This document discusses the isolation of lipoprotein(a) [Lp(a)] from human serum and characterization of its sialic acid concentration. Key points:
- Lp(a) was isolated from human serum through multiple centrifugation and chromatography steps.
- Like LDL, Lp(a) contains significant amounts of sialic acid, which can be completely digested by neuraminidase enzyme treatment for 24-48 hours.
- Treated Lp(a) samples showed negligible sialic acid and decreased electrophoretic mobility compared to untreated samples, indicating sialic acid affects mobility.
- The results suggest sialic acid plays a role in Lp(a) similar to LDL,
This document describes a study that developed a method to determine cabergoline and L-dopa levels in human plasma using liquid chromatography coupled with tandem mass spectrometry (LC-MS/MS). The researchers were able to directly analyze deproteinized plasma samples for both cabergoline and L-dopa. Calibration curves for each drug showed good linearity over specific concentration ranges. The method was then applied to analyze plasma samples from patients taking these drugs, with coefficients of variation ranging from 2.9-10.5% indicating good precision. The LC-MS/MS method proved suitable for routine therapeutic drug monitoring of cabergoline and L-dopa.
The document summarizes an experiment on acid-base titrations. Sodium hydroxide was standardized and used to titrate hydrochloric acid, phosphoric acid, and an unknown acid. Titrations of hydrochloric acid showed the equivalence point was pH 7. Phosphoric acid titration showed two equivalence points, determining its two pKa values. Titration of the unknown acid found its molecular weight to be 76.09 g/mol, identifying it as tartaric acid. A final potassium hydrogen phthalate titration found the experimental curve nearly identical to the theoretical curve.
The document provides information on solubility constants (Ksp) and equilibria involving complex ions and slightly soluble ionic compounds in aqueous solutions. It discusses solubility product principles, writing ion-product expressions, determining Ksp values from solubility data and vice versa, the effect of common ions on solubility, and how pH affects the solubility of salts containing anions of weak acids. Sample problems demonstrate calculating solubility, Ksp, and the impact of adding acids or salts containing common ions. Key concepts covered include solubility constants, solubility product expressions, and factors that influence the solubility of sparingly soluble salts.
The document describes several experiments related to protein analysis:
1. It provides procedures for preparing phosphate and acetate buffers, estimating protein concentration using Lowry's method, determining the absorption maximum of riboflavin, performing paper chromatography to separate amino acids, and titrating acetic acid to find its pKa value.
2. It also outlines protocols for purifying protein from a sample via ammonium sulfate precipitation and estimating the precipitated protein, separating amino acids from a mixture using ion exchange chromatography, and performing SDS-PAGE to separate proteins by molecular weight.
3. Reagents, materials, and step-by-step procedures are described for each experiment to analyze different properties of proteins and separate mixtures of
The document summarizes an experiment to identify an unknown weak acid through titration with a standardized sodium hydroxide solution and measurement of pH. Key results include:
- The unknown acid was titrated and a titration curve was generated, from which two pKa values were estimated: 3.73 and 4.79.
- Calculations based on the titration data determined the molecular weight of the unknown to be 172.782 g/mol.
- Based on the pKa values and molecular weight, the unknown acid was identified as phthalic acid.
This document discusses pKa (acid dissociation constant) and log P (partition coefficient).
It defines pKa as the negative base 10 logarithm of Ka, which is a quantitative measurement of acid strength. Common methods to determine pKa include potentiometric titration and spectrophotometric methods. Log P is the ratio of concentrations of a compound between octanol and water, and is a measure of lipophilicity. Common methods to determine log P include the shake flask method and HPLC. Both pKa and log P values are important parameters in drug design and development as they influence absorption, distribution, and other pharmacokinetic properties.
This document describes a study that developed a method to determine cabergoline and L-dopa levels in human plasma using liquid chromatography coupled with tandem mass spectrometry (LC-MS/MS). The researchers were able to directly analyze deproteinized plasma samples for both cabergoline and L-dopa. Calibration curves for each drug showed good linearity over specific concentration ranges. The method was then applied to analyze plasma samples from patients taking these drugs, with coefficients of variation ranging from 2.9-10.5% indicating good precision. The LC-MS/MS method proved suitable for routine therapeutic drug monitoring of cabergoline and L-dopa.
The document summarizes an experiment on acid-base titrations. Sodium hydroxide was standardized and used to titrate hydrochloric acid, phosphoric acid, and an unknown acid. Titrations of hydrochloric acid showed the equivalence point was pH 7. Phosphoric acid titration showed two equivalence points, determining its two pKa values. Titration of the unknown acid found its molecular weight to be 76.09 g/mol, identifying it as tartaric acid. A final potassium hydrogen phthalate titration found the experimental curve nearly identical to the theoretical curve.
The document provides information on solubility constants (Ksp) and equilibria involving complex ions and slightly soluble ionic compounds in aqueous solutions. It discusses solubility product principles, writing ion-product expressions, determining Ksp values from solubility data and vice versa, the effect of common ions on solubility, and how pH affects the solubility of salts containing anions of weak acids. Sample problems demonstrate calculating solubility, Ksp, and the impact of adding acids or salts containing common ions. Key concepts covered include solubility constants, solubility product expressions, and factors that influence the solubility of sparingly soluble salts.
The document describes several experiments related to protein analysis:
1. It provides procedures for preparing phosphate and acetate buffers, estimating protein concentration using Lowry's method, determining the absorption maximum of riboflavin, performing paper chromatography to separate amino acids, and titrating acetic acid to find its pKa value.
2. It also outlines protocols for purifying protein from a sample via ammonium sulfate precipitation and estimating the precipitated protein, separating amino acids from a mixture using ion exchange chromatography, and performing SDS-PAGE to separate proteins by molecular weight.
3. Reagents, materials, and step-by-step procedures are described for each experiment to analyze different properties of proteins and separate mixtures of
The document summarizes an experiment to identify an unknown weak acid through titration with a standardized sodium hydroxide solution and measurement of pH. Key results include:
- The unknown acid was titrated and a titration curve was generated, from which two pKa values were estimated: 3.73 and 4.79.
