The document provides an overview of capillary electrophoresis (CE) and its application in drug discovery. It discusses key CE principles such as electroosmotic flow, electrophoretic mobility, separation efficiency, and injections. Safety considerations for operating a CE system are also covered. The document demonstrates how CE can be used in drug discovery by one company to screen thousands of compounds daily. It concludes by explaining how to set up and run samples on a Beckman Coulter CE system in a classroom exercise.
Capillary electrophoresis is a separation technique that uses charged molecules' differential migration in response to an applied electric field. Key components include a capillary, buffers, and detectors. Molecules are separated based on their charge and size. There are several modes, including capillary zone electrophoresis which separates based on charge and size, and micellar electrokinetic capillary chromatography which uses micelles to separate charged and neutral molecules. Capillary electrophoresis provides high resolution, efficiency, and versatility in analyzing various molecules like proteins, nucleic acids, and inorganic ions.
This document summarizes a study that evaluated different air-liquid interface cell exposure systems and their ability to deliver gases and particles to cells under identical conditions. The study found that (1) systems relying only on sedimentation or diffusion delivered particles poorly, (2) applying an external force like electrostatic precipitation or thermophoresis improved particle delivery, and (3) the Gillings sampler was most effective at delivering both gases and particles to cells. Proper characterization of exposure system performance is important for understanding their advantages and limitations.
This document discusses basic techniques used in clinical chemistry laboratories, including automation, photometry, electrophoresis, chromatography, and various types of analyzers. The key points are:
- Automation has improved productivity and quality by allowing high-throughput analysis with minimal human intervention using integrated computer systems.
- Common techniques include photometry, turbidimetry, nephelometry, fluorometry, electrophoresis, chromatography, and various types of automated analyzers like batch, random-access, and semi-auto analyzers.
- Photometry techniques like colorimetry and spectrophotometry quantify the absorbance or transmittance of light as it passes through a sample, allowing concentration measurements based on Beer's
All you need to know about the HPLC Chromatography!
HPLC (High-performance liquid chromatography) also referred as high-pressure liquid chromatography, is a technique used in analytical labs to separate, identify, and quantify each component in a mixture. HPLC separates and purifies compounds according to their polarity. I have tried to simplify the whole process of HPLC Chromatography and explain in simple terms. Let’s look at the main components involved in HPLC in this video.
I discussed about Normal Phase Chromatography, Reverse Phase Chromatography and their comparison and differences. Watch this video for more details.
If you have any doubts: Please leave a comment in the comments section below.
High performance liquid chromatography (HPLC) is an improved form of liquid chromatography that forces solvent through a column at high pressure. It separates mixtures by interacting differently with stationary and mobile phases in the column based on molecular structure. HPLC uses pumps to push solvent through an injector, column, detector, and recorder/computer. The column contains porous particles that substances differentially bind to. Detectors identify separated substances and recorders display chromatograms showing separation and quantification. HPLC has many applications like pharmaceutical quality control and forensic drug analysis due to its accuracy, precision, and versatility.
This content is suitable for medical technologists/technicians/lab assistants/scientists writing the SMLTSA board exam. The content is also suitable for biomedical technology students and people also interested in learning about test methodologies used in medical technology. This chapter describes test methodologies and their uses. Please note that these notes are a collection I used to study for my board exam and train others who got distinctions using these.
Disclaimer: Credit goes to those who wrote the notes and the examiners of each exam question. Please use only as a reference guide and use your prescribed textbook for the latest and most accurate notes and ranges. The material here is not referenced as it is a collection of pieces of study notes from multiple people, and thus will not be held viable for any misinterpretations. Please use at your own discretion.
Detection Method for Low Level of Potent ToxinsShreyas Patel
This document discusses various analytical methods for detecting low levels of potent toxins and impurities. It describes gas chromatography, high-performance liquid chromatography, and thin-layer chromatography techniques. For GC, it outlines instrumentation components like injectors, detectors, and sample introduction methods. For HPLC, it discusses separation modes, detectors, and instrumentation. TLC is described as using capillary action to separate components based on their affinity for the stationary phase. Various compounds are listed along with example separation and detection methods used for their analysis in different matrices.
Electrophoresis is a process that separates and isolates different compounds based on their charge and size. It works by applying an electric current to a medium containing charged particles, causing the particles to migrate. The key factors affecting particle movement are net charge, size, strength of the electric field, and properties of the supporting medium. Electrophoresis can be used to identify, isolate, and determine the molecular weight of many charged biomolecules like proteins, DNA, and hemoglobin. Common applications include polyacrylamide gel electrophoresis and hemoglobin electrophoresis.
Capillary electrophoresis is a separation technique that uses charged molecules' differential migration in response to an applied electric field. Key components include a capillary, buffers, and detectors. Molecules are separated based on their charge and size. There are several modes, including capillary zone electrophoresis which separates based on charge and size, and micellar electrokinetic capillary chromatography which uses micelles to separate charged and neutral molecules. Capillary electrophoresis provides high resolution, efficiency, and versatility in analyzing various molecules like proteins, nucleic acids, and inorganic ions.
