The document discusses guidelines for analyzing viscous samples using micro-flow imaging (MFI) particle characterization. It finds that MFI can accurately measure particle size, concentration, and morphology in samples with viscosities up to 20 centipoise without needing to dilute. Analysis of calibration standards from 1-20 cP showed recovered particle concentrations and sizes were within specification. Morphology parameters like intensity, circularity and aspect ratio were also unaffected by viscosity. MFI thus provides an advantage over other techniques that require diluting viscous samples.
Analyzing Aggregates of Different Sizes and Types: SEC vs. Analytical Ultrace...KBI Biopharma
This document discusses different methods for analyzing protein aggregates of various sizes and types, including size exclusion chromatography (SEC), analytical ultracentrifugation, and light scattering. It notes some limitations of SEC, including that changes in solvent conditions can alter aggregate distributions and larger aggregates may be unresolved. Analytical ultracentrifugation methods like sedimentation velocity and equilibrium are presented as alternatives that are sensitive to all aggregate types and sizes. Sedimentation velocity provides high resolution separation while equilibrium can determine equilibrium constants and stoichiometry of reversible associations. On-line light scattering with SEC is also discussed.
AATCC GWP Orlando Daniel Shook 06-13-01Daniel Shook
This document describes a test method for evaluating the effectiveness of cellulases for reducing surface hair on fabrics using a tergotometer. A tergotometer simulates a home washing machine. The test fabric is a red cotton interlock knit dyed with Direct Red 80. Three cellulases (A, B, C) are tested at different amounts. Results show that cellulase B reduces surface hair and weight loss twice as effectively as cellulase A at the same dosage, indicating it has twice the strength. Cellulase C performs similarly to cellulase A, suggesting it has comparable strength. The test provides a standardized method for comparing cellulase products.
Modified Methylene Blue (MMB) Test ProcedureYasin Engin
This document provides detailed procedures for performing the Modified Methylene Blue (MMB) test to determine the amount of methylene blue adsorbed by aggregate fines. The test involves mixing a sample with methylene blue solution, filtering the mixture, diluting it, and measuring absorbance with a colorimeter. A higher methylene blue value indicates higher clay content, which is undesirable for construction materials. Research has shown a strong correlation between methylene blue value and properties like strength reduction and shrinkage in Portland cement concrete containing clay-contaminated aggregates.
The document summarizes a study that prepared solid dispersions of the drug griseofulvin with the polymer HPMCAS using ball milling. Three surfactants - SDS, DTAB, and Pluronic F127 - were incorporated at 1% and the dispersions characterized. SDS produced the smallest particle sizes and highest drug solubility. DSC showed decreased melting temperature with increased milling time, indicating higher amorphous content. SEM images showed differences in particle size and morphology between samples. In conclusion, ball milling was effective for preparing solid dispersions and surfactant incorporation improved drug solubility and stability.
A Novel Method for Measuring the Sizes and Concentrationskanomaxfmt
The document describes a new liquid nanoparticle sizing (LNS) system that can measure the sizes and concentrations of particles as small as 5 nm in colloidal suspensions. The system works by nebulizing the liquid suspension into an aerosol and then using scanning mobility particle sizing to analyze the particles. The LNS provides direct measurement of particle number concentrations and can detect small changes in particle size distributions. It has demonstrated accurate sizing for particles from 5-500 nm and measurement of filter retention for particles as small as 10 nm. The system allows detailed analysis of polishing slurries and effects of handling on particle size distributions.
Sensitivity and Reproducibility of Protein Aggregate Analysis by Sedimentatio...KBI Biopharma
Sedimentation velocity provides highly sensitive and reproducible analysis of protein aggregates. Replicate samples show the standard deviation for the main peak is typically 0.3% or less, and individual aggregate peaks can be detected below 0.1% and reproducibly quantitated. While data analysis resembles chromatography, the nature of noise is different, so minor peak positions may shift and peaks near detection thresholds may appear or disappear between replicates. Systematic errors from detergents, excipients, UV exposure, or sample loss can also impact reproducibility and limit sensitivity.
The document discusses how operating conditions like pump flow rate and templating agents affect the surface area of highly porous food powders produced via spray drying. It finds that glucose at a high pump flow rate has the greatest change in surface area over temperature changes, while sucrose at a low pump flow rate has the lowest change. The choice of templating agent also impacts properties like crystallinity, glass transition temperature, and surface area due to different self-assembly mechanisms. Characterization of the powders involved analyzing yield, moisture content, sorption behavior, and BET surface area to understand these effects of production conditions.
UV spectrophotometric method development and validation for quantitative esti...Sagar Savale
UV Spectrophotometric Method Development and Validation for quantitative estimation of Ondansetron
Hydrochloride (HCL). U.V Spectrophotometric method have been widely employed in determination of
individual components in a mixture or fixed dose combination. Our aim is to develop spectroscopic method for
estimation of the Ondansetron HCL in ternary mixture by using U.V spectrophotometry. The method was
validated as per ICH guidelines. The recovery studies confirmed the accuracy and precision of the method. It was
successfully applied for the analysis of the drug in bulk and could be effectively used for the routine analysis.
Analyzing Aggregates of Different Sizes and Types: SEC vs. Analytical Ultrace...KBI Biopharma
This document discusses different methods for analyzing protein aggregates of various sizes and types, including size exclusion chromatography (SEC), analytical ultracentrifugation, and light scattering. It notes some limitations of SEC, including that changes in solvent conditions can alter aggregate distributions and larger aggregates may be unresolved. Analytical ultracentrifugation methods like sedimentation velocity and equilibrium are presented as alternatives that are sensitive to all aggregate types and sizes. Sedimentation velocity provides high resolution separation while equilibrium can determine equilibrium constants and stoichiometry of reversible associations. On-line light scattering with SEC is also discussed.
