Paper chromatography was invented in 1944 by British biochemists Martin and Synge. They developed the technique to separate and analyze amino acids, winning the 1952 Nobel Prize. Paper chromatography works on the principle of partition between two liquid phases - water held in paper pores and a mobile solvent phase. There are various modes like ascending, descending, and radial. Qualitative analysis involves choosing a stationary/mobile phase, developing the chromatogram, detecting spots, and calculating Rf values. Quantitative analysis can extract and analyze spots or use in situ methods like densitometry. Paper chromatography found many applications and was crucial to discoveries like DNA structure.
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 an overview of instrument validation and calibration techniques. It discusses the definitions of calibration and validation, highlighting that calibration demonstrates an instrument produces results within specified limits compared to a reference standard, while validation establishes that an analytical procedure meets requirements for intended use. The need for regular calibration of instruments is explained, including after installation, time periods, shocks, or questionable observations. Methods for calibrating an infrared spectrophotometer are presented, including verifying wave numbers and resolution performance against tolerance limits. Applications of infrared spectroscopy like structure determination and identification are also mentioned.
Multi component analysis (uv visible spectroscopy) by mr. pradeep swarnkar)Pradeep Swarnkar
This document discusses multi-component analysis using UV-Visible spectroscopy. It describes using the absorbance values of multiple components at different wavelengths to determine concentration ratios between the components known as Q values. The Q values can then be used to determine the individual concentrations of each component in a mixture.
This document discusses evaluation parameters for gastroretentive drug delivery systems (GRDDS). It describes several approaches to gastroretention including floating systems, swelling systems, mucoadhesive systems, and others. It also discusses pre-compression parameters like angle of repose and compressibility index, and post-compression parameters like tablet shape, dimensions, hardness, friability, weight variation, and density that are used to evaluate GRDDS tablets. The document provides information on the anatomy and physiology factors influencing gastric retention and emptying.
Gastro retentive drug delivery system (GRDDS)Shweta Nehate
This document discusses gastro-retentive drug delivery systems (GRDDS), which aim to prolong the gastric residence time of drugs and target drug release in the upper gastrointestinal tract. It describes the physiology of the gastrointestinal tract and potential drug candidates for GRDDS. Various approaches for GRDDS are covered, including floating, high density, bioadhesive, swelling, and superporous hydrogel systems. Evaluation parameters, applications, marketed formulations, and conclusions about GRDDS are also summarized.
This document discusses various in vitro, in vivo, and in situ methods used to study drug absorption. It describes several in vitro techniques like partition coefficient studies, artificial membrane models, and cell culture models. It also explains various in vivo methods like direct blood/urine sampling and indirect pharmacological response studies in animals. Finally, it covers in situ techniques like intestinal perfusion that better simulate in vivo conditions compared to in vitro models. The document provides detailed descriptions of specific methods like the everted sac technique, diffusion cell method, and stomach perfusion studies in rats.
The document discusses various spectrophotometric methods for analyzing drug samples containing multiple absorbing components, including simultaneous equation method. The simultaneous equation method allows determination of concentration of two drugs (X and Y) that absorb at each other's wavelength maxima. It requires knowing the absorptivity of each drug at the two wavelengths and the sample absorbances at those wavelengths to set up two equations and solve for the unknown concentrations. Certain criteria on absorbance ratios must be met for precise results.
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 an overview of instrument validation and calibration techniques. It discusses the definitions of calibration and validation, highlighting that calibration demonstrates an instrument produces results within specified limits compared to a reference standard, while validation establishes that an analytical procedure meets requirements for intended use. The need for regular calibration of instruments is explained, including after installation, time periods, shocks, or questionable observations. Methods for calibrating an infrared spectrophotometer are presented, including verifying wave numbers and resolution performance against tolerance limits. Applications of infrared spectroscopy like structure determination and identification are also mentioned.
Multi component analysis (uv visible spectroscopy) by mr. pradeep swarnkar)Pradeep Swarnkar
This document discusses multi-component analysis using UV-Visible spectroscopy. It describes using the absorbance values of multiple components at different wavelengths to determine concentration ratios between the components known as Q values. The Q values can then be used to determine the individual concentrations of each component in a mixture.
This document discusses evaluation parameters for gastroretentive drug delivery systems (GRDDS). It describes several approaches to gastroretention including floating systems, swelling systems, mucoadhesive systems, and others. It also discusses pre-compression parameters like angle of repose and compressibility index, and post-compression parameters like tablet shape, dimensions, hardness, friability, weight variation, and density that are used to evaluate GRDDS tablets. The document provides information on the anatomy and physiology factors influencing gastric retention and emptying.
Gastro retentive drug delivery system (GRDDS)Shweta Nehate
This document discusses gastro-retentive drug delivery systems (GRDDS), which aim to prolong the gastric residence time of drugs and target drug release in the upper gastrointestinal tract. It describes the physiology of the gastrointestinal tract and potential drug candidates for GRDDS. Various approaches for GRDDS are covered, including floating, high density, bioadhesive, swelling, and superporous hydrogel systems. Evaluation parameters, applications, marketed formulations, and conclusions about GRDDS are also summarized.
This document discusses various in vitro, in vivo, and in situ methods used to study drug absorption. It describes several in vitro techniques like partition coefficient studies, artificial membrane models, and cell culture models. It also explains various in vivo methods like direct blood/urine sampling and indirect pharmacological response studies in animals. Finally, it covers in situ techniques like intestinal perfusion that better simulate in vivo conditions compared to in vitro models. The document provides detailed descriptions of specific methods like the everted sac technique, diffusion cell method, and stomach perfusion studies in rats.
The document discusses various spectrophotometric methods for analyzing drug samples containing multiple absorbing components, including simultaneous equation method. The simultaneous equation method allows determination of concentration of two drugs (X and Y) that absorb at each other's wavelength maxima. It requires knowing the absorptivity of each drug at the two wavelengths and the sample absorbances at those wavelengths to set up two equations and solve for the unknown concentrations. Certain criteria on absorbance ratios must be met for precise results.
Ultraviolet-visible spectroscopy instrumentation, choice of solvents, and solvent effects are discussed. Key points include:
UV-Vis spectrophotometers measure light intensity as a function of wavelength using sources, wavelength selectors, sample containers, detectors, and readout devices. Common sources include tungsten-halogen lamps, deuterium lamps, and LEDs. Wavelength selectors include filters and monochromators using prisms or gratings. Choice of solvent is important to avoid absorption in the detection region, with 95% ethanol commonly used. Solvent effects can cause absorption band shifts due to changes in solute-solvent interactions between ground and excited states.
