New generation of sequential injection analysis (SIA) called sequential injection chromatography (SIC) has already been consolidated as a good alternative of high performance liquid chromatography (HPLC) for fast analysis of simple samples. Benefits of flow methods are automation, miniaturization and low sample and mobile phase consumption. Implementation of short monolithic chromatographic column into SIA opens new area-on-line chromatographic separation of multi-compound sample in low-pressure flow system, with the advantage of flow programming and possibility of sample manipulation. In the presented review the potential of SIC and its comparison with HPLC for determination of pharmaceutical mixtures is discussed and outlines past and recent trends focused on separation with SIC.

• MeSH
• chromatografie metody   přístrojové vybavení   MeSH
• financování organizované MeSH
• lékové formy MeSH
• lidé MeSH
• průtoková injekční analýza metody   přístrojové vybavení   MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• přehledy MeSH

Currently, for Sequential Injection Chromatography (SIC), only reversed phase C18 columns have been used for chromatographic separations. This article presents the first use of three different stationary phases: three core-shell particle-packed reversed phase columns in flow systems. The aim of this work was to extend the chromatographic capabilities of the SIC system. Despite the particle-packed columns reaching system pressures of ≤ 610 PSI, their conditions matched those of a commercially produced and optimised SIC system (SIChrom™ (FIAlab(®), USA)) with a 8-port high-pressure selection valve and medium-pressure Sapphire™ syringe pump with a 4 mL reservoir and maximum system pressure of ≤ 1000 PSI. The selectivity of each of the tested columns, Ascentis(®) Express RP-Amide, Ascentis(®) Express Phenyl-Hexyl and Ascentis(®) Express C18 (30 mm × 4.6mm, core-shell particle size 2.7 μm), was compared by their ability to separate seven phenolic acids that are secondary metabolite substances widely distributed in plants. The separations of all of the components were performed by isocratic elution using binary mobile phases composed of acetonitrile and 0.065% phosphoric acid at pH 2.4 (a specific ratio was used for each column) at a flow-rate of 0.60 mL/min. The volume of the mobile phase was 3.8 mL for each separation. The injection volume of the sample was 10 μL for each separation. The UV detection wavelengths were set to 250, 280 and 325 nm. The RP-Amide column provided the highest chromatographic resolution and allowed for complete baseline separation of protocatechuic, syringic, vanillic, ferulic, sinapinic, p-coumaric and o-coumaric acids. The Phenyl-Hexyl and C18 columns were unable to completely separate the tested mixture, syringic and vanillic acid and ferulic and sinapinic acids could not be separated from one another. The analytical parameters were a LOD of 0.3 mg L(-1), a LOQ of 1.0 mg L(-1), a calibration range of 1.0-50.0 (100.0) mg L(-1) (r>0.997) and a system precision of 10 mg L(-1) with a RSD ≤ 1.65%. The high performance of the chromatography process with the RP-Amide column under optimised conditions was highlighted and well documented (HETP values ≤ 10 μm, peak symmetry ≤ 1.33, resolution ≥ 1.87 and time for one analysis <8.0 min). The results of these experiments confirmed the benefits of extending chromatographic selectivity using core-shell particle column technology in a SIC manifold.

• MeSH
• chromatografie s reverzní fází * MeSH
• hydroxybenzoáty analýza   MeSH
• injekce MeSH
• kalibrace MeSH
• molekulární struktura MeSH
• spektrofotometrie ultrafialová MeSH
• velikost částic MeSH
• vysokoúčinná kapalinová chromatografie * MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

On-line sample pretreatment (clean-up and analyte preconcentration) is for the first time coupled to sequential injection chromatography. The approach combines anion-exchange solid-phase extraction and the highly effective pentafluorophenylpropyl (F5) fused-core particle column for separation of eight sulfonamide antibiotics with similar structures (sulfathiazole, sulfanilamide, sulfacetamide, sulfadiazine, sulfamerazine, sulfadimidine, sulfamethoxazole and sulfadimethoxine). The stationary phase was selected after a critical comparison of the performance achieved by three fused-core reversed phase columns (Ascentis(®) Express RP-Amide, Phenyl-Hexyl, and F5) and two monolithic columns (Chromolith(®) High Resolution RP-18 and CN). Acetonitrile and acetate buffer pH 5.0 at 0.60 mL min(-1) were used as mobile phase to perform the separations before spectrophotometric detection. The first mobile phase was successfully used as eluent from SPE column ensuring transfer of a narrow zone to the chromatographic column. Enrichment factors up to 39.2 were achieved with a 500 µL sample volume. The developed procedure showed analysis time <10.5 min, resolutions >1.83 with peak symmetry ≤1.52, LODs between 4.9 and 27 µg L(-1), linear response ranges from 30.0 to 1000.0 µg L(-1) (r(2)>0.996) and RSDs of peak heights <2.9% (n=6) at a 100 µg L(-1) level and enabled the screening control of freshwater samples contaminated at the 100 µg L(-1) level. The proposed approach expanded the analytical potentiality of SIC and avoided the time-consuming batch sample pretreatment step, thus minimizing risks of sample contamination and analyte losses.