AATCC GWP Orlando Daniel Shook 06-13-01Daniel Shook
This document describes a test method for evaluating the effectiveness of cellulases for reducing surface hair on fabrics using a tergotometer. A tergotometer simulates a home washing machine. The test fabric is a red cotton interlock knit dyed with Direct Red 80. Three cellulases (A, B, C) are tested at different amounts. Results show that cellulase B reduces surface hair and weight loss twice as effectively as cellulase A at the same dosage, indicating it has twice the strength. Cellulase C performs similarly to cellulase A, suggesting it has comparable strength. The test provides a standardized method for comparing cellulase products.
Modified Methylene Blue (MMB) Test ProcedureYasin Engin
This document provides detailed procedures for performing the Modified Methylene Blue (MMB) test to determine the amount of methylene blue adsorbed by aggregate fines. The test involves mixing a sample with methylene blue solution, filtering the mixture, diluting it, and measuring absorbance with a colorimeter. A higher methylene blue value indicates higher clay content, which is undesirable for construction materials. Research has shown a strong correlation between methylene blue value and properties like strength reduction and shrinkage in Portland cement concrete containing clay-contaminated aggregates.
The document summarizes a study that prepared solid dispersions of the drug griseofulvin with the polymer HPMCAS using ball milling. Three surfactants - SDS, DTAB, and Pluronic F127 - were incorporated at 1% and the dispersions characterized. SDS produced the smallest particle sizes and highest drug solubility. DSC showed decreased melting temperature with increased milling time, indicating higher amorphous content. SEM images showed differences in particle size and morphology between samples. In conclusion, ball milling was effective for preparing solid dispersions and surfactant incorporation improved drug solubility and stability.
A Novel Method for Measuring the Sizes and Concentrationskanomaxfmt
The document describes a new liquid nanoparticle sizing (LNS) system that can measure the sizes and concentrations of particles as small as 5 nm in colloidal suspensions. The system works by nebulizing the liquid suspension into an aerosol and then using scanning mobility particle sizing to analyze the particles. The LNS provides direct measurement of particle number concentrations and can detect small changes in particle size distributions. It has demonstrated accurate sizing for particles from 5-500 nm and measurement of filter retention for particles as small as 10 nm. The system allows detailed analysis of polishing slurries and effects of handling on particle size distributions.
Sensitivity and Reproducibility of Protein Aggregate Analysis by Sedimentatio...KBI Biopharma
Sedimentation velocity provides highly sensitive and reproducible analysis of protein aggregates. Replicate samples show the standard deviation for the main peak is typically 0.3% or less, and individual aggregate peaks can be detected below 0.1% and reproducibly quantitated. While data analysis resembles chromatography, the nature of noise is different, so minor peak positions may shift and peaks near detection thresholds may appear or disappear between replicates. Systematic errors from detergents, excipients, UV exposure, or sample loss can also impact reproducibility and limit sensitivity.
The document discusses how operating conditions like pump flow rate and templating agents affect the surface area of highly porous food powders produced via spray drying. It finds that glucose at a high pump flow rate has the greatest change in surface area over temperature changes, while sucrose at a low pump flow rate has the lowest change. The choice of templating agent also impacts properties like crystallinity, glass transition temperature, and surface area due to different self-assembly mechanisms. Characterization of the powders involved analyzing yield, moisture content, sorption behavior, and BET surface area to understand these effects of production conditions.
UV spectrophotometric method development and validation for quantitative esti...Sagar Savale
UV Spectrophotometric Method Development and Validation for quantitative estimation of Ondansetron
Hydrochloride (HCL). U.V Spectrophotometric method have been widely employed in determination of
individual components in a mixture or fixed dose combination. Our aim is to develop spectroscopic method for
estimation of the Ondansetron HCL in ternary mixture by using U.V spectrophotometry. The method was
validated as per ICH guidelines. The recovery studies confirmed the accuracy and precision of the method. It was
successfully applied for the analysis of the drug in bulk and could be effectively used for the routine analysis.
Introduction to Powder Size Analysis for Cell Culture MediaKurt Sundgren
This technical bulletin discusses particle size analysis in dry powder cell culture media production. It provides an overview of SAFC Biosciences' particle size analysis capabilities using sieving and laser diffraction, which measure particle size distributions from 10 nm to 3000 μm. It also discusses how particle size variability can affect material properties like flowability, mixing, and solubility. Statistical analysis of particle size distributions is important to fully characterize samples and communicate results. Understanding these relationships helps ensure a consistent, high-quality product.
Analytical method Development and Validation for the estimation of Pioglitazo...SriramNagarajan15
This paper describes the analytical method suitable for validation of Pioglitazone hydrochloride by UV Spectrophotometric method. The method utilized UV spectroscopy and the solvent system was consists of 6 N Glacial acetic acid at wave length 270 nm. Validation experiments were performed to demonstrate Specificity, Precision, Linearity, Accuracy, ruggedness. The method was linear over the concentration range of 10-50 µg/ml. The Proposed method was simple, sensitive & reliable with good Precise, Accurate, and Reproducible and rapid for the determination of Pioglitazone. The commercial formulations are estimated without interference. Hence this method can be used for routine determination of Pioglitazone hydrochloride in bulk and their pharmaceutical dosage forms.
This document provides an overview of sample preparation techniques for X-ray fluorescence (XRF) analysis. It discusses preparation of metal, powder, and liquid samples. For metals, the sample surface must be ground flat to remove impurities and obtain consistent roughness. Powder samples are pressed into pellets after pulverization to reduce heterogeneity effects. Liquid samples can be analyzed directly in sample cells or by drying microdroplets on filter paper to measure lighter elements. Proper sample preparation is crucial for obtaining accurate and reproducible XRF analysis results.