- Calculations based on the titration data determined the molecular weight of the unknown to be 172.782 g/mol.
- Based on the pKa values and molecular weight, the unknown acid was identified as phthalic acid.
This document discusses pKa (acid dissociation constant) and log P (partition coefficient).
It defines pKa as the negative base 10 logarithm of Ka, which is a quantitative measurement of acid strength. Common methods to determine pKa include potentiometric titration and spectrophotometric methods. Log P is the ratio of concentrations of a compound between octanol and water, and is a measure of lipophilicity. Common methods to determine log P include the shake flask method and HPLC. Both pKa and log P values are important parameters in drug design and development as they influence absorption, distribution, and other pharmacokinetic properties.
Sipma, 2004, Effect Of Carbon Monoxide, Hydrogen And Sulfate On Thermophilic ...roelmeulepas
This document summarizes a study on the conversion of carbon monoxide (CO) by two anaerobic sludge samples at 55°C. The study aimed to elucidate the conversion routes and determine the effect of substrate (CO) concentration and the presence of hydrogen gas. Inhibition experiments showed CO conversion occurred via a hydrogenogenic population producing hydrogen and carbon dioxide, with the products then used by methanogens, acetogens, or sulfate reducers depending on sludge source and inhibitors. Both sludges could produce hydrogen from CO, indicating potential for biological hydrogen production from synthesis gas containing CO. The paper mill sludge was also capable of sulfate reduction using hydrogen produced from high CO concentrations, showing CO-rich synthesis gas can efficiently
Abraham model correlations for ionic liquid solvents computational methodolog...Bihan Jiang
The document describes a computational methodology for updating existing Abraham model ion-specific equation coefficients using new experimental solubility and partition coefficient data for ionic liquid solvents. Specifically, it illustrates updating the coefficients for the trifluoroacetate anion based on 51 data points from three ionic liquid solvents containing that anion. The updated coefficients have significantly smaller standard errors and are able to better predict solubility and partition behavior in the three ionic liquids based on the increased data. The methodology allows coefficients to be refined as new data becomes available without needing to re-regress the entire Abraham model data set.
The document discusses solubility equilibria and solubility product constants (Ksp). It defines Ksp as the product of ion concentrations raised to their coefficients in the solubility reaction equation. It provides examples of using Ksp to calculate solubility from solubility data and vice versa. Additionally, it discusses factors that affect solubility such as common ion effect, pH, and complexation.
This document provides an overview of acid-base titration and volumetric analysis. It defines key terms like titration, indicator, equivalence point, and standardization. It describes different types of titrations including direct, indirect, and back titration. Acid-base concepts are explained based on Arrhenius, Bronsted-Lowry, and Lewis theories. The document also discusses the ionic product of water, common ion effect, classification of indicators, and theories of indicators including Ostwald and chromophore theories.
This document discusses pKa values, which refer to the acid dissociation constant of acids. It defines pKa and explains how pKa values can be used to understand and predict the behavior of acids and bases in solution. The document covers several key topics in pKa including the factors that influence pKa values, common types of acids like mono- and polyprotic acids, and the significance and applications of pKa in fields like chemistry, biology, and medicine.
Acid-base titration is a technique used to determine the concentration of an acid or base by neutralizing a solution of unknown concentration with a solution of known concentration. The reaction is monitored using an indicator that changes color at the endpoint of the titration. The document outlines the basic procedure which involves transferring a sample of the solution of unknown concentration into a flask, adding an indicator, and then titrating with the solution of known concentration until the endpoint is reached as indicated by the color change of the indicator. The volume added is used to calculate the concentration of the original unknown solution.
Computational and Experimental Studies of MTO Catalyzed Olefin HydrogenationKaram Idrees
The poster that I presented at the 253rd American Chemical Society National Meeting and Exposition in San Francisco,
CA. It highlights some of my REU research at North Carolina State University under the mentorship of Dr. Elon Ison.
Activity coefficients at infinite dilution for organic solutes dissolved in t...Bihan Jiang
This document discusses measuring activity coefficients and gas-to-liquid partition coefficients for 45 organic solutes dissolved in two 1,2,3-tris(diethylamino)cyclopenylium based room temperature ionic liquids. Abraham model correlations were derived to predict solute transfer into the ionic liquids from water and the gas phase. The correlations described the observed partition coefficients to within 0.13 log units or less. Measured activity coefficients were used to calculate selectivity values for potential separations like hexane/benzene and hexane/pyridine.
Learning objectives
Introduction
Preparation of a standard solution used for redox titration
Oxidizing and reducing agents used in volumetric analysis
N/10 potassium permanganate preparation
N/10 potassium dichromate preparation
N/10 Iodine solution preparation
Examples of redox titrations
Conclusion
References
This document provides an introduction to analytical methods used in pharmaceutical analysis. It discusses various classical analytical methods like titrimetric methods including acid-base titrations, precipitation titrations, complexometric titrations and instrumental methods. It also summarizes different types of analytical techniques classified as classical methods, separation methods, spectroscopic methods, electrochemical methods and thermal methods. Specific techniques discussed in detail include acid-base titrations, indicators used in titrations, and types of complexometric titrations. The document provides an overview of key concepts and methods in pharmaceutical analytical chemistry.
- Chemical equilibrium is reached when the rates of the forward and reverse reactions of a reversible reaction are equal. At equilibrium, the concentrations of reactants and products no longer change over time.
- The equilibrium constant, K, is a measure of the extent to which a reaction favors products or reactants at equilibrium. If K is large, products are favored. If K is small, reactants are favored.
- Le Chatelier's principle states that if a system at equilibrium is disturbed, the equilibrium will shift in a direction that counteracts the applied change.
Displacement titration as analytical techniqueP.K. Mani
This document discusses displacement titration, specifically titrations of anions of weak acids (Bronsted bases) with strong acids. It provides examples of titrating salts of weak acids like sodium acetate, potassium cyanide, and borax with hydrochloric acid. The pH at the endpoint is calculated based on the concentration of the weak acid produced. Indicators that can be used for different titrations are also discussed based on the pH range at the endpoint.
This document provides an overview of acid-base titration including definitions, concepts, and procedures. It discusses the Arrhenius, Bronsted-Lowry, and Lewis definitions of acids and bases. It explains the process of ionization and factors that influence it such as relative permittivity. Key aspects of acid-base titration covered include types of reactions that can occur, use of indicators, and standards. The document also discusses acid and base ionization constants and how they relate to strength. Examples are provided to illustrate acid strength calculations and indicator color changes corresponding to pH.
Ion specific equation coefficient version of the Abraham model for ionic liqu...Bihan Jiang
This document summarizes research determining ion-specific equation coefficients for the Abraham solvation parameter model for several ionic liquid solvents. The researchers compiled partition coefficient data for solutes dissolved in five ionic liquids and correlated the data using the Abraham model. They were able to describe the partition coefficients within 0.13 log units. Cation-specific coefficients were then calculated for each ionic liquid, allowing predictions of solute partitioning in 76 ionic liquid solvents using previously reported anion coefficients.
The document discusses electrolyte balance and acid-base balance in the body. It provides details on various electrolytes like sodium, potassium, calcium salts and their role in maintaining balance. It also discusses the buffer systems and mechanisms involved in regulating pH of blood and treatment of acid-base imbalances. Specifically, it summarizes commonly used pharmaceutical compounds for correcting acid-base imbalances like sodium bicarbonate, sodium acetate, potassium acetate and their properties, methods of preparation, uses and official preparations.
The document discusses the concept of pH, which was developed in 1909 by Sorensen to measure the acidity of beer. pH is defined as the negative logarithm of the hydrogen ion concentration and measures the acidity or basicity of an aqueous solution on a scale from 0 to 14. The pH of blood, cytoplasm, and other bodily fluids is tightly regulated within a narrow range important for biological functions. Key factors discussed include acid-base equilibria, acid dissociation constants (Ka), and the Henderson-Hasselbalch equation relating pH to Ka for weak acids and bases.
* HCl is a strong acid and will titrate first
* Its equivalence point was at 35.00 mL of NaOH
* NaOH concentration is 0.100 M
* Moles of NaOH used = Volume x Concentration
= 0.03500 L x 0.100 mol/L = 0.003500 mol
* Moles of HCl = Moles of NaOH used = 0.003500 mol
* H3PO4 is a weak acid and will titrate second
* Its equivalence point was at 50.00 mL of NaOH
* Additional NaOH used = 50.00 mL - 35.00 mL = 15.00 mL
* Moles of additional Na
The document provides information about acid-base titrations including Bronsted-Lowry and Lewis acid-base theories, the self-ionization of water, and examples of water acting as an acid or base. It also discusses acid-base indicators and how they can be used to detect the equivalence point during titrations. Examples are given for titrations involving strong acid-strong base, weak acid-strong base, and weak base-strong acid. The dependence of the titration curve on concentration is illustrated and factors that can affect the choice of indicator are described.
Activity coefficients at infinite dilution for organic solutes dissolved in t...Bihan Jiang
This document reports on activity coefficients and gas-to-liquid partition coefficients for organic solutes dissolved in two ionic liquid solvents containing quinuclidinium cations with alkyl chains of six and eight carbons. Inverse gas chromatography was used to measure the partition coefficients of 47 organic probes in one ionic liquid and 45 probes in the other between 313-353 K. The data were analyzed using Abraham solvation models to derive mathematical correlations allowing prediction of partition coefficients for additional solutes in the two ionic liquids.
This study examined the individual metabolism of apolipoprotein (a) [apo(a)] and apolipoprotein B-100 [apoB-100] within lipoprotein (a) [Lp(a)] particles in the blood. Researchers found that the rate at which apo(a) is cleared from the blood is about half the rate at which apoB-100 is cleared when using an immunoprecipitation method to separate the proteins, whereas earlier studies found similar clearance rates using ultracentrifugation. The differences in clearance rates depending on the separation technique suggest that apo(a) and apoB-100 within Lp(a) particles may not be catabolized together in the non-fasting state. The study helps
Evaluation of LdhIsozymes Following the Treatment of Methyl Parathion in the ...inventionjournals
Lactate dehydrogenase converts lactate into pyruvate and has a very important role in carbohydrate metabolism. LDH activity depends on its isozymes and their activities change under pathological conditions. The increase in LDH activity suggests the increased anaerobic conditionsunder the influence of methylparathion to meet the energy demand when the aerobic oxidation is lowered. Following the treatment of methyl parathion there was a differential percentage in increase or decrease of different isozymes in the fish Labeo rohita. There were three distinct bands during the 96h of treatment almost parallel to LDH5 band of human serum suggesting liver damage. This was also confirmed by histopathological studies.
Evaluation of LdhIsozymes Following the Treatment of Methyl Parathion in the ...inventionjournals
The document evaluates changes in lactate dehydrogenase (LDH) isozyme levels in the fish Labeo rohita following exposure to the organophosphate pesticide methyl parathion. LDH isozymes were separated and quantified using electrophoresis. Exposure resulted in changes to the percentages of different LDH isozymes over time. Three distinct bands appeared after 96 hours of exposure, similar to a human LDH isozyme indicative of liver damage. Histological examination of liver tissue showed damage including disorganized hepatic cords and central veins, ruptured blood vessels, and necrotic changes over time with exposure. The study demonstrates methyl parathion exposure can alter LDH isozyme levels and cause liver damage in L. ro
Sipma, 2004, Effect Of Carbon Monoxide, Hydrogen And Sulfate On Thermophilic ...roelmeulepas
This document summarizes a study on the conversion of carbon monoxide (CO) by two anaerobic sludge samples at 55°C. The study aimed to elucidate the conversion routes and determine the effect of substrate (CO) concentration and the presence of hydrogen gas. Inhibition experiments showed CO conversion occurred via a hydrogenogenic population producing hydrogen and carbon dioxide, with the products then used by methanogens, acetogens, or sulfate reducers depending on sludge source and inhibitors. Both sludges could produce hydrogen from CO, indicating potential for biological hydrogen production from synthesis gas containing CO. The paper mill sludge was also capable of sulfate reduction using hydrogen produced from high CO concentrations, showing CO-rich synthesis gas can efficiently
Abraham model correlations for ionic liquid solvents computational methodolog...Bihan Jiang
The document describes a computational methodology for updating existing Abraham model ion-specific equation coefficients using new experimental solubility and partition coefficient data for ionic liquid solvents. Specifically, it illustrates updating the coefficients for the trifluoroacetate anion based on 51 data points from three ionic liquid solvents containing that anion. The updated coefficients have significantly smaller standard errors and are able to better predict solubility and partition behavior in the three ionic liquids based on the increased data. The methodology allows coefficients to be refined as new data becomes available without needing to re-regress the entire Abraham model data set.
The document discusses solubility equilibria and solubility product constants (Ksp). It defines Ksp as the product of ion concentrations raised to their coefficients in the solubility reaction equation. It provides examples of using Ksp to calculate solubility from solubility data and vice versa. Additionally, it discusses factors that affect solubility such as common ion effect, pH, and complexation.
This document provides an overview of acid-base titration and volumetric analysis. It defines key terms like titration, indicator, equivalence point, and standardization. It describes different types of titrations including direct, indirect, and back titration. Acid-base concepts are explained based on Arrhenius, Bronsted-Lowry, and Lewis theories. The document also discusses the ionic product of water, common ion effect, classification of indicators, and theories of indicators including Ostwald and chromophore theories.
This document discusses pKa values, which refer to the acid dissociation constant of acids. It defines pKa and explains how pKa values can be used to understand and predict the behavior of acids and bases in solution. The document covers several key topics in pKa including the factors that influence pKa values, common types of acids like mono- and polyprotic acids, and the significance and applications of pKa in fields like chemistry, biology, and medicine.
Acid-base titration is a technique used to determine the concentration of an acid or base by neutralizing a solution of unknown concentration with a solution of known concentration. The reaction is monitored using an indicator that changes color at the endpoint of the titration. The document outlines the basic procedure which involves transferring a sample of the solution of unknown concentration into a flask, adding an indicator, and then titrating with the solution of known concentration until the endpoint is reached as indicated by the color change of the indicator. The volume added is used to calculate the concentration of the original unknown solution.
Computational and Experimental Studies of MTO Catalyzed Olefin HydrogenationKaram Idrees
The poster that I presented at the 253rd American Chemical Society National Meeting and Exposition in San Francisco,
CA. It highlights some of my REU research at North Carolina State University under the mentorship of Dr. Elon Ison.
Activity coefficients at infinite dilution for organic solutes dissolved in t...Bihan Jiang
This document discusses measuring activity coefficients and gas-to-liquid partition coefficients for 45 organic solutes dissolved in two 1,2,3-tris(diethylamino)cyclopenylium based room temperature ionic liquids. Abraham model correlations were derived to predict solute transfer into the ionic liquids from water and the gas phase. The correlations described the observed partition coefficients to within 0.13 log units or less. Measured activity coefficients were used to calculate selectivity values for potential separations like hexane/benzene and hexane/pyridine.
Learning objectives
Introduction
Preparation of a standard solution used for redox titration
Oxidizing and reducing agents used in volumetric analysis
N/10 potassium permanganate preparation
N/10 potassium dichromate preparation
N/10 Iodine solution preparation
Examples of redox titrations
Conclusion
References
This document provides an introduction to analytical methods used in pharmaceutical analysis. It discusses various classical analytical methods like titrimetric methods including acid-base titrations, precipitation titrations, complexometric titrations and instrumental methods. It also summarizes different types of analytical techniques classified as classical methods, separation methods, spectroscopic methods, electrochemical methods and thermal methods. Specific techniques discussed in detail include acid-base titrations, indicators used in titrations, and types of complexometric titrations. The document provides an overview of key concepts and methods in pharmaceutical analytical chemistry.
- Chemical equilibrium is reached when the rates of the forward and reverse reactions of a reversible reaction are equal. At equilibrium, the concentrations of reactants and products no longer change over time.
- The equilibrium constant, K, is a measure of the extent to which a reaction favors products or reactants at equilibrium. If K is large, products are favored. If K is small, reactants are favored.
- Le Chatelier's principle states that if a system at equilibrium is disturbed, the equilibrium will shift in a direction that counteracts the applied change.
Displacement titration as analytical techniqueP.K. Mani
This document discusses displacement titration, specifically titrations of anions of weak acids (Bronsted bases) with strong acids. It provides examples of titrating salts of weak acids like sodium acetate, potassium cyanide, and borax with hydrochloric acid. The pH at the endpoint is calculated based on the concentration of the weak acid produced. Indicators that can be used for different titrations are also discussed based on the pH range at the endpoint.
This document provides an overview of acid-base titration including definitions, concepts, and procedures. It discusses the Arrhenius, Bronsted-Lowry, and Lewis definitions of acids and bases. It explains the process of ionization and factors that influence it such as relative permittivity. Key aspects of acid-base titration covered include types of reactions that can occur, use of indicators, and standards. The document also discusses acid and base ionization constants and how they relate to strength. Examples are provided to illustrate acid strength calculations and indicator color changes corresponding to pH.
Ion specific equation coefficient version of the Abraham model for ionic liqu...Bihan Jiang
This document summarizes research determining ion-specific equation coefficients for the Abraham solvation parameter model for several ionic liquid solvents. The researchers compiled partition coefficient data for solutes dissolved in five ionic liquids and correlated the data using the Abraham model. They were able to describe the partition coefficients within 0.13 log units. Cation-specific coefficients were then calculated for each ionic liquid, allowing predictions of solute partitioning in 76 ionic liquid solvents using previously reported anion coefficients.
The document discusses electrolyte balance and acid-base balance in the body. It provides details on various electrolytes like sodium, potassium, calcium salts and their role in maintaining balance. It also discusses the buffer systems and mechanisms involved in regulating pH of blood and treatment of acid-base imbalances. Specifically, it summarizes commonly used pharmaceutical compounds for correcting acid-base imbalances like sodium bicarbonate, sodium acetate, potassium acetate and their properties, methods of preparation, uses and official preparations.
The document discusses the concept of pH, which was developed in 1909 by Sorensen to measure the acidity of beer. pH is defined as the negative logarithm of the hydrogen ion concentration and measures the acidity or basicity of an aqueous solution on a scale from 0 to 14. The pH of blood, cytoplasm, and other bodily fluids is tightly regulated within a narrow range important for biological functions. Key factors discussed include acid-base equilibria, acid dissociation constants (Ka), and the Henderson-Hasselbalch equation relating pH to Ka for weak acids and bases.
* HCl is a strong acid and will titrate first
* Its equivalence point was at 35.00 mL of NaOH
* NaOH concentration is 0.100 M
* Moles of NaOH used = Volume x Concentration
= 0.03500 L x 0.100 mol/L = 0.003500 mol
* Moles of HCl = Moles of NaOH used = 0.003500 mol
* H3PO4 is a weak acid and will titrate second
* Its equivalence point was at 50.00 mL of NaOH
* Additional NaOH used = 50.00 mL - 35.00 mL = 15.00 mL
* Moles of additional Na
The document provides information about acid-base titrations including Bronsted-Lowry and Lewis acid-base theories, the self-ionization of water, and examples of water acting as an acid or base. It also discusses acid-base indicators and how they can be used to detect the equivalence point during titrations. Examples are given for titrations involving strong acid-strong base, weak acid-strong base, and weak base-strong acid. The dependence of the titration curve on concentration is illustrated and factors that can affect the choice of indicator are described.
Activity coefficients at infinite dilution for organic solutes dissolved in t...Bihan Jiang
This document reports on activity coefficients and gas-to-liquid partition coefficients for organic solutes dissolved in two ionic liquid solvents containing quinuclidinium cations with alkyl chains of six and eight carbons. Inverse gas chromatography was used to measure the partition coefficients of 47 organic probes in one ionic liquid and 45 probes in the other between 313-353 K. The data were analyzed using Abraham solvation models to derive mathematical correlations allowing prediction of partition coefficients for additional solutes in the two ionic liquids.
This study examined the individual metabolism of apolipoprotein (a) [apo(a)] and apolipoprotein B-100 [apoB-100] within lipoprotein (a) [Lp(a)] particles in the blood. Researchers found that the rate at which apo(a) is cleared from the blood is about half the rate at which apoB-100 is cleared when using an immunoprecipitation method to separate the proteins, whereas earlier studies found similar clearance rates using ultracentrifugation. The differences in clearance rates depending on the separation technique suggest that apo(a) and apoB-100 within Lp(a) particles may not be catabolized together in the non-fasting state. The study helps
Evaluation of LdhIsozymes Following the Treatment of Methyl Parathion in the ...inventionjournals
Lactate dehydrogenase converts lactate into pyruvate and has a very important role in carbohydrate metabolism. LDH activity depends on its isozymes and their activities change under pathological conditions. The increase in LDH activity suggests the increased anaerobic conditionsunder the influence of methylparathion to meet the energy demand when the aerobic oxidation is lowered. Following the treatment of methyl parathion there was a differential percentage in increase or decrease of different isozymes in the fish Labeo rohita. There were three distinct bands during the 96h of treatment almost parallel to LDH5 band of human serum suggesting liver damage. This was also confirmed by histopathological studies.
Evaluation of LdhIsozymes Following the Treatment of Methyl Parathion in the ...inventionjournals
The document evaluates changes in lactate dehydrogenase (LDH) isozyme levels in the fish Labeo rohita following exposure to the organophosphate pesticide methyl parathion. LDH isozymes were separated and quantified using electrophoresis. Exposure resulted in changes to the percentages of different LDH isozymes over time. Three distinct bands appeared after 96 hours of exposure, similar to a human LDH isozyme indicative of liver damage. Histological examination of liver tissue showed damage including disorganized hepatic cords and central veins, ruptured blood vessels, and necrotic changes over time with exposure. The study demonstrates methyl parathion exposure can alter LDH isozyme levels and cause liver damage in L. ro
Evaluation of LdhIsozymes Following the Treatment of Methyl Parathion in the ...inventionjournals
Lactate dehydrogenase converts lactate into pyruvate and has a very important role in carbohydrate metabolism. LDH activity depends on its isozymes and their activities change under pathological conditions. The increase in LDH activity suggests the increased anaerobic conditionsunder the influence of methylparathion to meet the energy demand when the aerobic oxidation is lowered. Following the treatment of methyl parathion there was a differential percentage in increase or decrease of different isozymes in the fish Labeo rohita. There were three distinct bands during the 96h of treatment almost parallel to LDH5 band of human serum suggesting liver damage. This was also confirmed by histopathological studies.
This study investigated the polymerization of lactic acid as a model for prebiotic peptide formation via ester-amide exchange. Lactic acid was polymerized in a closed system at 85°C over various time points. HPLC and 1H-NMR were used to analyze the polymers and determine degree of polymerization (DP) and total lactic acid units. DP was found to increase with time while total units decreased, showing polymer regeneration. Methods showed consistent results within 10-15% error. Further studies will compare kinetics to a computer simulation to determine rate constants and model polymerization from various monomers.
The researchers aimed to purify cellular retinol binding protein (CRBP) from bovine liver. Through a process involving homogenization, centrifugation, cation exchange chromatography, gel filtration, and concentration, they obtained a final product. However, characterization through SDS-PAGE and absorption spectroscopy identified the protein as catalase rather than CRBP. Despite initial absorption at 350nm for CRBP, the maximal absorption and thermal/pH profiles matched those of catalase. The purification resulted in the isolation of catalase rather than the intended CRBP.
- A low-volume ELISA assay was developed that can specifically quantify active GLP-1 (7-36 amide) using only 10 μL of sample. It showed excellent sensitivity below 0.5 pM and accuracy as demonstrated by spike and recovery studies in human, mouse, and rat plasma samples. The assay can distinguish between fasted and fed states, showing on average a 4.5-fold change in GLP-1 levels.
DEVELOPMENT AND VALIDATION OF AN RP-HPLC METHOD FOR SIMULTANEOUS DETERMINATIO...rahul ampati
This document describes the development and validation of an RP-HPLC method for the simultaneous determination of ramipril and amlodipine in tablets. The method utilizes a gradient elution with a C18 column, mobile phase of acetonitrile and sodium perchlorate buffer, and UV detection. The method was validated per ICH guidelines and showed good linearity, accuracy, precision, specificity and robustness. Forced degradation studies demonstrated the method can separate ramipril, amlodipine and their degradation products. The method was successfully applied to determine the content of ramipril and amlodipine in three tablet batches.
Crystalloids are electrolyte solutions that can freely diffuse throughout the extracellular space. The principal crystalloid is isotonic saline (0.9% NaCl), which expands the interstitial space rather than plasma volume. Ringer's lactate is also commonly used as it more closely matches plasma composition. Dextrose 5% in water (D5W) is hypotonic and expands both intra and extracellular spaces, providing calories but not electrolytes. Each crystalloid has different indications and disadvantages to consider when selecting the appropriate fluid for treatment.
This document presents the development and validation of an RP-HPLC method for the simultaneous determination of ramipril and amlodipine in tablets. The method utilizes a C18 column with a mobile phase of acetonitrile, sodium perchlorate buffer and triethylamine at pH 2.6 and 55°C. Ramipril and amlodipine were well resolved with retention times of 5.2 and 9.8 minutes respectively. The method was validated per ICH guidelines and demonstrated selectivity, linearity, accuracy, precision, robustness and ability to detect ramipril and amlodipine in the presence of degradation products. Forced degradation studies showed the drugs were stable except under acidic and alkaline conditions
1. The document discusses a study investigating the effect of thrombin on serotonin transporter (SERT) activity and the Na+/K+ ATPase pump in rat brain synaptosomes.
2. The study found that thrombin inhibits serotonin uptake in purified synaptosomes, likely by inhibiting the Na+/K+ ATPase pump, thereby reducing the sodium gradient required for SERT-mediated serotonin transport.
3. Further experiments examined the effects of thrombin on lipid raft fractions containing SERT and ATPase subunits, and on ATPase pump activity, finding thrombin reduced ATPase activity in crude and purified synaptosomes.
This study examined the interaction between the Abl kinase domain and the cancer drug Gleevec (imatinib) using site-directed mutagenesis to introduce a mutation (S417Y) associated with drug resistance. Researchers used PCR, protein purification techniques, and kinase assays to compare the inhibitory effect of Gleevec on wild-type Abl and the S417Y mutant. Their results supported the hypothesis that the S417Y mutation decreases Gleevec's ability to inhibit Abl kinase activity, though protein impurities limited the strength of the conclusions. The findings help explain why some cancers become resistant to Gleevec treatment.
This document summarizes in vitro experiments evaluating the anti-diabetic effects of alkaloidal fractions from Tinospora cordifolia and pentacyclic acid triterpenoids. Rat insulinoma cells were treated with fractions to measure insulin secretion. The fractions inhibited PTP-1B enzyme activity and glucose production in hepatocytes in a concentration-dependent manner, indicating anti-diabetic effects. Molecular docking suggested the compounds bind to a secondary site on PTP-1B, inhibiting the enzyme by a mixed inhibition mechanism. The results suggest pentacyclic triterpenoids may have potential as insulin sensitizers for treating type 2 diabetes.
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Isolation of lp (a) from human serum lipoproteins and its sialic acid concentration
1. International Journal of Science and Research (IJSR), India Online ISSN: 2319 – 7064
Volume 1 Issue 1, October 2012
www.ijsr.net
Isolation of LP (a) from Human Serum
Lipoproteins and its Sialic Acid Concentration
Gayatri Prakash1
Dept. of Zoology, Daulat Ram College, University of Delhi,
Delhi-110007, India
gayatriprakash@hotmail.com
Abstract: Lp(a), a genetic variant of β-lipoproteins (LDL) was isolated from human serum and was characterized to study its behavior
and sialic acid concentration. Like LDL, Lp (a) also contains significant amounts of sialic acid which can be digested completely by
neuraminidase enzyme treatment for 24 hours at 370C. Thus, Lp (a) samples treated with neuraminidase for 24 or 48 hours at 370C
showed negligible amounts of sialic acid. The electrophoretic mobility of these Lp (a) samples decreased in comparison to the native or
untreated Lp (a) samples. Presence of high amounts of sialic acid in Lp (a) like that in LDL suggest that this carbohydrate moiety may
play some role in these lipoproteins.
Keywords: Lipoproteins, Sialic acid, Neuraminidase enzyme, Lp (a)
Introduction
Lipoproteins are complexes of lipids and proteins having
the solubility characteristics of proteins. Human serum
lipoproteins can be isolated into a number of fractions
depending on their electrophoretic mobility and their
hydrated densities. These are high density lipoproteins
(HDL or α-lipoproteins), low density lipoproteins (LDL or
β-lipoproteins), very low density lipoproteins (VLDL or
pre- β -lipoproteins) and chilomicrons (exogenous
particles). In addition, another class of lipoproteins,
designated as Lp (a), can be isolated from human serum.
It represents genetic variant of β-lipoproteins and is
composed of β -lipoproteins, varying amounts of adsorbed
lipoproteins or albumin and the specific Lp (a) fraction.
Lp (a) has apo B as the major protein constituent and the
lipid moiety is indistinguishable from LDL [1] and [2].
The characteristic feature of Lp (a) is the presence of an
additional apoprotein (a-protein) which is distinct from all
other serum proteins and apoproteins. It is believed that
Lp (a) is an additional risk factor for atherosclerosis and
myocardial infarction independently from all other serum
lipoproteins [3] and [4]. The later assumption is based on
the finding that Lp (a) is synthesized differently from
other apo B containing lipoproteins [5]. Also, it has been
shown that the catabolism of Lp (a) proceeds via the same
routes as LDL since it is bound to the B/E receptor in
cultured human fibroblasts [6]. Further, the protein
moiety of low density lipoproteins has been
shown to contain 5-9 % carbohydrate
consisting of various sugars including sialic
acid [7], [8], [9], [10], and [11]. Therefore, it was of
interest to us to isolate Lp (a) from the human serum,
study its behavior in terms of electrophoretic mobility and
also to measure its contents of sialic acid.
1. Materials and Methods
Isolation of Lp (a): Human serum from twenty different
individuals was tested for Lp (a) positive nature against
Lp (a) antibody horse no. XI by radial immunodiffusion
using 1 % Latex agarose prepared in Rocket buffer. Two
samples showing positive antigen-antibody precipitation
reaction of 4-5mm diameter corresponding to
approximately 60-70 µg/µl Lp (a) were used for the
preparation of Lp (a).
The pooled serum (=380 ml) was centrifuged in Sorvall
centrifuge at slow speed (3000 RPM) to free it from blood
cells. The supernatant was brought to a density 1,055 with
sodium chloride and was then ultra centrifuged at 50,000
RPM for 24 hours using Beckman quick seal tubes. After
centrifugation, the tubes were sliced at the clear zone. The
bottom fraction (=200 ml) was collected while the top
fraction was discarded. The density of the bottom fraction
was then brought to 1,125 with sodium chloride and it was
ultra centrifuged again at 50,000 RPM for 24 hours. The
tubes were sliced as before at the clear zone just below the
cap and the top or upper Lp (a) rich fraction was collected
with the help of syringe (=38 ml). The Lp (a) rich fraction
was then dialyzed against glycine buffer (0.9% Nacl +
0.05 M glycine + 0.1% EDTA and 0.1% Sodium acid, pH
8.2), with pressure to concentrate it from 38 ml to 5.2 ml.
This concentrated Lp (a) sample was given to the column
once the later was ready.
2. Preparation of the column
The chromatographic column (100x2.5 cm) was prepared
by packing it with Biogel A5M. The column was washed
for the first time with 50 ml of urea solution (7.5M urea +
1.5% Nacl + 0.05 M glycine, pH=8) and subsequently
with glycine buffer (0.9% Nacl+ 0.05M glycine+ 0.1%
EDTA+ 0.1% sodium acid). The pressure in the column
was always adjusted to nearly 50 cm. Once the column
was washed two to three times its volume with glycine
buffer, it was ready for use.
The Lp (a) fraction was applied to the column across its
walls at a time when buffer layer was nearly one mm over
the gel in the column. When the sample moved into the
gel, small amount of buffer was added over the gel layer
and waited till this buffer also moved into the gel. Then
filled the column with buffer and allowed the sample to
1
2. International Journal of Science and Research (IJSR), India Online ISSN: 2319 – 7064
Volume 1 Issue 1, October 2012
www.ijsr.net
move down through the whole length of the column. The
outlet of the column was connected to the fraction
collector which was adjusted to 20 drops/tube. All the
fractions coinciding all the peaks on the chromatographic
column were tested for Lp (a) positive nature by
immunodiffusion and disc electrophoresis using 7% urea.
Eleven fractions (Tube Nos. 32 to 41) constituting the first
peak on the column showed Lp (a) positive reaction with
Lp (a) antibody Horse no. XI. Therefore, these fractions
were pooled together and were concentrated with pressure
to nearly 4ml against glycine buffer (0.9% Nacl+0.05M
glycine + 0.1% EDTA+ 0.1% Sodium acid, pH-8. 2).
Once the sample was concentrated, it was dialyzed in the
same buffer for two hours with vigorous shaking to mix
the precipitate /contents of the sample thoroughly. The
amount of lipoprotein in the prepared Lp (a) sample was
measured at 280µ against 0.001N NaOH.
3. Incubation of Lp (a) with Neuraminidase:
The prepared Lp (a) was treated with known amounts of
neuraminidase from Clostridium perfringens (Type IX,
Sigma). For this, two samples of 250 µl each were kept at
370
C with 5 µl neuraminidase for 24 and 48 hours
respectively. One sample of 250 µl was also kept at 370
C
for 48 hours without neuraminidase to serve as control for
the treated samples. After the neuraminidase treatment,
the samples were tested for :-
1. Cellulose acetate electrophoresis,
2. SDS Electrophoresis with 3.5 and 5% gels,
and
3. Concentration of Sialic acid.
The amount of N-acetyl neuraminic acid (Sialic acid) in
Lp(a) fraction of serum lipoprotein with and without the
neuraminidase treatment was determined by the
thiobarbituric acid procedure of Warren [12]. Protein
determinations were done by the Lowry method [13].
Three samples of 250 µL each of Lp (a), pH-8.2, (referred
as 2, 3 and 4) were used for these estimations. In sample
numbers 3 and 4, 5µl neuraminidase (0.5 Units) was
added separately with the help of micro-pipette and the
contents after mixing thoroughly were kept in the oven at
370
C for 24 and 48 hours respectively. The sample 2 was
also kept in the oven along with the samples 3 and 4 at
370
C for 48 hours but without neuraminidase to serve as
control for the treated samples. In addition, another
untreated Lp (a) control (sample 1) which was not kept in
the oven was used to compare with sample 2. Following
neuraminidase treatment, these samples were used for the
estimation of sialic acid.
4. Lipid Electrophoresis
Another aliquot of 0.8 ml Lp (a) was treated with 16 µl
(1.6 Units) neuraminidase (Type IX, same as before) at
370
C for 24 hours. The reaction mixture was then
dialyzed against sodium bromide sodium of density 1,125
for 24 hours. The sample was ultra centrifuged for 24
hours at 45,000 RPM using 50.3 Rotar with sodium
bromide solution of density 1,125. After centrifugation,
the top fraction of the sample was collected with the help
of syringe from the Beckman ultracentrifuge tube. Four
aliquots of 10 µl each of this treated samples of Lp (a)
were used for SDS electrophoresis while the remaining
sample was dialyzed against glycine buffer (0.9%
NaCl+O.05M glycine+1% EDTA+1% Na acid, pH-8.2)
for overnight. This neuraminidase free de-sialated Lp (a)
was used for lipid electrophoresis along with the native or
untreated Lp (a) and the normal serum.
5. Results
Results from Cellulose acetate electrophoresis studies
showed that neuraminidase treatment decreased the
mobility of Lp (a) in comparison to untreated/control
samples (Figure I). However, there was no difference in
the mobility pattern of Lp (a) treated with neuraminidase
for 24 and 48 hours respectively. Mobility pattern of Lp
(a) kept in the oven at 370
C for 48 hours without
neuraminidase to serve as control for the treated samples
did not change and was similar to that of the native Lp(a)
which was not kept in the oven (Figure I).
Figure I: Results of Cellulose acetate electrophoresis
1. Native/Untreated Lp (a), not kept in the oven;
2. Untreated Lp (a) kept in the oven for 48 hours at
370
C without the addition of neuraminidase;
3. Treated Lp (a) with neuraminidase for 24 hours
at 370
C;
4. Treated Lp (a) with neuraminidase for 48 hours
at 370
C.
The results of lipid electrophoresis are shown in Figure II.
These indicate that composition of Lp (a) resembles that
of LDL as the electrophoretic band of Lp (a) sample
corresponds with that of LDL band of the normal serum.
However, the electrophoretic mobility of the
neuraminidase treated Lp (a) samples decreased in
comparison to the untreated Lp (a) sample.
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3
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3. International Journal of Science and Research (IJSR), India Online ISSN: 2319 – 7064
Volume 1 Issue 1, October 2012
www.ijsr.net
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Figure II: Results of Lipid electrophoresis
2. Normal serum;
3. Native or Untreated Lp (a);
4. Treated Lp (a) with neuraminidase for 24 hours
at 370
C.
The amount of lipoprotein in the prepared Lp (a) sample
was 11.6 mg/ml. The amount of sialic acid in various Lp
(a) samples as calculated from the mean values obtained
from the optical densities of two parallel samples and
estimated according to the method of Warren (12) were as
follows:-
Sample 1: Native Lp (a) contained 84.56 µgm sialic
acid/mg protein
Sample 2: Untreated control contained 84.47µgm sialic
acid / mg protein
Samples 3 and 4: Neuraminidase treated Lp (a) for 24 and
48 hours respectively showed negligible amounts of sialic
acid.
The amount of sialic acid estimated in two LDL samples
was 9.77 and 10.76 µgm/mg protein.
Discussion
The results obtained from Cellulose acetate
electrophoresis (Figure I) and estimation of sialic acid
indicate that most of the sialic acid from Lp (a) was
removed with 24 hours neuraminidase treatment and 48
hours neuraminidase treatment did not cleave off sialic
acid any further. A decrease in the mobility pattern of
neuraminidase treated Lp (a) in comparison to native Lp
(a) as seen in the cellulose acetate electrophoresis may be
accounted for a decrease in the negative charge on Lp (a)
due to the removal of negatively charged sialic acid.
Treated Lp (a) with neuraminidase for 48 hours at 370
C.
The results of lipid electrophoresis as seen in Figure II
indicate that composition of Lp (a) resembles that of LDL
as the electrophoretic band of Lp (a) sample corresponds
with that of LDL band of the normal serum. However, the
electrophoretic mobility of the neuraminidase treated Lp
(a) samples decreased in comparison to the untreated Lp
(a) sample. This once again shows a decrease in the net
negative charge on Lp (a) due to the removal of negatively
charged sialic acid following the treatment with
neuraminidase.
Estimation of sialic acid indicates that Lp (a) contains this
in large amounts which gets digested completely with 24
hours treatment of neuraminidase and because of this
digestion, neuraminidase treated Lp (a) for 24 and 48
hours respectively showed negligible amounts of sialic
acid. Two LDL samples analyzed for the amount of sialic
acid showed that these contained 9.77 and 10.76 µgm/mg
protein sialic acid respectively thus suggesting
resemblance of Lp (a) with LDL. These results confirm
the studies which showed that the protein moiety of
low density lipoproteins contain 5-9 %
carbohydrate consisting of various sugars
such as galactose, mannose, glucosamine
and sialic acid [7], [8], [9], [10], and [11].
However, Swaminathan and Aladjem [14] reported a
marked variation in sialic acid values ranging from 6 to 17
µgm/mg protein in LDL. Further, our studies are in
conformation with the results of Margolis and Langdon
[15] who suggested that sialic acid residues could be
removed from native LDL with sialidase treatment
without affecting the lipid binding.
The presence of large amount of sialic acid in Lp (a) is not
clear at present. A number of possible functions of
carbohydrate moiety in glycoprotein have been proposed
by many scientists. According to them, it may be
involved in the secretion from the cell [16], or in the
regulation of their catabolism [17]. It has been shown that
the removal of sialic acid from glycoproteins decreases
their biological half-life [17]. However, similar studies
using native and desialyzed iodide labeled LDL did not
show significant differences in their respective rates of
disappearance [10]. Sialic acid may also play a role in
membrane permeability [18]. The presence of LDL in
atherosclerosis plaques [19], [20] has been demonstrated
and also uptake of LDL by cultured fibroblasts has been
reported by Brown and Goldstein [21]. These processes
might involve the interaction of carbohydrate moiety of
LDL with cell membranes. Since human LDL has been
demonstrated to be taken up specifically and degraded by
human aortic smooth muscle cells [22] and cultured
fibroblasts [23] and has been shown to regulate the
content of free and esterified cholesterol in human
fibroblasts [24], the relative atherogenecity of LDL has
been suggested to depend upon the carbohydrate moiety
[14]. This may also be true for Lp (a) as it resembles with
LDL in its composition and also contains large amounts of
sialic acid.
Acknowledgements
These studies have been supported by grants from the
Osterreichische Fonds zur Forderung der
wissenschaftlichen Forschung. The anti-Lp (a) from
sheep was a gift from Immuno A.G. Vienna, Austria.
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