• MeSH
• antibakteriální látky analýza   izolace a purifikace   MeSH
• chemické látky znečišťující vodu analýza   izolace a purifikace   MeSH
• design vybavení MeSH
• extrakce na pevné fázi přístrojové vybavení   MeSH
• limita detekce MeSH
• průtoková injekční analýza přístrojové vybavení   MeSH
• řeky chemie   MeSH
• sulfonamidy analýza   izolace a purifikace   MeSH
• vysokoúčinná kapalinová chromatografie přístrojové vybavení   MeSH
• Publikační typ
• časopisecké články MeSH
• hodnotící studie MeSH
• práce podpořená grantem MeSH

Dasatinib is a novel oral prescription drug proposed for treating adult patients with chronic myeloid leukemia. Three analytical methods, namely ultra high performance liquid chromatography, capillary zone electrophoresis, and sequential injection analysis, were developed, validated, and compared for determination of the drug in the tablet dosage form. The total analysis time of optimized ultra high performance liquid chromatography and capillary zone electrophoresis methods was 2.0 and 2.2 min, respectively. Direct ultraviolet detection with detection wavelength of 322 nm was employed in both cases. The optimized sequential injection analysis method was based on spectrophotometric detection of dasatinib after a simple colorimetric reaction with folin ciocalteau reagent forming a blue-colored complex with an absorbance maximum at 745 nm. The total analysis time was 2.5 min. The ultra high performance liquid chromatography method provided the lowest detection and quantitation limits and the most precise and accurate results. All three newly developed methods were demonstrated to be specific, linear, sensitive, precise, and accurate, providing results satisfactorily meeting the requirements of the pharmaceutical industry, and can be employed for the routine determination of the active pharmaceutical ingredient in the tablet dosage form.

• MeSH
• dasatinib analýza   MeSH
• elektroforéza kapilární * MeSH
• farmaceutická chemie metody   normy   MeSH
• lidé MeSH
• limita detekce MeSH
• reprodukovatelnost výsledků MeSH
• spektrofotometrie * MeSH
• tablety chemie   normy   MeSH
• vysokoúčinná kapalinová chromatografie * MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

Sequential Injection Chromatography (SIC) evolved from fast and automated non-separation Sequential Injection Analysis (SIA) into chromatographic separation method for multi-element analysis. However, the speed of the measurement (sample throughput) is due to chromatography significantly reduced. In this paper, a sub-1min separation using medium polar cyano monolithic column (5mm×4.6mm) resulted in fast and green separation with sample throughput comparable with non-separation flow methods The separation of three synthetic water-soluble dyes (sunset yellow FCF, carmoisine and green S) was in a gradient elution mode (0.02% ammonium acetate, pH 6.7 - water) with flow rate of 3.0mLmin-1corresponding with sample throughput of 30h-1. Spectrophotometric detection wavelengths were set to 480, 516 and 630nm and 10Hz data collection rate. The performance of the separation was described and discussed (peak capacities 3.48-7.67, peak symmetries 1.72-1.84 and resolutions 1.42-1.88). The method was represented by validation parameters: LODs of 0.15-0.35mgL-1, LOQs of 0.50-1.25mgL-1, calibration ranges 0.50-150.00mgL-1(r>0.998) and repeatability at 10.0mgL-1of RSD≤0.98% (n=6). The method was used for determination of the dyes in "forest berries" colored pharmaceutical cough-cold formulation. The sample matrix - pharmaceuticals and excipients were not interfering with vis determination because of no retention in the separation column and colorless nature. The results proved the concept of fast and green chromatography approach using very short medium polar monolithic column in SIC.