This document describes procedures for isolating and characterizing chloroplasts and photosynthetic pigments from plant tissues. Cellular components are isolated using differential centrifugation after homogenization. Chloroplasts are isolated from spinach leaves and their chlorophyll content is estimated spectrophotometrically. Photosynthetic pigments are separated from crude extracts via paper chromatography and identified based on their retention factor (Rf) values and absorbance spectra, which are compared to reference spectra. The document provides methods for isolating and analyzing subcellular structures and pigments from plant samples.
Module 4 isolation of chloroplasts and characterization of photosynthetic pi...Hara O.
This document describes procedures for isolating chloroplasts from plant leaves and characterizing the photosynthetic pigments contained within them. The key steps are:
1. Isolating intact chloroplasts from spinach or lettuce leaves through cell disruption and differential centrifugation.
2. Separating the photosynthetic pigments within the chloroplasts using paper chromatography and determining their Rf values.
3. Eluting the separated pigment bands and analyzing their absorbance spectra from 350-700nm using a spectrophotometer to identify the specific pigments.
The procedures allow for studying the structure and function of chloroplasts as well as characterizing their light-absorbing pigments like chlorophylls and
Micromeritics is the study of small particle characteristics like size, shape, density, and how they impact properties. Particle size and shape most affect flow properties - spherical particles between 75-250 microns flow freely while smaller or nonspherical particles can cause issues. Other factors like density, porosity, surface roughness and packing arrangement also influence flow. Common techniques to determine particle size include microscopy, sieving, and sedimentation methods. Proper characterization of particle properties is important for developing effective dosage forms and ensuring therapeutic effectiveness.
Reversible Association of a Monoclonal Antibody Studied by Concentration-Grad...KBI Biopharma
1) Initial sedimentation velocity experiments and size exclusion chromatography of monoclonal antibody mAb-X suggested it exhibits reversible self-association below a concentration of 1 mg/mL.
2) Concentration-gradient multi-angle light scattering experiments on mAb-X revealed an increase in apparent molar mass with concentration, indicating the presence of oligomers larger than dimers near 1.5 mg/mL.
3) Fitting the light scattering data to a monomer-dimer-tetramer association model returned dissociation constants of 15 μM for dimers and 3.5 μM for tetramers, providing evidence of a specific reversible oligomerization pathway for mAb-X.
The document discusses gel permeation chromatography (GPC), which separates molecules by size as they pass through a column of porous beads. It describes the basic components and working principles of GPC, including how larger molecules have shorter residence times in the column than smaller molecules. Applications include determining relative molecular weight and molecular weight distribution of polymer samples, and separating substances like sugars, polypeptides, proteins, and polymers.
This presentation discusses micromeritics, which involves the study of small particles around a few micrometers in size. It summarizes several key methods for analyzing particle size, shape, and distribution, including optical microscopy, sieving, sedimentation, and conductivity. It also covers techniques for measuring surface area, such as adsorption and air permeability methods. Derived powder properties like density, bulk density, tapped density, porosity and their importance are also highlighted.
Micromeritics is the study of the fundamental properties of small particles. Particle size and distribution impact many physical properties of pharmaceuticals like bulk density, flow properties, dissolution rate, chemical reactivity, and drug release characteristics. Key properties of particle collections include size, shape, volume, number, and surface area. Particle size can be determined through microscopic, sieving, and sedimentation techniques. Flow properties are influenced by particle size, shape, surface forces, and moisture content and can be improved by altering these characteristics.
UV Spectrophotometric Method Development and Validation for Quantitative Esti...Sagar Savale
UV Spectrophotometric Method Development and Validation for quantitative estimation of Miconazole nitrate
(MIC). U.V Spectrophotometric method have been widely employed in determination of individual components in
a mixture or fixed dose combination. Our aim is to develop spectroscopic method for estimation of the Miconazole
nitrate (MIC) in ternary mixture by using U.V spectrophotometry. The method was validated as per ICH
guidelines. The recovery studies confirmed the accuracy and precision of the method. It was successfully applied
for the analysis of the drug in bulk and could be effectively used for the routine analysis.
Lectut btn-202-ppt-l16. recovery of dna from agarose gelRishabh Jain
This document describes several methods for extracting DNA that has been separated via agarose gel electrophoresis. The most common methods include organic extraction using phenol/chloroform, which involves melting the agarose slice and extracting the DNA into an aqueous phase. Electroelution places gel slices in dialysis bags through which an electric current is passed to elute DNA. Enzymatic digestion uses the enzyme agarase to break down the agarose, releasing DNA. A freeze-squeeze method involves freezing and thawing gel slices in buffer to extract DNA.
UV Spectrophotometric Method Development and Validation for Quantitative Esti...Sagar Savale
Aim: UV Spectrophotometric Method Development and Validation for quantitative estimation of
Diclofenac Sodium. Objective: U.V Spectrophotometric method have been widely employed for
determination of analyte in a mixture. Our aim is to develop spectroscopic method for estimation of the
diclofenac sodium in ternary mixture by using U.V spectrophotometry. Methodology: The method was
validated as per ICH guidelines. The recovery studies confirmed the accuracy and precision of the method.
Conclusion: It was successfully applied for the analysis of the drug in bulk and could be effectively used for
the routine analysis.
Physical pharmacy and drug manufacturer guestionsMINANI Theobald
The document contains questions and answers related to physical pharmacy concepts. It discusses:
1) Formulas for arithmetic and geometric equivalent diameter, and methods to determine particle size like sieving, sedimentation, microscopic analysis, and laser diffraction.
2) Using the Edmundson equation to calculate dvs and dln for particle size distribution.
3) Calculating the number of particles in a given mass by finding the volume and density of individual particles.
4) The importance of surface area in drug manufacture for properties like absorption.
5) Distinguishing between bulk and true density, and the meaning of granular porosity.