This document summarizes a study that evaluated different air-liquid interface cell exposure systems and their ability to deliver gases and particles to cells under identical conditions. The study found that (1) systems relying only on sedimentation or diffusion delivered particles poorly, (2) applying an external force like electrostatic precipitation or thermophoresis improved particle delivery, and (3) the Gillings sampler was most effective at delivering both gases and particles to cells. Proper characterization of exposure system performance is important for understanding their advantages and limitations.
This document discusses basic techniques used in clinical chemistry laboratories, including automation, photometry, electrophoresis, chromatography, and various types of analyzers. The key points are:
- Automation has improved productivity and quality by allowing high-throughput analysis with minimal human intervention using integrated computer systems.
- Common techniques include photometry, turbidimetry, nephelometry, fluorometry, electrophoresis, chromatography, and various types of automated analyzers like batch, random-access, and semi-auto analyzers.
- Photometry techniques like colorimetry and spectrophotometry quantify the absorbance or transmittance of light as it passes through a sample, allowing concentration measurements based on Beer's
All you need to know about the HPLC Chromatography!
HPLC (High-performance liquid chromatography) also referred as high-pressure liquid chromatography, is a technique used in analytical labs to separate, identify, and quantify each component in a mixture. HPLC separates and purifies compounds according to their polarity. I have tried to simplify the whole process of HPLC Chromatography and explain in simple terms. Let’s look at the main components involved in HPLC in this video.
I discussed about Normal Phase Chromatography, Reverse Phase Chromatography and their comparison and differences. Watch this video for more details.
If you have any doubts: Please leave a comment in the comments section below.
High performance liquid chromatography (HPLC) is an improved form of liquid chromatography that forces solvent through a column at high pressure. It separates mixtures by interacting differently with stationary and mobile phases in the column based on molecular structure. HPLC uses pumps to push solvent through an injector, column, detector, and recorder/computer. The column contains porous particles that substances differentially bind to. Detectors identify separated substances and recorders display chromatograms showing separation and quantification. HPLC has many applications like pharmaceutical quality control and forensic drug analysis due to its accuracy, precision, and versatility.
This content is suitable for medical technologists/technicians/lab assistants/scientists writing the SMLTSA board exam. The content is also suitable for biomedical technology students and people also interested in learning about test methodologies used in medical technology. This chapter describes test methodologies and their uses. Please note that these notes are a collection I used to study for my board exam and train others who got distinctions using these.
Disclaimer: Credit goes to those who wrote the notes and the examiners of each exam question. Please use only as a reference guide and use your prescribed textbook for the latest and most accurate notes and ranges. The material here is not referenced as it is a collection of pieces of study notes from multiple people, and thus will not be held viable for any misinterpretations. Please use at your own discretion.
Detection Method for Low Level of Potent ToxinsShreyas Patel
This document discusses various analytical methods for detecting low levels of potent toxins and impurities. It describes gas chromatography, high-performance liquid chromatography, and thin-layer chromatography techniques. For GC, it outlines instrumentation components like injectors, detectors, and sample introduction methods. For HPLC, it discusses separation modes, detectors, and instrumentation. TLC is described as using capillary action to separate components based on their affinity for the stationary phase. Various compounds are listed along with example separation and detection methods used for their analysis in different matrices.
Electrophoresis is a process that separates and isolates different compounds based on their charge and size. It works by applying an electric current to a medium containing charged particles, causing the particles to migrate. The key factors affecting particle movement are net charge, size, strength of the electric field, and properties of the supporting medium. Electrophoresis can be used to identify, isolate, and determine the molecular weight of many charged biomolecules like proteins, DNA, and hemoglobin. Common applications include polyacrylamide gel electrophoresis and hemoglobin electrophoresis.
This document discusses isoelectric focusing, a technique used to separate proteins based on their isoelectric point (PI). Proteins are subjected to an electric field within a pH gradient, which causes them to migrate to the point in the gradient where their net charge is zero (their PI). Different proteins have different PIs and will therefore migrate to distinct positions in the gel. Isoelectric focusing provides high resolution separation and is useful for research applications such as taxonomy, cytology and immunology.
Capillary electrophoresis is a separation technique that uses narrow bore capillaries. Charged molecules migrate through the capillary under the influence of an applied electric field and separate based on their charge and size. The principle involves electrostatic forces moving molecules toward the electrode of opposite charge, as well as electroosmotic flow dragging buffer molecules. Capillary electrophoresis has various modes of operation and is used to separate and analyze biological samples in clinical and diagnostic applications.
‘Separation of sample components after their distribution between two phases.’’ - IUPAC definition
Ion Chromatography (IC) was introduced in 1975 by Small, Stevens and Baumann as a new analytical method for sensitive detection of ions via their electrical conductance. Chromatography is a separation technique that is used for separation a sample mixture into its constituents or components.
