This document provides an introduction and review of ultra performance liquid chromatography (UPLC). UPLC uses sub-2 micrometer particles and high pressures to achieve dramatic increases in resolution, sensitivity, and speed compared to traditional HPLC. To fully realize the benefits of small particles, instrumentation had to be developed that can operate at pressures over 15,000 psi with minimal band broadening from injection, pumping, or detection. The first commercially available UPLC system was able to separate pharmaceutical compounds faster than HPLC.
UPLC is a chromatographic technique that can separate mixtures faster and with higher resolution than HPLC using columns packed with 1.7μm particles operating at pressures up to 15,000 psi. It provides increased speed, sensitivity, and resolution for applications in various fields including drug discovery, toxicology, and cancer research. Key components of a UPLC system include binary solvent manager pumps, sample manager, column heater, and detectors. UPLC's use of small particle sizes and high pressures allows for rapid analysis times, less solvent usage, and improved performance over traditional HPLC.
Ultra pressure liquid chromatography(uplc)Pradeep Pal
Ultra performance liquid chromatography (UPLC) is an LC technique that uses sub-2 μm particle columns to improve speed, resolution, and sensitivity compared to HPLC. UPLC works on the principle that decreasing particle size increases efficiency according to the van Deemter equation. The instrumentation includes injection systems for low volume samples, 1.7μm particle packed columns, high pressure pumps, and detectors like UV/VIS, RI, and fluorescence. UPLC provides faster run times, uses less solvent, and maintains resolution compared to HPLC. Applications include analysis of natural products, metabolites, bioanalysis, impurity profiling, and dissolution testing.
This document provides an overview of ultra-performance liquid chromatography (UPLC). UPLC utilizes columns packed with 1-2 micrometer particles and operates at ultra-high pressures to improve resolution, speed, and sensitivity compared to traditional high performance liquid chromatography. Key aspects of UPLC instrumentation include high-pressure pumps, optimized sample injection, and specialized sub-2 micrometer particle columns. UPLC provides benefits like reduced run times, lower solvent consumption, and faster analytical results. It has applications in areas like natural products analysis, metabolite identification, and dissolution testing.
This document discusses Ultra Performance Liquid Chromatography (UPLC). UPLC uses smaller particle sizes of 1.7μm in columns compared to 4μm in HPLC, which allows for faster separations, higher resolution, and shorter run times. UPLC provides improvements in speed, sensitivity and resolution compared to HPLC. It also uses higher pressures up to 15,000 psi compared to HPLC. The document compares various parameters of HPLC and UPLC and discusses their instrumentation including columns, pumps, detectors and applications in pharmaceutical analysis.
UHPLC/UPLC: Ultra High Performance Liquid ChromatographyDarewin Mendonsa
Chromatography Techniques mainly include two basic sub-divisions: Separation Science and Analytical Science.
In 2004, separation science was revolutionized with the introduction of ‘Ultra High-Performance Liquid Chromatography which provides improved resolution, increased separation efficiency, shorter analysis time and lower operating costs.
It uses HPLC columns with a mean particle size less than 2μm and pressures up to 15,000 psi which drastically increases the number of theoretical plates of the column and results in enhanced column efficiency.
Ultra high performance liquid chromatography (UHPLC) provides faster, more sensitive and higher resolution separations compared to traditional high performance liquid chromatography (HPLC). UHPLC uses columns packed with smaller particles less than 2um in diameter which allows for higher pressures and flow rates. This leads to significantly shorter run times, lower detection limits, and better resolution of peaks. The key components of a UHPLC system include pumps that can handle higher pressures, injection systems with low dwell volumes, specialized columns, and detectors capable of measuring small changes. UHPLC has applications in areas like pharmaceutical analysis, metabolomics, and impurity profiling where high resolution and sensitivity are important.
This document summarizes recent advances in HPLC and GC techniques presented by Sajan Maharjan. It describes Ultra Performance Liquid Chromatography (UPLC) which achieves higher resolution, speed, sensitivity and separation efficiency than HPLC using columns with smaller particles (<1.7μm) and higher mobile phase pressures (>15,000psi). Two-dimensional GC (GCxGC) is discussed, which uses two columns in series with a modulator to further separate complex mixtures. A miniaturized GC system is also presented, featuring low power requirements, direct injection, portable design, and the ability to use room air as the carrier gas.
UPLC is an improved version of HPLC that provides higher resolution, speed, and sensitivity. It uses smaller particle sizes of 1.7μm in its columns compared to 4μm in HPLC columns. This allows for faster separations using shorter columns or higher flow rates. UPLC also uses less solvent and reduces analysis times. It has various applications like analysis of natural products, metabolites, bioanalysis, ADME screening, dissolution testing, method development and validation, forced degradation studies, impurity profiling, and analysis in manufacturing and quality control.
UPLC is a chromatographic technique that can separate mixtures faster and with higher resolution than HPLC using columns packed with 1.7μm particles operating at pressures up to 15,000 psi. It provides increased speed, sensitivity, and resolution for applications in various fields including drug discovery, toxicology, and cancer research. Key components of a UPLC system include binary solvent manager pumps, sample manager, column heater, and detectors. UPLC's use of small particle sizes and high pressures allows for rapid analysis times, less solvent usage, and improved performance over traditional HPLC.
Ultra pressure liquid chromatography(uplc)Pradeep Pal
Ultra performance liquid chromatography (UPLC) is an LC technique that uses sub-2 μm particle columns to improve speed, resolution, and sensitivity compared to HPLC. UPLC works on the principle that decreasing particle size increases efficiency according to the van Deemter equation. The instrumentation includes injection systems for low volume samples, 1.7μm particle packed columns, high pressure pumps, and detectors like UV/VIS, RI, and fluorescence. UPLC provides faster run times, uses less solvent, and maintains resolution compared to HPLC. Applications include analysis of natural products, metabolites, bioanalysis, impurity profiling, and dissolution testing.
This document provides an overview of ultra-performance liquid chromatography (UPLC). UPLC utilizes columns packed with 1-2 micrometer particles and operates at ultra-high pressures to improve resolution, speed, and sensitivity compared to traditional high performance liquid chromatography. Key aspects of UPLC instrumentation include high-pressure pumps, optimized sample injection, and specialized sub-2 micrometer particle columns. UPLC provides benefits like reduced run times, lower solvent consumption, and faster analytical results. It has applications in areas like natural products analysis, metabolite identification, and dissolution testing.
This document discusses Ultra Performance Liquid Chromatography (UPLC). UPLC uses smaller particle sizes of 1.7μm in columns compared to 4μm in HPLC, which allows for faster separations, higher resolution, and shorter run times. UPLC provides improvements in speed, sensitivity and resolution compared to HPLC. It also uses higher pressures up to 15,000 psi compared to HPLC. The document compares various parameters of HPLC and UPLC and discusses their instrumentation including columns, pumps, detectors and applications in pharmaceutical analysis.
UHPLC/UPLC: Ultra High Performance Liquid ChromatographyDarewin Mendonsa
Chromatography Techniques mainly include two basic sub-divisions: Separation Science and Analytical Science.
In 2004, separation science was revolutionized with the introduction of ‘Ultra High-Performance Liquid Chromatography which provides improved resolution, increased separation efficiency, shorter analysis time and lower operating costs.
It uses HPLC columns with a mean particle size less than 2μm and pressures up to 15,000 psi which drastically increases the number of theoretical plates of the column and results in enhanced column efficiency.
Ultra high performance liquid chromatography (UHPLC) provides faster, more sensitive and higher resolution separations compared to traditional high performance liquid chromatography (HPLC). UHPLC uses columns packed with smaller particles less than 2um in diameter which allows for higher pressures and flow rates. This leads to significantly shorter run times, lower detection limits, and better resolution of peaks. The key components of a UHPLC system include pumps that can handle higher pressures, injection systems with low dwell volumes, specialized columns, and detectors capable of measuring small changes. UHPLC has applications in areas like pharmaceutical analysis, metabolomics, and impurity profiling where high resolution and sensitivity are important.
This document summarizes recent advances in HPLC and GC techniques presented by Sajan Maharjan. It describes Ultra Performance Liquid Chromatography (UPLC) which achieves higher resolution, speed, sensitivity and separation efficiency than HPLC using columns with smaller particles (<1.7μm) and higher mobile phase pressures (>15,000psi). Two-dimensional GC (GCxGC) is discussed, which uses two columns in series with a modulator to further separate complex mixtures. A miniaturized GC system is also presented, featuring low power requirements, direct injection, portable design, and the ability to use room air as the carrier gas.
UPLC is an improved version of HPLC that provides higher resolution, speed, and sensitivity. It uses smaller particle sizes of 1.7μm in its columns compared to 4μm in HPLC columns. This allows for faster separations using shorter columns or higher flow rates. UPLC also uses less solvent and reduces analysis times. It has various applications like analysis of natural products, metabolites, bioanalysis, ADME screening, dissolution testing, method development and validation, forced degradation studies, impurity profiling, and analysis in manufacturing and quality control.
This document compares HPLC and UPLC and provides an overview of UPLC. UPLC uses smaller particle sizes of 1.75-1.8 μm compared to 3-10 μm for HPLC, which allows for higher pressures over 15,000 psi, improved precision, faster analysis times, and higher resolution. The key factors that enable improved performance in UPLC are increased efficiency through smaller particle sizes and improved selectivity, retentivity, and efficiency. UPLC also provides increased sensitivity, faster run times, and the ability to analyze more samples due to its enhanced resolving power. The document discusses UPLC stationary phases, columns, instrumentation including pumps, columns, and detectors, as well as applications of UPLC in areas like method
UPLC provides faster, more sensitive and higher resolution chromatography compared to HPLC. It uses smaller particle sizes (<2um) which allows for better separation of compounds at higher pressures. This leads to reduced analysis times, solvent usage and improved productivity. Key aspects of UPLC include specialized instrumentation like injection valves and detectors adapted for high pressure, as well as shorter narrow-bore columns packed with smaller particles. It has applications in fields like pharmaceutical analysis, food testing and forensic toxicology.
Ultra Performance Liquid Chromatography (UPLC) is an analytical technique that improves chromatographic resolution, speed, and sensitivity compared to traditional HPLC. It uses columns packed with smaller particles less than 2.5 μm. This allows for higher pressures, faster flow rates, and shorter run times. UPLC provides increased peak capacity and sensitivity for applications like metabolite identification, impurity profiling, and dissolution testing in pharmaceutical analysis and quality control.
Ultra Performance Liquid Chromatography (UPLC) is a newer chromatographic technique that uses smaller particle sizes (1.7-1.8 μm) and higher pressures (>1000 bar) than High Performance Liquid Chromatography (HPLC) to improve resolution, sensitivity, and speed of analysis. UPLC provides faster separations, higher peak capacity, and uses less solvent than HPLC. It allows for more complex sample separations to be achieved in shorter time frames. UPLC instrumentation is designed to withstand the higher back pressures and uses specialized analytical columns packed with smaller 1.7 μm particles.
This presentation covers an introduction to UPLC, its general chemistry, and laws behind it. It also discusses the instrumentation of UPLC, advantages, disadvantages, and application of UPLC.
This document provides an overview of ultra performance liquid chromatography (UPLC). It begins with an introduction to chromatography and defines UPLC. The main advantages of UPLC are listed as decreased run time, increased sensitivity, and maintaining resolution. The document outlines the basic principles, instrumentation, and differences between UPLC and HPLC. Applications discussed include drug discovery, herbal medicine analysis, and metabolomics studies. The document is presented by Mr. Atish Khilari and provides references for additional information.
UPLC uses smaller particle sizes (<2 microns) than HPLC (3-5 microns) which allows it to operate at higher pressures and provide faster, more sensitive and selective separations compared to HPLC. Some key differences are that UPLC uses smaller diameter columns, achieves higher resolution and plate counts, reduces run times, solvent consumption and cost of operation compared to HPLC. However, the smaller particles used in UPLC columns have a narrower usable lifespan than HPLC columns.
ultra high performance liquid chromatographyacademic
This document provides an overview of ultra high performance liquid chromatography (UHPLC). It begins with an introduction defining UHPLC and explaining that it improves resolution, speed and sensitivity compared to traditional HPLC. The document then covers the principles of UHPLC, provides a comparison of UHPLC and HPLC parameters, describes UHPLC instrumentation including pumps, columns and detectors, and discusses applications and advantages/disadvantages of UHPLC.
Hplc instrumentation in detail (Practical) Hplc pump inj_columnPratikShinde120
This document discusses HPLC instrumentation and techniques. It describes the key components of an HPLC system including the solvent delivery system, pumps, sample introduction methods, and detectors. For solvent delivery, it explains the mobile phase reservoirs, degassing, and tubing used. It discusses different types of pumps like reciprocating, syringe, and dual piston pumps. For sample introduction, it covers manual injection methods like septum and valve, as well as automated injection. It also provides details on various detectors like UV-Vis, fluorescence, refractive index, and conductivity.
UPLC provides improved resolution, speed, and sensitivity compared to HPLC by using smaller particle sizes of 1.7μm instead of 3-5μm. This leads to higher column efficiency and lower retention times while maintaining resolution. UPLC utilizes the same principles as HPLC but with smaller particles, increased efficiency, and faster analysis times. It has various applications like pharmaceutical development, metabolite identification, bioanalysis, impurity profiling, and stressed degradation studies due to its high resolution and sensitivity.
UPLC provides faster, more sensitive chromatographic separations compared to HPLC. It works by using smaller particle sizes (<2.5um) in the column packing which allows for higher pressure and flow rates based on the van Deemter equation. This provides benefits like reduced run times, decreased sample volume needs, and improved resolution. However, it also requires more robust instrumentation to handle the increased pressures and columns have reduced lifespan. UPLC has applications in fields like pharmaceutical analysis, metabolomics, and impurity profiling due to its enhanced resolution and sensitivity capabilities.
This document discusses nano liquid chromatography. It begins by describing different types of liquid chromatography techniques including rapid resolution liquid chromatography, ultra performance liquid chromatography, ultra fast liquid chromatography, and nano liquid chromatography. It then focuses on nano liquid chromatography, explaining that it uses capillary columns with diameters of 75 micrometers or less and flow rates between 10-700 nanoliters per minute. The document outlines the advantages of nano LC including reduced solvent usage and improved sensitivity. It also reviews the instrumentation involved including pumps, injection systems, columns, and detectors such as diode array detection. Finally, it discusses applications of nano LC in fields like food analysis and proteomics and concludes by noting its increasing use.
This document provides an overview of high performance liquid chromatography (HPLC). It discusses various modes of separation in HPLC including normal phase, reversed phase, ion exchange, and size exclusion chromatography. The document also describes HPLC instrumentation components such as solvent delivery systems, pumps, sample injection systems, chromatographic columns, and detectors. It provides details on the development, optimization, and validation of HPLC methods.
The document presents information on a seminar about Ultra Performance Liquid Chromatography (UPLC). It discusses the principles, instrumentation, advantages, and applications of UPLC. UPLC uses columns packed with particles less than 2 μm to provide improved resolution, speed, and sensitivity compared to traditional HPLC. The seminar covers topics like pump design to withstand high pressures, types of detectors used, applications in pharmaceutical analysis and natural products, and advantages like faster run times and higher sensitivity.
This document discusses new developments in high performance liquid chromatography (HPLC) and the role of ultra performance liquid chromatography (UPLC) and nano liquid chromatography in pharmaceutical analysis. It begins with an introduction to HPLC and lists some new developments including UHPLC, HPLC-MS systems, and HPLC chips. It then focuses on UPLC, explaining that it can acquire results faster with more resolution compared to traditional HPLC. UPLC allows for more samples to be analyzed per system and scientist. The document concludes by listing references for further information.
Many people pursue ideas of “efficiency” as an ideal for daily life; the same can be true in the HPLC laboratory. In this work, we demonstrate the efficiency, throughput, and reliability of a dual injection system for finished pharmaceutical products and in-process active pharmaceutical ingredients
UPLC provides faster, more sensitive and efficient separations compared to HPLC by using sub-2 μm particles. It operates at higher pressures of up to 100 MPa. Smaller particle sizes and columns allow for shorter run times, lower detection limits, less solvent usage and improved resolution. Key aspects of UPLC instrumentation include high-pressure binary pumps, low-volume injection systems, 1.7 μm particle columns, and detectors adapted for smaller flow cells like tunable UV detectors. UPLC finds applications in pharmaceutical analysis, natural products analysis, and metabolomics studies.
This document provides an overview of ultra performance liquid chromatography (UPLC). UPLC uses sub-2 μm particles and operates at higher pressures than HPLC to dramatically increase resolution, sensitivity, and speed of analysis while maintaining practicality. Key advantages of UPLC include decreased run times, increased sensitivity and throughput, and reduced solvent consumption compared to HPLC. The document discusses UPLC instrumentation including pumps, injectors, columns, detectors, and applications such as amino acid analysis and analysis of natural products and herbal medicines.
This document compares HPLC and UPLC and provides an overview of UPLC. UPLC uses smaller particle sizes of 1.75-1.8 μm compared to 3-10 μm for HPLC, which allows for higher pressures over 15,000 psi, improved precision, faster analysis times, and higher resolution. The key factors that enable improved performance in UPLC are increased efficiency through smaller particle sizes and improved selectivity, retentivity, and efficiency. UPLC also provides increased sensitivity, faster run times, and the ability to analyze more samples due to its enhanced resolving power. The document discusses UPLC stationary phases, columns, instrumentation including pumps, columns, and detectors, as well as applications of UPLC in areas like method
UPLC provides faster, more sensitive and higher resolution chromatography compared to HPLC. It uses smaller particle sizes (<2um) which allows for better separation of compounds at higher pressures. This leads to reduced analysis times, solvent usage and improved productivity. Key aspects of UPLC include specialized instrumentation like injection valves and detectors adapted for high pressure, as well as shorter narrow-bore columns packed with smaller particles. It has applications in fields like pharmaceutical analysis, food testing and forensic toxicology.
Ultra Performance Liquid Chromatography (UPLC) is an analytical technique that improves chromatographic resolution, speed, and sensitivity compared to traditional HPLC. It uses columns packed with smaller particles less than 2.5 μm. This allows for higher pressures, faster flow rates, and shorter run times. UPLC provides increased peak capacity and sensitivity for applications like metabolite identification, impurity profiling, and dissolution testing in pharmaceutical analysis and quality control.
Ultra Performance Liquid Chromatography (UPLC) is a newer chromatographic technique that uses smaller particle sizes (1.7-1.8 μm) and higher pressures (>1000 bar) than High Performance Liquid Chromatography (HPLC) to improve resolution, sensitivity, and speed of analysis. UPLC provides faster separations, higher peak capacity, and uses less solvent than HPLC. It allows for more complex sample separations to be achieved in shorter time frames. UPLC instrumentation is designed to withstand the higher back pressures and uses specialized analytical columns packed with smaller 1.7 μm particles.
This presentation covers an introduction to UPLC, its general chemistry, and laws behind it. It also discusses the instrumentation of UPLC, advantages, disadvantages, and application of UPLC.
This document provides an overview of ultra performance liquid chromatography (UPLC). It begins with an introduction to chromatography and defines UPLC. The main advantages of UPLC are listed as decreased run time, increased sensitivity, and maintaining resolution. The document outlines the basic principles, instrumentation, and differences between UPLC and HPLC. Applications discussed include drug discovery, herbal medicine analysis, and metabolomics studies. The document is presented by Mr. Atish Khilari and provides references for additional information.
UPLC uses smaller particle sizes (<2 microns) than HPLC (3-5 microns) which allows it to operate at higher pressures and provide faster, more sensitive and selective separations compared to HPLC. Some key differences are that UPLC uses smaller diameter columns, achieves higher resolution and plate counts, reduces run times, solvent consumption and cost of operation compared to HPLC. However, the smaller particles used in UPLC columns have a narrower usable lifespan than HPLC columns.
ultra high performance liquid chromatographyacademic
This document provides an overview of ultra high performance liquid chromatography (UHPLC). It begins with an introduction defining UHPLC and explaining that it improves resolution, speed and sensitivity compared to traditional HPLC. The document then covers the principles of UHPLC, provides a comparison of UHPLC and HPLC parameters, describes UHPLC instrumentation including pumps, columns and detectors, and discusses applications and advantages/disadvantages of UHPLC.
Hplc instrumentation in detail (Practical) Hplc pump inj_columnPratikShinde120
This document discusses HPLC instrumentation and techniques. It describes the key components of an HPLC system including the solvent delivery system, pumps, sample introduction methods, and detectors. For solvent delivery, it explains the mobile phase reservoirs, degassing, and tubing used. It discusses different types of pumps like reciprocating, syringe, and dual piston pumps. For sample introduction, it covers manual injection methods like septum and valve, as well as automated injection. It also provides details on various detectors like UV-Vis, fluorescence, refractive index, and conductivity.
UPLC provides improved resolution, speed, and sensitivity compared to HPLC by using smaller particle sizes of 1.7μm instead of 3-5μm. This leads to higher column efficiency and lower retention times while maintaining resolution. UPLC utilizes the same principles as HPLC but with smaller particles, increased efficiency, and faster analysis times. It has various applications like pharmaceutical development, metabolite identification, bioanalysis, impurity profiling, and stressed degradation studies due to its high resolution and sensitivity.
UPLC provides faster, more sensitive chromatographic separations compared to HPLC. It works by using smaller particle sizes (<2.5um) in the column packing which allows for higher pressure and flow rates based on the van Deemter equation. This provides benefits like reduced run times, decreased sample volume needs, and improved resolution. However, it also requires more robust instrumentation to handle the increased pressures and columns have reduced lifespan. UPLC has applications in fields like pharmaceutical analysis, metabolomics, and impurity profiling due to its enhanced resolution and sensitivity capabilities.
This document discusses nano liquid chromatography. It begins by describing different types of liquid chromatography techniques including rapid resolution liquid chromatography, ultra performance liquid chromatography, ultra fast liquid chromatography, and nano liquid chromatography. It then focuses on nano liquid chromatography, explaining that it uses capillary columns with diameters of 75 micrometers or less and flow rates between 10-700 nanoliters per minute. The document outlines the advantages of nano LC including reduced solvent usage and improved sensitivity. It also reviews the instrumentation involved including pumps, injection systems, columns, and detectors such as diode array detection. Finally, it discusses applications of nano LC in fields like food analysis and proteomics and concludes by noting its increasing use.
This document provides an overview of high performance liquid chromatography (HPLC). It discusses various modes of separation in HPLC including normal phase, reversed phase, ion exchange, and size exclusion chromatography. The document also describes HPLC instrumentation components such as solvent delivery systems, pumps, sample injection systems, chromatographic columns, and detectors. It provides details on the development, optimization, and validation of HPLC methods.
The document presents information on a seminar about Ultra Performance Liquid Chromatography (UPLC). It discusses the principles, instrumentation, advantages, and applications of UPLC. UPLC uses columns packed with particles less than 2 μm to provide improved resolution, speed, and sensitivity compared to traditional HPLC. The seminar covers topics like pump design to withstand high pressures, types of detectors used, applications in pharmaceutical analysis and natural products, and advantages like faster run times and higher sensitivity.
This document discusses new developments in high performance liquid chromatography (HPLC) and the role of ultra performance liquid chromatography (UPLC) and nano liquid chromatography in pharmaceutical analysis. It begins with an introduction to HPLC and lists some new developments including UHPLC, HPLC-MS systems, and HPLC chips. It then focuses on UPLC, explaining that it can acquire results faster with more resolution compared to traditional HPLC. UPLC allows for more samples to be analyzed per system and scientist. The document concludes by listing references for further information.
Many people pursue ideas of “efficiency” as an ideal for daily life; the same can be true in the HPLC laboratory. In this work, we demonstrate the efficiency, throughput, and reliability of a dual injection system for finished pharmaceutical products and in-process active pharmaceutical ingredients
UPLC provides faster, more sensitive and efficient separations compared to HPLC by using sub-2 μm particles. It operates at higher pressures of up to 100 MPa. Smaller particle sizes and columns allow for shorter run times, lower detection limits, less solvent usage and improved resolution. Key aspects of UPLC instrumentation include high-pressure binary pumps, low-volume injection systems, 1.7 μm particle columns, and detectors adapted for smaller flow cells like tunable UV detectors. UPLC finds applications in pharmaceutical analysis, natural products analysis, and metabolomics studies.
This document provides an overview of ultra performance liquid chromatography (UPLC). UPLC uses sub-2 μm particles and operates at higher pressures than HPLC to dramatically increase resolution, sensitivity, and speed of analysis while maintaining practicality. Key advantages of UPLC include decreased run times, increased sensitivity and throughput, and reduced solvent consumption compared to HPLC. The document discusses UPLC instrumentation including pumps, injectors, columns, detectors, and applications such as amino acid analysis and analysis of natural products and herbal medicines.
Enhancing Sensitivities and Peak Capacities for UHPLC-MS Fast Gradient Analys...Sandy Simmons
When compared to 1.7 μm fully porous materials, the ultra-high
efficiency and low backpressures provided by Kinetex core-shell
2.6 μm columns, provides users opportunities to go beyond what
is traditionally accepted for UHPLC runs
The document evaluates using the Halo TM fused-core column to reduce bioanalytical run times for pharmacokinetic screening samples from 4 minutes to 2.25 minutes. This faster analysis allowed an entire pharmacokinetic study involving 72 plasma samples and 4 brain samples to be analyzed in 5.5 hours instead of longer. The Halo column provided consistent results with no significant suppression and allowed a 40% reduction in run time without new equipment.
HPTLC is an improved version of TLC that provides better resolution and allows for quantitative analysis. It uses plates with finer silica gel particles between 5-7 micrometers compared to 10-25 micrometers for regular TLC. This allows for faster development times of 3-20 minutes for HPTLC versus 30-200 minutes for TLC. HPTLC also has automated instrumentation for precise sample application and development as well as densitometric scanning for quantification. It has various applications in pharmaceutical analysis, clinical analysis, food and environmental testing by providing fingerprints to identify compounds and allowing quantification of biomarkers.
Life Cycle Management of Chromatographic Methods for BiopharmaceuticalsWaters Corporation
The development and manufacture of biopharmaceuticals is a dynamic and rapidly growing industry. By the nature of their production, biopharmaceuticals are highly complex heterogeneous mixtures that require many analytical techniques for characterization and routine testing. As a result, many manufacturers incorporate life cycle management into their respective workflows to take advantage of newer technologies and methodologies to ensure efficacy and patient safety.
In this presentation, we will address the range of chromatographic categories – HPLC, UHPLC, and UPLC – and define the characteristics associated with each. The discussion will continue with several examples of methods transferred from legacy HPLC instrumentation to modern UHPLC and UPLC instruments. We will compare qualitative and quantitative data across each chromatographic class. Resolution, sensitivity, and overall run time will be used as metrics to assess the success of the method transfer to the respective LC platform, to ensure the transferred methods are in line with current acceptance criteria.
Learn:
- The importance of selecting the correct instrumentation to meet user needs.
- Which parameters influence method transfer from one LC platform to another.
- How workflows can benefit from features such as Multi-flow path technology and Gradient SmartStart when transitioning methods.
Interested in more detail? Watch the related on-demand webinar: http://view6.workcast.net/register?pak=3479247014905635
Webinar - Pharmacopeial Modernization: How Will Your Chromatography Workflow ...Waters Corporation
In this webinar, Dr. Leonel Santos and Dr. Horacio Pappa from the United States Pharmacopeia (USP) will provide an overview of its pharmacopeial harmonization and modernization efforts. The pair will also review changes described in the pending USP General Chapter <621> on liquid chromatography (LC), which will provide increased flexibility for gradient methods.