The MEA 2 Personal Purification System for enhanced automated protein purific...Anne Burke
The document introduces the MEA 2 Personal Purification System, which utilizes high-throughput robotics and PhyTip columns for automated sample preparation and purification. Key features of the PhyTip columns include flexible resin bed volumes, back-and-forth flow for increased throughput, and placement of resin and screens for low dead volume and high concentration elution. The MEA 2 system is equipped for automated processing of PhyTip columns, accommodating tips, reservoirs, and plates with a simple interface compared to FPLC systems. It allows for methods development, screening, scale-up, and various downstream applications such as crystallization and glycan analysis.
UV Spectrophotometric Method Development And Validation For Quantitative Esti...Sagar Savale
U.V Spectrophotometric method have been widely employed in determination of Halcinonide in a mixture or fixed dose combination. For the ternary mixture containing Halcinonide, no spectrophotometric method for evaluation has been reported so far. Thus our aim is to develop method for Halcinonide estimation in ternary mixture using U.V spectrophotometry.
Method development and validation for the estimation of metronidazole in tabl...pharmaindexing
This document describes the development and validation of two spectrophotometric methods for the estimation of metronidazole in tablet dosage forms. The methods utilize UV spectroscopy and first derivative spectroscopy. Metronidazole showed maximum absorbance at 313nm in methanol:water for UV spectroscopy and a minimum at 298nm for derivative spectroscopy. Both methods were linear between 4-12μg/ml and were validated according to ICH guidelines. The methods were found to be accurate, precise and reproducible for the analysis of metronidazole in pure form and pharmaceutical formulations.
Este documento apresenta o código de conduta ética da Universidade do Minho. Estabelece valores como a transparência, integridade e respeito pela dignidade humana. Define deveres para a comunidade académica como promover o interesse público, respeitar os outros e os bens da universidade. Também descreve situações de conduta imprópria e as possíveis consequências.
Introduction to Powder Size Analysis for Cell Culture MediaKurt Sundgren
This technical bulletin discusses particle size analysis in dry powder cell culture media production. It provides an overview of SAFC Biosciences' particle size analysis capabilities using sieving and laser diffraction, which measure particle size distributions from 10 nm to 3000 μm. It also discusses how particle size variability can affect material properties like flowability, mixing, and solubility. Statistical analysis of particle size distributions is important to fully characterize samples and communicate results. Understanding these relationships helps ensure a consistent, high-quality product.
Analytical method Development and Validation for the estimation of Pioglitazo...SriramNagarajan15
This paper describes the analytical method suitable for validation of Pioglitazone hydrochloride by UV Spectrophotometric method. The method utilized UV spectroscopy and the solvent system was consists of 6 N Glacial acetic acid at wave length 270 nm. Validation experiments were performed to demonstrate Specificity, Precision, Linearity, Accuracy, ruggedness. The method was linear over the concentration range of 10-50 µg/ml. The Proposed method was simple, sensitive & reliable with good Precise, Accurate, and Reproducible and rapid for the determination of Pioglitazone. The commercial formulations are estimated without interference. Hence this method can be used for routine determination of Pioglitazone hydrochloride in bulk and their pharmaceutical dosage forms.
This document provides an overview of sample preparation techniques for X-ray fluorescence (XRF) analysis. It discusses preparation of metal, powder, and liquid samples. For metals, the sample surface must be ground flat to remove impurities and obtain consistent roughness. Powder samples are pressed into pellets after pulverization to reduce heterogeneity effects. Liquid samples can be analyzed directly in sample cells or by drying microdroplets on filter paper to measure lighter elements. Proper sample preparation is crucial for obtaining accurate and reproducible XRF analysis results.
This document describes procedures for isolating and characterizing chloroplasts and photosynthetic pigments from plant tissues. Cellular components are isolated using differential centrifugation after homogenization. Chloroplasts are isolated from spinach leaves and their chlorophyll content is estimated spectrophotometrically. Photosynthetic pigments are separated from crude extracts via paper chromatography and identified based on their retention factor (Rf) values and absorbance spectra, which are compared to reference spectra. The document provides methods for isolating and analyzing subcellular structures and pigments from plant samples.
Module 4 isolation of chloroplasts and characterization of photosynthetic pi...Hara O.
This document describes procedures for isolating chloroplasts from plant leaves and characterizing the photosynthetic pigments contained within them. The key steps are:
1. Isolating intact chloroplasts from spinach or lettuce leaves through cell disruption and differential centrifugation.
2. Separating the photosynthetic pigments within the chloroplasts using paper chromatography and determining their Rf values.
3. Eluting the separated pigment bands and analyzing their absorbance spectra from 350-700nm using a spectrophotometer to identify the specific pigments.
The procedures allow for studying the structure and function of chloroplasts as well as characterizing their light-absorbing pigments like chlorophylls and
Micromeritics is the study of small particle characteristics like size, shape, density, and how they impact properties. Particle size and shape most affect flow properties - spherical particles between 75-250 microns flow freely while smaller or nonspherical particles can cause issues. Other factors like density, porosity, surface roughness and packing arrangement also influence flow. Common techniques to determine particle size include microscopy, sieving, and sedimentation methods. Proper characterization of particle properties is important for developing effective dosage forms and ensuring therapeutic effectiveness.
Reversible Association of a Monoclonal Antibody Studied by Concentration-Grad...KBI Biopharma
1) Initial sedimentation velocity experiments and size exclusion chromatography of monoclonal antibody mAb-X suggested it exhibits reversible self-association below a concentration of 1 mg/mL.
2) Concentration-gradient multi-angle light scattering experiments on mAb-X revealed an increase in apparent molar mass with concentration, indicating the presence of oligomers larger than dimers near 1.5 mg/mL.
3) Fitting the light scattering data to a monomer-dimer-tetramer association model returned dissociation constants of 15 μM for dimers and 3.5 μM for tetramers, providing evidence of a specific reversible oligomerization pathway for mAb-X.
The document discusses gel permeation chromatography (GPC), which separates molecules by size as they pass through a column of porous beads. It describes the basic components and working principles of GPC, including how larger molecules have shorter residence times in the column than smaller molecules. Applications include determining relative molecular weight and molecular weight distribution of polymer samples, and separating substances like sugars, polypeptides, proteins, and polymers.
