An off-the-shelf Agilent 7100 capillary electrophoresis (CE) instrument was employed for the automated processing and analysis of dried blood spots (DBSs) collected by Capitainer®B volumetric devices. Solutions for DBS elutions were transferred directly into CE vials through a separation capillary by the application of an auxiliary nitrogen gas connected to the external pressure line of the CE instrument. This allowed for liquid handling at pressures up to 15 bar and enabled the use of a single capillary for rapid DBS processing and efficient CE separations. The resulting DBS eluates were at-line injected into a short capillary end, which served for improved instrumental simplicity and short CE analysis times. The current set-up necessitated neither hardware nor software adjustments of the CE instrument, except for the connection of a gas cylinder to an in-built connector. The novel features presented in this study (DBSs with exact blood volumes, high external pressures, and short-end injections) were used for the automated determination of clinically relevant markers, phenylalanine (Phe) and tyrosine (Tyr), in DBS samples. Sensitive and selective Phe and Tyr quantification was achieved by CE-UV in 375 mM formic acid and 0.01 % (v/v) Tween 20 (pH 2.09) as a background electrolyte. The total processing and analysis times per one DBS were <1.5 and 4.5 min, respectively, in a sequence of 36 DBSs, and resulted in a sample throughput of >10 DBSs per hour. The intra- and inter-day repeatability values were better than 5.9 and 1.1 % RSD for peak areas and migration times, respectively, and calibration curves were linear in the 20-3000 μM (Phe) and 20-250 μM (Tyr) range (R2 ≥ 0.9973). The limits of detection were ≤2 μM and enabled the determination of endogenous Phe and Tyr concentrations as well as elevated Phe concentrations and Phe/Tyr ratios, which are the typical markers for neonatal phenylketonuria screening.

• Klíčová slova
• Amino acids, Automation, Capillary electrophoresis, Dried blood spots, Rapid analysis, Short-end injection, Volumetric microsampling,
• MeSH
• automatizace * MeSH
• časové faktory MeSH
• elektroforéza kapilární * metody   MeSH
• fenylalanin * krev   MeSH
• lidé MeSH
• limita detekce MeSH
• test suché kapky krve * metody   MeSH
• tyrosin * krev   MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• fenylalanin * MeSH
• tyrosin * MeSH

Electroextraction (EE) encompasses a range of sample preparation methods whose effectiveness, selectivity, and efficiency are significantly influenced by the physical-chemical characteristics of analytes, samples, and instrumental conditions. This article explores, for the first time, various strategies aimed at enhancing the extraction efficiency of a recent approach of EE utilizing a paper point (PP) combined with a hollow fiber (HF) (abbreviated as PP-HF-EE) to extract various cationic and anionic model compounds from water samples. The study also explores, experimentally, the impact of agitation, organic filter composition, PP diameter, and PP brand on extraction performance, and proves that all these factors are quite important, especially when digital image analysis is utilized for determination. Furthermore, this work demonstrates the ease and feasibility of simultaneously extracting cations and anions using PP-HF-EE and proposes a straightforward method to enhance analyte concentration on the vertex of the PP through a base-to-vertex focusing. Lastly, it is demonstrated, using capillary electrophoresis coupled to a UV-Vis detector, that for PP-HF-EE, the extraction efficiency and pre-concentration factor are less dependent on other parameters when multiple PPs per sample are utilized, with signal enhancement values of up to 111 and 339 for nortriptyline and haloperidol, respectively. All the findings and strategies presented herein constitute significant contributions that can facilitate future research in method development, particularly in the utilization of PP-HF-EE and similar EE approaches.

• Klíčová slova
• concentration focusing, environmental analysis, green sample preparation, multiphase electroextraction,
• Publikační typ
• časopisecké články MeSH

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.

• Klíčová slova
• Automation, Capacitively coupled contactless conductivity detection, Capillary electrophoresis, Creatinine, Dried blood spot, Volumetric microsampling,
• MeSH
• automatizace MeSH
• elektroforéza kapilární * metody   MeSH
• kreatinin krev   MeSH
• lidé MeSH
• test suché kapky krve * metody   přístrojové vybavení   MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• kreatinin MeSH

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.

• Klíčová slova
• Complex matrices, Cost-effective detection, Green sample preparation, Low-voltage electroextraction,
• Publikační typ
• časopisecké články MeSH

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.

• Klíčová slova
• 3D printing, Fluidic devices, Methylene blue, Microelectromembrane extraction, Prototyping, Stereolithography, Wastewater,
• Publikační typ
• časopisecké články MeSH

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.

• MeSH
• aminokyseliny * MeSH
• elektrická vodivost MeSH
• elektroforéza kapilární * metody   MeSH
• kapiláry MeSH
• lidé MeSH
• test suché kapky krve metody   MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• aminokyseliny * MeSH

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.

• Klíčová slova
• Automation, Capillary electrophoresis, Dried blood spot, Uric acid,
• MeSH
• cetrimonium MeSH
• elektroforéza kapilární * metody   MeSH
• kyselina močová * MeSH
• lidé MeSH
• odběr vzorku krve MeSH
• test suché kapky krve metody   MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• cetrimonium MeSH
• kyselina močová * MeSH

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%.

• Klíčová slova
• Acidic drugs, Basic drugs, Capillary electrophoresis, Creatinine, Dried urine spot, Inorganic cations, Quantitative analysis, Uric acid,
• MeSH
• lidé MeSH
• odběr biologického vzorku * MeSH
• tělesné tekutiny * MeSH
• test suché kapky krve metody   MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH

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.

• Klíčová slova
• Acidic drugs, Capillary electrophoresis, Dried blood spots, Hollow fiber liquid-phase microextraction, Soluble polymeric foams,
• MeSH
• elektroforéza kapilární metody   MeSH
• ibuprofen MeSH
• ketoprofen * MeSH
• naproxen * MeSH
• rozpouštědla MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• ibuprofen MeSH
• ketoprofen * MeSH
• naproxen * MeSH
• rozpouštědla MeSH

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%.

• Klíčová slova
• automation, diclofenac, electrochemical sensing, nonsupported electrically driven extraction, sequential injection analysis,
• MeSH
• diklofenak * MeSH
• lidé MeSH
• membrány umělé * MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• diklofenak * MeSH
• membrány umělé * MeSH