Enhancing the Concentration Capability of Nonsupported Electrically Driven Liquid-Phase Microextraction through Programmable Flow Using an All-In-One 3D-Printed Optosensor: A Proof of Concept.

A versatile millifluidic 3D-printed inverted Y-shaped unit (3D-YSU) was prototyped to ameliorate the concentration capability of nonsupported microelectromembrane extraction (µ-EME), exploiting optosensing detection for real-time monitoring of the enriched acceptor phase (AP). Continuous forward-flow and stop-and-go flow modes of the donor phase (DP) were implemented via an automatic programmable-flow system to disrupt the electrical double layer generated at the DP/organic phase (OP) interface while replenishing the potentially depleted layers of analyte in DP. To further improve the enrichment factor (EF), the organic holding section of the OP/AP channel was bifurcated to increase the interfacial contact area between the DP and the OP. Exploiting the synergistic assets of (i) the continuous forward-flow of DP (1050 µL), (ii) the unique 3D-printed cone-shaped pentagon cross-sectional geometry of the OP/AP channel, (iii) the bifurcation of the OP that creates an inverted Y-shape configuration, and (iv) the in situ optosensing of the AP, a ca. 24 EF was obtained for a 20 min extraction using methylene blue (MB) as a model analyte. The 3D-YSU was leveraged for the unsupervised µ-EME and the determination of MB in textile dye and urban wastewater samples, with relative recoveries ≥88%. This is the first work toward analyte preconcentration in µ-EME with in situ optosensing of the resulting extracts using 3D-printed millifluidic platforms.

Automated analyses of dried blood spots collected by volumetric microsampling devices.

BACKGROUND: Dried blood spot (DBS) sampling on cellulose cards suffers from varying blood haematocrit levels and from chromatographic effects, which have a direct impact on quantitative DBS analyses. Commercial volumetric microsampling devices were, therefore, introduced to mitigate these effects, however, these devices are not compatible with automated DBS processing systems and must be processed manually. RESULTS: Capillary electrophoresis (CE) instruments use fused-silica (FS) capillaries for precise and accurate liquid handling as well as for injection, separation, and quantitative analyses of liquid samples. These inherent features of an Agilent 7100 CE instrument were employed for the automated processing (elution and homogenization) of DBSs collected by hemaPEN® volumetric devices (2.74 µL of capillary blood per spot). The hemaPEN® samples were processed directly in CE vials by consecutive transfers of 56 µL of methanol and 14 µL of deionized water through the FS capillary in a sequence of 39 DBSs with repeatability of the liquid transfers better than 1.4 %. The resulting DBS eluates were homogenized by a quick air flush through the capillary and analyzed by the same capillary and CE instrument. Creatinine was selected as a clinically relevant model analyte and its endogenous concentrations in DBSs were determined by CE with capacitively coupled contactless conductivity detection (CE-C4D) in a background electrolyte solution consisting of 50 mM acetic acid and 0.1 % (v/v) Tween 20 (pH 3.0). The overall repeatability of the automated DBS processing and CE-C4D analyses of 39 DBSs was ≤7.1 % (peak areas) and ≤0.6 % (migration times), the calibration curve was linear in the 25-500 µM range (R2 = 0.9993) and covered all endogenous blood creatinine levels, the limit of detection was 5.0 µM, and sample throughput was >12 DBSs per hour. DBS ageing for 60 days and varying blood haematocrit levels (20-70 %) did not affect creatinine quantitative results (≤6.9 % for peak areas). Inter-capillary and inter-instrument repeatability was ≤7.7 % (peak areas) and ≤3.4 % (migration times) and demonstrated an excellent transferability of the proposed analytical concept among laboratories. SIGNIFICANCE AND NOVELTY: This contribution is the first-ever report on the use of a single off-the-shelf analytical instrument for fully automated analyses of DBSs collected by commercial volumetric microsampling devices and holds great promise for future unmanned quantitative DBS analyses.