This presentation discusses micromeritics, which involves the study of small particles around a few micrometers in size. It summarizes several key methods for analyzing particle size, shape, and distribution, including optical microscopy, sieving, sedimentation, and conductivity. It also covers techniques for measuring surface area, such as adsorption and air permeability methods. Derived powder properties like density, bulk density, tapped density, porosity and their importance are also highlighted.
Micromeritics is the study of the fundamental properties of small particles. Particle size and distribution impact many physical properties of pharmaceuticals like bulk density, flow properties, dissolution rate, chemical reactivity, and drug release characteristics. Key properties of particle collections include size, shape, volume, number, and surface area. Particle size can be determined through microscopic, sieving, and sedimentation techniques. Flow properties are influenced by particle size, shape, surface forces, and moisture content and can be improved by altering these characteristics.
UV Spectrophotometric Method Development and Validation for Quantitative Esti...Sagar Savale
UV Spectrophotometric Method Development and Validation for quantitative estimation of Miconazole nitrate
(MIC). U.V Spectrophotometric method have been widely employed in determination of individual components in
a mixture or fixed dose combination. Our aim is to develop spectroscopic method for estimation of the Miconazole
nitrate (MIC) in ternary mixture by using U.V spectrophotometry. The method was validated as per ICH
guidelines. The recovery studies confirmed the accuracy and precision of the method. It was successfully applied
for the analysis of the drug in bulk and could be effectively used for the routine analysis.
Lectut btn-202-ppt-l16. recovery of dna from agarose gelRishabh Jain
This document describes several methods for extracting DNA that has been separated via agarose gel electrophoresis. The most common methods include organic extraction using phenol/chloroform, which involves melting the agarose slice and extracting the DNA into an aqueous phase. Electroelution places gel slices in dialysis bags through which an electric current is passed to elute DNA. Enzymatic digestion uses the enzyme agarase to break down the agarose, releasing DNA. A freeze-squeeze method involves freezing and thawing gel slices in buffer to extract DNA.
UV Spectrophotometric Method Development and Validation for Quantitative Esti...Sagar Savale
Aim: UV Spectrophotometric Method Development and Validation for quantitative estimation of
Diclofenac Sodium. Objective: U.V Spectrophotometric method have been widely employed for
determination of analyte in a mixture. Our aim is to develop spectroscopic method for estimation of the
diclofenac sodium in ternary mixture by using U.V spectrophotometry. Methodology: The method was
validated as per ICH guidelines. The recovery studies confirmed the accuracy and precision of the method.
Conclusion: It was successfully applied for the analysis of the drug in bulk and could be effectively used for
the routine analysis.
Physical pharmacy and drug manufacturer guestionsMINANI Theobald
The document contains questions and answers related to physical pharmacy concepts. It discusses:
1) Formulas for arithmetic and geometric equivalent diameter, and methods to determine particle size like sieving, sedimentation, microscopic analysis, and laser diffraction.
2) Using the Edmundson equation to calculate dvs and dln for particle size distribution.
3) Calculating the number of particles in a given mass by finding the volume and density of individual particles.
4) The importance of surface area in drug manufacture for properties like absorption.
5) Distinguishing between bulk and true density, and the meaning of granular porosity.
The MEA 2 Personal Purification System for enhanced automated protein purific...Anne Burke
The document introduces the MEA 2 Personal Purification System, which utilizes high-throughput robotics and PhyTip columns for automated sample preparation and purification. Key features of the PhyTip columns include flexible resin bed volumes, back-and-forth flow for increased throughput, and placement of resin and screens for low dead volume and high concentration elution. The MEA 2 system is equipped for automated processing of PhyTip columns, accommodating tips, reservoirs, and plates with a simple interface compared to FPLC systems. It allows for methods development, screening, scale-up, and various downstream applications such as crystallization and glycan analysis.
UV Spectrophotometric Method Development And Validation For Quantitative Esti...Sagar Savale
U.V Spectrophotometric method have been widely employed in determination of Halcinonide in a mixture or fixed dose combination. For the ternary mixture containing Halcinonide, no spectrophotometric method for evaluation has been reported so far. Thus our aim is to develop method for Halcinonide estimation in ternary mixture using U.V spectrophotometry.
Method development and validation for the estimation of metronidazole in tabl...pharmaindexing
This document describes the development and validation of two spectrophotometric methods for the estimation of metronidazole in tablet dosage forms. The methods utilize UV spectroscopy and first derivative spectroscopy. Metronidazole showed maximum absorbance at 313nm in methanol:water for UV spectroscopy and a minimum at 298nm for derivative spectroscopy. Both methods were linear between 4-12μg/ml and were validated according to ICH guidelines. The methods were found to be accurate, precise and reproducible for the analysis of metronidazole in pure form and pharmaceutical formulations.
Este documento apresenta o código de conduta ética da Universidade do Minho. Estabelece valores como a transparência, integridade e respeito pela dignidade humana. Define deveres para a comunidade académica como promover o interesse público, respeitar os outros e os bens da universidade. Também descreve situações de conduta imprópria e as possíveis consequências.
The main product is a local newspaper that uses appropriate color schemes, fonts, and layout conventions. The ancillary texts are a poster and radio advertisement that feature the same slogan ("THE BEST NEWS AT THE RIGHT PRICE") and color scheme as the newspaper to maintain consistency across products. By keeping the logo, slogan, and colors the same, the combination of products is effective at making the brand more recognizable and familiar to people.
This document is a transcript of a podcast discussion on the barriers that prevent women from achieving leadership positions and gender equality. It explores economic, social, and regulatory obstacles such as laws limiting women's work opportunities, gender pay gaps, lack of childcare, and discrimination. Participants from various countries and organizations discuss how poverty, unemployment, social conditioning, and lack of female role models hold women back. Improving access to education, empowering girls, engaging men, and establishing strong legal frameworks for gender equality through the UN Sustainable Development Goals are proposed as ways to advance more women into power and break through the glass ceiling.
This presentation is for the UK Association of Directors of Public Health policy workshop 2016 and looks at how Public Health can support and lead health approaches to Housing strategy and delivery. It takes a number of examples and case studies and identifies 7 key policy and strategy principles
Le montant des dons à des organisations caritatives effectués par des foyers assujettis à l'ISF a baissé de 9% par rapport à 2015, selon une étude de la fondation des Apprentis d'Auteuil. Cette tendance risque de durer."
The document discusses several new trends in computing including cloud computing, surface computing, cyborg computing, voice recognition, and Google Glass. Cloud computing allows data to be stored virtually online and accessed from anywhere. Surface computing uses touchscreens without physical input devices. Cyborg computing involves users controlling computers through their own body parts. Voice recognition and Google Glass enable interaction through sound and voice commands.
Sucrose low in nanoparticulate impurities for biopharmaceutical formulationsMilliporeSigma
1) The document describes a study to develop a purification process to reduce nanoparticle impurities (NPIs) in sucrose and analyze differences between NPIs from beet- and cane-derived sucrose.
2) Methods like DLS, NTA, and zeta potential analysis show that beet-derived NPIs have a more negative charge and greater impact on protein stability than cane-derived NPIs.
3) A purification method was successfully developed to reduce NPIs in sucrose and launch Sucrose Emprove® Expert, shown to significantly decrease particle concentration compared to non-purified raw materials.
Final submission –Pay attention to APA formatting, spelling, andChereCheek752
Final submission –
Pay attention to APA formatting, spelling, and grammar. Your similarity index/plagiarism score must be below 10%. Higher scores may impact your grade.
The final submission is the combination of the other four phases into one paper. You will combine Phase I, Phase II, Phase III, and Phase IV to make Phase V. You are responsible for editing and formatting your paper so that your paper will flow for the reader. This paper will need to be corrected with all the feedback provided from previous papers. Include conclusion and learning experiences from the essentials and from the class. Do not forget to document limitations and implications for future research/practice. Please review the PowerPoint prior to submitting your assignment, thank you.
Amino Acids and Proteins
Structure of -amino acids
The 20 Amino Acids Found in Proteins
Formation of a Peptide
Polypeptide backbone
9.bin
10.bin
Proteins are made of 20 amino acids linked by peptide bonds
Polypeptide backbone is the repeating sequence of the N-C-C-N-C-C… in the peptide bond
The side chain or R group is not part of the backbone or the peptide bond
ProteinsMake up about 15% of the cellHave many functions in the cellEnzymesStructuralTransportMotorStorageSignalingReceptorsGene regulationSpecial functions
Motor- myosin
Storage- ferritin, transport- hemoglobyn
*
Importance of ProteinsMain catalysts in biochemistry: enzymes (involved in virtually every biochemical reaction)Structural components of cells (both inside and outside of cells in tissues)Regulatory functions (if/when a cell divides, which genes are expressed, etc.)Carrier and transport functions (ions, small molecules)
Levels of Protein StructurePrimary Structure - amino acid sequence in a polypeptide
Secondary Structure - local spatial arrangement of a polypeptide’s backbone atoms (without regard to
side chain conformation)
Tertiary Structure - three-dimensional structure of entire polypeptide
Quaternary Structure - spatial arrangement of subunits of proteins composed of multiple polypeptides (protein complexes)
3-D Structure of Myoglobin
People with proteinuria have urine containing an abnormal amount of protein. The condition is often a sign of kidney disease.
Healthy kidneys do not allow a significant amount of protein to pass through their filters. Kidney disease often has no early symptoms. One of its first signs may be proteinuria that's discovered by a urine test done during a routine physical exam. Blood tests will then be done to see how well the kidneys are working.
Both diabetes and high blood pressure can cause damage to the kidneys, which leads to proteinuria.
Proteinuria (Protein in Urine)
Proteinuria (Protein in Urine)
Methods of Protein Estimation
Quantitative
Biruet methodBradford methodFolin-Lowry methodKjeldahl methodBicinchoninic acid method (BCA method)UV methodFlourimetric methodMass spectrometry
Protein Determination assay
Bicinch ...
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Similar to AppNote_MFI Manual Viscosity Study (20)
Practical 3 Quantitative determination of protein concentration using spectro...
AppNote_MFI Manual Viscosity Study
1. application note
Our Tips for Viscous Sample Introduction
Following these simple guidelines will get you the best
data for viscous samples:
Flushing — The most important thing when it comes
to viscous samples is to equilibrate your flow cell.
Not doing it properly can lead to the formation of
Schlieren lines which can cause issues with particle
count and morphology. Flush the flow cell either with
the sample itself or a buffer with similar optical and
physical properties until no Schlieren lines (Figure 1)
appear along the edge of the flow cell. The volume
needed mainly depends on your solution’s viscosity,
and for very viscous samples you’ll need higher flush
volumes to equilibrate. We chose 5 mL of flush volume
for our experiments here because it worked for all our
conditions. But you can often go lower for less viscous
samples.
Mixing — Mix your samples really well. A homogenous
sample is crucial for accurate sample characterization.
Easy Particle Analysis for Viscous Samples with MFI
Introduction
Many protein therapeutics in development today are viscous because
they contain excipients for stability or have a high concentration
of the protein itself. If you use particle analysis techniques like light
obscuration, the only way to analyze these samples accurately is to
dilute them. But dilution can change the characteristics of the sample,
which means more comparability studies to validate the dilution
effect. Why make your work harder? MFI lets you skip the whole
dilution process.
We studied the MFI 5200 manual system performance across a broad
range of sample viscosities and you guessed it — we got precise
and quantitative characterization of particle size, concentration, and
morphology with even the most viscous solutions. And while the data
in this application note is from the MFI 5200 system, we also looked at
the MFI 5100 system and saw the same great results: robust particle
measurement using neat samples. So if you want out of the dilution
boat, read on!
