Future and potential of Countercurrent Chromatography (CCC) from preparative isolation of compounds to the production of Knock-out Extracts.
Can CCC become a mainstream technique?
Listing and details on the different elution methods (e.g., EECCC, BECCC, Dual mode, recycling mode) that can be implemented in countercurrent chromatography.
1. The document provides information about an upcoming conference on solvent system selection for countercurrent chromatography (CCC) to be held in Chicago in August 2016.
2. It discusses various topics that will be covered at the conference including historic and current solvent systems used in CCC, instrumentation, empirical and rational approaches for solvent system selection, and guidelines for selecting solvent-stationary phase combinations.
3. References are provided for papers on selecting solvent systems for natural product separations by CCC and strategies for solvent system selection.
The different types of instruments are compared in terms of capacity (column volume), resolution, and duration required for the separation of targeted compounds.
This workshop presentation was prepared by Dr. Friesen (http://www.dom.edu/departments/physicalsciences/faculty/j-brent-friesen).
This document provides instructions for representing chromatographic data using reciprocal symmetry (ReS) and reciprocal symmetry scaling (ReSS). It explains how to calculate partition coefficients (K) from chromatograms and plot the data in ReS or ReSS format. Guidelines are given for selecting the midline position and adjusting the x-axis to fit the data appropriately. Examples show how these techniques can be used to compare chromatographic methods and solvent systems.
How to perform partitioning experiments to calculate the partition coefficient K, and ultimately identify or selection the optimal solvent system for countercurrent chromatography.
1. pH-zone refining countercurrent chromatography (CCC) can be used to separate acidic compounds. It utilizes differences in partition coefficients (K values) of analytes between basic (Kbase) and acidic (Kacid) conditions.
2. Key steps include testing Kbase and Kacid values of analytes using different pH solvent systems to determine suitability for separation. A suitable system yields Kbase << 1 and Kacid >> 1.
3. Separation of D&C Orange No. 5 is demonstrated using a diethyl ether-acetonitrile-aqueous ammonium acetate solvent system at basic pH as stationary phase and acidic mobile phase.
Future and potential of Countercurrent Chromatography (CCC) from preparative isolation of compounds to the production of Knock-out Extracts.
Can CCC become a mainstream technique?
Listing and details on the different elution methods (e.g., EECCC, BECCC, Dual mode, recycling mode) that can be implemented in countercurrent chromatography.
1. The document provides information about an upcoming conference on solvent system selection for countercurrent chromatography (CCC) to be held in Chicago in August 2016.
2. It discusses various topics that will be covered at the conference including historic and current solvent systems used in CCC, instrumentation, empirical and rational approaches for solvent system selection, and guidelines for selecting solvent-stationary phase combinations.
3. References are provided for papers on selecting solvent systems for natural product separations by CCC and strategies for solvent system selection.
The different types of instruments are compared in terms of capacity (column volume), resolution, and duration required for the separation of targeted compounds.
This workshop presentation was prepared by Dr. Friesen (http://www.dom.edu/departments/physicalsciences/faculty/j-brent-friesen).
This document provides instructions for representing chromatographic data using reciprocal symmetry (ReS) and reciprocal symmetry scaling (ReSS). It explains how to calculate partition coefficients (K) from chromatograms and plot the data in ReS or ReSS format. Guidelines are given for selecting the midline position and adjusting the x-axis to fit the data appropriately. Examples show how these techniques can be used to compare chromatographic methods and solvent systems.
How to perform partitioning experiments to calculate the partition coefficient K, and ultimately identify or selection the optimal solvent system for countercurrent chromatography.
1. pH-zone refining countercurrent chromatography (CCC) can be used to separate acidic compounds. It utilizes differences in partition coefficients (K values) of analytes between basic (Kbase) and acidic (Kacid) conditions.
2. Key steps include testing Kbase and Kacid values of analytes using different pH solvent systems to determine suitability for separation. A suitable system yields Kbase << 1 and Kacid >> 1.
