New trends in fast and high-resolution liquid chromatography: a critical comparison of existing approaches (original) (raw)
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Journal of Chromatography A, 2007
In this study, ultra performance liquid chromatography (UPLC) using pressures up to 1000 bar and columns packed with sub-2 m particles has been combined with high temperature mobile phase conditions (up to 90 • C). By using high temperature ultra performance liquid chromatography (HT-UPLC), it is possible to drastically decrease the analysis time without loss in efficiency. The stability and chromatographic behavior of sub-2 m particles were evaluated at high temperature and high pressure. The chromatographic support remained stable after 500 injections (equivalent to 7500 column volumes) and plate height curves demonstrated the capability of HT-UPLC to obtain fast separations. For example, a separation of nine doping agents was performed in less than 1 min with sub-2 m particles at 90 • C. Furthermore, a shorter column (30 mm length) was used and allowed a separation of eight pharmaceutical compounds in only 40 s.
European Journal of Advances in Engineering and Technology, 2021
High-Performance Liquid Chromatography (HPLC) stands as a cornerstone technique in analytical chemistry, offering unparalleled precision and versatility in the separation, identification, and quantification of chemical compounds. This abstract provides a comprehensive overview of the fundamental principles, applications, and advancements in HPLC. HPLC operates on the principles of chromatographic separation, utilizing a liquid mobile phase and a solid stationary phase to separate analytes based on their differential interactions with the stationary phase. The methodology encompasses various modes of chromatography, including reversed-phase, normal-phase, ion-exchange, size exclusion, and affinity chromatography, each tailored to specific analyte characteristics and separation requirements. Applications of HPLC span across diverse scientific disciplines, from pharmaceutical analysis, environmental monitoring, and food safety to clinical diagnostics, forensic analysis, and material science. Its high sensitivity, resolution, and reproducibility make it indispensable in both qualitative and quantitative analysis, offering insights into complex mixtures and trace-level components. Furthermore, advancements in HPLC instrumentation, including column technology, detectors, and data analysis software, continue to enhance its capabilities and applicability. This abstract aims to elucidate the pivotal role of HPLC in analytical chemistry, highlighting its significance in scientific research, quality control, and industrial applications, while also discussing future perspectives and emerging trends in the field.
New Trends in Fast Liquid Chromatography
CHIMIA International Journal for Chemistry, 2007
Analytical laboratories are currently interested in enhancing overall productivity by increasing sample throughput and reducing analysis time. Different approaches are proposed in liquid chromatography (LC) to perform fast or ultra-fast separations with cycle times of less than 5 or 1 min, respectively. Among these approaches, the use of monolithic supports, high temperature LC (HTLC), short columns and ultra-performance LC (UPLC) are described and compared in this study. A comparison of the above LC approaches is presented through Knox curves and pressure plots based on experimental data. Fast separations of pharmaceutical compounds are presented in order to illustrate the interest of these techniques and compare them with conventional LC separations.
Journal of Chromatography A, 2012
The separation range of superficially porous particles (Fused-core®) has been extended by design of particles with 160 Å pores. These particles show superior kinetics (lower resistance to mass transfer), allowing fast separations of peptides and small proteins (molecular weights of <15,000). The high efficiency and relatively low back pressure of these 2.7 μm fused-core particles has been maintained so that separations can be performed with conventional HPLC instruments. Longer columns can be used for higher resolution of complex mixtures of peptides, such as proteolytic digests. Highly reproducible separations of peptides at elevated temperatures with low pH mobile phases are maintained as a result of a stable bonded stationary phase. The utility of such highly stable materials is exemplified by separations of problematic amyloid peptides at low pH (TFA mobile phase) at an operational temperature of 100 °C. To address the issue of poor peptide peak shape in formic acid-containing mobile phases we show that the addition of 10-20 mM ammonium formate improves peak shape, retention and load tolerance of peptides. Use of the Fused-core particle materials for separations of synthetic peptides and tryptic digests yields peak capacities that are comparable to those obtained using columns packed with sub-2-μm particles, but with less than one-half of the operating back pressure. A peak capacity of 530 was obtained in 150 minutes on coupled columns of HALO Peptide ES-C18 with a combined length of 250 mm.
High-Performance Liquid Chromatography: Comprehensive Techniques and Cutting-Edge Innovations
European Journal of Advances in Engineering and Technology, 2023
High-Performance Liquid Chromatography (HPLC) has emerged as a pivotal analytical technique, revolutionizing the field of chemical analysis through its precision, versatility, and efficiency. This paper provides a comprehensive overview of HPLC, detailing its fundamental principles, advanced techniques, and wide-ranging applications. We delve into the core components and operational mechanisms of HPLC, elucidating the intricacies of column selection, mobile phase composition, and detection methods. Furthermore, the discussion extends to recent technological innovations that enhance HPLC performance, such as ultra-high-performance liquid chromatography (UHPLC), multidimensional chromatography, and novel stationary phases. The practical applications of HPLC in pharmaceuticals, environmental analysis, food safety, and biomedical research are explored, demonstrating its critical role in ensuring quality control, regulatory compliance, and scientific discovery. By examining both the theoretical underpinnings and cutting-edge advancements, this paper aims to provide a holistic understanding of HPLC, highlighting its continued importance and potential for future developments in analytical science.