• MeSH
• barvicí látky MeSH
• chromatografie MeSH
• farmaceutická chemie metody   MeSH
• příprava léků MeSH
• voda MeSH
• Publikační typ
• časopisecké články MeSH

The use of small scale renewable sorbent material for automated solid phase extraction of multi-residue pharmaceuticals in environmental samples exploiting the sequential injection analysis-bead injection with direct coupling to liquid chromatography-electrospray ionization tandem mass spectrometry (SIA-BI-μSPE-LC-ESI-MS/MS) is presented to determine beta-blockers, namely atenolol, sotalol, pindolol, acebutolol, timolol, metoprolol, labetalol, carazolol, propranolol and betaxolol. These compounds yielded the same product ions, therefore were affected in terms of quantification when flow injection analysis-mass spectrometry (FIA-MS) was used. Thus, analytes and matrix present in the sample travel together into the ionization source which can seriously affect the ionization efficiency and analyte signals due to monitoring over a short time period. Graphical abstract A two-dimensional analysis involving a time dimension (retention time) and an m/z dimension (fragmentation ion) is promising for the various sample types. Using the developed method, absolute recoveries percentages of 10 mL of sample loading volume were >91% for all β-blockers with enrichment factor of 62-74, limits of detection of 0.005-0.07 μg L(-1), limits of quantification of 0.01-0.23 μg L(-1), enrichment factor of 62-72 and repeatability within range 7-12%. This developed method is suggested to be used as quantitative screening technique for drugs of abuse or persistent contamination using different kinds of sorbent materials and complex matrix such as biological fluid sample as well.

• MeSH
• chromatografie kapalinová metody   MeSH
• extrakce na pevné fázi metody   MeSH
• hmotnostní spektrometrie s elektrosprejovou ionizací metody   MeSH
• limita detekce MeSH
• reprodukovatelnost výsledků MeSH
• tandemová hmotnostní spektrometrie metody   MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

This paper deals with a novel approach to separate two analytes with different chemical properties and different lipophilicity. The newly described methodology is based on the two column system that was used for isocratic separation of two analytes with very different lipophilicity-dexamethasone and cinchocaine. Simultaneous separation of model compounds cinchocaine and dexamethasone was carried under the following conditions in two-column sequential injection chromatography system (2-C SIC). A 25×4.6 mm C-18 monolithic column was used in the first dimension for retention and separation of dexamethasone with mobile phase acetonitrile:water 30:70 (v/v), flow rate 0.9 mL min(-1) and consumption of 1.7 mL. A 10×4.6 mm C-18 monolithic column with 5×4.6 mm C-18 precolumn was used in the second dimension for retention and separation of cinchocaine using mobile phase acetonitrile:water 60:40 (v/v), flow rate 0.9 mL min(-1) and consumption 1.5 mL. Whole analysis time including both mobile phase's aspirations and both column separations was performed in less than 4 min. The method was fully validated and used for determination of cinchocaine and dexamethasone in pharmaceutical otic drops. The developed 2-C SIC method was compared with HPLC method under the isocratic conditions of separation on monolithic column (25×4.6 mm C-18). Spectrophotometric detection of both compounds was performed at wavelength 240 nm. System repeatability and method precision were found in the range (0.39-3.12%) for both compounds. Linearity of determination was evaluated in the range 50-500 μg mL(-1) and coefficients of determination were found to be r(2)=0.99912 for dexamethasone and r(2)=0.99969 for cinchocaine.

• MeSH
• anestetika lokální analýza   chemie   MeSH
• antiflogistika analýza   chemie   MeSH
• chromatografie metody   MeSH
• cinchokain analýza   chemie   MeSH
• dexamethason analýza   chemie   MeSH
• lékové roztoky MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

