivity. While the advances of various online preconcentration strategies in capillary and microchip employing aqueous background electrolytes are well-reviewed, there has been limited discussion of the feasible preconcentration techniques specifically developed for capillary and microchip using nonaqueous background electrolytes. This review provides the first consolidated overview of various online preconcentration techniques in nonaqueous capillary and microchip electrophoresis, covering the period of the last two decades. It covers developments in the field of sample stacking, isotachophoresis, and micellar-based stacking. Attention is also given to multi-stacking strategies that have been used for nonaqueous electrophoresis.

Capillary Zone Electrophoresis Determination of Five Trace Food Additives in Beverage Samples Using Counterflow Transient Isotachophoresis

Transient isotachophoresis (tITP) is a universal online sample preconcentration technology. It uses ITP to focus electrically charged analytes during the initial stage of capillary electrophoresis (CE) analysis, which can effectively improve the sensitivity of CE. In this work, an approach of tITP under counter-electroosmotic flow was developed for capillary zone electrophoresis (CZE) determination of five trace food additives which include benzoic acid, sorbic acid, sunset yellow, allura red, and amaranth in beverage samples. The parameters that affect the preconcentration effect of the system, such as the type of terminating electrolyte and its injection time, the preconcentration time of ITP, and the sample injection time, have been investigated in detail. The mechanism of counterflow-tITP was explored preliminarily. Under optimal conditions, the sensitivity of five food additives was improved 11.9, 11.5, 13.04, 10.05, and 15.3 times. Their limits of detection (LODs, S/N = 3) reached 0.15, 0.10, 0.38, 0.33, and 0.11 μg/ml, with good repeatability (peak area RSDs ≤ 6.9%) and calibration graph linearity (r ≥ 0.9992). Recoveries of five food additives in spiked beverage samples ranged from 92.0 to 100.8% with RSDs between 1.38 and 2.95%. This method is simple, rapid, and sensitive. It has been successfully applied to the separation and detection of trace food additives in cocktails, orange juice, and carbonated beverages. The feasibility of determination of trace additives in beverages by tITP-CZE under the condition of counterflow has been confirmed.

Recent Advances in Trace Bioanalysis by Capillary Electrophoresis.

Recently, single cell analysis is becoming more and more important to elucidate cellular heterogeneity. Except for nucleic acid that can be amplified by PCR, the required technical level for single cell analysis is extremely high and the appropriate design of sample preparation and a sensitive analytical system is necessary. Capillary/microchip electrophoresis (CE/MCE) can separate biomolecules in nL-scale solution with high resolution, and it is highly compatible with trace samples like a single cell. Coupled with highly sensitive detectors such as laser-induced fluorescence and nano-electrospray ionization-mass spectrometry, zmol level analytes can be detected. For further enhancing sensitivity, online sample preconcentration techniques can be employed. By integrating these high-sensitive techniques, single cell analysis of metabolites, proteins, and lipids have been achieved. This review paper highlights successful research on CE/MCE-based trace bioanalysis in recent 10 years. Firstly, an overview of basic knowledge on CE/MCE including sensitivity enhancement techniques is provided. Applications to trace bioanalysis are then introduced with discussion on current issues and future prospects.

Development of a titanium dioxide-assisted preconcentration/on-site vapor-generation chip hyphenated with inductively coupled plasma-mass spectrometry for online determination of mercuric ions in urine samples

In this study, a novel automatic analytical methodology using a titanium dioxide (TiO2)-assisted preconcentration/on-site vapor-generation (VG) chip hyphenated with inductively coupled plasma-mass spectrometry (ICP-MS) for online determination of mercuric ions (Hg2+) was developed. Interestingly, the TiO2 nanoparticle (nano-TiO2) coating on the channel surface acted not only as a sorbent for preconcentration but also as a catalyst for photocatalyst-assisted VG. Under optimum operation conditions, the developed method was validated by analyzing the certified reference material (CRM) Seronorm™ Trace Elements Urine L-2 (freeze-dried human urine). Based on the obtained results, the dramatic reduction of “hands-on” manipulation and the elimination of hazardous materials (e.g., sodium borohydride (NaBH4) and stannous chloride (SnCl2)) from the process enabled a simple and ultraclean procedure with an extremely low detection limit of 0.75 ng L−1 for Hg2+ in urine samples. To the best of our knowledge, this is the first study to report the direct exploitation of a nano-TiO2-coated microfluidic device for online sample preconcentration and on-site VG prior to ICP-MS measurement.

New Advances in Amino Acid Profiling in Biological Samples by Capillary Electrophoresis-Mass Spectrometry.