3. Separation of D&C Orange No. 5 is demonstrated using a diethyl ether-acetonitrile-aqueous ammonium acetate solvent system at basic pH as stationary phase and acidic mobile phase.
The 9th International CCC Event will be held August 1-3, 2016 at Dominican University in River Forest, Illinois. The event will include conferences on August 1-3 and workshops on July 30-31. Brent Friesen, a Chemistry Professor at Dominican University, is the contact for the event. Countercurrent separation is a type of support-free liquid chromatography that has various instrumentation types including countercurrent chromatography, centrifugal partition chromatography, and droplet countercurrent chromatography. It provides advantages such as minimal sample preparation, high mass resolution, no sample loss, reproducibility, flexibility, and mild separation conditions for sensitive molecules.
The presence of Per- and Polyfluorinated Alkyl Substances (PFAS) in drinking water is being thoroughly studied due to the persistence of these compounds in the environment and their potential health effects. However, there is limited knowledge about the occurrence of these chemicals in bottled water, despite the increasing concerns about PFAS in the food supply. This poster shows results from a fast and simple direct injection method similar to draft EPA method 8237, using the Shimadzu triple quad LCMS-8050 to analyze seven commercially available samples of bottled water for 24 PFAS.
This document evaluates the use of hydrogen (H2) carrier gas as an alternative to helium for the analysis of haloacetic acids (HAAs) according to EPA Method 552.3. Chromatograms of HAA standards using H2 were nearly identical to those using helium. Calibration curves for HAAs using H2 had coefficients of determination over 0.995. Accuracy and repeatability results using H2 met EPA requirements. Using H2 instead of helium reduces analysis costs by 2.69 to 6.15 times depending on gas purity, making H2 a suitable alternative carrier gas.
Over the past decade, there have been a growing number of mAb candidates entering the clinical pipeline. This results in a large increase on the demand for analytical characterization. This seminar discusses advances in analytical method development with analytical run times below 10 minutes for all routine methods with intelligent, integrated chromatography workflows. Orbitrap technology has been established as the most powerful MS technology for protein characterization. How this can be incorporated into a complete workflow for bio-pharma analysis is also discussed.
This presentation reports on the development of a GC FID method to accurately quantify ethanol and IPA concentrations in two hand sanitizer samples. By using nitrogen as the carrier gas, this method is cost-effective and ensures the product compliance with CDC and USP guidelines and regulations.
This document describes the development of a method to analyze 10 common seized drugs using a single quadrupole mass spectrometer. A LC-MS method was optimized to separate and detect the drugs simultaneously using selected ion monitoring and in-source collision-induced dissociation for identification. Linear calibration curves were obtained for each drug from 0.1-1 μg/mL. The method demonstrated reproducibility and accuracy for rapid screening of multiple drugs.
This document describes the separation and analysis of three samples sent to B&B Rod Co. by Mr. Smith using different chromatography techniques. For the first sample, caffeine content in an impure standard and coffee sample, HPLC was used. It found the impure standard contained 0.09967% caffeine with 55.66% error. For the second, TLC found an impure Allura Red AC sample contained 49.78% of the dye. For the third, GC found a mixture of alcohols contained 0.4786 mg/ml of Cetyl alcohol and 0.6177 mg/ml of Stearyl alcohol.
Automated SPE for Capillary Microsampling PosterRick Youngblood
1) An automated SPE method using small single-use cartridges was evaluated for plasma samples derived from capillary microsampling as an alternative to manual protein precipitation.
2) Results from the automated SPE method were comparable to manual protein precipitation in terms of accuracy, precision, calibration curves, and limits of quantification. The automated method provided equivalent results but with less hands-on time.
3) Further optimization is possible, including reducing the plasma sample volume and elution volume to allow analysis of samples from microsampling techniques using minimal volumes.