Multiphase electroextraction of malachite green from surface water and its determination using digital imaging and chemometric tools.

This study introduces a novel method for the quantification of malachite green (MG), a pervasive cationic dye, in surface water by synergizing multiphase electroextraction (MPEE) with digital image analysis (DIA) and partial least square discriminant analysis. Aimed at addressing the limitations of conventional DIA methods in terms of quantitation limits and selectivity, this study achieves a significant breakthrough in the preconcentration of MG using magnesium silicate as a novel sorbent. Demonstrating exceptional processing efficiency, the method allows for the analysis of 10 samples within 20 min, exhibiting remarkable sensitivity and specificity (over 0.95 and 0.90, respectively) across 156 samples in both training and test sets. Notably, the method detects MG at low concentrations (0.2 µg L-1) in complex matrices, highlighting its potential for broader application in environmental monitoring. This approach not only underscores the method's cost-effectiveness and simplicity but also its precision, making it a valuable tool for the preliminary testing of MG in surface waters. This study underscores the synergy among MPEE, DIA, and chemometric tools, presenting a cost-efficient and reliable alternative for the sensitive detection of water contaminants.

3D-printed stereolithographic fluidic devices for automatic nonsupported microelectromembrane extraction and clean-up of wastewater samples.

BACKGROUND: There is a quest of novel functional and reliable platforms for enhancing the efficiency of microextraction approaches in troublesome matrices, such as industrial wastewaters. 3D printing has been proven superb in the analytical field to act as the springboard of microscale extraction approaches. RESULTS: In this work, low-force stereolithography (SL) was exploited for 3D printing and prototyping bespoke fluidic devices for accommodating nonsupported microelectromembrane extraction (µEME). The analytical performance of 3D-printed µEME devices with distinct cross-sections, including square, circle, and obround, and various channel dimensions was explored against that of commonly used circular polytetrafluoroethylene (PTFE) tubing in flow injection systems. A computer-controlled millifluidic system was harnessed for the (i) automatic liquid-handling of minute volumes of donor, acceptor, and organic phases at the low µL level that spanned from 3 to 44 µL in this work, (ii) formation of three-phase µEME, (iii) in-line extraction, (iv) flow-through optical detection of the acceptor phase, and (v) solvent removal and regeneration of the µEME device and fluidic lines. Using methylene blue (MB) as a model analyte, experimental results evinced that the 3D-printed channels with an obround cross-section (2.5 mm × 2.5 mm) were the most efficient in terms of absolute extraction recovery (59%), as compared to PTFE tubing of 2.5 mm inner diameter (27%). This is attributed to the distinctive convex interface of the organic phase (1-octanol), with a more pronounced laminar pattern, in 3D-printed SL methacrylate-based fluidic channels against that of PTFE tubing on account of the enhanced 1-octanol wettability and lower contact angles for the 3D-printed devices. The devices with obround channels were leveraged for the automatic µEME and in-line clean-up of MB in high matrix textile dyeing wastewater samples with relative recoveries ≥81%, RSD% ≤ 17.1% and LOD of 1.3 mg L-1. The 3D-printed nonsupported µEME device was proven superb for the analysis of wastewater samples with an elevated ionic strength (0.7 mol L-1 NaCl, 5000 mg L-1 Na2CO3, and 0.013 mol L-1 NaOH) with recorded electric currents below 12 µA. NOVELTY: The coupling of 3D printing with nonsupported µEME in automatic flow-based systems is herein proposed for the first time and demonstrated for the clean-up of troublesome samples, such as wastewaters.

Electroextraction of methylene blue from aqueous environmental samples using paper points coupled with hollow fiber membranes.

This article introduces a novel approach by coupling paper points with hollow fiber membrane for electroextraction (PP-HF-EE). The method was innovatively applied to extract methylene blue (MB) from large water volumes (up to 580 mL). A comprehensive study of six key parameters - organic filter, acceptor and donor phase composition, extraction time, applied voltage, and sample volume - was conducted using conventional flatbed scanning and digital image analysis. Our results revealed that extraction performance was primarily influenced by time, with low voltages (50 V) and low-conductivity organic filters (1-decanol) yielding comparable results to higher settings (300 V or 1-pentanol). Under optimized conditions (50 V, 60 min, 1-decanol as the organic filter), analytical performance parameters were assessed, demonstrating acceptable precision (RSD <18% for intra- and inter-day measurements) within a linear range of 5-100 µg L-1 (r = 0.98). PP-HF-EE demonstrated reliability through stable and reproducible electric current measurements during all extraction studies. Utilizing an extremely cost-effective detection system, PP-HF-EE achieved detection limits in the low ppb range, highlighting its potential as a promising variation of electromembrane extraction for environmental sample analysis.

Electromembrane extraction - capillary zone electrophoresis for the quantitative determination of ß-lactam antibiotics in milk samples.

Three penicillin-based ß-lactam antibiotics (benzylpenicillin, amoxicillin, and ampicillin) were extracted by electromembrane extraction (EME) and determined in the resulting extracts by capillary zone electrophoresis (CZE) with UV-Vis detection. The EME was optimized for the simultaneous clean-up of complex samples and preconcentration of the three antibiotics and employed 1-octanol as the organic phase interface (impregnated in the pores of a hollow fiber), acidified donor solution (pH 3), and phosphate buffer (pH 5.6) as the acceptor solution. The EMEs were carried out for 20 min at 300 V and constant stirring (750 rpm) of the donor solution. At the optimized EME-CZE conditions, the sensitivity of the analytical method was sufficient for the determination of the three ß-lactam antibiotics in undiluted cow's milk at concentrations below the EU maximum residue limits (4 µg/L) in foodstuffs. The method was simple, rapid, and convenient and offered extraction recoveries of 13.5 - 87.3 %, enrichment factors of 23.6 - 152.8, repeatability (RSD values) better than 7.6 %, linear analytical response in the 1 - 100 µg/L (3 - 100 µg/L for benzylpenicillin) concentration range with correlation coefficients ≥ 0.9997, and limits of detection from 0.2 to 1.2 µg/L. The proposed analytical concept was used for the rapid control of milk quality (i.e. assessment of excessive use of antibiotics in dairy animals), moreover, it was further extended to the trace determination of ß-lactam antibiotics in other complex samples, such as in wastewater.

Capillary Electrophoresis with Interchangeable Cartridges for Versatile and Automated Analyses of Dried Blood Spot Samples.

A novel concept for highly versatile automated analyses of dried blood spot (DBS) samples by commercial capillary electrophoresis (CE) is presented. Two interchangeable CE cartridges with different fused-silica capillaries were used for the DBS elutions and the DBS eluate analyses, respectively. The application of one CE cartridge with a wide-bore capillary reduced DBS processing times to a minimum (1-2 min per sample) while fitting the other CE cartridge with a narrow-bore capillary served for highly efficient CE analyses. A comprehensive investigation of major variables affecting liquid handling in CE (capillary length, internal diameter, and temperature) was carried out with the aim of optimizing both procedures and enabling their maximum flexibility. The application of two CE cartridges also enabled the employment of CE detectors with different instrumental set-ups and/or principles as was demonstrated by the optical detection of nonsteroidal anti-inflammatory drugs (NSAIDs) and the conductivity detection of amino acids (AAs). The presented methods were optimized for the automated CE analyses of 36 DBS samples formed by a volumetric collection of 5 µL of capillary blood onto Whatman 903 discs and processed by direct in-vial elution using the CE instrument. The precision of liquid transfers for the automated DBS elutions was better than 0.9% and the precision of CE analyses did not exceed 5.1 and 12.3% for the determination of NSAIDs and AAs, respectively. Both methods were linear (R2 ≥ 0.996) over the therapeutic (NSAIDs) and the endogenous (AAs) concentration ranges, had limits of quantification below the typical analyte concentrations in human blood, and achieved sample throughputs of more than 6 DBSs per hour.