Avoid air bubbles — Air bubbles are more easily trapped
in viscous solutions than in aqueous ones. You can avoid
introducing them by gently mixing your samples using
end-over-end rotation. Avoid the forceful agitation route.
Pump Calibration — For sample viscosities up to 20 cP,
standard water calibration is fine, so you don’t have to
worry about recalibrating your pump.
Morphology — In our study, viscosity didn’t have a
strong effect on the data. But, if the viscosity of your
sample comes from a matrix with a significantly different
refractive index, you may see stronger effects.1
If that’s the
case, your best bet is to characterize NIST sizing beads
in your matrix to see if the sample’s refractive index is
impacted by solution viscosity.
2. application note
2
Easy Particle Analysis for Viscous Samples with MFI
For our study, we compared particle characterization
in viscous solutions to water on the MFI 5200 system.
The parameters we evaluated included sample volume
analyzed, particle count and particle size. We also looked
at important morphological parameters to confirm our
measurements weren’t affected by sample viscosity.
How We Analyzed Samples
For particle count and size, the analysis workflow
(Figure 2) and the Analysis Method (Figure 3) used
were the same. The only difference in the Analysis
Method was the application of Edge Particle Rejection.
For the counting analysis Edge Particle Rejection was
turned off, and for the sizing analysis it was turned on.
This allowed us to more accurately determine size and
count. All samples were introduced to the MFI 5200
system using a syringe barrel and analyzed in triplicate.
SAMPLE VOLUME ANALYSIS
We made samples with viscosities ranging from
1 centipoise (cP) to 20 cP using a series of polyethylene-
glycol (PEG) dilutions, then confirmed solution viscosity
using a Rheosense Inc. uVisc. Syringe Viscometer.
Viscous samples can be less mobile in the fluid path, so
we wanted to find out if this was a potential problem
area for the analysis volume. Before each sample was
analyzed, the MFI 5200 system was primed with the
appropriate PEG solution, and the method was set to
analyze 900 µL with 220 µL for optimize illumination and
no flush. We then recovered the analyzed volume from
the waste line with a pre-weighed Eppendorf tube. As
shown in Figure 4, the analyzed sample volumes were
consistent, regardless of sample viscosity. Because it
uses a slow analysis speed, the MFI flow cell precisely
analyzed the same amount of sample no matter if it
Figure 1. Schlieren line in priming view. Particles coincident with the Schlieren line are interpreted
as a single massive particle by the software.
Figure 2. Workflow used to analyze viscous samples.
Schlieren line
Remove syringe
barrel from
instrument and
wash with 20 mL of
particle-free water.
Reattach syringe
barrel and wash
flow cell with 10 mL
particle-free water.
Begin sample
analysis.
Flush 5 mL of
sample through
flow cell to get rid
of Schlieren lines.
Carefully load
sample into syringe
barrel. Take care to
minimize bubbles.
3. application note
3
Easy Particle Analysis for Viscous Samples with MFI
was an aqueous solution or a viscous (PEG) one. This
also means there isn’t any need to recalibrate the pump,
which is time saved during setup. How nice is that?
We all know the MFI 5200 system accurately and
precisely characterizes particles in the sub-visible range
(2 – 25 microns). So the next thing we looked at is how well it
stacked up with counting particles in viscous samples.
Same Great Data for Particle Size and
Concentration
For our concentration evaluation, NIST-certified 5 µm,
10 µm and 25 µm COUNT-CAL standards (ProteinSimple)
were mixed at a 1:1 ratio with the PEG solutions so the
final viscosities of the solutions would be at the intended
cP range. We confirmed viscosity with the viscometer,
Figure 3. Method used for sample analysis. The Total Available Volume was set to 0.90 mL, which translates to
an analyzed volume of 0.77 mL.
Figure 4. Sample analysis volumes are consistent across a range of viscosities. For all tested
samples, values for the analysis were within 5% of the nominal 900 µL, showing that even high
viscosity samples can be precisely measured through the flow cell. This precision over a wide
range of viscosities is key to accurately determining particle concentration.
10
800
820
840
860
880
900
920
940
960
980
1000
20 15 5 Water
VolumeRecovered(µL)
Viscosity (cP)
Analysis Volume at Different Viscosities
4. application note
4
Easy Particle Analysis for Viscous Samples with MFI
The Duke Size Standard analysis also showed viscosity
had no effect on data quality. Bead standards at all
viscosities were measured to within 5% of their expected
values (Figure 6), confirming that viscosity doesn’t impact
particle size measurements. So that means you’ll get
accurate sizing information with samples up to 20 cP. And,
replicates for each sample were super tight— variability
in the ECD measurements were <1% for the sample in
triplicate (Figure 6).
So What About Morphology?
Because we know it’ll help you identify the morphology
of your particles, we also analyzed common parameters
on top of the size and count metrics the standards are
designed to represent (Figure 7, 8, 9, 10). While the
data didn’t show strong differences in characterization
and then mixed samples by rotating end over end for
20 minutes before analysis.
Data for the COUNT-CAL bead analysis showed that you
can accurately determine particle concentration even
with samples as viscous as 20 cP (Figure 5). We analyzed
data with MVSS 4.0 using standard NIST filters (Table 1).
COUNT-CAL beads have a published concentration of
3000 particles per mL at their stock concentration, with
a variability of +/- 10%. After we diluted the bead stocks
1:1 with a PEG solution, the nominal concentration was
1500 particles per mL. Each condition was measured in
triplicate. The recovered concentrations for samples fell
within 90 – 110% of the expected 1500 particles/mL, and
CVs for the triplicate measurements were below 6%, right
within spec.
Figure 5. Recovered concentrations for samples over a range of
viscosities. Error bars represent standard deviations.
BEAD SIZE PARAMETER FILTER
5 µm ECD >3.0 µm
10 µm ECD >7.5 µm
25 µm ECD >15 µm
5 µm
10 µm
25 µm
Concentration(Particles/mL)
0
200
400
600
800
1000
1200
1400
1600
1800
2000
1 5 10 15 20
Concentration of COUNT-CAL Beads in Viscous Matrix
Viscosity (cP)
1 5 10 15 20
%ofNominalDiameter
Viscosity (cP)
Duke Sizing Standards Analyzed in Viscous Matrix
2 µm
5 µm
10 µm
25 µm
85
90
95
100
105
110
Figure 6. Size measurements from 1 – 20 cP. Bead standards were all
within 5% of the expected values. CVs for the triplicate measurements
were <1%.
Table 1. Standard NIST filters used to categorize
results.
5. application note
5
Easy Particle Analysis for Viscous Samples with MFI
results, statistical analysis did reveal subtle effects that the
sample matrix had on particle characterization. We chose
the 2 µm bead set for linear regression analysis to get a
read on any significant differences between the viscosity
conditions. And it turns out that morphologic measures
Figure 7. Measured particle intensity max. The maximum intensity
of the all pixels representing the particle.
Figure 8. Measured circularity. The circumference of an equivalent
area circle divided by the actual perimeter of the particle.
aren’t adversely impacted, so no significant changes
to intensity max, circularity, and aspect ratio (Table 2,
3, 4). We noticed only a very small change in pixel area
(Table 5).
0
100
200
300
400
500
600
700
800
900
1 5 10 15 20
AverageIntensityMax
Viscosity (cP)
Particle Intensity Max
2 µm
5 µm
10 µm
25 µm
2 µm
5 µm
10 µm
25 µm
1 5 10 15 20
Viscosity (cP)
0.91
0.90
0.89
0.88
0.87
0.86
0.85
0.84Circularity
Circularity
CIRCULARITY OF 2 µm BEADS COMPARED TO 1 cP CONDITION
VISCOSITY COEFFICIENTS STANDARD ERROR P-VALUE
Intercept 0.86 0.0003 2.35E-132
5 cP -0.0003 0.0003 0.27
10 cP -0.0003 0.0003 0.40
15 cP -0.0006 0.0003 0.06
20 cP -0.0003 0.0003 0.27
INTENSITY MAX OF 2µm BEADS COMPARED TO 1 cP CONDITION
VISCOSITY COEFFICIENTS STANDARD ERROR P-VALUE
Intercept 778.10 0.57 4.32E-116
5 cP -0.03 0.57 0.96
10 cP -0.01 0.57 0.99
15 cP 0.25 0.57 0.66
20 cP 0.69 0.57 0.23
Table 2. Particle intensity statistical analysis of 2 µm beads. Linear
regression confirmed there was no change in particle intensity max
that correlated with viscosity (Model R2
=0.999).
Table 3. Circularity statistical analysis of 2 µm beads. Linear
regression gave no significant difference in circularity for any of our
viscous conditions (Model R2
=0.998).
6. application note
6
Easy Particle Analysis for Viscous Samples with MFI
PIXEL AREA OF 2 µm BEADS COMPARED TO 1 cP CONDITION
VISCOSITY COEFFICIENTS STANDARD ERROR P-VALUE
Intercept 24.28 0.27 2.39E-57
5 cP 0.64 0.27 1.89E-02
10 cP 1.07 0.27 1.83E-04
15 cP 1.22 0.27 2.89E-05
20 cP 0.94 0.27 9.12E-04
But it gets even better! There were no adverse impacts
on morphologic parameters, and no significant changes
to aspect ratio, intensity max, or circularity — and only
a change of one pixel in particle area. Translation? In
a nutshell, you can run your viscous samples straight
up with MFI and still get precise and accurate particle
characterization.
Conclusion
MFI generates absolutely great data, even when sample
viscosity is as high as 20 cP. Now you can get the hands-
down most consistent measurement of count, size and
morphology without having to dilute a single sample!
Our analysis of NIST-certified COUNT-CAL concentration
standards from 5 to 25 microns in size proved we could
accurately determine particle concentrations to within
10% of the nominal value in viscosities ranging from
1 to 20 cP. And sizing data for NIST-certified Duke size
standards from 2 to 25 microns was within 5% of the
nominal value.
Table 4. Aspect ratio statistical analysis of 2 µm beads. Linear
regression showed a very small change in aspect ratio in viscous
conditions. This difference, though consistent, is less than a 1%
change in aspect ratio (Model R2
=0.997).
ASPECT RATIO OF 2 µm BEADS COMPARED TO 1 cP CONDITION
VISCOSITY COEFFICIENTS STANDARD ERROR P-VALUE
Intercept 0.8989 0.00036 3.93E-129
5 cP -0.0012 0.00036 2.07E-03
10 cP -0.0010 0.00036 7.53E-03
15 cP -0.0017 0.00036 2.52E-05
20 cP -0.0014 0.00036 2.48E-04
Table 5. Pixel area statistical analysis of 2 µm beads. Linear
regression showed a small but consistent increase in the pixel area
under viscous conditions. But we know from the calculated values
the change wasn’t sufficient enough to impact particle sizing
(Model R2
=0.999).
Figure 9. Measured aspect ratio. The ratio of the minor axis length
of the major axis length of an ellipse that has the same second
moments of the particle.
Figure 10. Measured pixel area. The number of pixels representing a
particle.
2 µm
5 µm
10 µm
25 µm
1 5 10 15 20
Viscosity (cP)
0.96
0.95
0.94
0.93
0.92
0.91
0.90
0.89
0.88
0.87
AspectRatio
Aspect Ratio
PixelArea
Pixel Area
450
400
350
300
250
200
150
100
50
0
1 5 10 15 20
Viscosity (cP)
2 µm
5 µm
10 µm
25 µm