This document discusses fluorescence spectroscopy. It begins by defining fluorescence as the emission of light by a substance when an electron returns to the ground state from an excited state. Factors that affect fluorescence include temperature, viscosity, oxygen concentration, pH, and molecular structure. Applications of fluorescence in pharmacy include determining inorganic substances, in nuclear research, as fluorescent indicators, in organic analysis, in liquid chromatography, and for determining vitamins B1 and B2. Instrumentation for fluorescence spectroscopy includes various light sources, filters, sample cells, and detectors such as photomultiplier tubes.
This document discusses factors that affect fluorimetry and quenching. It lists several factors that can influence fluorescence, including the nature of molecules, substituents, concentration, adsorption, light, oxygen, pH, temperature, and viscosity. It also describes different types of quenching such as self-quenching, chemical quenching, static quenching, and collisional quenching. Chemical quenching can occur due to changes in pH, presence of oxygen, or heavy metals. Static quenching involves complex formation between the fluorophore and quencher. Collisional quenching occurs through interactions between an excited fluorophore and quencher molecule.
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.
This document discusses factors that affect fluorescence intensity. It explains that fluorescence intensity is directly proportional to the rigidity of a structure and inversely proportional to temperature. Other factors that can decrease fluorescence intensity include oxygen, which can oxidize fluorescent substances, as well as electron withdrawing substituent groups. Fluorescence intensity is directly proportional to concentration at low concentrations but does not obey linearity at high concentrations. The presence of other non-fluorescent solutes can also impact intensity through inner filter effects. In conclusion, several physicochemical factors influence fluorescence intensity measurements.
This document provides an overview of flame photometry, which uses the intensity of light emitted from atoms in a flame to determine the concentration of certain metal ions in solutions. It describes the basic principles, where a sample is nebulized into a flame and the heat excites the metal atoms to emit light at specific wavelengths. It then discusses the key components of a flame photometer in detail, including the nebulizer, burners, mirrors, slits, and detectors used to measure the light intensities and determine concentrations. Limitations of the technique are that only certain elements can be analyzed as solutions that are volatile in flames.
This document discusses the instrumentation of UV spectrophotometry. It describes the key components which include sources of UV radiation like hydrogen discharge lamps, xenon discharge lamps, and mercury arc lamps. It also discusses monochromators like gratings to produce monochromatic light, and sample holders/cuvettes to hold liquid samples. Common detectors mentioned are barrier layer cells, phototubes, and photomultiplier tubes. Finally, it explains the basic setup of single beam and double beam UV spectrophotometers used for analysis.
Controlled release drug delivery system2Bansari Patel
This document provides an overview of controlled release drug delivery systems (CRDDS). It defines CRDDS as systems that provide some control over the temporal or spatial release of drugs. The key advantages of CRDDS are maintaining effective drug levels, decreasing dosing frequency and side effects, and improving patient compliance. Factors like drug properties, pharmacokinetics, and pharmacodynamics can affect CRDDS. Various approaches to designing CRDDS are discussed.
This document presents an overview of fluorimetry. It discusses that fluorimetry is the measurement of emitted fluorescence light. When certain substances are exposed to light, they emit visible light or radiation, known as fluorescence. Fluorescence occurs immediately after light absorption and stops when the light is removed. Phosphorescence is delayed fluorescence that continues after light removal. Fluorimetry works by exciting substances from their singlet ground state to a singlet excited state, then measuring the wavelength of light emitted as they return to the ground state.
This document discusses fluorescence spectroscopy. It begins by defining fluorescence as the emission of light by a substance when an electron returns to the ground state from an excited state. Factors that affect fluorescence include temperature, viscosity, oxygen concentration, pH, and molecular structure. Applications of fluorescence in pharmacy include determining inorganic substances, in nuclear research, as fluorescent indicators, in organic analysis, in liquid chromatography, and for determining vitamins B1 and B2. Instrumentation for fluorescence spectroscopy includes various light sources, filters, sample cells, and detectors such as photomultiplier tubes.
This document discusses factors that affect fluorimetry and quenching. It lists several factors that can influence fluorescence, including the nature of molecules, substituents, concentration, adsorption, light, oxygen, pH, temperature, and viscosity. It also describes different types of quenching such as self-quenching, chemical quenching, static quenching, and collisional quenching. Chemical quenching can occur due to changes in pH, presence of oxygen, or heavy metals. Static quenching involves complex formation between the fluorophore and quencher. Collisional quenching occurs through interactions between an excited fluorophore and quencher molecule.
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.
This document discusses factors that affect fluorescence intensity. It explains that fluorescence intensity is directly proportional to the rigidity of a structure and inversely proportional to temperature. Other factors that can decrease fluorescence intensity include oxygen, which can oxidize fluorescent substances, as well as electron withdrawing substituent groups. Fluorescence intensity is directly proportional to concentration at low concentrations but does not obey linearity at high concentrations. The presence of other non-fluorescent solutes can also impact intensity through inner filter effects. In conclusion, several physicochemical factors influence fluorescence intensity measurements.
This document provides an overview of flame photometry, which uses the intensity of light emitted from atoms in a flame to determine the concentration of certain metal ions in solutions. It describes the basic principles, where a sample is nebulized into a flame and the heat excites the metal atoms to emit light at specific wavelengths. It then discusses the key components of a flame photometer in detail, including the nebulizer, burners, mirrors, slits, and detectors used to measure the light intensities and determine concentrations. Limitations of the technique are that only certain elements can be analyzed as solutions that are volatile in flames.
This document discusses the instrumentation of UV spectrophotometry. It describes the key components which include sources of UV radiation like hydrogen discharge lamps, xenon discharge lamps, and mercury arc lamps. It also discusses monochromators like gratings to produce monochromatic light, and sample holders/cuvettes to hold liquid samples. Common detectors mentioned are barrier layer cells, phototubes, and photomultiplier tubes. Finally, it explains the basic setup of single beam and double beam UV spectrophotometers used for analysis.
Controlled release drug delivery system2Bansari Patel
This document provides an overview of controlled release drug delivery systems (CRDDS). It defines CRDDS as systems that provide some control over the temporal or spatial release of drugs. The key advantages of CRDDS are maintaining effective drug levels, decreasing dosing frequency and side effects, and improving patient compliance. Factors like drug properties, pharmacokinetics, and pharmacodynamics can affect CRDDS. Various approaches to designing CRDDS are discussed.