Getting Started with In-Vitro Blood Vessel ResearchInsideScientific
The goal of this webinar was to provide an overview of the essential tools and techniques required for successful isolation of blood vessels and study in pressurized arteriograph systems. Presenters review dissection and mounting techniques, equipment requirements, proper experimental design, and discuss the difference between pressurized cannulation systems and wire myograph/tissue bath systems. Dr. Scott Earley shares strategies for skills development, in particular starting research with 2nd order mesenteric artery sample and how to generate vessel constrictions and dilations. In addition, he discusses an advanced application involving the assessment of myogenic reactivity in a "blind-sac" vessel preparation using a cerebral parenchymal arteriole.
Key Topics:
- equipment needs and proper operation
- how to design an experiment to assess blood vessel function in-vitro
- techniques for vessel dissection, mounting, and assessing viability
- how to interpret experimental results
- studying myogenic responses using a cerebral parenchymal arteriole
This document discusses a steam and water analysis system (SWAS) for monitoring water quality in power plants. It describes the need for SWAS to prevent corrosion and optimize boiler performance. The key components of SWAS include sample conditioning equipment that cools, filters and regulates water samples, and various analyzers that measure critical water parameters like pH, conductivity, dissolved oxygen and silica. Maintaining high water quality through online monitoring with SWAS is essential for reducing shutdowns and maximizing power plant efficiency.
This document summarizes a capstone project to create a point-of-care device to continuously monitor urea and creatinine levels in urine samples from ICU patients. The device uses colorimetric reactions and a spectrophotometer to automate urine tests and provide results every 15 minutes. It incorporates pinch valves, reagent chambers, and electronics controlled by an Arduino board to process urine samples. Initial testing of the chemistry and prototype device demonstrated linear standard curves for urea and creatinine quantification. Future work includes further device miniaturization and clinical testing prior to regulatory approval and commercialization.
HPLC is a separation technique used in pharmaceutical analysis to separate, identify, and quantify components in mixtures. It works by pumping a mobile phase through a column containing adsorbent packing material. Samples are injected into the mobile phase and the components elute from the column at different rates depending on their interactions with the stationary and mobile phases. Detectors then convert the separated components into electrical signals to allow for qualitative and quantitative analysis. Common applications of HPLC include analysis of drugs and metabolites in biological samples.
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.
The document provides information on sample preparation and analysis methods for pharmaceutical dosage forms, biological samples, and proteins. It discusses sample extraction and processing steps like solvent extraction, centrifugation, filtration, and solid phase extraction for plasma and urine samples. For biological samples for protein analysis, it covers stabilization, solubilization, and desalting/buffer exchange using dialysis or diafiltration. It also describes hydrodynamic and electrokinetic injection methods for sample introduction in capillary electrophoresis and issues that can occur like peak migration problems, bubbles, and shadow peaks.
CE is an efficient separation technique that uses differences in charge and size to separate molecules in a capillary tube under the influence of an electric field. It provides high resolution separations using small sample volumes in short time frames. Variations include CZE which relies on differences in charge, CGE which separates by size in a gel, and CIEF which focuses molecules at their isoelectric point. CE has advantages over HPLC in speed, efficiency, and cost but lacks reproducibility and is not suitable for large-scale separations.
This document provides an overview of steam and water analysis systems (SWAS) used in power plants. SWAS continuously analyze boiler steam and water to ensure purity and prevent corrosion. It discusses the need for boiler water treatment, SWAS requirements, sample conditioning, important sampling system equipment like sample coolers and regulators, sample analysis including pH, conductivity, silica, and indications of problems from analysis results. The purpose is to maintain high purity feedwater and steam to prevent damage to turbines, boilers, and other equipment.
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.
Electrophoresis is a technique used to separate molecules based on their charge and size. It works by applying an electric field to move charged molecules through a medium such as a gel or paper. There are different types depending on the medium used, such as polyacrylamide gel electrophoresis and agarose gel electrophoresis. Electrophoresis has many applications including determining molecular weights, studying protein-protein interactions, and purifying proteins and DNA fragments.
High-performance liquid chromatography (HPLC) is an analytical chemistry technique used to separate, identify, and quantify components in mixtures. It works by forcing a pressurized liquid solvent through a column packed with adsorbent particles under high pressure. This allows for better separation than traditional column chromatography due to smaller particle sizes and detection methods. HPLC has applications in manufacturing, legal, research, and medical fields such as drug analysis, food testing, and pharmaceutical development.
This document provides information about an introductory chemistry class. It begins with contact information for the instructor and an overview of grading and expectations. It then details requirements for laboratory reports, including documentation of instruments, reagents, and chromatograms. The document outlines several lessons that will be covered, including HPLC components and operation, calibration, column installation and sample loops, and examples of HPLC use in the pharmaceutical industry for applications like content uniformity testing and dissolution. Maintaining detailed records and complying with GMP/GLP standards are emphasized throughout the document.
This document provides an overview of key terms and concepts related to clinical chemistry quality control. It defines common quality control terms like quality, control, standard, specificity, sensitivity, accuracy, and precision. It also describes different types of quality control, including intralaboratory quality control to detect random and systematic errors, and interlaboratory quality control to maintain long-term accuracy through comparison with other laboratories. Common quality control charts and rules for identifying errors are discussed. Key instrumentation and analytical techniques used in clinical chemistry are briefly outlined, including spectrophotometry, atomic absorption spectrophotometry, flame emission photometry, fluorometry, turbidimetry, nephelometry, potentiometry, electrophoresis, and chromatography.