Capillary electrophoresis-mass spectrometry (CE-MS) offers a high efficiency microseparation platform for amino acid profiling when analyzing volume-restricted biological samples, such as a dried blood spot punch. Direct analysis of amino acids and their analogs is routinely achieved using strongly acidic buffer conditions under positive-ion mode detection with a coaxial sheath liquid interface for electrospray ionization (ESI). New advances in online sample preconcentration, pre-column chemical derivatization, and/or low flow/sheathless CE-MS interface designs can further improve sensitivity while allowing for resolution of amino acid stereoisomers and labile aminothiols with low nanomolar detection limits. Additionally, multiplexed separations in CE-MS based on serial injection of seven or more samples within a single run greatly boosts sample throughput (<2-3min/sample) without added infrastructure costs while allowing for stringent quality control and signal batch correction. Accurate prediction of the electromigration behavior of amino acids and their analogs offers a convenient approach for structural elucidation that is complementary to high-resolution MS and MS/MS. Simultaneous analysis of amino acids together with other classes of ionic metabolites by CE-MS allows for comprehensive metabolomic screening as required for new advances in clinical medicine, nutritional sciences, and population health.

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.

Novel electrophoretic acetonitrile-based stacking for sensitive monitoring of the antiepileptic drug perampanel in human serum

Perampanel is a novel antiepileptic drug used in paediatric patients. Existing methods that determine serum perampanel are of limited practicability. We developed a novel capillary electrophoresis (CE) method using a new version of acetonitrile stacking for on-line sample pre-concentration, and fluorescence detection (FD). CE separations were performed in a fused-silica capillary where the electroosmotic flow was reduced by coating the inner surface using a INST coating solution. The optimised background electrolyte composition was 50 mM chloroacetic acid with addition of 0.5% m/v polyvinylalcohol (pH 2.15) and separation was driven by application of positive voltage + 30 kV. Serum samples (25 μL) treated by the addition of acetonitrile in a ratio of 1:3 v/v were each injected into the capillary at a large volume that corresponded to the length (129 mm) of the sample zone (hydrodynamic pressure impulse 6000 mbars). Acetonitrile stacking is based on the forcing the sample zone out of the capillary with simultaneous application of the separation voltage. Under such conditions, the enhancing factor achieves the value 57 for peak area compared to the small sample injection length (3.2 mm, hydrodynamic pressure impulse 150 mbar.s). A fluorescence detector with a broad excitation filter (240–400 nm) and an emission filter (495 nm) was used for visualisation of the native fluorescence of perampanel. The calibration dependence of the method was linear (in the range of 10–1000 ng mL−1), with adequate accuracy (99.8–103.3 %) and precision (13.1%). LOD and LOQ for perampanel were 2.9 ng mL-1 and 9.5 ng mL−1, respectively. Clinical applicability was validated using serum samples from patients treated with perampanel and the results corresponded with reference LC–MS/MS values. Our method offers a promising alternative for determining serum perampanel with several advantages. In particular, the low quantity of serum (25 μL) required means that testing can be performed on samples obtained for monitoring other antiepileptic medications, and thus reduces the test-burden on paediatric patients.

Simultaneous electrokinetic and hydrodynamic injection and sequential stacking featuring sweeping for signal amplification following MEKC during the analysis of rapamycin (sirolimus) in serum samples.

Simultaneous electrokinetic and hydrodynamic injection of rapamycin (sirolimus) with off-line and online sample preconcentration techniques and using MEKC has been studied. Compared to conventional hydrodynamic injection, a 168-fold improvement in the signal was obtained with a combination of simultaneous electrokinetic and hydrodynamic injectionand field enhanced sample injection in conjunction with a sweeping technique called sequential stacking featuring sweeping. However, the coupling of the developed electrophoretic method and solid-phase microextraction allowed the signal intensity to increase more than 231 times. In this approach, the injection of the sample at negative polarity (anode at the detector end) into the capillary and the MEKC separation was achieved within 5min using an electrolyte (composed of 10mM sodium tetraborate and 40mM SDS) when ultraviolet (UV) detection was performed at 280nm. Thus, by combining the application of the sequential stacking featuring sweeping supported by the solid-phase microextraction clean-up procedure, the detection limit (LOD) for rapamycin in a serum sample was significantly decreased, and was set at 25ng/mL. The proposed combined simultaneous electrokinetic and hydrodynamic injection with field enhanced sample injection -sweeping technique following MEKC separation of sirolimus in human serum could be an effective tool in biomedical and clinical applications.