Autonomous capillary electrophoresis processing and analysis of dried blood spots for high-throughput determination of uric acid.

A new set-up for fully autonomous and high-throughput capillary electrophoresis (CE) analyses of dried blood spot (DBS) samples is presented. The DBS samples were prepared by collecting exactly 5 µL of capillary blood from a finger-prick onto a pre-punched DBS disc in a disposable plastic CE vial and by in-vial blood drying. The vials with the DBS samples were then loaded into a commercial CE instrument for a fully unmanned sample processing and analysis. A fused-silica capillary of the CE instrument was first used for the transfer of 100 µL of elution solvent to each vial, in-vial DBS elution, and in-vial eluate homogenization. The same capillary was also used for at-line injection, separation, and selective analysis of the resulting eluates. Novel CE sequences were tailor-programmed for consecutive processing and analyses of multiple DBSs, which facilitated a fully autonomous determination of uric acid with a throughput of 240 DBS samples per day (24 h). The presented analytical protocol (using 100 µm i. d./30 cm capillary; 30 mM 2-(N-morpholino)-ethanesulfonic acid, 30 mM l-histidine, and 30 µM cetyltrimethylammonium bromide background electrolyte solution; and UV detection at 292 nm) provided excellent precision at endogenous and spiked uric acid concentrations with RSD values of peak areas below 3.2%. Calibration curves were linear over the 33.3 - 1200 µM range (R2 better than 0.998), limits of detection and quantification in the original capillary blood were 10 and 33.3 µM, respectively, and were well below the uric acid clinical range (140-420 µM). The stability of uric acid in DBS samples stored at laboratory temperature for up to 2 months was also excellent demonstrating less than a 4.2% decrease in uric acid concentrations. The actual set-up might thus be highly attractive for clinical subjects and laboratories because it is minimally invasive and requires minimum intervention from laboratory staff.

In-vial dried urine spot collection and processing for quantitative analyses.

Analysis of dried urine spots (DUSs) is becoming an emerging technique in clinical, toxicological, and forensic chemistry due to the fully non-invasive collection, facile transportation, and simple storage of DUS samples. Correct DUS collection and elution is of the utmost importance because inadequate DUS sampling/processing may have direct consequences on quantitative DUS analyses and these aspects were, for the first time, comprehensively investigated in this contribution. Various groups of endogenous and exogenous species were selected as model analytes and their concentrations were monitored in DUSs collected on standard cellulose-based sampling cards. Strong chromatographic effects were observed for most analytes having a crucial impact on their distribution within the DUSs during sampling. Concentrations of target analytes were up to 3.75-fold higher in the central DUS sub-punch in comparison to the liquid urine. Consequently, substantially reduced concentrations of these analytes were determined in peripheral DUS sub-punches demonstrating that sub-punching, often applied to dried material spots, is not acceptable for quantitative DUS analyses. Hence, a simple, rapid, and user-friendly procedure was suggested, which employed an in-vial collection of a known urine volume on a pre-punched sampling disc (using a low-cost micropipette designed for patient-centric clinical sampling) and in-vial processing of the whole DUS. Excellent accuracy (0.20%) and precision (0.89%) of liquid transfers were achieved by the micropipette, which was also applied to remote DUS collection by laic and expert users. The resulting DUS eluates were analysed by capillary electrophoresis (CE) for the determination of endogenous urine species. The CE results demonstrated no significant differences between the two user groups, elution efficiencies of 88-100% (in comparison to the liquid urine), and precision better than 5.5%.

Fully soluble polymeric foams for in-vial dried blood spot collection and analysis of acidic drugs by capillary electrophoresis.

Polymeric foams tailor-made of polyvinylpyrrolidone (PVP) and carboxymethylcellulose/oxidized 6-carboxycellulose (CMC07/OC) composite were proposed as suitable sorbents for the collection and analysis of dried blood spots (DBSs). The PVP and CMC07/OC foams were easy to prepare, enabled collection of minute volumes of capillary blood, and blood drying at ambient temperature. The resulting foams were prepared as small porous discs with uniform dimensions (approx. 6 × 3 mm) and were fully soluble in aqueous solutions. The DBSs were formed in standard capillary electrophoresis (CE) vials fitted with the soluble foam discs and enabled the direct in-vial DBS processing and at-line analysis by CE. The DBSs were pretreated with a simple process, which involved a complete dissolution of the foam disc in an acidic solution and a simultaneous hollow fiber liquid-phase microextraction (HF-LPME) in one step. The complete solubility of the foam disc with the DBS served for a quantitative transfer of all blood components into the eluate and a nearly exhaustive HF-LPME of target analytes, whereas the blood matrix and the polymeric foam components were efficiently retained by the organic solvent impregnated in the walls of the HF. The suitability of the PVP and CMC07/OC foams for the collection and the direct analysis of DBSs was demonstrated by the HF-LPME/CE determination of model acidic drugs (warfarin, ibuprofen, naproxen, ketoprofen, and diclofenac) at therapeutically relevant concentrations. Repeatability of the analytical method was better than 8.1% (RSD), extraction recoveries ranged from 70 to 99% (for PVP foam), calibration curves were linear over two orders of magnitude (R2 higher than 0.9991), and limits of detection were less than 44 µg/L (for concentrations in undiluted capillary blood). The soluble polymeric foams exhibited non-significant variations in analyte concentrations for DBSs prepared from blood samples with different hematocrit levels and for aged DBSs (less than 9.2%), moreover, they outperformed standard DBS sampling devices in terms of sample pretreatment time and extraction recovery.

Micro-electromembrane extraction through volatile free liquid membrane for the determination of ß-lactam antibiotics in biological and environmental samples.

Micro-electromembrane extraction (µ-EME) was presented for the selective extraction of four main ß-lactam antibiotics (penicillin, phenoxypenicillin, ampicillin, and amoxicillin) from complex samples. A volatile solvent (ethyl acetate or chloroform) was sandwiched between a plug of the complex sample and another plug of an aqueous acceptor solution in a transparent polymeric tube and formed the so-called free liquid membrane (FLM). The use of the FLM eliminated the evaporation of the solvent and enabled the µ-EME of the antibiotics, which was carried out by the application of DC voltage to the terminal aqueous solutions. The drugs in the complex sample were selectively transferred through the FLM to the acceptor solution, which was directly used for their determination by micellar electrokinetic chromatography with ultraviolet detection (MEKC-UV). The µ-EME was characterized by sub-µA electric currents, high elimination of matrix components, high stability of operational solutions, and suitability for extracting undiluted complex samples. The µ-EME/MEKC-UV method yielded good analytical repeatability (RSDs of peak areas ≤5%), extraction recoveries (40-84%), accuracy (92-105%) and linearity over one and a half order of magnitude (R2 ≥ 0.9998), and was applied to the determination of the four ß-lactam antibiotics in human serum and waste water at clinically and environmentally relevant concentration levels. Further improvement in the method sensitivity was achieved by changing the µ-EME tube geometry (conical shape) and increasing the complex sample volume (100 µL). The analytes were enriched by factors of 7.6-11.5, the limits of detection dropped down to less than 18 ng/mL, and the modified µ-EME/MEKC-UV method enabled the trace determination of ß-lactam antibiotics in complex samples.

Programmable Millifluidic Platform Integrating Automatic Electromembrane Extraction Cleanup and In-Line Electrochemical Detection: A Proof of Concept.