High-performance liquid chromatography (HPLC) involves forcing a pressurized liquid solvent through a column containing a stationary solid phase to separate and analyze compounds. It was developed in the 1960s and commercialized in the late 1960s and early 1970s. Key developments included the use of higher pressures up to 20,000 psi and particles sizes of 2 microns or less, allowing for faster separations. HPLC uses differences in how compounds partition between the liquid mobile phase and solid stationary phase to achieve separation. Common applications include pharmaceutical analysis, environmental testing, and forensic and clinical analysis.
HPLC involves injecting a liquid sample into a column packed with tiny adsorbent particles. Components are separated as they interact differently with the stationary phase and are eluted by the mobile phase. The separated components are then detected and analyzed. Key components of HPLC include the solvent reservoir, pump, injector, column, and various detectors. There are different modes of separation including reversed phase, normal phase, ion exchange, and size exclusion chromatography. Parameters like retention time, theoretical plate number, and resolution are used to characterize chromatographic separations.
High performance liquid chromatography (HPLC) is a widely used technique to separate mixtures of organic compounds. HPLC works by partitioning components of a mixture between a stationary phase made of small particles in a steel column and a mobile phase solvent. Pressure is required to force the mobile phase through the stationary phase due to the small particle size. HPLC provides enhanced separations in shorter time periods compared to traditional liquid chromatography. A variety of stationary and mobile phases can be used to optimize separations of different molecules.
Types Of Chromatography - liquid & Gas Chromatography(Mobile Phase).pptxPriyaDixit46
Liquid chromatography (LC) and gas chromatography (GC) are two common types of chromatography. In LC, the mobile phase is a liquid and separation is based on interactions between solutes and the mobile and stationary phases. GC uses an inert gas as the mobile phase, with separation dependent on solute boiling points. Both techniques can separate mixtures and are used in various applications like pharmaceutical analysis, environmental testing, and food and chemical quality control. However, GC is generally faster and provides better resolution than LC.
This document discusses isoelectric focusing, a technique used to separate proteins based on their isoelectric point (PI). Proteins are subjected to an electric field within a pH gradient, which causes them to migrate to the point in the gradient where their net charge is zero (their PI). Different proteins have different PIs and will therefore migrate to distinct positions in the gel. Isoelectric focusing provides high resolution separation and is useful for research applications such as taxonomy, cytology and immunology.
Capillary electrophoresis is a separation technique that uses narrow bore capillaries. Charged molecules migrate through the capillary under the influence of an applied electric field and separate based on their charge and size. The principle involves electrostatic forces moving molecules toward the electrode of opposite charge, as well as electroosmotic flow dragging buffer molecules. Capillary electrophoresis has various modes of operation and is used to separate and analyze biological samples in clinical and diagnostic applications.
‘Separation of sample components after their distribution between two phases.’’ - IUPAC definition
Ion Chromatography (IC) was introduced in 1975 by Small, Stevens and Baumann as a new analytical method for sensitive detection of ions via their electrical conductance. Chromatography is a separation technique that is used for separation a sample mixture into its constituents or components.
Getting Started with In-Vitro Blood Vessel ResearchInsideScientific
The goal of this webinar was to provide an overview of the essential tools and techniques required for successful isolation of blood vessels and study in pressurized arteriograph systems. Presenters review dissection and mounting techniques, equipment requirements, proper experimental design, and discuss the difference between pressurized cannulation systems and wire myograph/tissue bath systems. Dr. Scott Earley shares strategies for skills development, in particular starting research with 2nd order mesenteric artery sample and how to generate vessel constrictions and dilations. In addition, he discusses an advanced application involving the assessment of myogenic reactivity in a "blind-sac" vessel preparation using a cerebral parenchymal arteriole.
Key Topics:
- equipment needs and proper operation
- how to design an experiment to assess blood vessel function in-vitro
- techniques for vessel dissection, mounting, and assessing viability
- how to interpret experimental results
- studying myogenic responses using a cerebral parenchymal arteriole
This document discusses a steam and water analysis system (SWAS) for monitoring water quality in power plants. It describes the need for SWAS to prevent corrosion and optimize boiler performance. The key components of SWAS include sample conditioning equipment that cools, filters and regulates water samples, and various analyzers that measure critical water parameters like pH, conductivity, dissolved oxygen and silica. Maintaining high water quality through online monitoring with SWAS is essential for reducing shutdowns and maximizing power plant efficiency.
This document summarizes a capstone project to create a point-of-care device to continuously monitor urea and creatinine levels in urine samples from ICU patients. The device uses colorimetric reactions and a spectrophotometer to automate urine tests and provide results every 15 minutes. It incorporates pinch valves, reagent chambers, and electronics controlled by an Arduino board to process urine samples. Initial testing of the chemistry and prototype device demonstrated linear standard curves for urea and creatinine quantification. Future work includes further device miniaturization and clinical testing prior to regulatory approval and commercialization.