This document presents an overview of fluorimetry. It discusses that fluorimetry is the measurement of emitted fluorescence light. When certain substances are exposed to light, they emit visible light or radiation, known as fluorescence. Fluorescence occurs immediately after light absorption and stops when the light is removed. Phosphorescence is delayed fluorescence that continues after light removal. Fluorimetry works by exciting substances from their singlet ground state to a singlet excited state, then measuring the wavelength of light emitted as they return to the ground state.
The document discusses different types of gas chromatography columns. It describes packed columns which contain a solid support material that absorbs the stationary phase. Capillary or open columns have a very thin film coating on the inner wall that acts as the stationary phase. Key parameters that determine column selection and performance are discussed such as diameter, length, film thickness, and stationary phase properties. The advantages of capillary columns over packed columns are their higher efficiency and resolution due to higher ratio of stationary to mobile phase volumes.
This document discusses nepheloturbidometry, which uses light scattering measurements to determine the concentration of suspended particles in liquids. It can be used for both low concentrations (nephelometry) and high concentrations (turbidometry) by measuring either scattered light or transmitted light. A nepheloturbidimeter has detectors to measure both scattered light at 90 degrees and transmitted light at 180 degrees, allowing it to analyze suspensions of unknown concentration. Factors like particle properties, light source, sample cells, and detectors can affect light scattering measurements. Nepheloturbidimetry has various applications like analyzing water clarity, determining carbon dioxide, and quantifying ions at low levels.
This document discusses the principles and applications of fluorimetry. It defines fluorescence as the emission of light from excited electrons returning to ground state. Fluorimetry involves using ultraviolet light to excite sample molecules, which then emit light of lower energy. Factors that influence fluorescence intensity are discussed. Common instrumentation includes light sources, monochromators, cuvettes, detectors, and data analyzers. Applications include determining inorganic substances, proteins, and steroids. Two case studies on analyzing glucose in milk and silver in food/water samples using fluorimetry are presented.
Fluorimetry involves measuring fluorescence intensity at a particular wavelength using a fluorimeter or spectrofluorimeter. Fluorescence occurs when molecules absorb radiation and electrons are excited to a higher energy state. As electrons return to the ground state, they emit radiation. Factors like concentration, pH, and temperature can affect fluorescence intensity. Instrumentation includes a light source, filters/monochromators, sample cells, and detectors. Applications include determining inorganic/organic substances and compounds in pharmaceutical analysis.
The rate theory of chromatography proposes equations to describe the processes that contribute to band broadening in chromatography. These include eddy diffusion, longitudinal diffusion, and resistance to mass transfer in both the mobile and stationary phases. The key equation from rate theory is the Van Deemter equation, which relates plate height to the average linear velocity of the mobile phase and factors A, B, and C that describe the various broadening processes. The Van Deemter equation can be used to determine the optimum mobile phase flow rate.
Gas chromatography is a technique used to separate components in a mixture using an inert gas as the mobile phase and a stationary phase in the column. Key aspects of gas chromatography include the carrier gas, sample injection, columns with solid or liquid stationary phases, temperature programming, and detectors like FID, TCD, ECD that measure separated components. Gas chromatography provides sensitive, precise, and accurate analysis of mixtures like drugs, foods, pollutants, and more within a short time.
This document discusses Beer's law, which states that absorbance of a solution is directly proportional to the concentration of the absorbing material in the solution. It defines Beer's law, derives the mathematical equation, and lists some limitations and sources of deviation from the law, including high concentrations, dissociation/association reactions, use of polychromatic radiation, stray light, and mismatched sample cells. The derivation shows how the Beer's law equation is obtained based on probability of photon absorption in thin sections of the sample.
Spectrophotometric titration by Mr. Pradeep SwarnkarPradeep Swarnkar
Spectrophotometric titrations use changes in absorbance of light to determine the endpoint of a titration. There are two main types of spectrophotometric titration - acid-base and precipitation titrations. Acid-base titrations involve a color change of an indicator and precipitation titrations involve monitoring the formation of a precipitate.
The document discusses fluorescence spectroscopy. It defines fluorescence as emission of light that occurs when a substance absorbs light and returns to its ground state, emitting photons. Factors that affect fluorescence include the molecular structure, substituents, concentration, pH, temperature, and viscosity. Instrumentation for fluorescence spectroscopy includes a light source, filters, sample cells, and detectors such as photomultiplier tubes. Applications of fluorescence spectroscopy include determination of inorganic substances, use as fluorescent indicators, pharmaceutical analysis, and liquid chromatography.
This document provides calibration procedures for various instruments used in pharmaceutical analysis. It describes calibrating a UV-Vis spectrophotometer using holmium perchlorate and potassium dichromate primary standards to control wavelength and absorbance. It also provides procedures to calibrate using potassium chloride and toluene solutions. Regular calibration is important to ensure instruments produce accurate results, and should be performed when time periods elapse, operating hours change, a new instrument is used, or observations seem questionable.
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.
introduction
Interference is a phenomena
that leads to changes (either positive or negative) in intensity of the analyte signal in spectroscopy.
Interferences in atomic absorption spectroscopy fall into two basic categories, namely, non-spectral and spectral.
1. spectral 2. Non Spectral ( Matrix interference, chemical interference, ionization interference)
PRINCIPLE - atomic absorpion spectroscopy
Atoms of the analyte have a fixed number of electrons.
If the light of a specific wavelength is passed through a flame containing that atom, electrons present in different energy levels, known as orbitals, absorb a certain wavelength and excite to higher energy levels.
The extent of absorption ά the number of ground-state atoms in the flame.
Only for information- The ground state is more stable than the excited state. The electrons spontaneously return back to the ground state. It emits the same amount of radiant energy. This process is called fluorescence. Fluorescence is used in atomic emission spectroscopy.