This application note describes the methodology and use of the Shimadzu ICPMS-2030 ICP mass spectrometer for the analysis of trace elements in drinking and fresh waters following the EPA 200.8 method. This method is also used for analysis of wastewater. Here, we demonstrate the stability and sensitivity of the ICPMS-2030 for EPA 200.8 analyses.
Using THGA and Zeeman Background Correction for Blood-Lead Determination in C...PerkinElmer, Inc.
Validated applications determining whole blood levels are generally performed using graphite furnace atomic absorption spectroscopy (GFAAS). GFAAS is cost effective, allows for detection limits well under the blood-lead level action guideline, and requires less operator training than more advanced elemental techniques.2 In this study, we will demonstrate the applicability of the PerkinElmer® PinAAcle™ 900T atomic absorption spectrometer (Figure 1) using the stabilized temperature platform furnace (STPF) and transversely-heated graphite atomizer (THGA), for use in customer-validated applications to determine lead amounts in blood samples.
Learn more about our solutions: http://bit.ly/IG2kI1
This document analyzes and quantifies levels of heavy metals (lead, cadmium, mercury, and arsenic) in Cascade hops using graphite furnace atomic absorption and cold vapor techniques on a Shimadzu AA-7000 instrument. Dried and ground hops were digested in acids and diluted before analysis. Calibration curves for each metal showed good linearity above 0.999. Analysis of samples and spiked samples found metal levels within targeted ranges and recoveries of 90-110%, validating the method's accuracy and sensitivity at low concentration levels.
The growth of, and the confidence in, hemp products will require applicable testing to ensure product quality and safety. Chromatography technology will play a large role in this as the technique is used for potency testing. This study optimizes a quantitative chromatographic determination of 15 cannabinoids using the Shimadzu Hemp Analyzer.
This document summarizes a presentation on biological medicines and monoclonal antibody therapeutics. It discusses regulatory guidelines for biosimilars in Korea and ASEAN countries. The presentation provided an overview of global biosimilar guidelines, noting that the EU implemented the world's first well-organized biosimilar legislation and guidelines. It also noted that Korea has issued biosimilar guidelines since 2009, and Malaysia and Singapore are leading implementation of biosimilar guidelines in ASEAN countries.
Current sample preparation techniques for PFAS analysis are laborious and not easily automated. In this study, supercritical fluid extraction (SFE) was evaluated as an alternative sample preparation technique for the extraction of eighteen PFAS compounds from fish tissue, as a preconcentration step prior to their analysis by LC-MS/MS.
Drug discovery library design by biomimetic hplcKlara Valko
The slides explain the early drug discovery process and how the measurements of biomimetic properties can help to design the best ADME properties of compound libraries.
This is nice presentation given by Vishal Goyani, an Analytical student of National Institute Of Pharmaceutical education and research-INDIA.
mail your query at - vngoyanii@gmail.com
Biomimetic hplc methods to predict in vivo drug distributionCathe Barty
This document discusses how biomimetic HPLC methods can be used to predict in vivo drug distribution and improve drug candidate quality. It provides three key points:
1) HPLC retention times measured using stationary phases that mimic human serum albumin (HSA), alpha-1-acid glycoprotein (AGP), and artificial membranes can be used to estimate binding to these biomolecules and predict tissue distribution.
2) Chromatographic hydrophobicity indices (CHIs) measured at different pH values reveal a compound's acid/base properties without structural information.
3) HPLC methods allow rapid measurement of various physicochemical properties like lipophilicity, permeability, and solubility which influence drug absorption and efficacy.
There is high demand for oxysterol quantitation due to their correlation with neurodegenerative diseases. The ratios of various oxysterols in biological fluids are used by researchers to study disease states. This application presents a fast, sensitive LC-MS/MS method using the LCMS-8060, with detection quantitation limits determined using multiple reaction monitoring mode for each analyte.
This presentation evaluates ASTM D7979-16 for the “direct” analysis of 30 PFCs and compares data to the solid-phase extraction EPA drinking water Method 537.