This document discusses gastroretentive drug delivery systems (GRDDS), which are oral dosage forms designed to remain in the stomach for an extended period of time to prolong drug release. It covers the rationale for using GRDDS, factors controlling gastric residence time, and various approaches for prolonging gastric retention including floating systems, high-density systems, and bioadhesive or magnetic systems. Floating systems include non-effervescent and effervescent types that float due to low density or gas generation. High-density systems do not float but remain in the stomach through bioadhesion, magnetic forces, swelling to a large size, or raft formation on gastric fluids.
Application Of Polymer In Controlled Release FormulationAnindya Jana
Polymers are becoming increasingly important in the field of drug delivery. The pharmaceutical applications of polymers range from their use as binders in tablets to viscosity and flow controlling agents in liquids, suspensions and emulsions. Polymers can be used as film coatings to disguise the unpleasant taste of a drug, to enhance drug stability and to modify drug release characteristics.
As a consequence, increasing attention has been focused on methods of giving drugs continually for a prolonged time periods and in a controlled fashion.
This technology now spans many fields and includes pharmaceutical, food and agricultural applications, pesticides, cosmetics, and household products.
This document summarizes a seminar on gastroretentive drug delivery systems (GRDDS). GRDDS are designed to retain drugs in the stomach for prolonged periods of time to allow for sustained drug release. The seminar outlines various GRDDS technologies including floating, swelling, mucoadhesive, and high density systems. It also discusses candidate drugs for GRDDS, advantages like improved bioavailability, and evaluation methods like dissolution testing, floating time, and mucoadhesive strength testing. Limitations include instability at gastric pH and requirement of high fluid levels for floating systems.
This document summarizes a seminar on ion exchange chromatography. It introduces the topic, covering the basic principles of how ion exchange chromatography separates ions and polar molecules using stationary phases with positively or negatively charged groups. It then discusses the types of ion exchange resins, including classifications based on chemical nature and source. Examples of applications are given, such as separating similar ions, water softening, demineralization, separating sugars and amino acids. The document concludes by discussing some advantages and disadvantages of ion exchange chromatography.
Gas chromatography is a technique used to separate components of a vaporized sample. It works by partitioning the components between a mobile gaseous phase and a stationary phase within a column. The sample is injected and vaporized, then transported through the column by the mobile phase gas. As the components pass through the column they are separated and detected. Common detectors include the flame ionization detector (FID), thermal conductivity detector (TCD), and electron capture detector (ECD). The FID responds to organic compounds, the TCD is universal but less sensitive, and the ECD selectively detects halogen-containing compounds.
The document discusses the formulation and evaluation of gastro-retentive mucoadhesive microballoons of nizatidine for the management of peptic ulcer. It provides background information on peptic ulcers, drug delivery systems, gastroretentive drug delivery systems, and mechanisms of bioadhesion. The objective is to develop mucoadhesive microballoons of nizatidine to increase its retention time in the stomach and improve the treatment of peptic ulcers.
The document provides information on nasopulmonary drug delivery systems including nasal drug delivery and pulmonary drug delivery. It discusses the anatomy and physiology of the nasal cavity and respiratory tract. It also describes various formulation approaches for nasal delivery such as nasal gels, drops, sprays and powders. The document further explains dry powder inhalers, metered dose inhalers and nebulizers as pulmonary drug delivery systems along with their advantages and disadvantages. It also discusses some marketed products for nasal sprays, dry powder inhalers and metered dose inhalers.
This presentation discusses partition column chromatography. It begins with an introduction and presenter information. It then defines partition chromatography as a method of separation where components in a mixture distribute between two liquid phases due to differences in partition coefficients. The presentation notes that partition chromatography is the most widely used type of HPLC, where the stationary phase is a second immiscible liquid bonded to the column. It discusses the principles of how separation occurs due to differences in how components partition between the two phases. Finally, it provides examples of applications such as separation of compounds in biological and environmental samples and determination of water quality.
This document discusses common oral health problems and their causes, prevention, and treatment. It covers dental issues like dental plaque, calculus, tooth decay, dental hypersensitivity, dry mouth, as well as cosmetic concerns like dental staining and oral malodor. For each problem, it explains the causes such as bacteria, acid production, and dietary factors. Prevention methods include proper brushing, flossing, fluoride use, and limiting sugary foods. Treatment involves removing plaque and bacteria mechanically or chemically, using desensitizing toothpastes, artificial saliva, whitening agents, and masking odors. Maintaining good oral hygiene is key to preventing most oral health issues.
The document provides guidelines on validation of analytical procedures from the International Conference on Harmonisation (ICH) and the World Health Organization (WHO). It discusses validation characteristics like accuracy, precision, specificity, linearity, range, detection limit and quantitation limit that should be considered when validating identification tests, assays, and tests for impurities. It provides definitions for key terms and recommendations on how validation of these characteristics should be performed.
Approaches for the design of transdermal drug deliverykvineetha8
The document provides an introduction to transdermal drug delivery systems (TDDS). It discusses advantages like controlled delivery and avoidance of first-pass metabolism. Components are described, including rate-controlling polymers, drugs, permeation enhancers, adhesives and backing layers. Approaches to development include polymer membrane systems, polymer matrix systems, and adhesive or microreservoir systems. Recent approaches discussed are iontophoresis and electroporation, which use electric current to enhance skin permeation.
Principle and application of ptgc and isothermal programmingAthira39
Gas chromatography is the separation of gaseous and volatile substances which is achieved by employing gas as a mobile phase and moving it through a column containing stationary phase which could be a liquid or solid.
Two methods of temperature control are used during gas chromatography:
Isothermal operation and;
Temperature programming
Microencapsulation involves coating solid, liquid, or gaseous core materials in diameters between 1-1000 μm within an inert shell. This process isolates and protects core materials while controlling drug release. Methods like single emulsion, solvent evaporation, phase separation, and spray drying are used to prepare microspheres and microcapsules for applications like oral drug delivery, vaccines, gene delivery, and targeted therapies. Microencapsulation masks tastes, separates incompatible materials, and provides environmental protection or controlled release of core substances.
Transfer from R & D to production.pptxDipeshGamare
Transfer from R & D to production :
(Process, packaging and cleaning)
Granularity of TT Process :
(API, excipients, finished products, packaging materials)
Column chromatography is a separation technique that uses a column packed with a stationary phase. As a mixture passes through the column via a mobile phase, components separate based on how they partition between the stationary and mobile phases. Key factors that affect separation are the types of stationary and mobile phases used, as well as column characteristics like length and diameter. Common stationary phases include silica, alumina, and cellulose while mobile phases are selected based on solvent polarity and component solubility. Proper packing of the stationary phase into the column is also important for achieving efficient separation.