Amanda Dlugasch, from Waters Corporation, will follow with an illuminating case study, which leverages USP <621> allowable adjustments to illustrate the benefits of modernizing methods, including migrating HPLC methods to UHPLC or UPLC, without the need to revalidate.
Topics covered in this webinar will include:
- Pharmacopeial monograph modernization prioritization scheme
- Review of USP General Chapter <621> current allowable adjustments to validated chromatographic methods and forthcoming updates
- Case study on the migration of isocratic and gradient pharmacopeial methods to modern chromatography column technology, highlighting improved method performance and throughput
Replay the webinar, hosted by SelectScience:
https://www.selectscience.net/webinars/pharmacopeial-modernization-how-will-your-chromatography-workflow-benefit/?webinarID=1228
UPLC is an advance analytical technique where it takes advantage of innovation in various technologies such as instrumentation and particle size to achieve dramatic increases in resolution, speed and sensitivity of the liquid chromatography. It operates at higher pressure than that used in HPLC and uses fine particles (less than 2.5µm) & mobile phases at high linear velocities. It can be hyphenated with other techniques such as (MS), (IC), (NMR) and (IR) etc. This is used in all industries and has found application in various fields such as pharmaceutical, food, environmental, forensic, toxicology and pesticide.
This document summarizes a student's research project analyzing the quantitative analysis of the drug lamotrigine using high performance liquid chromatography (HPLC). The student aims to partially validate an HPLC technique for quantifying lamotrigine. The document provides background on HPLC, including its history, theory of operation, instrumentation such as columns, detectors, and pumps. It also discusses validation components like accuracy, precision, linearity and limits of detection and quantification that the student will analyze to partially validate the HPLC method for quantifying lamotrigine.
Plants contain thousands of compounds and are a valuable source of new drugs. High-performance thin-layer chromatography (HPTLC) is a simple and economical analytical method useful for characterizing herbal compounds. HPTLC provides better separation and repeatability compared to traditional thin-layer chromatography. It can be used to develop standardized herbal extracts, isolate pure compounds, and determine compound structures. HPTLC is also used to create chemical "fingerprints" of herbal products for quality evaluation and authentication. The HPTLC process involves selecting a stationary phase, applying samples, developing the plate, detecting and quantifying separated compounds, and documenting results.
Liquid chromatography still striving for high efficiencyguest63ff7d
The document discusses liquid chromatography and approaches to improve its efficiency. It provides an overview of the chromatographic process and factors that affect column efficiency such as mobile phase velocity, temperature, column length, particle size. Existing approaches to improve efficiency include high temperature liquid chromatography, monolithic stationary phases, fused-core particles, and sub-2-micron totally porous particles. Overall miniaturization shows potential for future efficiency gains in liquid chromatography.
We present a micro fluidic device that allows rapid and defined delivery of discrete and homogeneous reagents or samples to allow kinetic studies of surface-tethered biomolecules. We developed an Electro Osmotic Flow (EOF) based device consisting of an asymmetric Y-junction with an incorporated fast, pressure driven valve, two embedded measurement electrodes and reservoirs. The EOF is used to circumvent kinetic limitations on reagent transport to the surface of these tethered biomolecules due to the slow diffusion across parabolic concentration gradients in conventional pressure-driven flow devices. Using Fluorescein isothiocyanate (FITC) as the pH sensitive surface-immobilized biomolecules, we show that the reagent solution can be repeatedly exchanged within milliseconds, and that by using a synchronized triggering scheme we can monitor the reaction of our sensor biomolecules to the change of the environment on a time scale of 10ms.
36433 Topic HA W9 R1Number of Pages 1 (Double Spaced)N.docxrhetttrevannion
36433 Topic: HA W9 R1
Number of Pages: 1 (Double Spaced)
Number of sources: 2
Writing Style: APA
Type of document: Essay
Academic Level:Master
Category: Nursing
Language Style: English (U.S.)
Order Instructions: Attached
Shawna Harris
Wednesday Mar 6 at 3:38pm
Manage Discussion Entry
The U.S. Preventative Services Task Force has prostate screening recommendations. The U.S. Preventative Services Task Force suggests discussing with the patient the benefits and possible harm from obtaining a prostate specific antigen, PSA test (U.S. Preventative Services Task Force). There is a small percent of people for whom this test can correctly identify and thus reduce the risk of mortality from prostate cancer (U.S. Preventative Services Task Force). However, this test can often have false positives, which could result in obtaining an unnecessary biopsy (U.S. Preventative Services Task Force). Invasive procedures, such as biopsies always have risk factors of their own. Consequently, the U.S. Preventative Task Force recommends a PSA screening test for men ages fifty-five to sixty-nine, only if the patient is requesting this screening even after discussing benefits and possible harms from testing and biopsy. (U.S. Preventative Services Task Force).
The American Cancer Society, ACS, recommends that men age fifty and over discuss the benefits and risks of screening in order to make an informed decision with his provider (Wolf, Wender, Etzioni,….& Smith, 2010). ACS also recommends if a man is at a high risk, that this information is presented earlier than fifty (Wolf et al…2010). Those at a higher risk include African American men with a family history of prostate cancer occurring in a family member who is not elderly (Wolf et al…2010). ACS also does not recommend that men whose life expectancy is less than ten years be screened for prostate cancer (Wolf et al…2010). Providers need to provide men with the benefits of early detection and treatment with the risk factors of treatment for prostate cancer. The results of PSA testing are not conclusive and therefore, the ACS reiterates the importance of the patient having the knowledge and information to make an informed decision. The ACS provides educational brochures and handouts on PSA screening to help guide patients to a discussion of this subject with his provider.
References
U.S. Preventative Services Task Force, (Accessed March 2019). Screening Guidelines for Prostate. Retrieved from: https://www.uspreventiveservicestaskforce.org/Page/Document/RecommendationStatementFinal/prostate-cancer-screening1 (Links to an external site.)Links to an external site.
Wolf, A., Wender, R., Etzioni, R….& Smith, R. (2010). American Cancer Society Guideline for the Early Detection of Prostate Cancer: Update 2010. Retrieved from: https://onlinelibrary.wiley.com/doi/full/10.3322/caac.20066 (Links to an external site.)
** Provide response writing with references. All references must be in APA format and p.
Paper presenting practical explanation of kinetic energy separation technolog...Carroll Cobb
process sample conditioning, sample conditioning, sheffield separator, kinetic energy separation, ISA, process sample analyzer, petrochemical process sampling, petrochemical filters, petrochemical sample filters, petrochemical process sample conditioning, kinetic energy separation technology, kinetic energy, genie filter, centrifugal filter, petrochemical process filter, knockout filter, process sample conditioning, process sample filters, immiscible removal, condensate removal, contaminate removal, liquid process sample conditioning, gas process sample conditioning, sample cleaning
1) HPLC provides improved performance over classical column chromatography due to smaller particle sizes (<5 microns), higher operating pressures (>4000 psi), and higher column efficiencies (>100,000 theoretical plates per meter).