Profiling of N-linked glycans from 100 cells by capillary electrophoresis with large-volume dual preconcentration by isotachophoresis and stacking

Glycan structure is changed in response with pathogenesis like cancer. Profiling of glycans from limited number of pathogenetic cells in an early-stage tissue is essential for discovering effective drugs. For analyzing tiny biological samples, we developed sensitive, high-resolution, and salt-tolerant method for analyzing trace level of N-linked glycans by coupling capillary electrophoresis (CE), laser-induced fluorescence (LIF) detection, and a new online sample preconcentration (OSP) method named “large-volume dual preconcentration by isotachophoresis and stacking (LDIS)”, which is composed of two OSP methods, large-volume sample stacking (LVSS) and transient isotachophoresis (tITP). A typical LDIS-CE-LIF protocol was simple: a short-plug of leading electrolyte (LE) and large-volume sample solution were introduced to a capillary, followed by application of constant voltage. In the analysis of glucose ladder labeled with 8-aminopyrene-1,3,6-trisulfonic acid with 10 mM sodium chloride as LE, up to 2300-fold sensitivity increase was achieved with higher resolution than those in normal CE. By applying pressure assist during preconcentration, both viscous gel electrolyte and salty matrix of up to 10 mM NaCl were acceptable. Finally, N-glycans from approximately 100 cells (HeLa, MCF7, and HepG2) were analyzed as the model of localized tumor cells. From 30 to 40 glycans were successfully detected with almost same profile of large-scale sample. N-glycan structure could be predicted by searching glucose-unit value via Glycobase database, indicating that HepG2 expressed more sialylated glycans and MCF-7 expressed less glycans respectively, comparing with HeLa cells. It suggests the potential of LDIS-CE-LIF for discovery of disease-specific N-linked glycans in microscale environment.

Imidazolium ionic liquids as dynamic and covalent modifiers of electrophoretic systems for determination of catecholamines

The subject of this study is comparison of imidazolium based dynamic and covalent coatings of the quartz capillary wall on the example of catecholamines determination. A way of synthesis of covalent coatings was proposed. For the first time different type of on-line sample preconcentration techniques (field-amplified sample stacking (FASS), head-column field amplified sample stacking (HC FASS), electrostacking, sweeping) were performed for catecholamines determination in N-alkylsubstituted imidazolium coated capillary.It was found that long chain imidazolium ionic liquids (C12MImCl and C16MImCl) in background electrolyte content create a dynamic coating of the quartz capillary walls and generate anode electroosmotic flow. Same is true for covalent coatings based on N-alkylsubstituted imidazolium. Both coatings prevent sorption of catecholamines on the internal surface of the quartz capillary. As a result efficiency and peak symmetry are increased.The maximum stacking efficiency factor (SEF) values for dynamic coatings were in range of 70–85, and limits of detection (LODs) were about 0.05 µg/ml under sweeping condition (micelle forming agent – C16MImCl) when the electric conductivity of sample matrix was higher than electric conductivity of BGE. In case of covalent coating, the maximum SEF values were higher than 1000, and LODs were about 1–2 ng/ml under sweeping condition (micelle forming agent – sodium dodecyl sulphate) in combination with electrostacking.

Sensitive arsenic speciation by capillary electrophoresis using UV absorbance detection with on-line sample preconcentration techniques

The World Health Organization (WHO) guideline states that the total arsenic concentration in drinking water must not exceed 10 ppb. However, arsenic toxicity varies significantly, with inorganic arsenic species being more toxic than organic species. Arsenic speciation is therefore important for evaluating the health risks from arsenic-contaminated drinking water. Capillary electrophoresis provides the necessary high performance separation to determine arsenic species in water, but its sensitivity with absorbance detection is far below than needed. Using a coated capillary, several on-line sample preconcentration techniques such as large volume sample stacking with an electroosmotic flow pump, field amplified sample injection (FASI), transient isotachophoresis (tITP), electrokinetic supercharging (EKS) combining FASI and tITP, and counter flow (CF)-EKS, were therefore investigated. With CF-EKS using phosphate and N-cyclohexyl-2-aminoethanesulfonate as leading and terminating electrolytes, respectively, standard samples of arsenite, arsenate, monomethylarsonic acid, and dimethylarsinic acid were preconcentrated from 6,300- to 45,000-fold. The limits of detection obtained with UV absorbance detection were 0.08–0.3 ppb As. For a spring water sample spiked with the four arsenic species, LODs of 2–9 ppb As were obtained, which are lower than the WHO guideline of 10 ppb total As.