This document discusses derivatization techniques used to modify compounds to make them suitable for analysis by gas chromatography (GC) and high-performance liquid chromatography (HPLC). It describes how derivatization changes functional groups to alter chemical and physical properties without changing the overall chemical structure. Common derivatization methods include silylation, alkylation, acylation, and chiral derivatization. Silylation adds silyl groups to make compounds more volatile. Alkylation and acylation reduce polarity. Chiral derivatization forms diastereomers to separate enantiomers. Derivatization can occur before or after column separation in HPLC. Pre-column derivatization is manual while post-column occurs automatically via
Microspheres are solid spherical particles ranging from 1-1000μm that are used for drug delivery. They can be made of proteins or synthetic polymers. There are two main types - microcapsules which have a core and coating, and micromatrices which have a drug dispersed throughout the polymer matrix. Microspheres offer advantages like reduced dosing, constant drug levels, and protection of drugs. They are made using methods like solvent evaporation, emulsion techniques, and polymerization. Microspheres find applications in delivery to sites like the eyes, oral cavity, skin and more. Evaluation involves analyzing size, shape, drug content and release kinetics.
Mr. Darshan N U is studying for his M Pharmacy first semester in the Department of Pharmaceutical Chemistry. The document provides an introduction to paper chromatography, covering its history, principles, requirements, factors affecting it, applications, and advantages over other methods. Paper chromatography is defined as a technique where unknown substances are analyzed mainly by the flow of solvents on filter paper. It has been used to separate mixtures like amino acids, food colors, and biological components.
Paper chromatography is an analytical method used to separate coloured chemicals or substances.It is now primarily used as a teaching tool, having been replaced in the laboratory by other chromatography methods such as thin-layer chromatography (TLC).
Ultraviolet-visible spectroscopy instrumentation, choice of solvents, and solvent effects are discussed. Key points include:
UV-Vis spectrophotometers measure light intensity as a function of wavelength using sources, wavelength selectors, sample containers, detectors, and readout devices. Common sources include tungsten-halogen lamps, deuterium lamps, and LEDs. Wavelength selectors include filters and monochromators using prisms or gratings. Choice of solvent is important to avoid absorption in the detection region, with 95% ethanol commonly used. Solvent effects can cause absorption band shifts due to changes in solute-solvent interactions between ground and excited states.
This document discusses gastroretentive drug delivery systems (GRDDS), which are oral dosage forms designed to remain in the stomach for an extended period of time to prolong drug release. It covers the rationale for using GRDDS, factors controlling gastric residence time, and various approaches for prolonging gastric retention including floating systems, high-density systems, and bioadhesive or magnetic systems. Floating systems include non-effervescent and effervescent types that float due to low density or gas generation. High-density systems do not float but remain in the stomach through bioadhesion, magnetic forces, swelling to a large size, or raft formation on gastric fluids.
Application Of Polymer In Controlled Release FormulationAnindya Jana
Polymers are becoming increasingly important in the field of drug delivery. The pharmaceutical applications of polymers range from their use as binders in tablets to viscosity and flow controlling agents in liquids, suspensions and emulsions. Polymers can be used as film coatings to disguise the unpleasant taste of a drug, to enhance drug stability and to modify drug release characteristics.
As a consequence, increasing attention has been focused on methods of giving drugs continually for a prolonged time periods and in a controlled fashion.
This technology now spans many fields and includes pharmaceutical, food and agricultural applications, pesticides, cosmetics, and household products.
This document summarizes a seminar on gastroretentive drug delivery systems (GRDDS). GRDDS are designed to retain drugs in the stomach for prolonged periods of time to allow for sustained drug release. The seminar outlines various GRDDS technologies including floating, swelling, mucoadhesive, and high density systems. It also discusses candidate drugs for GRDDS, advantages like improved bioavailability, and evaluation methods like dissolution testing, floating time, and mucoadhesive strength testing. Limitations include instability at gastric pH and requirement of high fluid levels for floating systems.
This document summarizes a seminar on ion exchange chromatography. It introduces the topic, covering the basic principles of how ion exchange chromatography separates ions and polar molecules using stationary phases with positively or negatively charged groups. It then discusses the types of ion exchange resins, including classifications based on chemical nature and source. Examples of applications are given, such as separating similar ions, water softening, demineralization, separating sugars and amino acids. The document concludes by discussing some advantages and disadvantages of ion exchange chromatography.
Gas chromatography is a technique used to separate components of a vaporized sample. It works by partitioning the components between a mobile gaseous phase and a stationary phase within a column. The sample is injected and vaporized, then transported through the column by the mobile phase gas. As the components pass through the column they are separated and detected. Common detectors include the flame ionization detector (FID), thermal conductivity detector (TCD), and electron capture detector (ECD). The FID responds to organic compounds, the TCD is universal but less sensitive, and the ECD selectively detects halogen-containing compounds.
The document discusses the formulation and evaluation of gastro-retentive mucoadhesive microballoons of nizatidine for the management of peptic ulcer. It provides background information on peptic ulcers, drug delivery systems, gastroretentive drug delivery systems, and mechanisms of bioadhesion. The objective is to develop mucoadhesive microballoons of nizatidine to increase its retention time in the stomach and improve the treatment of peptic ulcers.
The document provides information on nasopulmonary drug delivery systems including nasal drug delivery and pulmonary drug delivery. It discusses the anatomy and physiology of the nasal cavity and respiratory tract. It also describes various formulation approaches for nasal delivery such as nasal gels, drops, sprays and powders. The document further explains dry powder inhalers, metered dose inhalers and nebulizers as pulmonary drug delivery systems along with their advantages and disadvantages. It also discusses some marketed products for nasal sprays, dry powder inhalers and metered dose inhalers.
This presentation discusses partition column chromatography. It begins with an introduction and presenter information. It then defines partition chromatography as a method of separation where components in a mixture distribute between two liquid phases due to differences in partition coefficients. The presentation notes that partition chromatography is the most widely used type of HPLC, where the stationary phase is a second immiscible liquid bonded to the column. It discusses the principles of how separation occurs due to differences in how components partition between the two phases. Finally, it provides examples of applications such as separation of compounds in biological and environmental samples and determination of water quality.
This document discusses common oral health problems and their causes, prevention, and treatment. It covers dental issues like dental plaque, calculus, tooth decay, dental hypersensitivity, dry mouth, as well as cosmetic concerns like dental staining and oral malodor. For each problem, it explains the causes such as bacteria, acid production, and dietary factors. Prevention methods include proper brushing, flossing, fluoride use, and limiting sugary foods. Treatment involves removing plaque and bacteria mechanically or chemically, using desensitizing toothpastes, artificial saliva, whitening agents, and masking odors. Maintaining good oral hygiene is key to preventing most oral health issues.
The document provides guidelines on validation of analytical procedures from the International Conference on Harmonisation (ICH) and the World Health Organization (WHO). It discusses validation characteristics like accuracy, precision, specificity, linearity, range, detection limit and quantitation limit that should be considered when validating identification tests, assays, and tests for impurities. It provides definitions for key terms and recommendations on how validation of these characteristics should be performed.