HPLC is a separation technique used in pharmaceutical analysis to separate, identify, and quantify components in mixtures. It works by pumping a mobile phase through a column containing adsorbent packing material. Samples are injected into the mobile phase and the components elute from the column at different rates depending on their interactions with the stationary and mobile phases. Detectors then convert the separated components into electrical signals to allow for qualitative and quantitative analysis. Common applications of HPLC include analysis of drugs and metabolites in biological samples.
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.
The document provides information on sample preparation and analysis methods for pharmaceutical dosage forms, biological samples, and proteins. It discusses sample extraction and processing steps like solvent extraction, centrifugation, filtration, and solid phase extraction for plasma and urine samples. For biological samples for protein analysis, it covers stabilization, solubilization, and desalting/buffer exchange using dialysis or diafiltration. It also describes hydrodynamic and electrokinetic injection methods for sample introduction in capillary electrophoresis and issues that can occur like peak migration problems, bubbles, and shadow peaks.
CE is an efficient separation technique that uses differences in charge and size to separate molecules in a capillary tube under the influence of an electric field. It provides high resolution separations using small sample volumes in short time frames. Variations include CZE which relies on differences in charge, CGE which separates by size in a gel, and CIEF which focuses molecules at their isoelectric point. CE has advantages over HPLC in speed, efficiency, and cost but lacks reproducibility and is not suitable for large-scale separations.
This document provides an overview of steam and water analysis systems (SWAS) used in power plants. SWAS continuously analyze boiler steam and water to ensure purity and prevent corrosion. It discusses the need for boiler water treatment, SWAS requirements, sample conditioning, important sampling system equipment like sample coolers and regulators, sample analysis including pH, conductivity, silica, and indications of problems from analysis results. The purpose is to maintain high purity feedwater and steam to prevent damage to turbines, boilers, and other equipment.
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.
Electrophoresis is a technique used to separate molecules based on their charge and size. It works by applying an electric field to move charged molecules through a medium such as a gel or paper. There are different types depending on the medium used, such as polyacrylamide gel electrophoresis and agarose gel electrophoresis. Electrophoresis has many applications including determining molecular weights, studying protein-protein interactions, and purifying proteins and DNA fragments.
High-performance liquid chromatography (HPLC) is an analytical chemistry technique used to separate, identify, and quantify components in mixtures. It works by forcing a pressurized liquid solvent through a column packed with adsorbent particles under high pressure. This allows for better separation than traditional column chromatography due to smaller particle sizes and detection methods. HPLC has applications in manufacturing, legal, research, and medical fields such as drug analysis, food testing, and pharmaceutical development.
This document provides information about an introductory chemistry class. It begins with contact information for the instructor and an overview of grading and expectations. It then details requirements for laboratory reports, including documentation of instruments, reagents, and chromatograms. The document outlines several lessons that will be covered, including HPLC components and operation, calibration, column installation and sample loops, and examples of HPLC use in the pharmaceutical industry for applications like content uniformity testing and dissolution. Maintaining detailed records and complying with GMP/GLP standards are emphasized throughout the document.
This document provides an overview of key terms and concepts related to clinical chemistry quality control. It defines common quality control terms like quality, control, standard, specificity, sensitivity, accuracy, and precision. It also describes different types of quality control, including intralaboratory quality control to detect random and systematic errors, and interlaboratory quality control to maintain long-term accuracy through comparison with other laboratories. Common quality control charts and rules for identifying errors are discussed. Key instrumentation and analytical techniques used in clinical chemistry are briefly outlined, including spectrophotometry, atomic absorption spectrophotometry, flame emission photometry, fluorometry, turbidimetry, nephelometry, potentiometry, electrophoresis, and chromatography.
High-performance liquid chromatography (HPLC) involves forcing a pressurized liquid solvent through a column containing a stationary solid phase to separate and analyze compounds. It was developed in the 1960s and commercialized in the late 1960s and early 1970s. Key developments included the use of higher pressures up to 20,000 psi and particles sizes of 2 microns or less, allowing for faster separations. HPLC uses differences in how compounds partition between the liquid mobile phase and solid stationary phase to achieve separation. Common applications include pharmaceutical analysis, environmental testing, and forensic and clinical analysis.
HPLC involves injecting a liquid sample into a column packed with tiny adsorbent particles. Components are separated as they interact differently with the stationary phase and are eluted by the mobile phase. The separated components are then detected and analyzed. Key components of HPLC include the solvent reservoir, pump, injector, column, and various detectors. There are different modes of separation including reversed phase, normal phase, ion exchange, and size exclusion chromatography. Parameters like retention time, theoretical plate number, and resolution are used to characterize chromatographic separations.
High performance liquid chromatography (HPLC) is a widely used technique to separate mixtures of organic compounds. HPLC works by partitioning components of a mixture between a stationary phase made of small particles in a steel column and a mobile phase solvent. Pressure is required to force the mobile phase through the stationary phase due to the small particle size. HPLC provides enhanced separations in shorter time periods compared to traditional liquid chromatography. A variety of stationary and mobile phases can be used to optimize separations of different molecules.