A fully automatic millifluidic sensing platform coupling in-line nonsupported microelectromembrane extraction (µ-EME) with electrochemical detection (ECD) is herein proposed for the first time. Exploiting the features of the second generation of flow analysis, termed sequential injection (SI), the smart integration of SI and µ-EME-ECD enables (i) the repeatable formation of microvolumes of phases for the extraction step in a membrane-less (nonsupported) arrangement, (ii) diverting the acceptor plug to the ECD sensing device, (iii) in-line pH adjustment before the detection step, and (iv) washing of the platform for efficient removal of remnants of wetting film solvent, all entirely unsupervised. The real-life applicability of the miniaturized sensing system is studied for in-line sample cleanup and ECD of diclofenac as a model analyte after µ-EME of urine as a complex biological sample. A comprehensive study of the merits and the limitations of µ-EME solvents on ECD is presented. Under the optimal experimental conditions using 14 µL of unprocessed urine as the donor, 14 µL of 1-nonanol as the organic phase, and 14 µL of 25 mM NaOH as the acceptor in a 2.4 mm ID PTFE tubing, an extraction voltage of 250 V, and an extraction time of 10 min, an absolute (mass) extraction recovery of 48% of diclofenac in urine is obtained. The proposed flow-through system is proven to efficiently remove the interfering effect of predominantly occurring organic species in human urine on ECD with RSD% less than 8.6%.

Automated Sequential Injection-Capillary Electrophoresis for Dried Blood Spot Analysis: A Proof-of-Concept Study.

A hyphenated analytical platform that enables fully automated analyses of dried blood spots (DBSs) is proposed by the at-line coupling of sequential injection (SI) to capillary electrophoresis (CE). The SI system, exploited herein for the first time for unattended DBS handling, serves as the "front end" mesofluidic platform for facilitating exhaustive elution of the entire DBS by flow programming. The DBS eluates are thus free from hematocrit and nonhomogeneity biases. The SI pump transfers the resulting DBS eluates into CE sample vials through an internal port of the CE instrument and homogenizes the eluates, whereupon the eluted blood compounds are automatically injected, separated, and quantified by the CE instrument. The SI and CE are commercially available off-the-shelf instruments and are interconnected through standard nuts, ferrules, and tubing without additional instrumental adjustments. They are controlled by dedicated software and are synchronized for a fully autonomous operation. The direct determination of endogenous (potassium and sodium) and exogenous (lithium as a model drug) inorganic cations in DBS samples has been used for the proof-of-concept demonstration. The hyphenated SI-CE platform provides excellent precision of the analytical method with relative standard deviation (RSD) values of peak areas below 1.5 and 3.5% for intraday and interday analyses, respectively, of the endogenous concentrations of the two inorganic cations. For the determination of lithium, calibration is linear in a typical clinical range of the drug (R2 better than 0.9993 for 2-20 mg/L), RSD values of peak areas are below 4.5% (in the entire calibration range), the limit of detection (0.4 mg/L) and the limit of quantification (1.3 mg/L) are well below the drug's minimum therapeutic concentration (4 mg/L), and total analysis time is shorter than 5 min. The SI-CE platform reflects the actual trends in the automation of analytical methods, offers rapid and highly flexible DBS elution/analysis processes, and might thus provide a general solution to modern clinical analysis as it can be applied to a broad range of analytes and dried biological materials.

At-line coupling of hollow fiber liquid-phase microextraction to capillary electrophoresis for trace determination of acidic drugs in complex samples.

Direct analysis of complex samples is demonstrated by the at-line coupling of hollow fiber liquid-phase microextraction (HF-LPME) to capillary electrophoresis (CE). The hyphenation of the preparative and the analytical technique is achieved through a 3D-printed microextraction device with an HF located in a sample vial of a commercial CE instrument. The internal geometry of the device guides the CE separation capillary into the HF and the CE injection of the HF-LPME extract is performed directly from the HF lumen. The 3D-printing process ensures uniform dimensions of the devices, their constant position inside the sample vial, and excellent repeatability of the HF-LPME as well as the CE injection. The devices are cheap (∼0.01 €) and disposable, thus eliminating any possible sample-carryover, moreover, the at-line CE analysis of the extract is performed fully autonomously with no need for operator's intervention. The developed HF-LPME/CE-UV method is applied to the determination of acidic drugs in dried blood spot and wastewater samples and is characterized by excellent repeatability (RSD, 0.6-9.6%), linearity (r2, 0.9991-0.9999), enrichment (EF, 29-97), sensitivity (LOD, 0.2-3.4 µg/L), and sample throughput (7 samples/h). A further improvement of selected characteristics of the analytical method is achieved by the at-line coupling of HF-LPME to capillary isotachophoresis (ITP) with electrospray ionization-mass spectrometry (ESI-MS). The HF-LPME/ITP-ESI-MS system facilitates enhanced selectivity, matrix-free analytical signals, and up to 34-fold better sensitivity due to the use of ESI-MS detection and additional on-capillary ITP preconcentration of the HF-LPME extracts.