Approaches for the design of transdermal drug deliverykvineetha8
The document provides an introduction to transdermal drug delivery systems (TDDS). It discusses advantages like controlled delivery and avoidance of first-pass metabolism. Components are described, including rate-controlling polymers, drugs, permeation enhancers, adhesives and backing layers. Approaches to development include polymer membrane systems, polymer matrix systems, and adhesive or microreservoir systems. Recent approaches discussed are iontophoresis and electroporation, which use electric current to enhance skin permeation.
Principle and application of ptgc and isothermal programmingAthira39
Gas chromatography is the separation of gaseous and volatile substances which is achieved by employing gas as a mobile phase and moving it through a column containing stationary phase which could be a liquid or solid.
Two methods of temperature control are used during gas chromatography:
Isothermal operation and;
Temperature programming
Microencapsulation involves coating solid, liquid, or gaseous core materials in diameters between 1-1000 μm within an inert shell. This process isolates and protects core materials while controlling drug release. Methods like single emulsion, solvent evaporation, phase separation, and spray drying are used to prepare microspheres and microcapsules for applications like oral drug delivery, vaccines, gene delivery, and targeted therapies. Microencapsulation masks tastes, separates incompatible materials, and provides environmental protection or controlled release of core substances.
Transfer from R & D to production.pptxDipeshGamare
Transfer from R & D to production :
(Process, packaging and cleaning)
Granularity of TT Process :
(API, excipients, finished products, packaging materials)
Column chromatography is a separation technique that uses a column packed with a stationary phase. As a mixture passes through the column via a mobile phase, components separate based on how they partition between the stationary and mobile phases. Key factors that affect separation are the types of stationary and mobile phases used, as well as column characteristics like length and diameter. Common stationary phases include silica, alumina, and cellulose while mobile phases are selected based on solvent polarity and component solubility. Proper packing of the stationary phase into the column is also important for achieving efficient separation.
This document discusses derivatization techniques used to modify compounds to make them suitable for analysis by gas chromatography (GC) and high-performance liquid chromatography (HPLC). It describes how derivatization changes functional groups to alter chemical and physical properties without changing the overall chemical structure. Common derivatization methods include silylation, alkylation, acylation, and chiral derivatization. Silylation adds silyl groups to make compounds more volatile. Alkylation and acylation reduce polarity. Chiral derivatization forms diastereomers to separate enantiomers. Derivatization can occur before or after column separation in HPLC. Pre-column derivatization is manual while post-column occurs automatically via
Microspheres are solid spherical particles ranging from 1-1000μm that are used for drug delivery. They can be made of proteins or synthetic polymers. There are two main types - microcapsules which have a core and coating, and micromatrices which have a drug dispersed throughout the polymer matrix. Microspheres offer advantages like reduced dosing, constant drug levels, and protection of drugs. They are made using methods like solvent evaporation, emulsion techniques, and polymerization. Microspheres find applications in delivery to sites like the eyes, oral cavity, skin and more. Evaluation involves analyzing size, shape, drug content and release kinetics.
Mr. Darshan N U is studying for his M Pharmacy first semester in the Department of Pharmaceutical Chemistry. The document provides an introduction to paper chromatography, covering its history, principles, requirements, factors affecting it, applications, and advantages over other methods. Paper chromatography is defined as a technique where unknown substances are analyzed mainly by the flow of solvents on filter paper. It has been used to separate mixtures like amino acids, food colors, and biological components.
Paper chromatography is an analytical method used to separate coloured chemicals or substances.It is now primarily used as a teaching tool, having been replaced in the laboratory by other chromatography methods such as thin-layer chromatography (TLC).
Paper chromatography is a technique used to separate mixtures based on differences in how components partition between a stationary phase (filter paper) and mobile phase (solvent). It involves applying a sample to a strip of paper and developing it by allowing the solvent to travel up the paper. This causes the different components to separate into bands at different distances based on how strongly they are retained by the paper. Paper chromatography is widely used for qualitative analysis and identification of organic and inorganic compounds in mixtures.
Paper chromatography (PC) was introduced in 1865 and is considered the simplest chromatographic technique. It involves using filter paper as the stationary phase and a solvent as the mobile phase. There are two types of PC - adsorption chromatography where the paper is coated with an adsorbent like silica or alumina, and partition chromatography where moisture in the paper acts as the stationary phase. Compounds are separated based on their partitioning between the stationary and mobile phases. PC is useful for separating mixtures and identifying compounds due to its low cost, simplicity, and good resolving power.
Chromatography is a technique used to separate mixtures by exploiting differences in how components interact with two phases - a stationary phase and a mobile phase. There are several types of chromatography including paper chromatography, thin layer chromatography, column chromatography, and ion exchange chromatography. Each type uses a different stationary phase and separation method to isolate analytes in a mixture.
This document discusses paper chromatography techniques. It begins with definitions of chromatography and a brief history. It then explains the principle of paper chromatography, which involves the separation of compounds through their partitioning between a stationary phase (filter paper) and mobile phase solvent. The document outlines the basic experimental procedure for paper chromatography and describes various development techniques, including ascending, descending, radial, and two-dimensional modes.
This document provides an overview of paper chromatography. It discusses the history and types of paper chromatography, including the principles of separation. It describes the practical requirements like the stationary and mobile phases used. It explains how to calculate Rf and Rx values and factors that can affect them. Various detection methods are outlined as well as sources of error. Finally, the applications and advantages of paper chromatography are highlighted.
Chromatography by narayan sarkar and simi baruah new versionNarayanSarkar6
Chromatography is a technique used to separate mixtures by exploiting differences in how components interact with two phases - a stationary phase and a mobile phase. The document provides an introduction to chromatography, including its history, principles, types (column, paper, thin layer, affinity), and applications. Column chromatography involves passing a mixture through a column containing a stationary phase to separate components based on differences in how strongly they adhere to the stationary phase.
Paper chromatography by Mr. Vinayak BodhankarMayur Bodhankar
Paper chromatography is a simple type of chromatography that uses specialized paper as the stationary phase. Components in a sample will separate as the mobile phase travels up the paper based on how strongly each component is absorbed to the stationary phase. Paper chromatography can be used to detect impurities, analyze food and drugs, and study biochemical reactions. It has advantages of being rapid, inexpensive, and requiring only small sample amounts. However, it is not effective for complex mixtures or quantitative analysis.
Review on Adopting Biophysical Chromatography TechniquesIJSRED
1. Chromatography is an important biophysical technique used for separating, identifying, and purifying the components of mixtures. It is widely used in industries like pharmaceutical production, environmental analysis, and food chemistry.