Types Of Chromatography - liquid & Gas Chromatography(Mobile Phase).pptxPriyaDixit46
Liquid chromatography (LC) and gas chromatography (GC) are two common types of chromatography. In LC, the mobile phase is a liquid and separation is based on interactions between solutes and the mobile and stationary phases. GC uses an inert gas as the mobile phase, with separation dependent on solute boiling points. Both techniques can separate mixtures and are used in various applications like pharmaceutical analysis, environmental testing, and food and chemical quality control. However, GC is generally faster and provides better resolution than LC.
This document provides an overview of wound healing, its functions, stages, mechanisms, factors affecting it, and complications.
A wound is a break in the integrity of the skin or tissues, which may be associated with disruption of the structure and function.
Healing is the body’s response to injury in an attempt to restore normal structure and functions.
Healing can occur in two ways: Regeneration and Repair
There are 4 phases of wound healing: hemostasis, inflammation, proliferation, and remodeling. This document also describes the mechanism of wound healing. Factors that affect healing include infection, uncontrolled diabetes, poor nutrition, age, anemia, the presence of foreign bodies, etc.
Complications of wound healing like infection, hyperpigmentation of scar, contractures, and keloid formation.
Leveraging Generative AI to Drive Nonprofit InnovationTechSoup
In this webinar, participants learned how to utilize Generative AI to streamline operations and elevate member engagement. Amazon Web Service experts provided a customer specific use cases and dived into low/no-code tools that are quick and easy to deploy through Amazon Web Service (AWS.)
Communicating effectively and consistently with students can help them feel at ease during their learning experience and provide the instructor with a communication trail to track the course's progress. This workshop will take you through constructing an engaging course container to facilitate effective communication.
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LAND USE LAND COVER AND NDVI OF MIRZAPUR DISTRICT, UPRAHUL
This Dissertation explores the particular circumstances of Mirzapur, a region located in the
core of India. Mirzapur, with its varied terrains and abundant biodiversity, offers an optimal
environment for investigating the changes in vegetation cover dynamics. Our study utilizes
advanced technologies such as GIS (Geographic Information Systems) and Remote sensing to
analyze the transformations that have taken place over the course of a decade.
The complex relationship between human activities and the environment has been the focus
of extensive research and worry. As the global community grapples with swift urbanization,
population expansion, and economic progress, the effects on natural ecosystems are becoming
more evident. A crucial element of this impact is the alteration of vegetation cover, which plays a
significant role in maintaining the ecological equilibrium of our planet.Land serves as the foundation for all human activities and provides the necessary materials for
these activities. As the most crucial natural resource, its utilization by humans results in different
'Land uses,' which are determined by both human activities and the physical characteristics of the
land.
The utilization of land is impacted by human needs and environmental factors. In countries
like India, rapid population growth and the emphasis on extensive resource exploitation can lead
to significant land degradation, adversely affecting the region's land cover.
Therefore, human intervention has significantly influenced land use patterns over many
centuries, evolving its structure over time and space. In the present era, these changes have
accelerated due to factors such as agriculture and urbanization. Information regarding land use and
cover is essential for various planning and management tasks related to the Earth's surface,
providing crucial environmental data for scientific, resource management, policy purposes, and
diverse human activities.
Accurate understanding of land use and cover is imperative for the development planning
of any area. Consequently, a wide range of professionals, including earth system scientists, land
and water managers, and urban planners, are interested in obtaining data on land use and cover
changes, conversion trends, and other related patterns. The spatial dimensions of land use and
cover support policymakers and scientists in making well-informed decisions, as alterations in
these patterns indicate shifts in economic and social conditions. Monitoring such changes with the
help of Advanced technologies like Remote Sensing and Geographic Information Systems is
crucial for coordinated efforts across different administrative levels. Advanced technologies like
Remote Sensing and Geographic Information Systems
9
Changes in vegetation cover refer to variations in the distribution, composition, and overall
structure of plant communities across different temporal and spatial scales. These changes can
occur natural.
Strategies for Effective Upskilling is a presentation by Chinwendu Peace in a Your Skill Boost Masterclass organisation by the Excellence Foundation for South Sudan on 08th and 09th June 2024 from 1 PM to 3 PM on each day.
it describes the bony anatomy including the femoral head , acetabulum, labrum . also discusses the capsule , ligaments . muscle that act on the hip joint and the range of motion are outlined. factors affecting hip joint stability and weight transmission through the joint are summarized.
1. So What’s the Big Deal with
Those Little Tubes?
Capillary Electrophoresis
Exercises for MSU Students
Prepared for Chemistry and Biochemistry Fall 2003
Kevin Olsen
2. Program
• General principles
• Safety
• Obtaining good results
• Example application, CE in drug discovery
• Running your samples on the Beckman
Coulter model P/ACE
7. Electroosmotic Flow
(EOF)
This flow is a phenomena resulting
when a solution is contained in a
capillary with fixed charges along its
wall. This is also known as the
Electroendosmotic Flow.