Comprehensive Investigation of the Utilization of SFC/ESI Positive Mode MS fo...Waters Corporation
This document discusses the use of single column convergence chromatography/mass spectrometry (UPC2/MS) for bioanalytical studies. It provides examples of how UPC2/MS can simplify workflows by reducing sample preparation times through direct injection of extracts and improving selectivity over reversed phase chromatography. UPC2/MS also allows for faster separation of challenging compound classes like isomers and lipids compared to traditional techniques like gas chromatography. The document concludes that UPC2/MS provides an orthogonal separation method and combines multiple techniques into one analytical platform for streamlining quantitative bioanalysis in drug discovery and development.
This document describes a fully automated method for analyzing 25-OH-Vitamin D3/D2 and 3-Epi-25-OH Vitamin D3/D2 in serum using LC-MS/MS. The method utilizes protein precipitation, solid phase extraction, and gradient elution on an Ascentis Express F5 column with MS/MS detection. Validation results show good linearity, accuracy, and precision for quantifying both the active and inactive vitamin D metabolites and their epimers. The fully automated system is capable of processing 96 samples in under 17 hours.
The 9th International CCC Event will be held August 1-3, 2016 at Dominican University in River Forest, Illinois. The event will include conferences on August 1-3 and workshops on July 30-31. Brent Friesen, a Chemistry Professor at Dominican University, is the contact for the event. Countercurrent separation is a type of support-free liquid chromatography that has various instrumentation types including countercurrent chromatography, centrifugal partition chromatography, and droplet countercurrent chromatography. It provides advantages such as minimal sample preparation, high mass resolution, no sample loss, reproducibility, flexibility, and mild separation conditions for sensitive molecules.
The presence of Per- and Polyfluorinated Alkyl Substances (PFAS) in drinking water is being thoroughly studied due to the persistence of these compounds in the environment and their potential health effects. However, there is limited knowledge about the occurrence of these chemicals in bottled water, despite the increasing concerns about PFAS in the food supply. This poster shows results from a fast and simple direct injection method similar to draft EPA method 8237, using the Shimadzu triple quad LCMS-8050 to analyze seven commercially available samples of bottled water for 24 PFAS.
This document evaluates the use of hydrogen (H2) carrier gas as an alternative to helium for the analysis of haloacetic acids (HAAs) according to EPA Method 552.3. Chromatograms of HAA standards using H2 were nearly identical to those using helium. Calibration curves for HAAs using H2 had coefficients of determination over 0.995. Accuracy and repeatability results using H2 met EPA requirements. Using H2 instead of helium reduces analysis costs by 2.69 to 6.15 times depending on gas purity, making H2 a suitable alternative carrier gas.
Over the past decade, there have been a growing number of mAb candidates entering the clinical pipeline. This results in a large increase on the demand for analytical characterization. This seminar discusses advances in analytical method development with analytical run times below 10 minutes for all routine methods with intelligent, integrated chromatography workflows. Orbitrap technology has been established as the most powerful MS technology for protein characterization. How this can be incorporated into a complete workflow for bio-pharma analysis is also discussed.
This presentation reports on the development of a GC FID method to accurately quantify ethanol and IPA concentrations in two hand sanitizer samples. By using nitrogen as the carrier gas, this method is cost-effective and ensures the product compliance with CDC and USP guidelines and regulations.
This document describes the development of a method to analyze 10 common seized drugs using a single quadrupole mass spectrometer. A LC-MS method was optimized to separate and detect the drugs simultaneously using selected ion monitoring and in-source collision-induced dissociation for identification. Linear calibration curves were obtained for each drug from 0.1-1 μg/mL. The method demonstrated reproducibility and accuracy for rapid screening of multiple drugs.