Volatile free liquid membranes for electromembrane extraction.

Volatile solvents are excellent extraction media for liquid-liquid extractions. However, their use in supported liquid membranes (SLMs) is limited by their evaporation from SLM and thus poor SLM stability and they have never been considered truly useful for electromembrane extraction (EME). In this contribution, volatile solvents were systematically investigated as liquid membranes for EME and their extraction characteristics were comprehensively examined for the first time. A short plug of a water immiscible volatile solvent (a free liquid membrane (FLM)) was sandwiched between two aqueous plugs (donor and acceptor solutions) in a narrow-bore polymeric tubing. Evaporation of the volatile FLM was thus completely avoided and excellent stability of the phase interface was ensured. Suitability of volatile FLMs for EMEs was justified by µ-EMEs of nortriptyline, haloperidol, loperamide and papaverine as model non-polar basic drugs. Extraction performance of µ-EME through ethyl acetate was comparable or better to that through standard non-volatile EME solvents and a high extraction selectivity was achieved for nortriptyline and haloperidol extracted through chloroform. µ-EMEs through the volatile FLMs were characterized by high extraction recoveries (62%-99% for standards and 40-89% for body fluids), low electric currents (10-1380 nA), no susceptibility to matrix ions and suitability for pretreatment of raw body fluids (human urine and serum). Resulting extracts were analysed by capillary electrophoresis with ultraviolet detection (CE/UV). Repeatability of the µ-EME-CE/UV method was excellent with intra-day and inter-day RSD values 0.8-3.2% and 1.8-4.6%, respectively. Further experiments demonstrated additional advantages of volatile FLMs by nearly exhaustive µ-EMEs of atenolol as the polar basic drug with no need for FLM modification by ionic carriers. The presented comprehensive examination of volatile solvents has broadened the range of liquid membranes suitable for EME and it is believed that this proof-of-concept study will stimulate further interest in a deeper investigation of volatile phase interfaces in EME.

Dried Blood Spot Self-Sampling with Automated Capillary Electrophoresis Processing for Clinical Analysis.

A simple and convenient concept of blood sampling followed by a fully automated analysis is presented. A disposable sampling kit is used for accurate self-sampling of capillary blood from a finger prick. A high-throughput blood sampling is thus enabled, which is essential in many clinical assays and considerably improves life quality and comfort of involved subjects. The collected blood samples are mailed to a laboratory for a fully automated dried blood spot (DBS) processing and analysis, which are performed with a commercial capillary electrophoresis instrument. Quantitative results are obtained within 20 min from the DBS delivery to the laboratory. The presented concept is exemplified by the determination of warfarin blood concentrations and demonstrates excellent analytical performance. Moreover, this concept is generally applicable to a wide range of endogenous and exogenous blood compounds and represents a novel and attractive analytical tool for personalized health monitoring.

Capacitively coupled contactless conductivity detection for analytical techniques - Developments from 2018 to 2020.