2. The document discusses various chromatography techniques including column chromatography, thin layer chromatography, gas chromatography, and liquid chromatography. It also covers the principles of chromatography, different types based on the stationary and mobile phases used, and applications in various fields.
3. Chromatography techniques are used in pharmaceutical industries for analysis, purification, and quality control of drugs and separation of compounds in research studies.
This document provides an overview of chromatography. It begins by defining chromatography as a technique used to separate components of a mixture based on their differential interactions with a mobile and stationary phase. It then discusses various types of chromatography, including paper chromatography, thin layer chromatography, and column chromatography. The key principle explained is that components with a higher affinity for the stationary phase will move more slowly through the system than those with a higher affinity for the mobile phase.
This document provides information about paper chromatography and column chromatography. It discusses the basic principles, instrumentation, steps, and applications of these chromatographic techniques. Paper chromatography involves using a paper sheet as the stationary phase, while column chromatography uses a packed column. Both techniques separate mixtures based on differences in how compounds partition between a stationary and mobile phase. Common applications include purification, analysis of drugs, foods, and other chemical and biological samples.
The document discusses paper chromatography, a technique used to separate mixtures by passing them through a paper medium. It describes the basic components and principles of paper chromatography, including the stationary and mobile phases. The document provides details on the procedure, such as sample preparation and development, and discusses using retention factor values. It outlines different types of paper chromatography and lists applications in areas like separating pigments and monitoring reactions. The document concludes by noting paper chromatography's initial use and expanded applications over time.
This document provides information on paper chromatography, including its history, key terms, processes, techniques, factors that affect separation, and applications. Paper chromatography is an analytical technique used to separate mixtures based on differences in how compounds partition between a stationary phase, such as the paper, and a mobile phase, such as a solvent. Key points covered include the different types of paper chromatography, factors that influence separation like the solvent used, and how paper chromatography can be used to analyze organic compounds.
Chromatographic techniques such as paper chromatography, thin layer chromatography, and column chromatography are used to separate mixtures based on differences in how components partition between a stationary and mobile phase. In paper chromatography, the stationary phase is water absorbed by a paper support and the mobile phase is a solvent such as ethanol or chloroform. Thin layer chromatography uses a finely divided substance coated on a glass plate as the stationary phase. Column chromatography packs the stationary phase into a glass or plastic column. Components separate as they migrate through the column at different rates based on their partitioning between the phases.
Chromatography is a technique used to separate components of a mixture. It works by distributing the components between two phases - a stationary phase and a mobile phase. Paper chromatography is a type of chromatography where the stationary phase is filter paper and the mobile phase is a liquid solvent. Key steps in paper chromatography include choosing a suitable filter paper and solvent, preparing and spotting the sample, developing the chromatogram by allowing the solvent to travel up or down the paper, visualizing the separated components, and calculating Rf values to identify substances.
Paper chromatography is a physical technique used to separate and analyze components in mixtures. It works by partitioning components between a stationary phase (paper) and mobile phase (solvent). Key steps include spotting the sample, developing the chromatogram by allowing the mobile phase to travel up the paper, drying the paper, and analyzing the separated components. Paper chromatography is useful for separating and identifying various substances like amino acids, drugs, and organic compounds. It has applications in fields like pharmacy, biochemistry, and research due to being inexpensive and easy to perform.
Chromatography is a technique used to separate mixtures by distributing components between two phases - a stationary phase and a mobile phase. Paper chromatography uses a cellulose paper as the stationary phase and a solvent as the mobile phase. Components separate based on how strongly they interact with each phase. The separation can be visualized by detecting spots, and identified using the retention factor (Rf) which characterizes how far each component travels up the paper relative to the solvent front. Paper chromatography is used to analyze, identify, purify, and quantify mixtures in applications like pharmaceutical analysis, forensics, and food and drug testing.
Paper chromatography is a technique used to separate mixtures into individual components. It works by allowing the molecules in a mixture to distribute themselves between a stationary phase (filter paper) and a mobile phase (solvent). Different molecules will travel different distances up the paper depending on how strongly they interact with each phase. Paper chromatography has been replaced in many applications by thin layer chromatography, which provides faster runs and better separation. However, paper chromatography is still useful for teaching chromatography principles in educational settings.
This document discusses fluorescence spectroscopy. It begins with definitions of fluorescence, phosphorescence, and luminescence. Fluorescence occurs when a substance emits light immediately upon absorption of radiation from an excited singlet state. Phosphorescence involves delayed emission of light from an excited triplet state.
The document then covers factors that affect fluorescence intensity such as molecular structure, substituents, concentration, pH, temperature, and viscosity. It also discusses fluorescence instrumentation including sources of light, filters, monochromators, sample cells, and detectors.
Finally, the document discusses applications of fluorescence spectroscopy in areas like inorganic analysis, nuclear research, organic analysis, pharmaceutical analysis, liquid chromatography, and vitamin analysis.
Spectroscopy is the measurement of electromagnetic radiation absorbed or emitted by molecules or atoms as they move between energy states. It is a powerful analytical technique used to study molecular structure. A spectrometer measures the spectrum of a sample, which is a graph of intensity versus wavelength or frequency. There are different types of spectroscopy depending on the type of radiation used, such as UV-visible, infrared, or mass spectroscopy. The interaction of matter with electromagnetic radiation follows Beer's law, which states that absorbance is directly proportional to concentration and path length.
This document discusses several antibiotics including clavulanic acid, sulbactum, tazobactum, and cefepime. It also mentions medicinal chemistry of macrolide antibiotics. The document was written by Bhoopendra Patidar and seems to cover different types of antibiotics used against bacterial infections.
STEREOISOMERISM by Mr Bhupendra patidarPrabhat Kumar
This document discusses different types of isomers, including constitutional isomers and stereoisomers. Stereoisomers differ in the spatial orientation of their atoms but have the same molecular formula and functional groups. There are several types of stereoisomers: enantiomers, which are non-superimposable mirror images; diastereomers, which are not mirror images; and meso compounds, which have an internal plane of symmetry and are optically inactive despite having multiple stereocenters. The document also discusses configuration, specific rotation, Fischer projections, and the distinguishing characteristics of enantiomers and diastereomers.
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1. The anatomy and physiology of the heart, including its chambers and layers. The heart pumps deoxygenated blood to the lungs and oxygenated blood to the body.
2. The three main types of blood circulation - pulmonary circulation between the heart and lungs, coronary circulation supplying the heart, and systemic circulation supplying the body. Blood vessels have three layers and blood flows through arteries, arterioles, capillaries and veins.
3. Key aspects of cardiac cycle including the phases of atrial and ventricular systole and diastole. Heart sounds and electrocardiograms are also summarized.
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With increasing population, people need to rely on packaged food stuffs. Packaging of food materials requires the preservation of food. There are various methods for the treatment of food to preserve them and irradiation treatment of food is one of them. It is the most common and the most harmless method for the food preservation as it does not alter the necessary micronutrients of food materials. Although irradiated food doesn’t cause any harm to the human health but still the quality assessment of food is required to provide consumers with necessary information about the food. ESR spectroscopy is the most sophisticated way to investigate the quality of the food and the free radicals induced during the processing of the food. ESR spin trapping technique is useful for the detection of highly unstable radicals in the food. The antioxidant capability of liquid food and beverages in mainly performed by spin trapping technique.
EWOCS-I: The catalog of X-ray sources in Westerlund 1 from the Extended Weste...Sérgio Sacani
Context. With a mass exceeding several 104 M⊙ and a rich and dense population of massive stars, supermassive young star clusters
represent the most massive star-forming environment that is dominated by the feedback from massive stars and gravitational interactions
among stars.
Aims. In this paper we present the Extended Westerlund 1 and 2 Open Clusters Survey (EWOCS) project, which aims to investigate
the influence of the starburst environment on the formation of stars and planets, and on the evolution of both low and high mass stars.
The primary targets of this project are Westerlund 1 and 2, the closest supermassive star clusters to the Sun.
Methods. The project is based primarily on recent observations conducted with the Chandra and JWST observatories. Specifically,
the Chandra survey of Westerlund 1 consists of 36 new ACIS-I observations, nearly co-pointed, for a total exposure time of 1 Msec.
Additionally, we included 8 archival Chandra/ACIS-S observations. This paper presents the resulting catalog of X-ray sources within
and around Westerlund 1. Sources were detected by combining various existing methods, and photon extraction and source validation
were carried out using the ACIS-Extract software.
Results. The EWOCS X-ray catalog comprises 5963 validated sources out of the 9420 initially provided to ACIS-Extract, reaching a
photon flux threshold of approximately 2 × 10−8 photons cm−2
s
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massive stars of the cluster, and we have collected over 71 000 photons from the magnetar CXO J164710.20-455217.
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Conducted over a period of >200 years, Thermodynamics R&D, and application, benefitted from the highest levels of professionalism, collaboration, and technical thoroughness. New layers of application, methodology, and practice were made possible by the progressive advance of technology. In turn, this has seen measurement and modelling accuracy continually improved at a micro and macro level.
Perhaps most importantly, Thermodynamics rapidly became a primary tool in the advance of applied science/engineering/technology, spanning micro-tech, to aerospace and cosmology. I can think of no better a story to illustrate the breadth of scientific methodologies and applications at their best.
Or: Beyond linear.
Abstract: Equivariant neural networks are neural networks that incorporate symmetries. The nonlinear activation functions in these networks result in interesting nonlinear equivariant maps between simple representations, and motivate the key player of this talk: piecewise linear representation theory.
Disclaimer: No one is perfect, so please mind that there might be mistakes and typos.
dtubbenhauer@gmail.com
Corrected slides: dtubbenhauer.com/talks.html
The technology uses reclaimed CO₂ as the dyeing medium in a closed loop process. When pressurized, CO₂ becomes supercritical (SC-CO₂). In this state CO₂ has a very high solvent power, allowing the dye to dissolve easily.
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Unlocking the mysteries of reproduction: Exploring fecundity and gonadosomati...AbdullaAlAsif1
The pygmy halfbeak Dermogenys colletei, is known for its viviparous nature, this presents an intriguing case of relatively low fecundity, raising questions about potential compensatory reproductive strategies employed by this species. Our study delves into the examination of fecundity and the Gonadosomatic Index (GSI) in the Pygmy Halfbeak, D. colletei (Meisner, 2001), an intriguing viviparous fish indigenous to Sarawak, Borneo. We hypothesize that the Pygmy halfbeak, D. colletei, may exhibit unique reproductive adaptations to offset its low fecundity, thus enhancing its survival and fitness. To address this, we conducted a comprehensive study utilizing 28 mature female specimens of D. colletei, carefully measuring fecundity and GSI to shed light on the reproductive adaptations of this species. Our findings reveal that D. colletei indeed exhibits low fecundity, with a mean of 16.76 ± 2.01, and a mean GSI of 12.83 ± 1.27, providing crucial insights into the reproductive mechanisms at play in this species. These results underscore the existence of unique reproductive strategies in D. colletei, enabling its adaptation and persistence in Borneo's diverse aquatic ecosystems, and call for further ecological research to elucidate these mechanisms. This study lends to a better understanding of viviparous fish in Borneo and contributes to the broader field of aquatic ecology, enhancing our knowledge of species adaptations to unique ecological challenges.
2. 2
CONTENTS
Introduction
Principle of Separation
Modes of Paper Chromatography
Experimental Details for Qualitative Analysis
Experimental Details for QuantitativeAnalysis
Applications
3. 3
The most dramatic advance in the history of chromatography
took place in 1944.
It was invented by two British biochemists, Archer John
Porter Martin and Richard Laurence Millington Synge.
In 1941 Martin and Synge began working together on
proteins, which are made up of chains of amino acids.
The duo was trying to characterize a particular protein by
determining the precise numbers of each amino acid present.
INTRODUCTION
4. 4
Amino acids are so similar to each other, however, that the
problem of separating them had defeated a whole generation of
biochemists.
Martin and Synge's development of paper
chromatography to solve this problem was an instant success.
It worked not only on amino acids but also on various other
mixtures. The two scientists were awarded the 1952 Nobel
Prize in chemistry for their work.
Synge determined the structure of an antibiotic peptide called
"Gramicidin-S.
5. 5
Frederick Sanger used paper chromatography to figure out
the structure of the insulin molecule. He determined the
number of amino acids in it as well as the order in which they
occurred.
Calvin discovered the complex series of reactions that enable
green plants to convert solar energy into the chemical
energy stored in food using paper chromatography in 1950s.
6. 6
It was also used by Austrian-American biochemist Erwin
Chargaff, who modified the technique to study the
components of the nucleic acid molecule. His research
revealed four components, or nitrogenous bases, that occur in
pairs.
British biochemists James Dewey Watson and Francis Harry
Compton Crick later used these results to work out the
structure of DNA(deoxyribonucleic acid).
7. 7
There are two types of paper chromatography, they are:
PAPER ADSORPTION CHROMATOGRAPHY
Paper impregnated with silica or alumina acts as adsorbent
(stationary phase) and solvent as mobile phase.
PAPER PARTITION CHROMATOGRAPHY
Moisture/ Water present in the pores of cellulose fibers
present in filter paper acts as stationary phase and solvent
used as mobile phase.
8. 8
PRINCIPLE OF SEPARATION
The principle involved is partition chromatography wherein
the substances are distributed or partitioned between liquid
phases. One phase is the water, which is held in the pores of
the filter paper used; and other is the mobile phase which
moves over the paper.
The compounds in the mixture get separated due to
differences in their affinity towards water (in stationary
phase) and mobile phase solvents during the movement of
mobile phase under the capillary action of pores in the paper.
9. 9
Principle of separation Cont.,
The principle can also be adsorption chromatography between
solid and liquid phases, wherein the stationary phase is the
solid surface of paper and the liquid phase is of mobile phase.
But most of the applications of paper chromatography work on
the principle of partition chromatography, i.e. partitioned
between to liquid phases.
10. 10
M
ODES OF PAPER CHROM
ATOGRAPHY
Based on the way the development of chromatogram on paper
is done in procedures, we have, broadly, five modes of
chromatography.
1. Ascending
2. Descending
3. Ascending-Descending
4. Circular/ Radial
5. Two-Dimensional
11. 1. Ascending Technique
When the development of the paper is done by allowing the solvent
to travel up the paper, it is known as ascending technique.
The chromatogram of this technique
ascends slowly due to the mobile phase
movement in a upward direction.
The solvent is kept at the bottom of the
filter paper or stationary phase with the
end of the filter paper dipped in.
The component mixture spot is kept
well above the solvent level and is not
allowed to touch the spot.
11
12. 2. Descending Technique
When the development of the paper is done by allowing the solvent
to travel down the paper, it is known as descending technique.
The mobile phase in this
type is kept at the top of
the chromatogram and the
components of the mixture
separates out downward
gravity and
action of the
due to
capillary
filter paper.
ADVANTAGE:
Development is faster
12
13. 3. Ascending-Descending Technique
Ahybrid of above two techniques is called ascending-descending
chromatography.
Only length of separation increased, first ascending takes place
followed by descending.
Overall shows a bi-directional movement of the mixture
components.
13
14. 4. Circular/ Radial Technique
14
In this type of paper chromatography the solvent moves from the
centre towards the peripheral regions of the filter paper. The
radiating mixture component is allowed to spread till all the
components have separated out. For precaution the entire system is
covered with the help of a Petri dish.
The centre of the paper is allowed to be dipped into the solvent
and the coloured components radiates out in concentric circles.
15. 5. Two- Dimensional Technique
The chromatogram in this type
develops at right angle to each other
and the filter paper is dipped at right
angle once the first chromatogram is
complete.
The second chromatogram then
develops at right angle to the first
one.
15
16. 16
1. Choice of Development Technique
2. Choice of Stationary Phase
3. Proper Developing Solvent System
4. Preparation of Sample
5. Application of Sample (Spotting)
6. Drying the Chromatogram
7. Detecting or Visualizing
8. Calculation of Rf Value
Experimental details for qualitative analysis
17. 17
1. Choice of Development Technique
The choice of technique depends
substances to separated.
upon the nature of the
Ascending
Descending
Ascending-Descending
Circular/ Radial
Two-Dimensional
18. 2. Choice of Stationary Phase
Chromatography makes use
stationary phase.
Paper essentially consists of
of paper which acts as a
cellulose fibers which are
polymers having -OH functional groups sticking out of the
polymer chains.
These groups lead to retention and separation of surface
absorbed molecules.
In practice the separating molecules equilibrate between the
layer of adsorbed water and the mobile phase solvent.
18
19. 19
2. Choice of Stationary Phase Cont.,
The choice of paper is dependent on the type of problem under
investigation. The prime factors, that govern the choice, are as
follows:
Whether the paper is being used for quantitative or
qualitative analysis;
Whether it is used for analytical or preparative
chromatography; or
Whether the substances used are hydropilic or lipophilic,
neutral or charged species.
20. 20
2. Choice of Stationary Phase Cont.,
Various types of Whatmann papers are available.
Rate of Flow
Fast Medium Slow
Thin Paper No. 4 No. 7 No. 2
No. 54 No. 1 No. 20
No. 540
Thick
Paper
No. 31 No. 3
No. 17 No. 3 MM
21. 21
2. Choice of Stationary Phase Cont.,
Whatmann filter papers commonly used for chromatographic
purposes have a content of 99% of α-cellulose. The rest is
mineral content.
For the efficient separation of polar substances, the exchange
capacity of the paper is increased by increasing the carboxyl
content (1.4%) by partial oxidation.
It is possible to increase the capillarity of the paper by partial
hydrolysis which is achieved by soaking the filter paper for 24 h
in 7% hydrochloric acid and washing successfully with water
and ethanol.
22. 22
2. Choice of Stationary Phase Cont.,
Cellulose paper is also used as a support for various adsorbents
like alumina, silica, zirconium oxide etc. which get precipitated
in the pores of the filter paper to produce a thin sheet of the
adsorbent with the flexibility of the paper but having adsorbent
characteristics of the precipitate.
Chelating agents such as 8-hydroxyquinoline, dimethyl
glyoxime and several other compounds impart papers with
special characteristics.
Papers have also been impregnated with powdered or liquid ion
exchangers to produce ion exchange papers.
23. 23
Modified Papers for Paper Chromatography
Type Mobile Phase
Carboxyl papers Cationic separation of protonated amines
and amino acids
Acetylated Papers RP chromatography of lipophilic substances
like steroids, insecticides and pigments and
also metal cations.
Kieselguhr papers,
Alumina papers,
Zirconia papers, Silica
papers
Separation of low polarity substances such as
amines, fatty acids, steroids, triglycerides,
vitamins and pesticides
Ion Exchange Papers Ion Exchange paper chromatography
2. Choice of Stationary Phase Cont.,
24. 24
3. Proper Developing Solvent System
Criteria for selection of a working solvent system:
Solvents should not be toxic or carcinogenic. In such cases
adequate handling care should be exercised.
The solvent mixture composition should not change with
time.
Solvent constituents should not chemically react with any of
the sample constituents.
Solvents should not interfere with the detection of spots on
the developed paper.
25. 25
Differences in Rf values of any two components should be at
least 0.05 for differentiation between two closely spaced spots.
The Rf values of the sample should lie between 0.05 and 0.85
in the system.
Suitable solvent systems for paper chromatography
Water as a stationary Phase
– Isopropanol: ammonia: water [9:1:2]
– N-butanol: glacial acetic acid: water [4:1:5]
– Phenol saturated with water
SP(dimethyl ether):- Cyclohexane
SP(kerosene) :- 70% isopropanol
26. 26
3. Proper Developing Mobile Phase cont.,
Commonly used solvent combinations:
Strongly polar compounds:
o ethyl acetate: n-butanol: acetic acid: water - 80:10:5:5
Polar compounds:
o 10% methanol or less in dichloromethane
Strongly basic compounds:
o 10% ammonium hydroxide in methanol and make 1-10%
mixture of this in dichloromethane
27. 27
4. Preparation of Sample
Pure solutions can be applied directly on the paper but solids
are always dissolved in small quantity of a suitable solvent.