Brief note on - Instrumentation
The basic components of atomic absorption are:
Light source
Chopper
Atomizer
Burners
flames
Monochromators
Detectors
Amplifier
Readout devices
WORKING PROCESS
Calibration
Quantitative analysis in AAS
Safety measures
Important questions and answer
This document discusses quality control tests for pharmaceutical packing materials. It describes the different types of primary, secondary, and tertiary packing materials used in the pharmaceutical industry. It focuses on quality control tests for various container materials like glass and plastic. Some key tests discussed for glass containers include chemical resistance testing, hydrolytic resistance testing, surface etching testing, and light transmission testing. For plastic containers, leakage testing, collapsibility testing, clarity of aqueous extract testing, and non-volatile residue testing are described. The document provides details on procedures and acceptance criteria for many of these important quality control tests.
the presentation gives knowledge about principle or fluorometry, factors that affect fluorescence including quenching instruments used in fluorometry, and the applications of fluorometry. added references in the end for more knowledge.
This document provides an overview of fluorescence spectroscopy. It defines luminescence as the emission of photons from electronically excited states. There are two main types of luminescence: fluorescence from singlet excited states and phosphorescence from triplet excited states. The document discusses instrumentation for fluorescence spectroscopy including light sources, wavelength selection devices like filters and monochromators, and detectors. It also covers factors that affect fluorescence intensity such as molecular structure, concentration, temperature, and pH.
Fluorimetry, principle, Concept of singlet,doublet,and triplet electronic sta...Vandana Devesh Sharma
This document discusses the principles and factors affecting fluorescence and fluorimetry. It begins by defining fluorescence as the emission of light by a substance that has absorbed light or other electromagnetic radiation. It then discusses various processes that can occur in excited molecules including fluorescence, phosphorescence, internal conversion, intersystem crossing, and collisional deactivation. The document also summarizes several factors that can influence fluorescence intensity, including molecular structure, temperature, viscosity, oxygen content, and pH. Structural factors discussed include conjugation, substituent groups, and molecular rigidity.
Counter current Extraction Solid phase extraction Gel filtrationPHARMA WORLD
This document provides an overview of counter current extraction, solid phase extraction, and gel filtration chromatography techniques. It discusses the principles, processes, instrumentation, and applications of each technique. Counter current extraction uses a series of tubes to separate mixtures into phases based on solubility differences. Solid phase extraction isolates analytes from samples using adsorption onto a solid phase. Gel filtration separates molecules based on size by allowing larger molecules to pass through column pores. Each technique has advantages for analytical separations and preparative isolation of compounds from complex mixtures.
This document discusses various topics related to UV-visible spectroscopy including:
1. Choice of solvents and their effects on UV-visible spectra. Polar solvents can cause red or blue shifts in absorption maxima depending on the solute.
2. Applications of UV-visible spectroscopy like quantitative analysis of single and multiple component samples and qualitative analysis through structural elucidation, detection of functional groups, and identification of compounds.
3. Difference spectroscopy, where the difference in absorbance between two samples is measured to improve selectivity in the presence of interfering absorbers.
Introduction to chromatography, Definition of Chromatography, Types of column chromatography, Theory of chromatography, Practical considerations in column chromatography , Factors affecting efficiency of a column, Applications.
Fluorimetry is a technique used in analytical chemistry and biochemistry to measure the concentration of a substance in a sample by analyzing the fluorescence it emits when exposed to specific wavelengths of light. This technique is based on the principle of fluorescence, which is the emission of light (or photons) by a molecule when it absorbs photons at a shorter wavelength.
Here's how fluorimetry works:
Excitation: A sample is exposed to a specific wavelength of light, known as the excitation wavelength, which is typically in the ultraviolet or visible range. This excitation light is absorbed by the molecules of interest in the sample, causing them to move to higher energy states.
Emission: After absorbing the excitation light, the molecules return to their ground state by releasing energy in the form of fluorescent light at longer wavelengths. The emitted light is typically at a longer wavelength than the excitation light, and it is specific to the particular molecule or compound being analyzed.
Detection: A detector, such as a photomultiplier tube or a photodiode, is used to measure the intensity of the emitted fluorescent light. The detector is sensitive to the specific wavelength of light emitted by the target molecules.
Data Analysis: The intensity of the emitted fluorescent light is correlated with the concentration of the substance in the sample. By comparing the intensity of the emitted light to a calibration curve or standard, the concentration of the substance can be determined.
Fluorimetry has various applications in chemistry and biology. It is commonly used for quantifying the concentration of fluorescent dyes, proteins, nucleic acids (e.g., DNA and RNA), and other biomolecules. It is also employed in environmental analysis, drug discovery, and medical diagnostics.
One of the advantages of fluorimetry is its high sensitivity, which allows for the detection of very low concentrations of analytes. Additionally, it offers high selectivity because the emitted fluorescence is specific to the target molecule.
Overall, fluorimetry is a valuable analytical tool that helps researchers and scientists measure and analyze a wide range of substances with high precision and sensitivity
fluorometry is used in pharmaceutical fields.An analytic method for detecting and measuring fluorescence in compounds that uses ultraviolet light stimulating the compounds, causing them to emit visible light. An important topic studied in instrumental analysis.
The document discusses different types of gas chromatography columns. It describes packed columns which contain a solid support material that absorbs the stationary phase. Capillary or open columns have a very thin film coating on the inner wall that acts as the stationary phase. Key parameters that determine column selection and performance are discussed such as diameter, length, film thickness, and stationary phase properties. The advantages of capillary columns over packed columns are their higher efficiency and resolution due to higher ratio of stationary to mobile phase volumes.
This document discusses nepheloturbidometry, which uses light scattering measurements to determine the concentration of suspended particles in liquids. It can be used for both low concentrations (nephelometry) and high concentrations (turbidometry) by measuring either scattered light or transmitted light. A nepheloturbidimeter has detectors to measure both scattered light at 90 degrees and transmitted light at 180 degrees, allowing it to analyze suspensions of unknown concentration. Factors like particle properties, light source, sample cells, and detectors can affect light scattering measurements. Nepheloturbidimetry has various applications like analyzing water clarity, determining carbon dioxide, and quantifying ions at low levels.
This document discusses the principles and applications of fluorimetry. It defines fluorescence as the emission of light from excited electrons returning to ground state. Fluorimetry involves using ultraviolet light to excite sample molecules, which then emit light of lower energy. Factors that influence fluorescence intensity are discussed. Common instrumentation includes light sources, monochromators, cuvettes, detectors, and data analyzers. Applications include determining inorganic substances, proteins, and steroids. Two case studies on analyzing glucose in milk and silver in food/water samples using fluorimetry are presented.