This document describes the separation and analysis of three samples sent to B&B Rod Co. by Mr. Smith using different chromatography techniques. For the first sample, caffeine content in an impure standard and coffee sample, HPLC was used. It found the impure standard contained 0.09967% caffeine with 55.66% error. For the second, TLC found an impure Allura Red AC sample contained 49.78% of the dye. For the third, GC found a mixture of alcohols contained 0.4786 mg/ml of Cetyl alcohol and 0.6177 mg/ml of Stearyl alcohol.
Automated SPE for Capillary Microsampling PosterRick Youngblood
1) An automated SPE method using small single-use cartridges was evaluated for plasma samples derived from capillary microsampling as an alternative to manual protein precipitation.
2) Results from the automated SPE method were comparable to manual protein precipitation in terms of accuracy, precision, calibration curves, and limits of quantification. The automated method provided equivalent results but with less hands-on time.
3) Further optimization is possible, including reducing the plasma sample volume and elution volume to allow analysis of samples from microsampling techniques using minimal volumes.
This application note describes the methodology and use of the Shimadzu ICPMS-2030 ICP mass spectrometer for the analysis of trace elements in drinking and fresh waters following the EPA 200.8 method. This method is also used for analysis of wastewater. Here, we demonstrate the stability and sensitivity of the ICPMS-2030 for EPA 200.8 analyses.
Using THGA and Zeeman Background Correction for Blood-Lead Determination in C...PerkinElmer, Inc.
Validated applications determining whole blood levels are generally performed using graphite furnace atomic absorption spectroscopy (GFAAS). GFAAS is cost effective, allows for detection limits well under the blood-lead level action guideline, and requires less operator training than more advanced elemental techniques.2 In this study, we will demonstrate the applicability of the PerkinElmer® PinAAcle™ 900T atomic absorption spectrometer (Figure 1) using the stabilized temperature platform furnace (STPF) and transversely-heated graphite atomizer (THGA), for use in customer-validated applications to determine lead amounts in blood samples.
Learn more about our solutions: http://bit.ly/IG2kI1
This document analyzes and quantifies levels of heavy metals (lead, cadmium, mercury, and arsenic) in Cascade hops using graphite furnace atomic absorption and cold vapor techniques on a Shimadzu AA-7000 instrument. Dried and ground hops were digested in acids and diluted before analysis. Calibration curves for each metal showed good linearity above 0.999. Analysis of samples and spiked samples found metal levels within targeted ranges and recoveries of 90-110%, validating the method's accuracy and sensitivity at low concentration levels.
The growth of, and the confidence in, hemp products will require applicable testing to ensure product quality and safety. Chromatography technology will play a large role in this as the technique is used for potency testing. This study optimizes a quantitative chromatographic determination of 15 cannabinoids using the Shimadzu Hemp Analyzer.
This document summarizes a presentation on biological medicines and monoclonal antibody therapeutics. It discusses regulatory guidelines for biosimilars in Korea and ASEAN countries. The presentation provided an overview of global biosimilar guidelines, noting that the EU implemented the world's first well-organized biosimilar legislation and guidelines. It also noted that Korea has issued biosimilar guidelines since 2009, and Malaysia and Singapore are leading implementation of biosimilar guidelines in ASEAN countries.
Current sample preparation techniques for PFAS analysis are laborious and not easily automated. In this study, supercritical fluid extraction (SFE) was evaluated as an alternative sample preparation technique for the extraction of eighteen PFAS compounds from fish tissue, as a preconcentration step prior to their analysis by LC-MS/MS.
Drug discovery library design by biomimetic hplcKlara Valko
The slides explain the early drug discovery process and how the measurements of biomimetic properties can help to design the best ADME properties of compound libraries.
This is nice presentation given by Vishal Goyani, an Analytical student of National Institute Of Pharmaceutical education and research-INDIA.
mail your query at - vngoyanii@gmail.com
Biomimetic hplc methods to predict in vivo drug distributionCathe Barty
This document discusses how biomimetic HPLC methods can be used to predict in vivo drug distribution and improve drug candidate quality. It provides three key points:
1) HPLC retention times measured using stationary phases that mimic human serum albumin (HSA), alpha-1-acid glycoprotein (AGP), and artificial membranes can be used to estimate binding to these biomolecules and predict tissue distribution.