Since its inception, sequential injection chromatography (SIC) has evolved through several stages. Key moments including introduction of the novel technique combining sequential injection analysis and monolithic column, the first generation of commercial SIC system employing robust pump, the utilization of columns packed with fused-core particles, the on-line hyphenation of extraction and separation steps in SIC, are now followed by the second generation of commercial SIC system employing stainless steel syringe pump and parts optimized for chromatographic separation. The key developments always mean acceleration of the evolution by opening new avenues and reduction of compromises in automated analytical methods based on the flow analysis. The updates, new features, and prospects of the novel instrument are described and discussed on perspective of the method developed for extraction and separation of selected phenolic acids (gallic, protocatechuic, caffeic, p-coumaric and ferulic). The method hyphenates miniaturized on-line solid phase extraction using strong anion exchange sorbent in commercial cartridge for HPLC (20 × 1 mm) and liquid chromatography using chromatographic column (C18 50 × 4.6 mm, 5 μm particles) packed with fused-core particles in the SIC manifold. The separation in gradient mode used acetonitrile: aqueous formic acid pH 2.0 mobile phase and spectrophotometric detection at 270, 300, and 320 nm. Injected sample volumes were 200 and 500 μL. The performance of the extraction step was characterized by the recovery 94.0-107.8%, enrichment factors about 20 or 50, and the separation by peak capacities 13-34, peak symmetries 1.17-1.64, and resolutions 0.82-3.75). While using a sample volume of 200 μL, our method was characterized by the following validation parameters: LODs of 0.0075-0.03 mg L-1, LOQs of 0.025-0.10 mg L-1, calibration ranges 0.025-2.50 mg L-1 (r > 0.999), repeatability of signal at 0.50 mg L-1of RSD ≤ 1.46% (n = 6), and overall time of analysis 7.1 min. The results including pilot analysis of white and red wines demonstrated the capability of novel SIC instrument to enable fast, selective, and sensitive analysis.

• Publikační typ
• časopisecké články MeSH

In this work, the applicability of Sequential Injection Chromatography for the determination of transition metals in water is evaluated for the separation of copper(II), zinc(II), and iron(II) cations. Separations were performed using a Dionex IonPAC™ guard column (50mm×2mm i.d., 9 µm). Mobile phase composition and post-column reaction were optimized by modified SIMPLEX method with subsequent study of the concentration of each component. The mobile phase consisted of 2,6-pyridinedicarboxylic acid as analyte-selective compound, sodium sulfate, and formic acid/sodium formate buffer. Post-column addition of 4-(2-pyridylazo)resorcinol was carried out for spectrophotometric detection of the analytes׳ complexes at 530nm. Approaches to achieve higher robustness, baseline stability, and detection sensitivity by on-column stacking of the analytes and initial gradient implementation as well as air-cushion pressure damping for post-column reagent addition were studied. The method allowed the rapid separation of copper(II), zinc(II), and iron(II) within 6.5min including pump refilling and aspiration of sample and 1mmol HNO3 for analyte stacking on the separation column. High sensitivity was achieved applying an injection volume of up to 90µL. A signal repeatability of<2% RSD of peak height was found. Analyte recovery evaluated by spiking of different natural water samples was well suited for routine analysis with sub-micromolar limits of detection.

• MeSH
• chemické látky znečišťující vodu analýza   MeSH
• chromatografie metody   MeSH
• formiáty chemie   MeSH
• měď analýza   MeSH
• minerální vody analýza   MeSH
• pitná voda analýza   MeSH
• pyridiny chemie   MeSH
• resorcinoly chemie   MeSH
• sírany chemie   MeSH
• sladká voda analýza   MeSH
• železo analýza   MeSH
• zinek analýza   MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

The paper deals with the concept of simple automated creation of gradient profile of the mobile phase for gradient-elution sequential injection chromatography (GE-SIC). The feasibility and merits of this concept are demonstrated on the separation and simultaneous assay of indomethacin as active principle and of its two degradation products (5-methoxy-2-methylindoleacetic acid and 4-chloro-benzoic acid) in a topical pharmaceutical formulation. The GE-SIC separation was performed with a FIAlab(®) 3000 SIC set-up (USA) equipped with an Onyx™ Monolithic C18 (25 mm × 4.6mm, Phenomenex(®)) column, a six-port selection valve, a 5-mL syringe pump and a fiber-optics UV CCD detector. Ketoprofen was used as an internal standard (IS). The gradient elution was achieved by automated reproducible mixing of acetonitrile and aqueous 0.2% phosphoric acid in the holding coil of the SIC system. Different profiles of the gradient elution were tested. The optimal gradient using two mobile phases 30:70 and 50:50 of acetonitrile/0.2% phosphoric acid (v/v) was achieved under the optimum flow rate 1.2 mL min(-1). The chromatographic resolution R between the peaks of all solutes (including the IS) was >2.00. The repeatability of retention times was characterized by the RSD values 0.18-0.30% (n=6). Net separation time was 3.5 min and the mobile phase consumption was 4.5 mL for a single GE-SIC assay. The figures of merit of the novel GE-SIC method compared well with those of conventional HPLC.

• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH