This document provides an introduction to high performance liquid chromatography (HPLC). It describes the basic components of an HPLC system including the reservoir, pump, injector, separation column, and detector. Compounds are separated by injecting a sample mixture onto the column, where the different components pass through at different rates due to differences in how they partition between the mobile and stationary phases. There are four main types of liquid chromatography: partition, adsorption, ion exchange, and size exclusion. HPLC is used for analyzing complex mixtures, purifying compounds, and quality control in various fields such as chemistry, biochemistry, environmental analysis, and pharmaceuticals.
Ind-Swift Laboratories Ltd. is a USD ~200 million pharmaceutical company based in India with manufacturing sites across the country and an R&D center. The company produces APIs and offers contract manufacturing services. It has capabilities for reactions like Grignard, Friedel-Crafts, hydrogenation, and others. Major products include clarithromycin, roxithromycin, azithromycin, ezetimibe, and others. The company has units for R&D, quality control using HPLC, GC, and microbiology, and production. HPLC and GC are used in the quality control unit to separate and analyze compound mixtures.
The document provides an overview of high performance liquid chromatography (HPLC). It begins with defining HPLC and explaining the basic principles of chromatography. It then describes the different types of HPLC based on the mode and principle of separation. The document also discusses HPLC instrumentation, including the solvent reservoir, pump, injector, column, and various detectors. It concludes by outlining some common applications of HPLC and discussing quantitative and qualitative analysis.
Its very effective presentation of HPLC technique with VWD detectors.we also demonstrate the Force degradation Study,Stability indicating Methods, in Different Environment Conditions.
Hplc (basic principles, operation and maintenance)alim125135
This document provides an overview of high performance liquid chromatography (HPLC), including basic principles, instrumentation, separation modes, detectors, maintenance, troubleshooting, and good laboratory practices. It discusses reversed phase and normal phase HPLC systems. Key topics covered include column configuration, peak parameters, solvent selection, converting between normal and reversed phase, various detectors such as UV-Vis and ELSD, individual module maintenance, common problems, and filtration best practices. The document emphasizes the importance of proper solvent and buffer preparation, filtration, and storage to minimize contamination and maximize system performance.
Today’s analytical laboratory is faced with tight deadlines to produce results from testing environmental samples. Too often, solid-phase extraction (SPE) presents a bottleneck in the analytical testing process and may cause poor analyte recoveries and highly variable. Despite advances in analytical instrumentation, sample prep often relies on tedious, manual, and expensive techniques such as liquid-liquid extraction.
Sample preparation of environmental water samples can be automated, however.. Use of automated sample preparation addresses the many challenges that laboratories face when preparing samples and can help improve sample processing turnaround times.
Chromatography presentation goes with this free on-demand webinar. Link to webinar: https://event.on24.com/eventRegistration/EventLobbyServlet?target=registration.jsp&eventid=832348&sessionid=1&key=7401504685427A0804ABBD1F956E617C&partnerrefthermo=undefined&sourcepage=register
HPLC stands for high-performance liquid chromatography, a technique used to separate components of a mixture. It works by pumping a mobile phase and sample at high pressure through a column containing a stationary phase. Components have different interaction strengths with the phases, allowing separation as they travel through the column at different speeds. An HPLC system includes a pump, injector, column, detector, and data processing unit. It has various applications in fields like pharmaceuticals, environmental analysis, and food and flavor testing. Advantages are its ability to analyze complex mixtures accurately and quickly, while disadvantages include potential co-elution issues and high costs.
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.
Ind-Swift Laboratories Ltd. is a USD ~200 million pharmaceutical company based in India with manufacturing sites across the country and an R&D center. The company produces APIs and offers contract manufacturing services. It has capabilities for reactions like Grignard, Friedel-Crafts, hydrogenation, and others. Major products include clarithromycin, roxithromycin, azithromycin, ezetimibe, and others. The company has units for R&D, quality control using HPLC, GC, and microbiology, and production. HPLC and GC are used in the quality control unit to separate and analyze compound mixtures.
The document provides an overview of high performance liquid chromatography (HPLC). It begins with defining HPLC and explaining the basic principles of chromatography. It then describes the different types of HPLC based on the mode and principle of separation. The document also discusses HPLC instrumentation, including the solvent reservoir, pump, injector, column, and various detectors. It concludes by outlining some common applications of HPLC and discussing quantitative and qualitative analysis.
Its very effective presentation of HPLC technique with VWD detectors.we also demonstrate the Force degradation Study,Stability indicating Methods, in Different Environment Conditions.
Hplc (basic principles, operation and maintenance)alim125135
This document provides an overview of high performance liquid chromatography (HPLC), including basic principles, instrumentation, separation modes, detectors, maintenance, troubleshooting, and good laboratory practices. It discusses reversed phase and normal phase HPLC systems. Key topics covered include column configuration, peak parameters, solvent selection, converting between normal and reversed phase, various detectors such as UV-Vis and ELSD, individual module maintenance, common problems, and filtration best practices. The document emphasizes the importance of proper solvent and buffer preparation, filtration, and storage to minimize contamination and maximize system performance.
Today’s analytical laboratory is faced with tight deadlines to produce results from testing environmental samples. Too often, solid-phase extraction (SPE) presents a bottleneck in the analytical testing process and may cause poor analyte recoveries and highly variable. Despite advances in analytical instrumentation, sample prep often relies on tedious, manual, and expensive techniques such as liquid-liquid extraction.
Sample preparation of environmental water samples can be automated, however.. Use of automated sample preparation addresses the many challenges that laboratories face when preparing samples and can help improve sample processing turnaround times.
Chromatography presentation goes with this free on-demand webinar. Link to webinar: https://event.on24.com/eventRegistration/EventLobbyServlet?target=registration.jsp&eventid=832348&sessionid=1&key=7401504685427A0804ABBD1F956E617C&partnerrefthermo=undefined&sourcepage=register
HPLC stands for high-performance liquid chromatography, a technique used to separate components of a mixture. It works by pumping a mobile phase and sample at high pressure through a column containing a stationary phase. Components have different interaction strengths with the phases, allowing separation as they travel through the column at different speeds. An HPLC system includes a pump, injector, column, detector, and data processing unit. It has various applications in fields like pharmaceuticals, environmental analysis, and food and flavor testing. Advantages are its ability to analyze complex mixtures accurately and quickly, while disadvantages include potential co-elution issues and high costs.
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.
This is a PPT for HPLC which I made for presenting my assigned topic for Practice School during 7 sem of my graduation . Hope it is useful for you guys :)
HPLC
Chromatography
Mobile Phase & Stationary Phase
CLASSIFICATION OF CHROMATOGRAPHY
Characteristics of HPLC
Purpose
Superiority of HPLC
TYPES OF HPLC TECHNIQYES
Principle
PHASING SYSTEM & (normal vs reversed phase)
INSTRUMENTATION
Flow diagram of HPLC instrument
Advantages of HPLC
HPLC is now a powerful analytical technique used for separating compounds soluble in solvents. It works by pumping a pressurized mobile phase through a column containing a stationary phase, which separates the compounds in a sample based on their interactions. The basic components of an HPLC system are a solvent reservoir, pump, injector, analytical column, detector, and recorder. HPLC can be used for quality control in pharmaceuticals and environmental and forensic applications.
The document discusses high performance liquid chromatography (HPLC). It describes how HPLC provides higher separation and reduced analysis time compared to liquid chromatography through the use of smaller particle sizes, high pressure pumps, and specialized equipment. Some key advantages of HPLC include high separation capacity, moderate analytical conditions, and generally high sensitivity. HPLC is used in fields like biogenic substances, medical products, food products, environmental samples, and organic industrial products.
HPLC is a chromatographic technique used to separate components in a mixture. It works by pumping a pressurized mobile liquid phase through a column containing a stationary phase, which causes the components in a sample to separate as they are transported through the column at different rates. Key components of an HPLC system include solvent reservoirs, pumps to precisely deliver the mobile phase, an injector to introduce samples, columns for separation, detectors, and a data system to analyze results. HPLC offers advantages like high separation capacity, reproducibility, and ability to analyze a wide range of biological, medical, food, environmental, and industrial samples.
Pharmagupshup supercritical fluid chrometography and flash chromatography p...Hitesh Katariya
The document discusses supercritical fluid chromatography (SFC), including its principles and applications. SFC uses supercritical fluids like carbon dioxide as the mobile phase. It provides advantages over gas chromatography and liquid chromatography like being faster, more cost effective, and allowing better separation of some compounds. The document explains the instrumentation, process, and properties of supercritical fluids. It also gives examples of SFC applications in analyzing natural products, drugs, pesticides, lipids, and other compounds.
This document provides an overview of high performance liquid chromatography (HPLC). It discusses the key components of an HPLC system including the solvent rack, pump, injector, separation column, and detector. It also describes different chromatography techniques such as normal phase chromatography, reversed phase chromatography, and ion exchange chromatography which separate compounds based on polarity, electrical charge, and molecular size. Finally, it discusses factors that influence separation such as stationary phase particle size and eluent composition.
Solid phase extraction is the very popular technique currently available for rapid and selective sample preparation. The versatility of SPE allows use of this technique for many purposes, such as purification, trace enrichment, desalting, and class fractionation and etc.
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
HPLC: Principle and Maintenance with Applicationijtsrd
High performance liquid chromatography (HPLC) is a significant qualitative and quantitative technique, usually used for the estimation of pharmaceutical and biological samples. The chromatography term is derived from the Greek words namely chroma (colour) and graphein (to write). The chromatography is very accepted technique and it is mostly used analytically. It is the most resourceful, safest, reliable and fastest chromatographic technique for the quality control of drug components. This technique involves 2 phases"™ stationary and mobile phases. There are different types of chromatographic techniques. The separation of constituents is based on the variation between the partition coefficients of the two phases. This article is primed with an aim to review different aspects of HPLC, such as principle, types, instrumentation and application with maintenance. Yogesh Kumar | Sayed Md Mumtaz | Mustaq Ahmad"HPLC: Principle and Maintenance with Application" Published in International Journal of Trend in Scientific Research and Development (ijtsrd), ISSN: 2456-6470, Volume-2 | Issue-5 , August 2018, URL: http://www.ijtsrd.com/papers/ijtsrd17134.pdf http://www.ijtsrd.com/pharmacy/pharmacology-/17134/hplc-principle-and-maintenance-with-application/yogesh-kumar
HPLC is a liquid chromatography technique used to separate mixtures of compounds. It involves injecting a sample into a column packed with porous particles, through which a liquid mobile phase is pumped under high pressure. The separated components interact differently with the stationary phase and elute from the column at different retention times, producing a chromatogram. HPLC is used for qualitative and quantitative analysis of compounds, identification of unknown mixtures, and preparation of pure samples. It is well-suited for separation of non-volatile and thermally-fragile compounds across various application areas.
High performance liquid chromatography (HPLC) is summarized as follows:
HPLC is a technique used to separate mixtures by distributing the components between a stationary and mobile phase. It can be used for both qualitative and quantitative analysis. HPLC utilizes high pressure pumps to pass a mobile phase through a column packed with adsorbent particles, allowing separation of components based on differences in their partitioning behavior between the mobile and stationary phases. Common detectors used in HPLC include UV/Vis, refractive index, fluorescence, and mass spectrometry.
High performance liquid chromatography (HPLC) is a technique used to separate components in a mixture. It works by pumping a sample mixture through a column containing chromatographic packing material at high pressure. The sample components interact differently with the stationary phase in the column, causing them to elute out at different rates and allowing separation. HPLC has many applications in fields like pharmaceuticals, environmental analysis, food and flavors testing, clinical testing, and forensics. It provides a powerful analytical tool for identifying and quantifying compounds in samples.
Flash chromatography is a technique that uses compressed gas to rapidly push solvent through a column packed with adsorbent material like silica gel. This allows for faster separations compared to traditional gravity chromatography. The document discusses the principles, common solvent systems, packing columns, instrumentation used like pumps, fraction collectors, and detectors, and applications of flash chromatography.
High Performance Liquid Chromatography HPLC is a process of separating components in a liquid mixture. A liquid sample is injected into a stream of solvent mobile phase flowing through a column packed with a separation medium stationary phase . Sample components separate from one another by a process of differential migration as they flow through the column.As bands emerge from the column, flow carries them to one or more detectors which deliver a voltage response as a function of time. This is called a chromatogram. For each peak, the time at which it emerges identifies the sample constituent with respect to a standard. The peak’s area represents the quantity .HPLC provides a highly specific, reasonably precise, and fairly rapid analytical method for a plethora of complicated samples.This is difficult in detecting compounds. Low sensitivity of some compounds towards the stationary phase in the columns is difficult. Mohd Ali | Panjak Chasta | Dr. Kausal Kishore Chandrul "High Performance Liquid Chromatography (HPLC)" Published in International Journal of Trend in Scientific Research and Development (ijtsrd), ISSN: 2456-6470, Volume-5 | Issue-5 , August 2021, URL: https://www.ijtsrd.com/papers/ijtsrd45146.pdf Paper URL: https://www.ijtsrd.com/pharmacy/other/45146/high-performance-liquid-chromatography-hplc/mohd-ali
This document provides an overview of high performance liquid chromatography (HPLC). It discusses the basic principles of chromatography and various HPLC modes, including normal phase, reversed phase, ion exchange, and size exclusion chromatography. The document also describes HPLC system components like pumps, injectors, columns, ovens, and detectors. It explains isocratic and gradient elution modes and discusses the instrumentation, functions, and advantages of various HPLC detectors.
This document outlines the five main steps for developing an analytical HPLC method: 1) selecting the initial HPLC method and conditions, 2) selecting the initial chromatographic conditions, 3) optimizing selectivity, 4) optimizing system parameters, and 5) validating the method. Key aspects of each step are discussed, including selecting the type of chromatography, column, detector, and mobile phase based on the analytes. The goal is to develop a validated method that provides adequate resolution and selectivity within a desired analysis time.
This document provides an overview of high performance liquid chromatography (HPLC). It describes the basic components of an HPLC system including the solvent delivery system, injector, column, and detector. It explains how samples are separated based on the differences in how components partition between the mobile and stationary phases. Various types of chromatography are also summarized, including partition, adsorption, ion exchange, and size exclusion. Factors that can affect separations like column parameters, instrument parameters, and sample parameters are outlined.
This document provides an overview of high performance liquid chromatography (HPLC). It describes the key components of an HPLC system, including the pump, injector, separation column, and detector. It explains how the different components of a mixture are separated through their interaction with the mobile and stationary phases in the column. The document also discusses various types of columns, detectors, and applications of HPLC in chemistry, biochemistry, and quality control.
This is a PPT for HPLC which I made for presenting my assigned topic for Practice School during 7 sem of my graduation . Hope it is useful for you guys :)
HPLC
Chromatography
Mobile Phase & Stationary Phase
CLASSIFICATION OF CHROMATOGRAPHY
Characteristics of HPLC
Purpose
Superiority of HPLC
TYPES OF HPLC TECHNIQYES
Principle
PHASING SYSTEM & (normal vs reversed phase)
INSTRUMENTATION
Flow diagram of HPLC instrument
Advantages of HPLC
HPLC is now a powerful analytical technique used for separating compounds soluble in solvents. It works by pumping a pressurized mobile phase through a column containing a stationary phase, which separates the compounds in a sample based on their interactions. The basic components of an HPLC system are a solvent reservoir, pump, injector, analytical column, detector, and recorder. HPLC can be used for quality control in pharmaceuticals and environmental and forensic applications.
The document discusses high performance liquid chromatography (HPLC). It describes how HPLC provides higher separation and reduced analysis time compared to liquid chromatography through the use of smaller particle sizes, high pressure pumps, and specialized equipment. Some key advantages of HPLC include high separation capacity, moderate analytical conditions, and generally high sensitivity. HPLC is used in fields like biogenic substances, medical products, food products, environmental samples, and organic industrial products.
HPLC is a chromatographic technique used to separate components in a mixture. It works by pumping a pressurized mobile liquid phase through a column containing a stationary phase, which causes the components in a sample to separate as they are transported through the column at different rates. Key components of an HPLC system include solvent reservoirs, pumps to precisely deliver the mobile phase, an injector to introduce samples, columns for separation, detectors, and a data system to analyze results. HPLC offers advantages like high separation capacity, reproducibility, and ability to analyze a wide range of biological, medical, food, environmental, and industrial samples.
Pharmagupshup supercritical fluid chrometography and flash chromatography p...Hitesh Katariya
The document discusses supercritical fluid chromatography (SFC), including its principles and applications. SFC uses supercritical fluids like carbon dioxide as the mobile phase. It provides advantages over gas chromatography and liquid chromatography like being faster, more cost effective, and allowing better separation of some compounds. The document explains the instrumentation, process, and properties of supercritical fluids. It also gives examples of SFC applications in analyzing natural products, drugs, pesticides, lipids, and other compounds.
This document provides an overview of high performance liquid chromatography (HPLC). It discusses the key components of an HPLC system including the solvent rack, pump, injector, separation column, and detector. It also describes different chromatography techniques such as normal phase chromatography, reversed phase chromatography, and ion exchange chromatography which separate compounds based on polarity, electrical charge, and molecular size. Finally, it discusses factors that influence separation such as stationary phase particle size and eluent composition.
Solid phase extraction is the very popular technique currently available for rapid and selective sample preparation. The versatility of SPE allows use of this technique for many purposes, such as purification, trace enrichment, desalting, and class fractionation and etc.
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
HPLC: Principle and Maintenance with Applicationijtsrd
High performance liquid chromatography (HPLC) is a significant qualitative and quantitative technique, usually used for the estimation of pharmaceutical and biological samples. The chromatography term is derived from the Greek words namely chroma (colour) and graphein (to write). The chromatography is very accepted technique and it is mostly used analytically. It is the most resourceful, safest, reliable and fastest chromatographic technique for the quality control of drug components. This technique involves 2 phases"™ stationary and mobile phases. There are different types of chromatographic techniques. The separation of constituents is based on the variation between the partition coefficients of the two phases. This article is primed with an aim to review different aspects of HPLC, such as principle, types, instrumentation and application with maintenance. Yogesh Kumar | Sayed Md Mumtaz | Mustaq Ahmad"HPLC: Principle and Maintenance with Application" Published in International Journal of Trend in Scientific Research and Development (ijtsrd), ISSN: 2456-6470, Volume-2 | Issue-5 , August 2018, URL: http://www.ijtsrd.com/papers/ijtsrd17134.pdf http://www.ijtsrd.com/pharmacy/pharmacology-/17134/hplc-principle-and-maintenance-with-application/yogesh-kumar
HPLC is a liquid chromatography technique used to separate mixtures of compounds. It involves injecting a sample into a column packed with porous particles, through which a liquid mobile phase is pumped under high pressure. The separated components interact differently with the stationary phase and elute from the column at different retention times, producing a chromatogram. HPLC is used for qualitative and quantitative analysis of compounds, identification of unknown mixtures, and preparation of pure samples. It is well-suited for separation of non-volatile and thermally-fragile compounds across various application areas.
High performance liquid chromatography (HPLC) is summarized as follows:
HPLC is a technique used to separate mixtures by distributing the components between a stationary and mobile phase. It can be used for both qualitative and quantitative analysis. HPLC utilizes high pressure pumps to pass a mobile phase through a column packed with adsorbent particles, allowing separation of components based on differences in their partitioning behavior between the mobile and stationary phases. Common detectors used in HPLC include UV/Vis, refractive index, fluorescence, and mass spectrometry.
High performance liquid chromatography (HPLC) is a technique used to separate components in a mixture. It works by pumping a sample mixture through a column containing chromatographic packing material at high pressure. The sample components interact differently with the stationary phase in the column, causing them to elute out at different rates and allowing separation. HPLC has many applications in fields like pharmaceuticals, environmental analysis, food and flavors testing, clinical testing, and forensics. It provides a powerful analytical tool for identifying and quantifying compounds in samples.
Flash chromatography is a technique that uses compressed gas to rapidly push solvent through a column packed with adsorbent material like silica gel. This allows for faster separations compared to traditional gravity chromatography. The document discusses the principles, common solvent systems, packing columns, instrumentation used like pumps, fraction collectors, and detectors, and applications of flash chromatography.
High Performance Liquid Chromatography HPLC is a process of separating components in a liquid mixture. A liquid sample is injected into a stream of solvent mobile phase flowing through a column packed with a separation medium stationary phase . Sample components separate from one another by a process of differential migration as they flow through the column.As bands emerge from the column, flow carries them to one or more detectors which deliver a voltage response as a function of time. This is called a chromatogram. For each peak, the time at which it emerges identifies the sample constituent with respect to a standard. The peak’s area represents the quantity .HPLC provides a highly specific, reasonably precise, and fairly rapid analytical method for a plethora of complicated samples.This is difficult in detecting compounds. Low sensitivity of some compounds towards the stationary phase in the columns is difficult. Mohd Ali | Panjak Chasta | Dr. Kausal Kishore Chandrul "High Performance Liquid Chromatography (HPLC)" Published in International Journal of Trend in Scientific Research and Development (ijtsrd), ISSN: 2456-6470, Volume-5 | Issue-5 , August 2021, URL: https://www.ijtsrd.com/papers/ijtsrd45146.pdf Paper URL: https://www.ijtsrd.com/pharmacy/other/45146/high-performance-liquid-chromatography-hplc/mohd-ali
This document provides an overview of high performance liquid chromatography (HPLC). It discusses the basic principles of chromatography and various HPLC modes, including normal phase, reversed phase, ion exchange, and size exclusion chromatography. The document also describes HPLC system components like pumps, injectors, columns, ovens, and detectors. It explains isocratic and gradient elution modes and discusses the instrumentation, functions, and advantages of various HPLC detectors.
This document outlines the five main steps for developing an analytical HPLC method: 1) selecting the initial HPLC method and conditions, 2) selecting the initial chromatographic conditions, 3) optimizing selectivity, 4) optimizing system parameters, and 5) validating the method. Key aspects of each step are discussed, including selecting the type of chromatography, column, detector, and mobile phase based on the analytes. The goal is to develop a validated method that provides adequate resolution and selectivity within a desired analysis time.
This document provides an overview of high performance liquid chromatography (HPLC). It describes the basic components of an HPLC system including the solvent delivery system, injector, column, and detector. It explains how samples are separated based on the differences in how components partition between the mobile and stationary phases. Various types of chromatography are also summarized, including partition, adsorption, ion exchange, and size exclusion. Factors that can affect separations like column parameters, instrument parameters, and sample parameters are outlined.
This document provides an overview of high performance liquid chromatography (HPLC). It describes the key components of an HPLC system, including the pump, injector, separation column, and detector. It explains how the different components of a mixture are separated through their interaction with the mobile and stationary phases in the column. The document also discusses various types of columns, detectors, and applications of HPLC in chemistry, biochemistry, and quality control.
This document provides information about High Performance Liquid Chromatography (HPLC) and Gas Chromatography. It discusses the basic principles, instrumentation, and applications of HPLC, including the types of columns, mobile phases, pumps, injectors, detectors, and data acquisition systems used. It also summarizes the basic principles of Gas Chromatography, where a gas mobile phase is used to separate components of a vaporized sample based on interactions with a stationary phase. Key applications of HPLC mentioned include pharmaceutical analysis, environmental monitoring, clinical analysis, and food and flavor analysis.
HPLC Principle,Instrumentation and ApplicationAlakesh Pradhan
HPLC Chromatography and its principle
Liquid chromatography
High Performance Liquid Chromatography ( HPLC )
The components of the high performance liquid chromatograph (HPLC).
The separation process.
The chromatogram
High performance liquid chromatography (HPLC) is a technique used to separate compounds in a liquid mixture. An HPLC system consists of a solvent reservoir, pump, injector, separation column, and detector. The mixture is injected and the components separate as they pass through the column at different rates depending on how they partition between the mobile and stationary phases. HPLC is used for research, quality control, and regulatory purposes to separate, analyze, purify, and quantify complex mixtures like drugs, metabolites, polymers, proteins, lipids, and environmental pollutants.
This document discusses several chromatography techniques used in forensic science analysis, including high performance liquid chromatography (HPLC), gas chromatography (GC), and inductively coupled plasma mass spectrometry (ICP-MS). It describes the basic principles, instrumentation components, and applications of each technique. HPLC uses high pressure to separate mixtures based on interactions with a stationary and mobile liquid phase. GC separates volatile compounds using an inert gas mobile phase and liquid stationary phase. ICP-MS uses plasma to ionize elements and masses to identify unknown samples at very low concentrations.
Instrumentation of HPLC, principle by kk sahuKAUSHAL SAHU
INTRODUCTION
Instrumentation of HPLC
TYPES OF HPLC
PARAMETERS
APPLICATION
CONCLUSION
REFERENCE
High-performance liquid chromatography ( HPLC) is a specific form of column chromatography generally used in biochemistry and analysis to separate, identify, and quantify the active compounds.
HPLC mainly utilizes a column that holds packing material (stationary phase), a pump that moves the mobile phase(s) through the column, and a detector that shows the retention times of the molecules.
The document discusses High Performance Liquid Chromatography (HPLC), including its principles, types of separations, instrumentation, applications in fields like food and pharmaceuticals, and advantages like speed and accuracy over traditional chromatography techniques. HPLC uses high pressure to pass samples in liquid mobile phases through stationary phases in columns to separate mixtures based on properties like polarity, size, or ionic charge.
Bioanalysis using HPLC is crucial in the pharmaceutical industry to measure drug and metabolite levels. HPLC is a powerful and versatile liquid chromatography technique that forces samples through columns at high pressures for faster, more efficient separation of components. Key components of an HPLC system include columns, pumps, detectors, and data collection systems. HPLC follows the same principles as chromatography but provides higher speed and sensitivity through the use of high pressure. It has many applications including qualitative and quantitative analysis of chemicals, drugs, and biological compounds.
high performance liquid chromatography 22alishapep2020
HPLC is a form of column chromatography that separates compounds based on their interaction with a stationary and mobile phase. It works by pumping a mobile liquid phase through a column containing tightly packed microparticles that serve as the stationary phase. Samples are injected and different compounds are retained for different lengths of time as they pass through the column before being detected, usually with a UV detector. HPLC has various applications in fields like pharmaceuticals, forensics, environmental analysis, and more. It provides advantages of speed, sensitivity, and ability to separate complex mixtures over other techniques.
Chromatography is a technique used to separate mixtures by distributing compounds between a stationary and mobile phase. High-performance liquid chromatography (HPLC) is commonly used and separates compounds using a column with a stationary phase and liquid mobile phase. HPLC can identify, detect, quantify, and purify individual components in a mixture using an apparatus including a pump, injector, column, detector, and recorder. The separation occurs as the compounds interact differently with the stationary phase in the column.
This document discusses high performance liquid chromatography (HPLC) and its applications in forensic science. It begins with introducing HPLC and its principles, which involve separating components in a mixture using a stationary and mobile phase under high pressure. It then describes HPLC instrumentation, methodology, types based on separation mode (e.g. normal phase, reverse phase), columns, and forensic applications such as identifying drugs, dyes, and metabolites in samples. Specific examples given are using HPLC to estimate the age of a human bloodstain and matching a lipstick smear from a crime scene to a specific brand.
High performance liquid chromatography (HPLC) is described, including the basic principles and components of HPLC systems. HPLC uses high pressure to pass a liquid mobile phase through a column packed with solid adsorbent particles or porous beads. This allows for separation of mixtures based on differences in how components partition between the stationary and mobile phases. Key components reviewed are the solvent reservoirs, pump, injector, column, and detectors. Common applications of HPLC mentioned are qualitative and quantitative analysis of both volatile and non-volatile compounds.
High performance liquid chromatography (HPLC) is described, including the basic principles and components of HPLC systems. HPLC uses high pressure to pass a liquid mobile phase through a column packed with solid adsorbent particles or porous beads. This allows for separation of mixtures based on differences in how components partition between the stationary and mobile phases. Key components reviewed are the solvent reservoirs, pump, injector, column, and detectors. Common applications of HPLC mentioned are qualitative and quantitative analysis of both volatile and non-volatile compounds.
High Performance Liquid Chromatography (HPLC) is a form of column chromatography that pumps a sample mixture or analyte in a solvent (known as the mobile phase) at high pressure through a column with chromatographic packing material (stationary phase).
HPLC is a form of liquid chromatography used to separate compounds dissolved in solution using a pump to deliver a mobile phase through a column containing adsorbent particles. It can be used for qualitative analysis to identify compounds, quantitative analysis to measure concentrations, and trace analysis of low concentration compounds. The main components of an HPLC system are the pump, injector, column, detector and computer. Reversed phase chromatography is the most common separation mode using a non-polar stationary phase and polar mobile phase.
This document provides an overview of high performance liquid chromatography (HPLC). It describes chromatography as a separation technique using a mobile and stationary phase. It then discusses various types of chromatography including HPLC, and defines HPLC as high performance liquid chromatography. The document outlines the basic components of an HPLC system including the pump, injector, column, detector, and mobile phases. It provides details on each component and their functions. Various parameters used in HPLC are also defined, such as retention time, resolution, theoretical plates, and asymmetry factor. Finally, the document discusses some applications of HPLC like qualitative and quantitative analysis.
The document discusses High Performance Liquid Chromatography (HPLC). It provides background on the origins and development of HPLC from the early 1900s. By the 1980s, HPLC was commonly used for separation, identification, purification and quantification of chemical compounds. The document then discusses the basic principles, components, and instrumentation of HPLC systems. It explains how HPLC uses high pressure to force a mobile phase through a column containing a stationary phase to separate compounds based on their interactions with the phases. Common applications of HPLC across various fields like pharmaceuticals, environmental analysis, forensics and food are also summarized.
High performance liquid chromatography (hplc) by Muhammad ShakaibMuhammad Shakaib
High performance liquid chromatography (HPLC) is a technique used to separate components in a mixture. It works by differential migration of solutes through a column containing a stationary phase as a pressurized mobile phase flows through. Solutes are detected as they elute from the column. Key aspects of HPLC include the use of a high pressure pump to deliver the mobile phase, columns packed with microparticulate stationary phases, and detectors to identify eluted components. HPLC is useful for separating compounds that are not volatile enough for gas chromatography.
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Sectional dentures for microstomia patients.pptxSatvikaPrasad
Microstomia, characterized by an abnormally small oral aperture, presents significant challenges in prosthodontic treatment, including limited access for examination, difficulties in impression making, and challenges with prosthesis insertion and removal. To manage these issues, customized impression techniques using sectional trays and elastomeric materials are employed. Prostheses may be designed in segments or with flexible materials to facilitate handling. Minimally invasive procedures and the use of digital technologies can enhance patient comfort. Education and training for patients on prosthesis care and maintenance are crucial for compliance. Regular follow-up and a multidisciplinary approach, involving collaboration with other specialists, ensure comprehensive care and improved quality of life for microstomia patients.
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2. IntroductionIntroduction
HPLC is a form of liquid chromatography used toHPLC is a form of liquid chromatography used to
separate compounds that are dissolved in solution.separate compounds that are dissolved in solution.
HPLC instruments consist of a reservoir of mobile phase,HPLC instruments consist of a reservoir of mobile phase,
a pump, an injector, a separation column, and aa pump, an injector, a separation column, and a
detector.detector.
Compounds are separated by injecting a sample mixtureCompounds are separated by injecting a sample mixture
onto the column. The different component in the mixtureonto the column. The different component in the mixture
pass through the column at differentiates due topass through the column at differentiates due to
differences in their partition behavior between the mobiledifferences in their partition behavior between the mobile
phase and the stationary phase. The mobile phase mustphase and the stationary phase. The mobile phase must
be degassed to eliminate the formation of air bubbles.be degassed to eliminate the formation of air bubbles.
4. FOUR TYPES OF LIQUIDFOUR TYPES OF LIQUID
CHROMATOGRAPHYCHROMATOGRAPHY
Partition chromatographyPartition chromatography
Adsorption, or liquid-solidAdsorption, or liquid-solid
chromatographychromatography
Ion exchange chromatographyIon exchange chromatography
Size exclusion, or gel, chromatographySize exclusion, or gel, chromatography
5. COMPOSITION OF A LIQUIDCOMPOSITION OF A LIQUID
CHROMATOGRAPH SYSTEMCHROMATOGRAPH SYSTEM
SolventSolvent
Solvent Delivery System (Pump)Solvent Delivery System (Pump)
InjectorInjector
SampleSample
ColumnColumn
Detectors (Diode Array)Detectors (Diode Array)
Waste CollectorWaste Collector
Recorder (Data Collection)Recorder (Data Collection)
7. Uses of HPLCUses of HPLC
This technique is used for chemistry and biochemistryThis technique is used for chemistry and biochemistry
research analyzing complex mixtures, purifying chemicalresearch analyzing complex mixtures, purifying chemical
compounds, developing processes for synthesizing chemicalcompounds, developing processes for synthesizing chemical
compounds, isolating natural products, or predicting physicalcompounds, isolating natural products, or predicting physical
properties. It is also used in quality control to ensure the purityproperties. It is also used in quality control to ensure the purity
of raw materials, to control and improve process yields, toof raw materials, to control and improve process yields, to
quantify assays of final products, or to evaluate productquantify assays of final products, or to evaluate product
stability and monitor degradation.stability and monitor degradation.
In addition, it is used for analyzing air and water pollutants, forIn addition, it is used for analyzing air and water pollutants, for
monitoring materials that may jeopardize occupational safetymonitoring materials that may jeopardize occupational safety
or health, and for monitoring pesticide levels in theor health, and for monitoring pesticide levels in the
environment. Federal and state regulatory agencies use HPLCenvironment. Federal and state regulatory agencies use HPLC
to survey food and drug products, for identifying confiscatedto survey food and drug products, for identifying confiscated
narcotics or to check for adherence to label claims.narcotics or to check for adherence to label claims.
8. HPLC Chromatograph injectorsHPLC Chromatograph injectors
The function of the injector is to place the sample intoThe function of the injector is to place the sample into
the high-pressure flow in as narrow volume as possiblethe high-pressure flow in as narrow volume as possible
so that the sample enters the column as aso that the sample enters the column as a
homogeneous, low-volume plug. To minimize spreadinghomogeneous, low-volume plug. To minimize spreading
of the injected volume during transport to the column, theof the injected volume during transport to the column, the
shortest possible length of tubing should be used fromshortest possible length of tubing should be used from
the injector to the column.the injector to the column.
When an injection is started, an air actuator rotates theWhen an injection is started, an air actuator rotates the
valve: solvent goes directly to the column; and thevalve: solvent goes directly to the column; and the
injector needle is connected to the syringe. The airinjector needle is connected to the syringe. The air
pressure lifts the needle and the vial is moved intopressure lifts the needle and the vial is moved into
position beneath the needle. Then, the needle is loweredposition beneath the needle. Then, the needle is lowered
to the vial.to the vial.
9. HPLC columnsHPLC columns
The column is one of theThe column is one of the
most important componentsmost important components
of the HPLC chromatographof the HPLC chromatograph
because the separation ofbecause the separation of
the sample components isthe sample components is
achieved when thoseachieved when those
components pass throughcomponents pass through
the column. The Highthe column. The High
performance liquidperformance liquid
chromatography apparatuschromatography apparatus
is made out of stainlessis made out of stainless
steel tubes with a diametersteel tubes with a diameter
of 3 to 5mm and a lengthof 3 to 5mm and a length
ranging from 10 to 30cm.ranging from 10 to 30cm.
Normally, columns are filledNormally, columns are filled
with silica gel because itswith silica gel because its
particle shape, surfaceparticle shape, surface
properties, and pore structureproperties, and pore structure
help to get a goodhelp to get a good
separation. Silica is wetted byseparation. Silica is wetted by
nearly every potential mobilenearly every potential mobile
phase, is inert to mostphase, is inert to most
compounds and has a highcompounds and has a high
surface activity which can besurface activity which can be
modified easily with watermodified easily with water
and other agents. Silica canand other agents. Silica can
be used to separate a widebe used to separate a wide
variety of chemicalvariety of chemical
compounds, and itscompounds, and its
chromatographic behavior ischromatographic behavior is
generally predictable andgenerally predictable and
reproducible.reproducible.
13. Several column typesSeveral column types
(can be classified as )(can be classified as )
Normal phaseNormal phase
Reverse phaseReverse phase
Size exclusionSize exclusion
Ion exchangeIon exchange
14. Normal phaseNormal phase
In this column type, the retention isIn this column type, the retention is
governed by the interaction of the polargoverned by the interaction of the polar
parts of the stationary phase and solute.parts of the stationary phase and solute.
For retention to occur in normal phase, theFor retention to occur in normal phase, the
packing must be more polar than thepacking must be more polar than the
mobile phase with respect to the samplemobile phase with respect to the sample
15. Reverse phaseReverse phase
In this column the packing material is relativelyIn this column the packing material is relatively
nonpolar and the solvent is polar with respect tononpolar and the solvent is polar with respect to
the sample. Retention is the result of thethe sample. Retention is the result of the
interaction of the nonpolar components of theinteraction of the nonpolar components of the
solutes and the nonpolar stationary phase.solutes and the nonpolar stationary phase.
Typical stationary phases are nonpolarTypical stationary phases are nonpolar
hydrocarbons, waxy liquids, or bondedhydrocarbons, waxy liquids, or bonded
hydrocarbons (such as C18, C8, etc.) and thehydrocarbons (such as C18, C8, etc.) and the
solvents are polar aqueous-organic mixturessolvents are polar aqueous-organic mixtures
such as methanol-water or acetonitrile-water.such as methanol-water or acetonitrile-water.
16. Size exclusionSize exclusion
In size exclusion the HPLC column isIn size exclusion the HPLC column is
consisted of substances which haveconsisted of substances which have
controlled pore sizes and is able to becontrolled pore sizes and is able to be
filtered in an ordinarily phase according tofiltered in an ordinarily phase according to
its molecular size. Small moleculesits molecular size. Small molecules
penetrate into the pores within the packingpenetrate into the pores within the packing
while larger molecules only partiallywhile larger molecules only partially
penetrate the pores. The large moleculespenetrate the pores. The large molecules
elute before the smaller molecules.elute before the smaller molecules.
17. Ion exchangeIon exchange
In this column type the sampleIn this column type the sample
components are separated based uponcomponents are separated based upon
attractive ionic forces between moleculesattractive ionic forces between molecules
carrying charged groups of oppositecarrying charged groups of opposite
charge to those charges on the stationarycharge to those charges on the stationary
phase. Separations are made between aphase. Separations are made between a
polar mobile liquid, usually waterpolar mobile liquid, usually water
containing salts or small amounts ofcontaining salts or small amounts of
alcohols, and a stationary phasealcohols, and a stationary phase
containing either acidic or basic fixedcontaining either acidic or basic fixed
sites.sites.
18. Selectivity FactorSelectivity Factor
K’ values tell us where bands elute relativeK’ values tell us where bands elute relative
to the void volume. These values areto the void volume. These values are
unaffected by such variables as flow rateunaffected by such variables as flow rate
and column dimensions. The value tell usand column dimensions. The value tell us
where two peaks elute relative to eachwhere two peaks elute relative to each
other. This is referred to as the selectivityother. This is referred to as the selectivity
factor or separation factor (now and thenfactor or separation factor (now and then
as the chemistry factor).as the chemistry factor).