2) There are two main modes of HPLC separation - normal phase which uses a polar stationary phase and non-polar mobile phase, and reverse phase which uses a non-polar stationary phase and polar mobile phase.
3) Key components of an HPLC system include pumps to deliver the mobile phase at high pressure, injectors to introduce samples, columns packed with stationary phase to perform the separation, and detectors such as UV/Vis to identify eluted components.
Survey on Declining Curves of Unconventional Wells and Correlation with Key ...Salman Sadeg Deumah
The analysis of the decline curve is applied each year of production which gives the possibility to determine the average decline rate. The calculation of the correlation coefficient gives the possibility to link the different parameters.
New Technique for Measuring and Controlling the Permeability of Polymeric Mem...Editor IJCATR
Membranes have wide uses in industry and medicine applications. Polymer membranes are important materials because of their high chemical resistance, but they are of weak mechanical resistance against high pressures. Therefore, it was essential to modify a permeability measuring technique free from high pressure application. The current work represented a modification for the permeability measuring technique of membranes, where ionic salt was added with known concentration to water as common solvent and the electrolyte current was measured behind the membrane. The electrolysis current was correlated to the flow rate of water across a polyvinyl alcohol (PVA) membrane. Some other problems were raised such that polarization on electrodes and changes in electrolyte contents during the long time of the slow process. Pulsed potential on electrodes resolved these problems and other associated problems like rush in current and the double layer capacitance effect. An empirical equation was suggested to evaluate the permeability of polymer membranes by this modified method. Easy and accurate measurement of permeability helped authors to change the permeability of PVA membranes by adding copper nano particles in membrane to reduce its permeability, and adding silicone dioxide micro particles to the PVA membranes to increase its permeability. Authors suggested a mechanism for these permeability changes. Scanning electron microscope images for the filled PVA membranes supported the suggested mechanism.
New Technique for Measuring and Controlling the Permeability of Polymeric Mem...Editor IJCATR
Membranes have wide uses in industry and medicine applications. Polymer membranes are important materials
because of their high chemical resistance, but they are of weak mechanical resistance against high pressures. Therefore, it was
essential to modify a permeability measuring technique free from high pressure application. The current work represented a
modification for the permeability measuring technique of membranes, where ionic salt was added with known concentration
to water as common solvent and the electrolyte current was measured behind the membrane. The electrolysis current was
correlated to the flow rate of water across a polyvinyl alcohol (PVA) membrane. Some other problems were raised such that
polarization on electrodes and changes in electrolyte contents during the long time of the slow process. Pulsed potential on
electrodes resolved these problems and other associated problems like rush in current and the double layer capacitance effect.
An empirical equation was suggested to evaluate the permeability of polymer membranes by this modified method. Easy and
accurate measurement of permeability helped authors to change the permeability of PVA membranes by adding copper nano
particles in membrane to reduce its permeability, and adding silicone dioxide micro particles to the PVA membranes to
increase its permeability. Authors suggested a mechanism for these permeability changes. Scanning electron microscope
images for the filled PVA membranes supported the suggested mechanism
2. 1254 M. E. Swartz
For many years, researchers have looked at "fast LC" as a way to speed up
analyses.[2'31 The "need for speed" has been driven by the sheer numbers of
samples in some laboratories (particularly in drug discovery) and the
availability of affordable, easy to use mass spectrometers. Smaller columns
and faster flow rates (amongst other parameters) have been used. Elevated
temperature, having the dual advantages of lowering viscosity, and increasing
mass transfer by increasing the diffusivity of the analytes, has also been
investigated.M However, using conventional particle sizes and pressures, limit-
ations are soon reached and compromises must be made, sacrificing resolution
for time.
However, as illustrated in Figure 1, as the particle size decreases to less
than 2.5 urn, not only is there a significant gain in efficiency; but the efficiency
doesn't diminish at increased flow rates or linear velocities. By using
smaller particles, speed and peak capacity (number of peaks resolved per
unit time) can be extended to new limits, termed Ultra Performance Liquid
Chromatography, or UPLC . This introduction and review traces some of
the developments and technological advancements made in producing the
first commercially available UPLC instrument.
SMALL PARTICLE CHEMISTRY
The promises of the van Deemter equation cannot be fulfilled without smaller
particles than those traditionally used in HPLC. The design and development
10 Mm Porticle ,
HPLC
H
E'5
T
P .0
2 3 4 5
f law Kale [ml/mm]:
Linear Velocity [u, mm/sec]
10 - 1.0 mm 0.04 0.07 0.10 0.13 0.17 0.20 0.24
10 = 2.1 mm 0.15 0.3 0.45 0.6 0.75 0.9 1.05
!D » 4.6 mm 0.7 1.4 1.1 2.8 3.5 49
Figure 1. van Deemter plot, illustrating the evolution of particle sizes over the last
three decades.
3. UPLC : An Introduction and Review 1255
of sub-2 |jim particles is a significant challenge, and researchers have been
active in this area for some time to capitalize on their advantages.t5"71
Although high efficiency, non-porous 1.5|j,m particles are commercially
available, they suffer from poor loading capacity and retention due to low
surface area. To maintain retention and capacity similar to HPLC, UPLC
must use novel porous particles that can withstand high pressures. Silica
based particles have good mechanical strength, but can suffer from a
number of disadvantages, which include a limited pH range and tailing of
basic analytes. Polymeric columns can overcome pH limitations, but they
have their own issues, including low efficiencies and limited capacities.
In 2000, a first generation hybrid chemistry that took advantage of the best
of both the silica and polymeric column worlds was introduced.[8'9-1 Produced
using a classical sol-gel synthesis that incorporates carbon in the form of
methyl groups, these columns are mechanically strong, with high efficiency,
and operate over an extended pH range. But, in order to provide the kind of
enhanced mechanical stability required for UPLC, a second generation
bridged ethane hybrid (BEH) technology was developed.[10] These 1.7|JLm
particles derive their enhanced mechanical stability by bridging the methyl
groups in the silica matrix.
Packing 1.7 (Jim particles into reproducible and rugged columns was also
a challenge that needed to be overcome. Requirements include a smoother
interior surface of the column hardware, and re-designing the end frits
to retain the small particles and resist clogging. Packed bed uniformity is
also critical, especially if shorter columns are to maintain resolution while
accomplishing the goal of faster separations.
In addition, at high pressures, frictional heating of the mobile phase can
be quite significant and must be considered.1111 With column diameters
typically used in HPLC (3.0 to 4.6mm), a consequence of frictional heating
is the loss of performance due to temperature induced non uniform flow.
To minimize the effects of frictional heating, smaller diameter columns
(1-2.1 mm) are typically used for UPLC.[12'13]
CAPITALIZING ON SMALLER PARTICLES
Small particles alone do not make it possible to fulfill the promises of the van
Deemter equation (Figure 1). Instrument technology also had to keep pace
to truly take advantage of the increased speed, superior resolution, and
sensitivity afforded by smaller particles. Standard HPLC technology
(pumps, injectors, and detectors) simply doesn't have the horsepower to
take full advantage of sub-2 (Jim particles.
One-of-a-kind systems, capable of delivering the pressures required to
realize the potential of UPLC have been reported in the literature and
elsewhere.114"16]
4. 1256 M. E. Swartz
Lee et al. described the design of injection valves and separation reprodu-
cibility,[I4] and the use of a carbon dioxide enhanced slurry packing method on
the capillary scale for the separation of some benzodiazepines, herbicides, and
various pharmaceutical compounds.1171 Jorgenson et al. modified a commer-
cially available HPLC system to operate at 17,500 psi and used 22 cm long
capillaries packed with 1.5|jim Gig-modified particles for the analysis of
proteins.1-15-1
These reports illustrated that, to take full advantage of low dispersion and
small particle technology to achieve high peak capacity UPLC separations, a
greater pressure range than that achievable by today's HPLC instrumentation
was required. The calculated pressure drop at the optimum flow rate for
maximum efficiency across a 15cm long column packed with 1.7 [Am
particles is about 15,000 psi. Therefore, a pump capable of delivering
solvent smoothly and reproducibly at these pressures, that can compensate
for solvent compressibility, and can operate in both the gradient and
isocratic separation modes, was required.
Sample introduction is also critical. Conventional injection valves, either
automated or manual, are not designed and hardened to work at extreme
pressure. To protect the column from experiencing extreme pressure fluctu-
ations, the injection process must be relatively pulse-free. The swept
volume of the device also needs to be minimal to reduce potential band
spreading. A fast injection cycle time is needed to fully capitalize on the
speed afforded by UPLC which, in turn, requires a high sample capacity.
Low volume injections with minimal carryover are also required to realize
the increased sensitivity benefits.
With 1.7 (jum particles, half-height peak widths of less than one second
are obtained, posing significant challenges for the detector. In order to accu-
rately and reproducibly integrate an analyte peak, the detector sampling rate
must be high enough to capture enough data points across the peak. In
addition, the detector cell must have minimal dispersion (volume) to
preserve separation efficiency. Conceptually, the sensitivity increase for
UPLC detection should be 2-3 times higher than with HPLC separations,
depending on the detection technique that is used. Conventional absorbance-
based optical detectors are concentration sensitive detectors and, for UPLC
use, the flow cell volume would have to be reduced in standard UV/Visible
detectors to maintain concentration and signal, while avoiding Beers' Law
limitations.
In early 2004, the first commercially available UPLC system that
embodied these requirements was described for the separation of various
pharmaceutical related small organic molecules, proteins, and peptides; it is
called the ACQUITY UPLC™ System.[18-20]
Using UPLC, it is now possible to take full advantage of chromatographic
principles to run separations using shorter columns, and/or higher flow rates
for increased speed, with superior resolution and sensitivity. Figures 2 and 3
6. 0,08
004
002
0.08
0.06
002
0.00
0.00 5.00 10.00 15.00 20.00 25,00 30-00 35,00 40.00 45.00 50.00 55-00 60.00
Minutes
Figure 3. HPLC vs. UPLC peak capacity. In this gradient peptide map separation, the HPLC (top) separation (on a 5 ^.m CIS column) yields 70
V.
peaks, or a peak capacity of 143, while the UPLC separation (bottom) run under identical conditions yields 168 peaks, or a peak capacity of 360, a ^
2.5X increase. 3.
N
7. UPLC : An Introduction and Review 1259
illustrate UPLC in action. In Figure 2 a separation of eight diuretics is accom-
plished in under 1.6 minutes. The same separation on a 2.1 by 100mm 5 |a,m
Cig HPLC column yields comparable resolution, but takes over ten minutes.
For some analyses, however, speed is of secondary importance; peak
capacity and resolution take center stage. Figure 3 shows a peptide map
where the desired goal is to maximize the number of peaks. In this application,
the increased peak capacity (number of peaks resolved per unit time) of
UPLC dramatically improves the quality of the data resulting in a more
definitive map
APPLICATIONS
Chromatographers are accustomed to making compromises; one of the most
common scenarios involves sacrificing resolution for speed. In addition, for
complex samples like natural product extracts, added resolution can provide
more information in the form of additional peaks. Figure 4 shows an HPLC
0.8AUFS
HPLC
10jiL
Injection
Time in Minutes
2.1 AUFS
UPLC™
5^L Injection
Time in Minutes
Figure 4. Comparison HPLC and. UPLC for the separation of a ginger root extract.
HPLC conditions: Column: 2.1 by 100mm 5.0|a.m prototype BEH CIS at 28°C. A
25-96%B linear gradient over 10 minutes, at a flow rate of 1.0 mL/min was used.
Mobile phase A was water, B was acetonitrile. UV detection @ 230 nm, 10|j.L
injection. UPLC conditions: Column: 2.1 by 100mm 1.7|j.m ACQUITY BEH CIS
at 28°C. A 50-100%B linear gradient from 1.4 to 3.7 minutes, followed by a hold
until 6.0 minutes, at a flow rate of 0.3 mL/min was used. Mobile phase A was water,
B was acetonitrile. UV detection @ 230 nm, 5 (xL injection.
8. 1260 M. E. Swartz
versus UPLC separation comparison of a ginger root extract sample where
both speed and resolution are improved, as well as an increase in sensitivity.
DryLab software was used to model and redevelop the separation and
transfer it to the ACQUITY UPLC System and BEH chemistry.
Faster separations can lead to higher throughput and time savings when
running multiple samples. But, a significant amount of time can also be
consumed in developing the method in the first place. Faster, higher resolution
UPLC separations can cut method development time from days, to hours,
or even minutes. Figure 5 is an example of an UPLC separation of several
closely related coumarins and a metabolite that was developed in under an
hour, including UPLC scouting runs for gradient optimization, and individual
runs for elution order identification. These runs were performed in a fraction
of the time that would be necessary by conventional HPLC, resulting in
significant time savings in the method development laboratory.
As alluded to previously, mass spectrometry has gained widespread
acceptance as an analytical tool for the qualitative and quantitative analysis
0.03 3 4
AUFS
0.00 1.30
Figure 5. UPLC Separation of Seven Coumarins illustrating fast method develop-
ment. Column: 2.1 by 30mm 1.7 |xm ACQUITY UPLC BEH Cjg @ 35°C. A
20-40%B linear gradient over 1.0 minute, at a flow rate of 0.86mL/min was used.
Mobile phase A was 0.1 % formic acid, B was acetonitrile. UV detection @ 254 nm
and 40 pts/sec. Peaks are in order: 1: 7-hydroxycoumarin-glucuronide, 7-hydroxy-
coumarin, 4-hydroxycoumarin, coumarin, 7-methoxycoumarin, 7-ethoxycoumarin,
and 4-ethoxycoumarin.
9. • T0= M5 ES*
4 U-J BPI
I ,,14 41 ' 56e-l
13 68 PI
200 4 DO B [iD 8 [iD ID DO 12.DD 14 OD 1600 1600 20.00 22.00 2400 2G OD 2B OD 30 DO 3200 34 DO
Figure 6. Separation of rat bile following the administration of midolazam at 5 mg/kg: A) 30 minute separation on a 2.1 by 100mm 3.5 (xm CIS
HPLC Column and B) 30minute separation on a 2.1 by 100mm 1.7 |j.m CIS UPLC column. Reprinted by permission from reference 21.
10. i-iu-i wi. &. swartz
of many types of compounds. MS detection is significantly enhanced
by UPLC; increased peak concentrations with reduced chromatographic
dispersion at lower flow rates (no flow splitting) promotes increased source
ionization efficiencies. Jorgenson et al. have shown that higher chromato-
graphic efficiency, resulting from the use of UPLC, translates into better
resolution and higher peak capacity, which is particularly important for the
analysis of peptides and proteins.[15] The increased resolving power made
the resulting data easier to interpret, since more of the MS peaks consisted
of a single compound, and up to a 20-fold improvement in the quality of
the spectral information (vs. nanoelectrospray) was obtained. Lee et al. also
used MS detection for the analysis of low molecular weight compounds
similar to those that might comprise a combinatorial library.[14'17] It was
demonstrated that, in order to address the very narrow peaks produced by
UPLC, it is necessary to use a very high data capture rate MS such as a
TOP or quadrapole with fast scan rates. Lee et al. also pointed out that,
in some instances, related compounds of the same molecular weight
and similar structures could not be differentiated by MS, necessitating
chromatographic resolution on the UPLC time scaled1'
Plumb et al. have investigated the use of UPLC/MS for the analysis of
metabolites,121'22] and as a tool for differential metabolic pathway profiling
in functional genomic studies.[23] Their data illustrate the benefit obtained
from the extra resolution of UPLC, both in terms of specificity and spectral
quality, revealing new information and reducing the risk of not detecting
potentially important metabolites. Figure 6 shows the benefits of UPLC
versus HPLC for monitoring the in-vivo metabolism of midazolam (an anti-
convulsant) in rat liver bile. This is a challenging separation due to the high
concentration of bile salts that can interfere, and the presence of bilirubin
that can cause ion pairing. The resolution is dramatically improved, and
the number of discreet peaks has more than doubled.
CONCLUSION
UPLC presents the ability to extend and expand the utility of separation
science at a time when many scientists have reached separation barriers,
pushing the limits of conventional HPLC. New chemistry and instrumentation
technology can provide more information per unit of work as UPLC begins
to fulfill the promise of increased speed, resolution, and sensitivity
predicted for liquid chromatography.
ACKNOWLEDGMENTS
The author would like to acknowledge the contributions of the ACQUITY
program team at Waters, particularly Eric Grumbach, Michael Jones,
11. UPLC : An Introduction and Review 1263
Marianna Kele, Rob Plumb, and Jeff Mazzeo for their contributions to this
manuscript.
ACQUITY UPLC, UPLC, and Ultra Performance LC are trademarks of
Waters Corporation.
REFERENCES
1. van Deemter, J.J.; Zuiderweg, F.J.; Klinkenberg, A. Chem. Eng. Sci. 1956, 5, 271.
2. Cheng, Y.-F.; Lu, Z.; Neue, U. Rapid Commun. Mass Spectrom. 2001, 15, 141.
3. Tiller, P.R.; Romanyshyn, L.A.; Neue, U.D. Anal. Bioanal. Chem. 2003, 377, 788.
4. Neue, U.D.; Mazzeo, J.R. J. Sep. Sci. 2001, 24, 1.
5. Jerkovitch, A.D.; Mellors, J.S.; Jorgenson, J.W. LC-GC North Amer. 2003, 21,1.
6. Wu, N.; Lippert, J.A.; Lee, M.L. J. Chromotogr. 2001, 911, 1.
7. Unger, K.K.; Kumar, D.; Grun, M.; Buchel, G.; Ludtke, S.; Adam, Th.;
Scumacher, K.; Renker, S. J. Chromatogr. A 2000, 892, 47.
8. Neue, U.D.; Walter, T.H.; Alden, B.A.; Jiang, Z.; Fisk, R.P.; Cook, J.T.;
Close, K.H.; Carmody, J.L.; Grassi, J.M.; Cheng, Y.-F.; Lu, Z.; Crowley, R.J.
Amer. Lab. 1999, 31, 36.
9. Cheng, Y.-F.; Walter, T.H.; Lu, Z.; Iraneta, P.; Gendreau, C.; Neue, U.D.;
Grassi, J.M.; Carmody, J.L.; O'Gara, J.E.; Fisk, R.P. LC-GC 2000, 18, 1162.
10. Mazzeo, J.R. Anal. Chem. November 2004, In press.
11. Halasz, I.; Endele, R.; Asshauer, J. J. Chromatogr. 1975, 112, 37.
12. MacNair, J.E.; Lewis, K.C.; Jorgenson, J.W. Anal. Chem. 1997, 69, 983.
13. Colon, L.A.; Citron, J.M.; Anspach, J.A.; Fermier, A.M.; Swinney, K.A. Analyst
2004, 729, 503.
14. Wu, N.; Lippert, J.A.; Lee, M.L. J. Chromatogr. A 2001, 911, 1.
15. Tolley, L.; Jorgenson, J.W.; Mosely, M.A. Anal. Chem. 2001, 73, 2985.
16. Swinney, K.A.; Fermier, A.M.; Guo, Y.; Oyler, A.R.; Segmuller, B.E.; Dunphy, R.
Presentation #L1703 at the 28th Intl. Symp. on High Perform. Liq. Phase Sepns. &
Rel. Techn, Philadelphia, PA, June 2004.
17. Lippert, J.A.; Xin, B.; Wu, N.; Lee, M.L. J. Microcol. Sepn. 1997, 11, 631.
18. Swartz, M.E. Presentation at the Pitts, Conf. on Anal. Chem. Appl. Spectrosc,
Chicago, IL, 2004.
19. Swartz, M.E.; Murphy, B. Lab Plus Intl. 2004, 18, 6.
20. Swartz, M.E.; Murphy, B. Pharm.l Formul. Qual. November 2004, In press.
21. Plumb, R.S.; Castro-Perez, J.; Granger, J.H.; Beattie, I.; Joncour, K.; Wright, A.
Rapid Commun. Mass Spectrom. 2004, 18, 2331.
22. Castro-Perez, J.; Plumb, R.; Granger, J.H. Rapid Commun. Mass Spectrom.
October 2004, Submitted.
23. Wilson, I.D.; Nicholson, J.K.; Castro-Perez, J.; Granger, J.H.; Smith, B.W.;
Mazzeo, J.; Plumb, R.S. J. Proteome Res. October 2004, Submitted.
Received November 22, 2004
Accepted December 19, 2004
Manuscript 6549E