8. Where does the Electroosmotic flow come
from?
Anode
+
Cathode
-
Detector
9. Where does the Electroosmotic flow come
from?
Anode
+
Cathode
-
Applied Electric Field.
1000 Volts / Centimeter
Detector
10. Where does the Electroosmotic flow come
from?
Anode
+
Cathode
-
Detector
The interior wall of the capillary contains
charged sites that are created by the
ionization of Silanol groups on the fused
silica.
12. This is where the Electroosmotic flow
comes from.
What happens to the + cations when we turn on the power?
13. pH, Silanol Population, and the rate of
EOF flow.
• At very low pH, not many
silanols are ionized and
the EOF is slow.
• As pH increases the
number of ionized sites
also increases. The EOF
speed rises steadily.
• At very high pH values, a
maximum number of
ionized sites is reached.
The EOF speed also
reaches a maximum.
14. The apparent velocity of any analyte
(u) will be a combination of its
electrophoretic velocity and its
movement in response to the EOF.
u = (Uep + Ueo) E
15. How does apparent velocity help us?
Analytes with a net positive charge will move faster than EOF
EOF
Analytes with no net charge will move at the same speed as the EOF.
(This is a useful tool that helps us to measure the EOF.)
EOF
Analytes with a net negative charge will move slower than EOF
EOF
17. Separation Efficiency (Y) and Diffusion
Coefficient (X)
• Note the very dramatic
drop in separation
efficiency with
increasing diffusion
coefficient.
• This means that in
some cases, there is no
real advantage over
conventional HPLC
for smaller molecules.
18. Injections
There are two principle methods:
• Pressure differential works by applying a pressure across the
capillary while it it is dipping into the sample solution.
• Electrokinetic injection works by applying a voltage and
allowing ions to migrate into the capillary because of their
charge.
Injection volumes are typically very small:
• Typically if injection volumes exceed 1% of the column
volume, separation efficiency severely suffers.
• Sample volume can be increased by focusing the ions inside
the capillary. This technique uses a combination of
additives to the medium and selectively applied charges.
19. Preconcentration to Increase Sensitivity
• Attached to front of
column
• Contains a selective
binding agent
• Allows several
capillary volumes to
pass
• Analytes of interest
are then eluted
20. Pressure and Electrokinetic Injections
• One additional advantage of electrokinetic injections is that
if appropriate conditions are set, extended injection times
allow analytes to be concentrated without overloading the
column.
+ + +
+
+ +
+
-
-
+
21. Setting up the Capillary Column
• Cut the ends cleanly.
• Load capillary into
the cartridge
• Place the clear
portion in the
detector window.
22. The Capillary Column’s Cartridge
• Allows the column to
be moved from vial to
vial.
• Contains a cooling
medium.
• Contains gas and
vacuum connections.
• Holds electrodes that
place a charge on the
sample vials.
23. The Advantages of CE are:
• The number of theoretical plates is typically
in the hundreds of thousands.
• There is no mass transfer between mobile
and stationary phases as with HPLC and
GC, therefore the analytes remain in a
“plug” instead of spreading as a result of
laminar flow. (Peaks can still broaden
however.)
• Altering column conditions allows focusing
or concentration of samples.
24. Program
• General principles
• Safety
• Obtaining good results
• Example application, CE in drug discovery
• Running your samples on the Beckman
Coulter model P/ACE
25. SAFETY
• Chemical and Biological
• Remember that solvents will be flowing under high
pressure inside an electrically powered device.
• Aerosols may be generated, work in appropriate enclosure.
• Take all normal safety precautions when working with
toxic, pathogenic, or radioactive materials.
• Electrical
• Never remove covers and expose the electronics.
• Under certain conditions the chemist may have to be
grounded for protection against static electricity.
• Mechanical
• The CE unit features a robotic autosampler with many
moving parts and a sharp needle. Keep hands out of the
sample compartment while the unit is running.
26. Obtaining Reproducible (Good) Results
• Column condition.
• Composition and pH of the medium.
• Viscosity of the medium.
• Operating temperature.
• Adequate sample volume.
• Use of internal standards.
27. The pH must be tightly controlled to
obtain reproducible EOF flow.
• Remember that the
percentage of silanols that
are ionized is dependent
on the pH.
28. Column Condition
• As time goes on, certain
molecules will block or
otherwise neutralize the
ionized silanol sites.
This will change the
EOF and alter retention
times.
• It is also very important
to condition the column
properly before use.
Follow the directions in
the published method.
29. Internal Standards
• The main advantage of an internal standard is
that it is subject to the same conditions as the
analyte.
30. Program
• General principles
• Safety
• Obtaining good results
• Example application, CE in drug
discovery
• Running your samples on the Beckman
Coulter model P/ACE
32. Courtesy of Cetek Corporation
20,000 Compounds Tested per Day
6,000,000 Tested since 1998
33. Other Applications
• Analysis of molecules that are not suited to
HPLC.
• Chiral separations of enantiomers.
• Determination of drug molecules in biological
fluids.
• Separating bacteria.
• Expect the unexpected.