Fluorimetry involves measuring fluorescence intensity at a particular wavelength using a fluorimeter or spectrofluorimeter. Fluorescence occurs when molecules absorb radiation and electrons are excited to a higher energy state. As electrons return to the ground state, they emit radiation. Factors like concentration, pH, and temperature can affect fluorescence intensity. Instrumentation includes a light source, filters/monochromators, sample cells, and detectors. Applications include determining inorganic/organic substances and compounds in pharmaceutical analysis.
The rate theory of chromatography proposes equations to describe the processes that contribute to band broadening in chromatography. These include eddy diffusion, longitudinal diffusion, and resistance to mass transfer in both the mobile and stationary phases. The key equation from rate theory is the Van Deemter equation, which relates plate height to the average linear velocity of the mobile phase and factors A, B, and C that describe the various broadening processes. The Van Deemter equation can be used to determine the optimum mobile phase flow rate.
Gas chromatography is a technique used to separate components in a mixture using an inert gas as the mobile phase and a stationary phase in the column. Key aspects of gas chromatography include the carrier gas, sample injection, columns with solid or liquid stationary phases, temperature programming, and detectors like FID, TCD, ECD that measure separated components. Gas chromatography provides sensitive, precise, and accurate analysis of mixtures like drugs, foods, pollutants, and more within a short time.
This document discusses Beer's law, which states that absorbance of a solution is directly proportional to the concentration of the absorbing material in the solution. It defines Beer's law, derives the mathematical equation, and lists some limitations and sources of deviation from the law, including high concentrations, dissociation/association reactions, use of polychromatic radiation, stray light, and mismatched sample cells. The derivation shows how the Beer's law equation is obtained based on probability of photon absorption in thin sections of the sample.
Spectrophotometric titration by Mr. Pradeep SwarnkarPradeep Swarnkar
Spectrophotometric titrations use changes in absorbance of light to determine the endpoint of a titration. There are two main types of spectrophotometric titration - acid-base and precipitation titrations. Acid-base titrations involve a color change of an indicator and precipitation titrations involve monitoring the formation of a precipitate.
The document discusses fluorescence spectroscopy. It defines fluorescence as emission of light that occurs when a substance absorbs light and returns to its ground state, emitting photons. Factors that affect fluorescence include the molecular structure, substituents, concentration, pH, temperature, and viscosity. Instrumentation for fluorescence spectroscopy includes a light source, filters, sample cells, and detectors such as photomultiplier tubes. Applications of fluorescence spectroscopy include determination of inorganic substances, use as fluorescent indicators, pharmaceutical analysis, and liquid chromatography.
This document provides calibration procedures for various instruments used in pharmaceutical analysis. It describes calibrating a UV-Vis spectrophotometer using holmium perchlorate and potassium dichromate primary standards to control wavelength and absorbance. It also provides procedures to calibrate using potassium chloride and toluene solutions. Regular calibration is important to ensure instruments produce accurate results, and should be performed when time periods elapse, operating hours change, a new instrument is used, or observations seem questionable.
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.
introduction
Interference is a phenomena
that leads to changes (either positive or negative) in intensity of the analyte signal in spectroscopy.
Interferences in atomic absorption spectroscopy fall into two basic categories, namely, non-spectral and spectral.
1. spectral 2. Non Spectral ( Matrix interference, chemical interference, ionization interference)
PRINCIPLE - atomic absorpion spectroscopy
Atoms of the analyte have a fixed number of electrons.
If the light of a specific wavelength is passed through a flame containing that atom, electrons present in different energy levels, known as orbitals, absorb a certain wavelength and excite to higher energy levels.
The extent of absorption ά the number of ground-state atoms in the flame.
Only for information- The ground state is more stable than the excited state. The electrons spontaneously return back to the ground state. It emits the same amount of radiant energy. This process is called fluorescence. Fluorescence is used in atomic emission spectroscopy.
Brief note on - Instrumentation
The basic components of atomic absorption are:
Light source
Chopper
Atomizer
Burners
flames
Monochromators
Detectors
Amplifier
Readout devices
WORKING PROCESS
Calibration
Quantitative analysis in AAS
Safety measures
Important questions and answer
This document discusses quality control tests for pharmaceutical packing materials. It describes the different types of primary, secondary, and tertiary packing materials used in the pharmaceutical industry. It focuses on quality control tests for various container materials like glass and plastic. Some key tests discussed for glass containers include chemical resistance testing, hydrolytic resistance testing, surface etching testing, and light transmission testing. For plastic containers, leakage testing, collapsibility testing, clarity of aqueous extract testing, and non-volatile residue testing are described. The document provides details on procedures and acceptance criteria for many of these important quality control tests.
the presentation gives knowledge about principle or fluorometry, factors that affect fluorescence including quenching instruments used in fluorometry, and the applications of fluorometry. added references in the end for more knowledge.
This document provides an overview of fluorescence spectroscopy. It defines luminescence as the emission of photons from electronically excited states. There are two main types of luminescence: fluorescence from singlet excited states and phosphorescence from triplet excited states. The document discusses instrumentation for fluorescence spectroscopy including light sources, wavelength selection devices like filters and monochromators, and detectors. It also covers factors that affect fluorescence intensity such as molecular structure, concentration, temperature, and pH.
Fluorimetry, principle, Concept of singlet,doublet,and triplet electronic sta...Vandana Devesh Sharma
This document discusses the principles and factors affecting fluorescence and fluorimetry. It begins by defining fluorescence as the emission of light by a substance that has absorbed light or other electromagnetic radiation. It then discusses various processes that can occur in excited molecules including fluorescence, phosphorescence, internal conversion, intersystem crossing, and collisional deactivation. The document also summarizes several factors that can influence fluorescence intensity, including molecular structure, temperature, viscosity, oxygen content, and pH. Structural factors discussed include conjugation, substituent groups, and molecular rigidity.
Counter current Extraction Solid phase extraction Gel filtrationPHARMA WORLD
This document provides an overview of counter current extraction, solid phase extraction, and gel filtration chromatography techniques. It discusses the principles, processes, instrumentation, and applications of each technique. Counter current extraction uses a series of tubes to separate mixtures into phases based on solubility differences. Solid phase extraction isolates analytes from samples using adsorption onto a solid phase. Gel filtration separates molecules based on size by allowing larger molecules to pass through column pores. Each technique has advantages for analytical separations and preparative isolation of compounds from complex mixtures.