The developments of analytical contactless conductivity measurements based on capacitive coupling over the two years from mid-2018 to mid-2020 are covered. This mostly concerns applications of the technique in zone electrophoresis employing conventional capillaries and to a lesser extent lab-on-chip devices. However, its use for the detection in several other flow-based analytical methods has also been reported. Detection of bubbles and measurements of flow rates in two-phase flows are also recurring themes. A few new applications in stagnant aqueous samples, e.g. endpoint detection in titrations and measurement on paper-based devices, have been reported. Some variations of the design of the measuring cells and their read-out electronics have also been described.

Electromembrane extraction of highly polar bases from biological samples - Deeper insight into bis(2-ethylhexyl) phosphate as ionic carrier.

Similarly to many other sample extraction techniques, efficient extraction of very polar compounds with electromembrane extraction (EME) is difficult. To date, the best known strategy to improve the mass transfer of these compounds is the addition of an ionic carrier, often bis(2-ethylhexyl) phosphate (DEHP) to the supported liquid membrane (SLM). DEHP is known to work by providing ionic interactions with basic compounds, to improve the partitioning into the SLM. In this work, the behavior of DEHP during extractions was studied for the first time. Interestingly, substantial amounts of DEHP was found to leak from the SLM into the aqueous sample at pH > 4. Due to this leakage, the ion-pair formation between analytes and DEHP was moved from the sample/SLM interface (interfacial complexation) to the bulk of the sample solution (bulk-sample complexation), which improved the mass transfer of polar bases considerably. Based on this, an extraction procedure for eight polar bases with log P values from +0.7 to -5.9 was developed and optimized. The optimization demonstrated that extraction of more polar analytes was favored by bulk-sample complexation. With optimized conditions, extraction from biological samples such as urine, protein-precipitated plasma, and raw plasma were performed with recoveries >40%, except for a few analytes. In addition, the extraction system could be operated under robust conditions with relatively low current (<70 µA for plasma), and provided low variability (<16% RSD), as well as good clean-up efficiency. These findings are an important step in further scientific anchoring of EME, and development of the technique towards selective extraction of very polar substances from complex biological matrices.

Hollow Fiber Liquid-Phase Microextraction At-Line Coupled to Capillary Electrophoresis for Direct Analysis of Human Body Fluids.

A simple and cheap all-in-one concept for at-line coupling of hollow fiber liquid-phase microextraction (HF-LPME) to commercial capillary electrophoresis (CE) is demonstrated, which enables the direct analysis of complex samples. A disposable microextraction device compatible with injection systems of Agilent CE instruments is proposed, which consists of a short segment of a porous HF attached to a tapered polypropylene holder. The holder maintains a constant position of the HF in a CE vial during extraction and simultaneously guides the injection end of a separation capillary into the HF lumen for automated CE injection and analysis. In a typical analytical procedure, the HF is impregnated with a water-immiscible solvent, its lumen is filled with 5 µL of an aqueous acceptor solution, and the microextraction device is placed in a 2 mL glass CE vial containing 550 µL of a donor solution. The vial is agitated at 750 rpm for 10 min, and the resulting acceptor solution is injected directly from the HF lumen into the commercial CE. No additional manual handling is required, except for the transfer of the CE vial to the CE autosampler. Multiple complex samples can be simultaneously pretreated in a multiple-well plate format, thus significantly reducing the total analysis time. Suitability of the analytical method is demonstrated by the direct determination of model basic drugs (nortriptyline, haloperidol, loperamide, and papaverine) in physiological solutions, urine, and dried blood spot (DBS) samples. Repeatability of the method is better than 12.8% (%RSD), extraction recoveries range between 34 and 76%, and enrichment factors are 37-84. The method is linear in a range of 2 orders of magnitude (R2 ≥ 0.9977) with limits of detection of 0.7-1.55 µg/L. The method has a high potential for the direct analysis of DBS samples since DBS elution and HF-LPME are performed simultaneously during the 10 min agitation. The manual DBS handling is thus reduced to inserting the DBS punch into the CE vial only. Moreover, the universal character of the HF-LPME might extend the applicability of the method to a wide range of analytes/matrices, and combination with other commercial detectors might improve the selectivity/sensitivity of the CE analysis.