Biological tissues are treated with suitable solvents and their
extracts obtained.
The sample volume of 10-20 µL having as many µg of the
substances is that ideal quantity to be spotted.
28. Preparation of Paper
Cut the paper into desired
shape and size depending upon
work to be carried out.
The starting line is marked on
the paper with an ordinary
pencil 2 cm from the bottom
edge.
On the staring line marks are
made 2 cm apart from each
other.
5. Application of Sample (Spotting)
28
29. Spotting:
The sample to be applied is dissolved in the mobile phase and
applied as a small spot on the origin line, using capillary tube or
micropipette.
Very low concentration is used to avoid larger zone.
The sample is applied as a neat spot on a horizontal line drawn
with a pencil close to one edge.
The spot is dried on the filter paper and is placed in developing
chamber.
29
30. Developing:
Allow the spot to dry and then immerse the paper in the
developing chamber as per the selected technique with the marked
spot above the solvent level.
The solvent begins to move and draws the sample components
differentially along with it.
At the end of the development take out the paper and mark the
solvent front with another line.
30
31. 6. Drying of Chromatogram
They are dried by cold or hot air depending on volatility of
solvents. A simple hair dryer is a convenient device to dry
chromatograms.
31
32. 32
7. Detecting or Visualizing
Coloured spots are easily observed on developed
chromatograms.
However, different approaches need to be adopted when
colourless components are to be observed.
It is convenient to classify such methods as specific or
non-specific.
33. 33
Non-specific methods (Physical):
Iodine chamber: The developed plate is suspended in a
closed jar containing a few crystals of iodine for about a
vapour most organic
minute. In presence of iodine
compounds appear as brown spots.
UV viewing cabinet:
Majority of colourless compounds can be viewed under
illumination with UV light in a UV viewing cabinet.
Commonly the cabinets are equipped with a long
wavelength (366 nm) and short wavelength (254 nm)
light sources.
35. Rf VALUE (Retardation Factor)
The Rf defines the movement
of the substance relative to
the solvent front in a given
chromatographic system.
8. Calculation of Rf VALUE
35
36. 36
Factors affecting Rf VALUE
The temperature
The purity of the solvents used
The quality of the paper, adsorbents & impurities present in
the adsorbents
Chamber saturation techniques, method of drying &
development
The distance travelled by the solute & solvent
Chemical reaction between the substances being partitioned.
pH of the solution
37. 37
Rx VALUE
In many cases it has been observed that the solvent front is run
off the end of the paper.
It is the ratio of distance travelled by the sample and the
distance travelled by the standard.
Rx value is always closer to 1.
38. 38
For quantitative analysis, the preliminary separation is done
in the same way as in qualitative analysis. Then, the assay
can be performed either after extraction from the paper or
in situ on the paper.
1. Estimation after Extraction from the paper
2. In situ methods
Experimental details for Quantitative Analysis
39. 39
In this technique, the spots are cut into portions and eluted with
solvents. This solution can be analyzed by any techniques of
analysis like gravimetric estimation, spectrophotometry,
Colorimetry, Polarography, Flame photometry, etc.
In the assessment of the merits of any procedure, the following
information is required:
The nature of the substance to be assayed.
The scientific equipment available and its sensitivity.
The time available and
The alternative method, available, if any, and their relative
accuracies
1. Estimation after Extraction from the paper
40. 40
2. In situ methods
By Visual Assessment: The simplest procedure is to see the spot by the
human eye. However, it is not very accurate.
By Measurement of areas:
if the outline of the spots or zones are well defined, the size of the
spot (length or areas) may serve for determining the quantity of the
substances.
Then a linear relationship is obtained between the spot area and
amount of the substance present.
The random errors in the procedure are generally high as a results of
the variations of the spot shape during separation.
Other drawbacks: Volume of applied sample and speed of
application
41. 41
2. In situ methods cont.,
By Densitometer: It is a method whereby the intensity of the colour
of a substance is measured directly on the chromatogram.
By Potentiometer: Changes in the potential of a metal electrode in
contact with the filter paper is also utilised with quadrant
electrometer or electronic voltmeters.
Fluorimetry: The compound to be determined by fluorimetry must
be fluorescent or convertible into fluorescent derivatives.
Radiotracer Method: The compound containing radioactive
element is labeled and treated with locating reagent. Using Geiger
Muller counter.
42. 42
SOURCES of ERROR
Error during application of the spots:
o Apply minimum volume of the concentrated solution in order to
avoid diffusion through the paper which leads to poor separation
o Spots should be approximately of the same diameter.
Development:
o Improper adjustment of the paper in the tank leads to this error
so the paper should be held vertically.
o Do chamber saturation
Detection:
o The spraying methods affect the final result
43. 43
APPLICATIONS
Innumerable applications have been reported in analysis of
different classes of compounds such as:
Amino acids and organic acids
Alkaloids
Polysaccharides
Proteins and peptides
Natural and artificial pigments
Inorganic cations
Plant extracts
44. 44
Applications cont.,
Typical applications in key areas are briefly outlined here:
1. Reaction monitoring:
In a chemical reaction over a period of time the concentration of
reactants decreases whereas the concentration of products increases.
Developing the chromatogram over different time intervals by spotting
the reactants can give a fair idea on the progress of reaction.
Traditionally the technique was used for qualitative monitoring but
availability of densitometers made quantitative estimations possible.
45. 45
Applications cont.,
2. Isolation & Purification:
Paper chromatography has been put to use as a purification and
isolation technique for components of mixtures. The separated
components on the paper are cut, dissolved in suitable solvents and
their absorption characterised at specific wavelengths using
spectrophotometric methods.
3. Foods & Beverages:
Paper chromatography has been used for analysis of food colours in
synthetic drinks and beverages, ice creams, jams & jellies, sweets, etc.
46. 46
Applications cont.,
2. Isolation & Purification:
Paper chromatography has been put to use as a purification and
isolation technique for components of mixtures. The separated
components on the paper are cut, dissolved in suitable solvents and
their absorption characterised at specific wavelengths using
spectrophotometric methods.
3. Foods & Beverages:
Paper chromatography has been used for analysis of food colours in
synthetic drinks and beverages, ice creams, jams & jellies, sweets, etc.
47. 47
Applications cont.,
4. Forensics:
The major applications are for identification and comparison against
reference standards for drugs and their metabolites in viscera, explosive
residues from blast sites, inks used in forgery of documents and paint
pigments investigations in hit and run road accident cases.
5. Pharmaceuticals:
It offers a cost-effective alternative to monitoring the active ingredients
in drug forms administered in bioanalytical studies. Its main
contribution is when the quantity of sample available is in minute
amounts.