This document discusses various topics related to UV-visible spectroscopy including:
1. Choice of solvents and their effects on UV-visible spectra. Polar solvents can cause red or blue shifts in absorption maxima depending on the solute.
2. Applications of UV-visible spectroscopy like quantitative analysis of single and multiple component samples and qualitative analysis through structural elucidation, detection of functional groups, and identification of compounds.
3. Difference spectroscopy, where the difference in absorbance between two samples is measured to improve selectivity in the presence of interfering absorbers.
Introduction to chromatography, Definition of Chromatography, Types of column chromatography, Theory of chromatography, Practical considerations in column chromatography , Factors affecting efficiency of a column, Applications.
Fluorimetry is a technique used in analytical chemistry and biochemistry to measure the concentration of a substance in a sample by analyzing the fluorescence it emits when exposed to specific wavelengths of light. This technique is based on the principle of fluorescence, which is the emission of light (or photons) by a molecule when it absorbs photons at a shorter wavelength.
Here's how fluorimetry works:
Excitation: A sample is exposed to a specific wavelength of light, known as the excitation wavelength, which is typically in the ultraviolet or visible range. This excitation light is absorbed by the molecules of interest in the sample, causing them to move to higher energy states.
Emission: After absorbing the excitation light, the molecules return to their ground state by releasing energy in the form of fluorescent light at longer wavelengths. The emitted light is typically at a longer wavelength than the excitation light, and it is specific to the particular molecule or compound being analyzed.
Detection: A detector, such as a photomultiplier tube or a photodiode, is used to measure the intensity of the emitted fluorescent light. The detector is sensitive to the specific wavelength of light emitted by the target molecules.
Data Analysis: The intensity of the emitted fluorescent light is correlated with the concentration of the substance in the sample. By comparing the intensity of the emitted light to a calibration curve or standard, the concentration of the substance can be determined.
Fluorimetry has various applications in chemistry and biology. It is commonly used for quantifying the concentration of fluorescent dyes, proteins, nucleic acids (e.g., DNA and RNA), and other biomolecules. It is also employed in environmental analysis, drug discovery, and medical diagnostics.
One of the advantages of fluorimetry is its high sensitivity, which allows for the detection of very low concentrations of analytes. Additionally, it offers high selectivity because the emitted fluorescence is specific to the target molecule.
Overall, fluorimetry is a valuable analytical tool that helps researchers and scientists measure and analyze a wide range of substances with high precision and sensitivity
fluorometry is used in pharmaceutical fields.An analytic method for detecting and measuring fluorescence in compounds that uses ultraviolet light stimulating the compounds, causing them to emit visible light. An important topic studied in instrumental analysis.
Flourescence spectroscopy- instrumentation and applicationssinghsnehi01
This document discusses fluorescence and phosphorescence. It defines fluorescence as the emission of light that starts immediately upon absorption of light and stops when the light is removed. Phosphorescence is defined as delayed fluorescence where light continues to be emitted even after the absorbed light is removed. It discusses factors that affect fluorescence like concentration, quantum yield, incident light intensity, oxygen, pH, temperature, viscosity, photodecomposition, and quenchers. Instrumentation for fluorescence includes light sources, filters, sample cells, monochromators, and detectors like photomultiplier tubes. Applications include determination of metals in alloys and fluorescence-based assays.
Luminescence is the emission of light by a substance. It occurs when an electron returns to the electronic ground state from an excited state and loses its excess energy as a photon.
It is of 3 types.
Fluorescence spectroscopy.
Phosphorescence spectroscopy.
Chemiluminescence spectroscopy
Fluorescence spectroscopy. : When a beam of light is incident on certain substances they emit visible light or radiations. This is known as fluorescence. Fluorescence starts immediately after the absorption of light and stops as soon as the incident light is cut off. The substances showing this phenomenon are known as flourescent substances
Phosphorescence spectroscopy: When light radiation is incident on certain substances they emit light continuously even after the incident light is cut off.
This type of delayed fluorescence is called phosphorescence.
Substances showing phosphorescence are phosphorescent substances.
Chemiluminescence (also chemoluminescence) is the emission of light (luminescence) as the result of a chemical reaction. There may also be limited emission of heat
Fluorescence
Phosphorescence
Radiation less processes
Vibration relaxation
Internal conversion
External conversion
Intersystem crossing
Jablonski diagram is a graphical representation of the various transitions(electronic states, vibrational levels) that can occur after a molecule has been excited photochemically.
When a molecule is raised from its ground state to a higher state using light, photochemistry occurs.
The molecule in the excited state has a shorter lifetime and significantly more energy than the ground state from which it was formed.
As a result, molecules in the excited state are much more reactive.
A photochemical or photophysical process deactivates an excited state.
Therefore, the fate of the excited molecules is described by using the Jablonski diagram, which only focuses on the photophysical process occurring during the excitation and deactivation process.
Radiative transitions involve the absorption of a photon, if the transition occurs to a higher energy level, or the emission of a photon, for a transition to a lower level.
Nonradiative transitions arise through several different mechanisms, all differently labeled in the diagram. Relaxation of the excited state to its lowest vibrational level is called vibrational relaxation. This process involves the dissipation of energy from the molecule to its surroundings, and thus it cannot occur for isolated molecules. A second type of nonradiative transition is internal conversion (IC), which occurs when a vibrational state of an electronically excited state can couple to a vibrational state of a lower electronic state.
A third type is intersystem crossing (ISC); this is a transition to a state with a different spin multiplicity. In molecules with large spin-orbit coupling, intersystem crossing is much more important than in molecules that exhibit only small spin-orbit coupling. ISC can
This document discusses fluorimetry and phosphorimetry. It defines them as measurement techniques, with fluorimetry measuring fluorescence intensity at a particular wavelength, and phosphorimetry measuring phosphorescence in conjunction with pulsed radiation. It describes the principles behind photoluminescence, including fluorescence and phosphorescence. Factors affecting these processes and instrumentation used are summarized, including light sources, filters, monochromators, and detectors. Applications in pharmaceutical, clinical, environmental, and entertainment fields are also briefly outlined.
Fluorimetry involves the measurement of fluorescence from substances. When certain molecules absorb light, they emit light of a longer wavelength as they fall to the ground state. Factors like pH, oxygen, and temperature can affect fluorescence. Instruments used include single and double beam fluorimeters and spectrofluorimeters. Applications of fluorimetry include determination of inorganic ions, vitamins, and compounds in pharmaceutical and environmental analysis.
This document discusses fluorescence spectroscopy and its applications in pharmacy. It begins with definitions of fluorescence, phosphorescence, and chemiluminescence. It describes how fluorescent substances emit light when exposed to radiation and discusses factors that affect fluorescence like molecular structure, substituents, concentration, oxygen, pH, and temperature. The principles of fluorescence are explained using Jablonski diagrams. Instrumentation for fluorescence spectroscopy including light sources, filters, sample cells, and detectors are outlined. Finally, applications of fluorescence spectroscopy in inorganic analysis, organic analysis, liquid chromatography, and determination of vitamins and drugs are described.
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.
This document provides an overview of fluorimetry. It defines fluorimetry as the measurement of fluorescence with a spectrofluorimeter. Fluorescence occurs when a substance emits light after absorbing radiation. Factors that affect fluorescence include the nature of the molecule, substituents, concentration, oxygen, pH, temperature, and viscosity. The instrumentation involves a light source, filters, sample cells, and detectors like photomultiplier tubes. Applications of fluorimetry include determining inorganic substances, using fluorescent indicators, developing fluorescent reagents, organic analysis, pharmaceutical analysis, and liquid chromatography.
Fluorimetry involves measuring the fluorescence light emitted by a substance. A fluorimeter is used, which contains a light source, filters or monochromators to select excitation and emission wavelengths, sample cells, and a detector. Fluorescence occurs when molecules absorb light and emit light of a longer wavelength as they transition from an excited singlet state to the ground state. Factors like concentration, pH, and temperature can affect fluorescence. Fluorimetry is used in applications like determining ions and vitamins, and in organic analysis.
This document discusses fluorimetric analysis. It begins with defining fluorescence and the different types of luminescence. Excited electrons return to the ground state and emit photons. Factors affecting fluorescence include the nature of the molecule, substituents, concentration, pH, temperature, and viscosity. Instrumentation includes light sources, filters, monochromators, sample cells, and detectors like photomultiplier tubes. Applications of fluorescence include determining inorganic substances, using fluorescent indicators, organic analysis, pharmaceutical analysis, and liquid chromatography.
Fluorimetry is a technique that measures fluorescence intensity at a particular wavelength using a fluorimeter or spectrofluorimeter. It works by exciting a molecule's electrons with radiation, causing them to emit radiation upon returning to the ground state. Factors like concentration, quantum yield, incident light intensity, pH, temperature and presence of quenchers can affect fluorescence. Fluorimetry has advantages like high sensitivity, precision and specificity and is useful for determining various inorganic/organic substances and compounds.
This document provides an overview of spectrofluorimetry. It begins with an introduction that defines fluorescence and phosphorescence as types of photoluminescence that occur when electrons return to the ground state from an excited state. It then discusses the principle, theory, instrumentation, factors affecting fluorescence, and applications of spectrofluorimetry. The instrumentation section describes the main components, including a light source, excitation and emission monochromators, sample holder, detector, and readout device. Common factors that can affect fluorescence intensity are concentration, incident light intensity, quantum yield, absorption, pH, oxygen, temperature, viscosity, and scatter. Applications include chemical modification of compounds, identification of compounds based on excitation and emission spectra, and assays of vitamins
This document provides an overview of spectrofluorimetry and fluorescence. It begins by explaining the principles behind fluorescence - how absorption of UV or visible light causes electrons to transition to an excited state and then emit light as they fall back down. It then discusses fluorescence and fluorimetry in more detail. The rest of the document covers the Jablonski diagram, factors that affect fluorescence, types of quenching, instrumentation used including light sources, filters, sample cells and detectors, and applications of fluorescence including pharmaceutical analysis.
spectrofluorometer is the instrument for recording fluorescence emission and absorption spectra When a beam of light is incident on certain substances they emit visible light or radiations. This is known as fluorescence. Fluorescence starts immediately after the absorption of light and stops as soon as the incident light is cut off. The substances showing this phenomenon are known as flourescent substances.
This document discusses fluorescence spectroscopy. It begins with an introduction and definition of luminescence as the emission of light by a substance when an electron returns to the ground state. It then describes the three main types of luminescence: fluorescence, phosphorescence, and chemiluminescence. Factors affecting fluorescence are covered such as temperature, viscosity, oxygen, pH, and chemical structure. Applications in pharmacy and instrumentation are also summarized.
This document discusses spectrofluorimetry, which involves using fluorescence spectroscopy to study the emission of radiation from molecules. It begins by explaining fluorescence, where molecules absorb UV or visible light and emit light of a longer wavelength as they relax to the ground state. The document then defines key terms like singlet and triplet states. It describes the instrumentation used, factors that influence fluorescence intensity, and applications of spectrofluorimetry like determining organic and inorganic substances. Examples are given of using it to analyze minerals, plant pigments, and food and pharmaceutical samples.
This document provides an overview of UV-visible spectroscopy. It discusses the electromagnetic spectrum and how UV-visible spectroscopy involves absorption of UV or visible light by molecules, promoting electrons to excited states. Beer-Lambert's law describes the relationship between light absorption and analyte concentration. Chromophores and auxochromes determine peak absorption wavelengths. Factors like pH and solvents affect absorption spectra. UV-visible spectroscopy has applications in determining molecular structures, concentrations, and impurities. Instrumentation includes sources like hydrogen lamps, monochromators, sample holders, detectors like photomultiplier tubes, and single or double beam spectrophotometers.
Similar to fluorimetry.pptx FINAL YEAR B PHARM SEVENTH SEMESTER PCI PATTERN (20)
This slide is special for master students (MIBS & MIFB) in UUM. Also useful for readers who are interested in the topic of contemporary Islamic banking.
How to Setup Warehouse & Location in Odoo 17 InventoryCeline George
In this slide, we'll explore how to set up warehouses and locations in Odoo 17 Inventory. This will help us manage our stock effectively, track inventory levels, and streamline warehouse operations.
Strategies for Effective Upskilling is a presentation by Chinwendu Peace in a Your Skill Boost Masterclass organisation by the Excellence Foundation for South Sudan on 08th and 09th June 2024 from 1 PM to 3 PM on each day.
বাংলাদেশের অর্থনৈতিক সমীক্ষা ২০২৪ [Bangladesh Economic Review 2024 Bangla.pdf] কম্পিউটার , ট্যাব ও স্মার্ট ফোন ভার্সন সহ সম্পূর্ণ বাংলা ই-বুক বা pdf বই " সুচিপত্র ...বুকমার্ক মেনু 🔖 ও হাইপার লিংক মেনু 📝👆 যুক্ত ..
আমাদের সবার জন্য খুব খুব গুরুত্বপূর্ণ একটি বই ..বিসিএস, ব্যাংক, ইউনিভার্সিটি ভর্তি ও যে কোন প্রতিযোগিতা মূলক পরীক্ষার জন্য এর খুব ইম্পরট্যান্ট একটি বিষয় ...তাছাড়া বাংলাদেশের সাম্প্রতিক যে কোন ডাটা বা তথ্য এই বইতে পাবেন ...
তাই একজন নাগরিক হিসাবে এই তথ্য গুলো আপনার জানা প্রয়োজন ...।
বিসিএস ও ব্যাংক এর লিখিত পরীক্ষা ...+এছাড়া মাধ্যমিক ও উচ্চমাধ্যমিকের স্টুডেন্টদের জন্য অনেক কাজে আসবে ...
This presentation includes basic of PCOS their pathology and treatment and also Ayurveda correlation of PCOS and Ayurvedic line of treatment mentioned in classics.
How to Manage Your Lost Opportunities in Odoo 17 CRMCeline George
Odoo 17 CRM allows us to track why we lose sales opportunities with "Lost Reasons." This helps analyze our sales process and identify areas for improvement. Here's how to configure lost reasons in Odoo 17 CRM
हिंदी वर्णमाला पीपीटी, hindi alphabet PPT presentation, hindi varnamala PPT, Hindi Varnamala pdf, हिंदी स्वर, हिंदी व्यंजन, sikhiye hindi varnmala, dr. mulla adam ali, hindi language and literature, hindi alphabet with drawing, hindi alphabet pdf, hindi varnamala for childrens, hindi language, hindi varnamala practice for kids, https://www.drmullaadamali.com
The simplified electron and muon model, Oscillating Spacetime: The Foundation...RitikBhardwaj56
Discover the Simplified Electron and Muon Model: A New Wave-Based Approach to Understanding Particles delves into a groundbreaking theory that presents electrons and muons as rotating soliton waves within oscillating spacetime. Geared towards students, researchers, and science buffs, this book breaks down complex ideas into simple explanations. It covers topics such as electron waves, temporal dynamics, and the implications of this model on particle physics. With clear illustrations and easy-to-follow explanations, readers will gain a new outlook on the universe's fundamental nature.
A workshop hosted by the South African Journal of Science aimed at postgraduate students and early career researchers with little or no experience in writing and publishing journal articles.
3. FLUORESCENCE
It is a phenomenon of emission of
radiation when the molecules are
exited by radiation at certain
wavelength.
4. FLUORIMETRY:- It is measurement
of fluorescence intensity at a particular
wavelength wit the help of a filter
fluorimeter or a spectrofluorimeter.
5. PRINCIPLE:- Molecule contains electrons,
electrons and non bonding (n) electron.
The electrons may be present in bonding molecular
orbital. It is called as highest occupied molecular
orbital (HOMO).It has lest energy and more stable.
When the molecules absorbs radiant energy
from a light source, the bonding electrons may be
promoted to anti bonding molecular orbital (LUMO).
It has more energy and hence less stable.
6. The process of promotion of electrons from
HOMO to LUMO with absorption of energy
is called as excitation.
Singlet state:-a state in which all the
electrons in a molecule are paired
Doublet state:- a state in which un paired
electrons is present or
Triplet state:- a state in which unpaired
electrons of same spin present
Singlet excited state:- a state in which
electrons are unpaired but of opposite spin
like (un paired and opposite spin)
7. When light of appropriate wavelength is
absorbed by a molecule the electrons are
promoted from singlet ground state to
singlet excited state. once the molecule is in
this excited state relaxation can occur via
several process. For ex by emission of
radiation . The process can be the following
1) Collisional deactivation
2)Fluorescence
3)Phosphorescence.
9. Collisional de activation :- In which entire energy
lost due to collision de activation and no radiation
emitted.
Fluorescence:-excited singlet state is highly
unstable. Relaxation of electrons from excited
singlet to singlet ground state with emission of light.
Phosphorescence:-At favorable condition like low
temperature and absence of oxygen there is
transition from excited singlet state to triplet state
which is called as inner system crossing. The
emission of radiation when electrons undergo
transition from triplet state to singlet ground state is
called as phosphorescence.
11. Fluorescence intensity is praportnal to
concentration of substance only when the
absorbance is less than 0.02
A=log IoIt or A= abc
Io=intensity of incident light
a= absorptivity of constant
b= Pathlength
c= concentration
Concentration:-
12. ()=NUMBER OF PHOTONS EMITTEDNUMBER OF
PHOTONS ABSORBEDS
It Is Always Less Than 1.0 Since Some
Energy Is Lost By Radiation less Pathways
(Collisional, Intersystem Crossing, Vibrational
Relaxation)
13. Increase In The Intensity Of Incident Light On The
Sample Fluorescence Intensity Also Increases.
Adsorption Of Sample Solution In The Container
May Leads T
o A Serious Problem.
OXYGEN:-
Oxidation of fluorescent species to a non
fluorescent species, quenches fluorescent substance.
Ph:-
Alteration of ph of a solution will have significant
effect on fluorescence
For ex Aniline in alkali medium gives visible
fluorescence but in acidic condition gives fluorescence
in visible region.
14. Temperature and viscosity:-
Temperature Increases Can Increase the
collisional de activation, and reduce fluorescent
intensity.
If viscosity of solution is more the frequency of
collisions are reduced and increase in fluorescent
intensity.
Photochemical decomposition:-
Absorption of intense radiation leads to photochemical
decomposition of a fluorescent substance to less
fluorescent or non fluorescent substance.
15. Quenchers:-
Quenching is the reduction of fluorescence intensity
by the presence of substance in the sample other
than the fluorescent analyte.
Quenching is following types:-
16. INNER FLUORESCENT EFFECT:- Absorption Of
Incident (uv) Light Or Emitted (fluorescent) Light By
Primary And Secondary Filters Leads To Decrease
In Fluorescence intensity.
SELF QUENCHING:-At Low Concentration
Linearity Is Observed, At High Concentration Of
The Same Substance Increase In Fluorescent
Intensity Is Observed. This phenomena is called
self quenching.
17. COLLISONAL QUENCHING:- Collisions between
the fluorescent substance and halide ions leads to
reduction in fluorescence intensity.
STATIC QUENCHING:- This occurs because of
complex formation between the fluorescent
molecule and other molecules.
Ex: caffeine reduces fluorescence of riboflavin.
18. Scatter is mainly due to colloidal
particles in solution. Scattering of incident light
after passing through the sample leads to
decrease in fluorescence intensity.
22. 1)SOURCE OF LIGHT:-
mercury vapour lamp: Mercury vapour at high
pressure give intense lines on continuous
background above 350nm.low pressure mercury
vapour gives an additional line at 254nm.it is used
in filter fluorimeter.
FIGURE 3
23. xenon arc lamp: It give more intense radiation
than mercury vapour lamp. it is used in
spectrofluorimeter.
FIGURE 4
tungsten lamp:- If excitation has to be done in
visible region this can be used. It is used in low cost
instruments.
FIGURE 5
24. 2) FILTERS AND MONOCHROMATORS:-
Filters: these are nothing but optical filters works
on the principle of absorption of unwanted light and
transmitting the required wavelength of light. In
inexpensive instruments fluorimeter primary filter
and secondary filter are present.
Primary filter:-absorbs visible radiation
and transmit UV radiation.
Secondary filter:-absorbs UV
radiation and transmit visible
radiation.
FIGURE 6
25. Monochromators: they
convert polychromatic light
into monochromatic light.
They can isolate a specific
range of wavelength or a
particular wavelength of
radiation from a source.
Excitation
monochromators:-provides
suitable radiation for
excitation of molecule .
Emission monochromators:-
isolate only the radiation
emitted by the fluorescent
molecules.
FIGURE 7
26. 3) Sample cells: These are ment for holding liquid
samples. These are made up of quartz and can
have various shapes ex: cylindrical or rectangular
etc.
Barrier layer cell/Photo voltaic cells
Photomultiplier cells
FIGURE 8
4) Detectors: Photometric detectors are used
they are
27. For ex:
1. Barrier layer /photovoltaic cell:
it is employed in inexpensive instruments.
Filter Fluorimeter.
It consists of a copper plate coated with a thin layer of
cuprous oxide (Cu2o). A semi transparent film of silver
is laid on this plate to provide good contact.
When external light falls on the oxide layer, the
electrons emitted from the oxide layer move into the
copper plate.
Then oxide layer becomes positive and copper plate
becomes negative.
28. Hence an emf develops between the oxide layer
and copper plate and behaves like a voltaic cell. So
it is called photovoltaic cell..
A galvanometer is connected externally between
silver film and copper plate and the deflection in the
galvanometer shows the current flow through it.
The amount of current is found to be proportional to
the intensity of incident light
30. 2. Photomultiplier tubes:
These are incorporated in expensive instruments
like spectrofluorimeter. Its sensitivity is high due to
measuring weak intensity of light.
The principle employed in this detector Is that,
multiplication of photoelectrons by secondary
emission of electrons.
This is achieved by using a photo cathode and a
series of anodes (Dyanodes). Up to 10 dyanodes
are used. Each dyanode is maintained at 75-
100Vhigher than the preceding one.
31. At each stage, the electron emission is multiplied by
a factor of 4 to 5 due to secondary emission of
electrons and hence an overall factor of 106 is
achieved.
.
PMT can detect very weak signals, even 200 times
weaker than that could be done using photovoltaic
cell. Hence it is useful in fluorescence
measurements.
PMT should be shielded from stray light in order to
have accurate results.
33. INSTRUMENTS
The most common types are:-
Single beam (filter) fluorimeter
Double beam (filter )fluorimeter
Spectrofluorimeter(double beam)
34. Instruments:-
It contains tungsten lamp as a source of light
and has an optical system consists of primary filter.
The emitted radiations is measured at 900 by
using a secondary filter and detector. Primary filter
absorbs visible radiation and transmit uv radiation
which excites the molecule present in sample cell.
In stead of 90 if we use 180 geometry as in
colorimetry secondary filter has to be highly efficient
other wise both the unabsorbed uv radiation and
fluorescent radiation will produce detector response
and give false result.
35. Single beam instruments are simple in construction
cheaper and easy to operate.
FIGURE 11
36. it is similar to single
beam except that the two incident beams from a
single light source pass through primary filters
separately and fall on the another reference
solution. Then the emitted radiations from the
sample or reference sample pass separately
through secondary filter and produce response
combinly on a detector.
38. In spectrofluorimeter:-
In this primary filter in double beam fluorimeter is
replaced by excitation monochromator and the
secondary filter is replaced by emission
monochromator.
Incident beam is split into sample and
reference beam by using beam splitter.
40. APPLICATIONS
1)Determination of inorganic substances.
Al3+,Li+,ZN2+
2)Determination of thiamine Hcl.
3)Detemination of phenytoin.
4)Determination of indoles, phenols, &
phenothiazines
5) Determination of napthols, proteins, plant pigments
and steroids.
6)Fluorimetry ,nowadays can be used in detection of
impurities in nanogram level better than
absorbance spectrophotometer with special
emphasis in determining components of sample at
the end of chromatographic or capillary column.
41. Determination of ruthenium ions in presence of other
platinum metals.
Determination of boron in steel, aluminum in alloys,
manganese in steel.
Determination of boron in steel by complex formed with
benzoin.
Estimation of cadmium with 2-(2 hydroxyphenyl)
benzoxazole in presence of tartarate .
Respiratory tract infections.