SilicaROD� � A new challenge in fast high-performance liquid chromatography separations
Trac, 1998
High performance liquid chromatography (HPLC) has become one of the most used methods for the analysis of compound mixtures in industry, especially for the quality control of products. Nowadays productivity is the major and dominant upcoming issue, i.e. the goal is to drastically reduce the analysis time and cost per analysis. The solution of the task is higher throughput and faster HPLC methods. Here we describe a new monolithic type of HPLC column, the SilicaROD™ column, which permits the fast HPLC separation of compound mixtures within a few minutes.
Perspectives on Recent Advances in the Speed of High-Performance Liquid Chromatography
Analytical Chemistry, 2011
Perhaps the most consistent trend in the development of HPLC since its inception in the 1960's has been the continuing reach for ever faster analyses. The pioneering work of Knox, Horvath, Halasz, and Guiochon set forth a theoretical framework that was used early on to improve the speed of HPLC, primarily through the commercialization of smaller and smaller particles. Over the past decade, approaches to improving the speed of HPLC have become more diverse, and now practitioners of HPLC are faced with the difficult task of deciding which of these approaches will lead them to the fastest analysis for their application. Digesting the rich literature on the optimization of HPLC is a difficult task in itself, which is further complicated by contradictory marketing messages from competing commercial outlets for HPLC technology. In this perspectives article we provide an overview of the theoretical and practical aspects of the principal modern approaches to improving the speed of HPLC. We present a straightforward theoretical basis, informed by decades of literature on the problem of optimization, that is useful for comparing different technologies for improving the speed of HPLC. Through mindful optimization of conditions high performance separations on the sub-minute timescale are now possible and becoming increasingly common under both isocratic and gradient elution conditions, and the continued development of ultrafast separations will play an important role in the development of two-dimensional HPLC separations. Despite the relatively long history of HPLC as an analytical technique, there is no sign of a slowdown in the development of novel HPLC technologies.
Core-shell particles lead the way to renewing high-performance liquid chromatography
TrAC Trends in Analytical Chemistry, 2015
In this review, we present the latest highlights and trends in the use of core-shell particles as stationary phases in columns for liquid chromatography (LC). These highly homogeneous particles have a diameter of 1.3-5 μm with a solid silica core and a porous surface that make their performance excellent compared to fully-porous particles. Use of these columns has been growing exponentially since their first largescale manufacture in 2006, and they are an emerging trend in the analysis of biological, toxicological and pharmaceutically interesting compounds. We review the main theoretical aspects responsible for their surprisingly good chromatographic behavior and analytical features. We also summarize state-ofthe-art analytical applications taking advantage of this column technology that is leading the current revolution in LC.
Polymers
Separation with high efficiency and good resolution is constantly in demand in the pharmaceutical industry. The fast and efficient separation of complex samples such as peptides and proteins is a challenging task. To achieve high efficiency with good resolution, chromatographers are moving towards small particles packed into narrow-bore columns. Silica monolith particles (sub-2 µm) were derivatized with chlorodimethyl octadecyl silane (C18) and packed into stainless steel columns (100 mm × 1.8 mm i.d) by a slurry-packing method. The developed columns were used for the separation of peptides and proteins. A separation efficiency (N) of 40,000 plates/column (400,000 plates/m) was achieved for the mixture of five peptides. Similarly, the fast separation of the peptides was carried out using a high flow rate, and the separation of the five peptides was achieved in one minute with high efficiency (N ≅ 240,000 plates/m). The limit of detection (DL) and the limit of quantification (QL) for...
Journal of Separation Science, 2007
The application of small porous particles, high temperatures, and high pressures to generate very high resolution LC and LC/MS separations The effect of combining sub-2 lm porous particles with elevated operating temperatures on chromatographic performance has been investigated in terms of chromatographic efficiency, productivity, peak elution order, and observed operating pressure. The use of elevated temperature in LC does not increase the obtainable performance but allows the same performance to be obtained in less time. Increasing the column temperature did allow the use of longer columns, generating column efficiencies in excess of 100 000 plates and gradient peak capacities approaching 1000. Raising the temperature increased the optimal mobile phase linear velocity, negating somewhat the pressure benefits observed by reducing the solvent viscosity. When operating at higher temperature the analyte retention is not only reduced, but the order of elution will also often change. High temperature separations allowed exotic organic modifiers such as isopropanol to be exploited for alternative selectivity and faster analysis. Finally, care must be taken when using high temperature separations to ensure that the narrow peak widths produced do not compromise the quality of data obtained from detectors such as high resolution mass spectrometers.