34. Program
• General principles
• Safety
• Obtaining good results
• Example application, CE in drug discovery
• Running your samples on the Beckman
Coulter model P/ACE
35.
36. Setting Up Methods
• Use File|Method|New from the menu bar to
create your method.
• When finished use File|Method|Save as from
the menu bar.
38. A1 B1 C1 D1 E1 F1
A1 B1 C1 D1 E1 F1
A1 B1 C1 D1 E1 F1
A1 B1 C1 D1 E1 F1
A1 B1 C1 D1 E1 F1
A1 B1 C1 D1 E1 F1
A1 B1 C1 D1 E1 F1
A1 B1 C1 D1 E1 F1
A1 B1 C1 D1 E1 F1
A1 B1 C1 D1 E1 F1
A1 B1 C1 D1 E1 F1
A1 B1 C1 D1 E1 F1
Manipulating Tray Layouts
39. A1 B1 C1 D1 E1 F1
A1 B1 C1 D1 E1 F1
A1 B1 C1 D1 E1 F1
A1 B1 C1 D1 E1 F1
A1 B1 C1 D1 E1 F1
A1 B1 C1 D1 E1 F1
Manipulating Tray Layouts
Blue = Rinse
Green = Multiple use
Orange = Fraction
collection
Dark purple = Sample
Light purple = Other
injection
Red = Selected vial
Yellow = Separation
40. Manipulating Tray Layouts
• Vials positions can be designated under
“instrument set-up”. Afterwards, you will be
prompted to save the changes to your method.
• Vial positions can also be designated while setting
up the “Timed events” table in your method.
• In either case, the software commands and
procedures are the same.
41. P/ACE System Control
• Select your method from the drop-dow method
under FILE
• You may then change the tray layouts but all
changes become part of the method.
• In this class, we will have two control options:
1. Direct Control
2. Single Run
42. THE EXERCISES
• Each team will create one method that will rinse
and condition the column, then inject a sample.
• Each team’s method parameters will be slightly
different.
• Your instructor will string the methods together
for an overnight run.
• When the class meets again, the results will be
compared. Each student will submit a report
comparing and contrasting the results.
43. Generic Test Solution Method
• Rinse: 0.5 minutes, Regenerator sol’n A, 20 psi.
Destination = waste vial.
• Rinse: 1.5 minutes, Run buffer A, 20 psi.
Destination = waste vial.
• Inject: 10 Seconds, Test mix, 0.5 psi. Destination
= Run buffer A on outlet tray.
• Separate: 7 minutes, Run buffer A, 25 kV. Ramp
time 0.2 minutes. Destination = Run buffer A on
outlet tray.
44. Team 1, Variation
• Rinse: 0.5 minutes, Regenerator sol’n A, 20 psi.
Destination = waste vial.
• Rinse: 1.5 minutes, Run buffer A, 20 psi.
Destination = waste vial.
• Inject: 10 Seconds, Test mix, 0.5 psi. Destination
= Run buffer A on outlet tray.
• Separate: 7 minutes, Run buffer A, 25 kV. Ramp
time 0.2 minutes. Destination = Run buffer A on
outlet tray.
45. Team 2 Variation
• Rinse: 0.5 minutes, Regenerator sol’n A, 20 psi.
Destination = waste vial.
• Rinse: 1.5 minutes, Run buffer A, 20 psi.
Destination = waste vial.
• Inject: 10 Seconds, Test mix, 0.5 psi. Destination
= Run buffer A on outlet tray.
• Separate: 7 minutes, Run buffer A, 25 kV. Ramp
time 0.2 minutes. Destination = Run buffer A on
outlet tray.
46. Team 3 Variation
• Rinse: 0.5 minutes, Regenerator sol’n A, 20 psi.
Destination = waste vial.
• Rinse: 1.5 minutes, Run buffer A, 20 psi.
Destination = waste vial.
• Inject: 10 Seconds, Test mix, 0.5 psi. Destination
= Run buffer A on outlet tray.
• Separate: 7 minutes, Run buffer A, 25 kV. Ramp
time 0.2 minutes. Destination = Run buffer A on
outlet tray.
47. Team 4 Variation
• Rinse: 0.5 minutes, Regenerator sol’n A, 20 psi.
Destination = waste vial.
• Rinse: 1.5 minutes, Run buffer A, 20 psi.
Destination = waste vial.
• Inject: 10 Seconds, Test mix, 0.5 psi. Destination
= Run buffer A on outlet tray.
• Separate: 7 minutes, Run buffer A, 25 kV. Ramp
time 0.2 minutes. Destination = Run buffer A on
outlet tray.
48. Team 5 Variation
• Rinse: 0.5 minutes, Regenerator sol’n A, 20 psi.
Destination = waste vial.
• Rinse: 1.5 minutes, Run buffer A, 20 psi.
Destination = waste vial.
• Inject: 10 Seconds, Test mix, 0.5 psi. Destination
= Run buffer A on outlet tray.
• Separate: 7 minutes, Run buffer A, 25 kV. Ramp
time 0.2 minutes. Destination = Run buffer A on
outlet tray.