2) Chromatographic hydrophobicity indices (CHIs) measured at different pH values reveal a compound's acid/base properties without structural information.
3) HPLC methods allow rapid measurement of various physicochemical properties like lipophilicity, permeability, and solubility which influence drug absorption and efficacy.
There is high demand for oxysterol quantitation due to their correlation with neurodegenerative diseases. The ratios of various oxysterols in biological fluids are used by researchers to study disease states. This application presents a fast, sensitive LC-MS/MS method using the LCMS-8060, with detection quantitation limits determined using multiple reaction monitoring mode for each analyte.
This presentation evaluates ASTM D7979-16 for the “direct” analysis of 30 PFCs and compares data to the solid-phase extraction EPA drinking water Method 537.
Comprehensive Investigation of the Utilization of SFC/ESI Positive Mode MS fo...Waters Corporation
This document discusses the use of single column convergence chromatography/mass spectrometry (UPC2/MS) for bioanalytical studies. It provides examples of how UPC2/MS can simplify workflows by reducing sample preparation times through direct injection of extracts and improving selectivity over reversed phase chromatography. UPC2/MS also allows for faster separation of challenging compound classes like isomers and lipids compared to traditional techniques like gas chromatography. The document concludes that UPC2/MS provides an orthogonal separation method and combines multiple techniques into one analytical platform for streamlining quantitative bioanalysis in drug discovery and development.
This document describes a fully automated method for analyzing 25-OH-Vitamin D3/D2 and 3-Epi-25-OH Vitamin D3/D2 in serum using LC-MS/MS. The method utilizes protein precipitation, solid phase extraction, and gradient elution on an Ascentis Express F5 column with MS/MS detection. Validation results show good linearity, accuracy, and precision for quantifying both the active and inactive vitamin D metabolites and their epimers. The fully automated system is capable of processing 96 samples in under 17 hours.
The document discusses new innovations in UHPLC and LC-MS workflows for characterizing monoclonal antibodies and antibody drug conjugates. It summarizes the analytical challenges in characterizing modifications, impurities, immunogenicity, efficacy, and drug-antibody ratios. The Thermo Scientific Vanquish UHPLC system provides high-performance separations, reproducibility, and sensitivity needed for comprehensive characterization within short timeframes.
This presentation reviews the results of a study in which the authors investigated the effects of poly-diallydimethylammonium chloride (pDADMAC) flocculation and clarification on the performance and longevity of protein A resin.
To learn more about this topic or collaborate with our technical experts, schedule an in-person or remote visit at our M Lab™ Collaboration Centers: http://www.merckmillipore.com/mlab
This presentation reviews the results of a study in which the authors investigated the effects of poly-diallydimethylammonium chloride (pDADMAC) flocculation and clarification on the performance and longevity of protein A resin.
To learn more about this topic or collaborate with our technical experts, schedule an in-person or remote visit at our M Lab™ Collaboration Centers: http://www.emdmillipore.com/mlab
New Software Methods Enhance Sedimentation Velocity Analysis of Protein Aggre...KBI Biopharma
1) New software methods have improved the resolution and sensitivity of sedimentation velocity analysis, making it useful for comparability testing, formulation development, and quality control.
2) A method developed by Peter Schuck increases resolution of multiple components and detects very minor components below 1%. It provides higher resolution than SEC columns.
3) Another new method allows obtaining conformation and mass information at extremely low concentrations below 1 microgram total protein.
Purification optimization and characterization of protease from Bacillus va...Vaibhav Maurya
This presentation is a research work carried out by me in B.Tech 8 semester. and gives an idea about purification, optimization and characterization of protease from Bacillus Valismortis
This document provides example calculations for chlorine dosage, sodium hypochlorite (NaOCl) solution strength, and chemical feed rate for water disinfection. It includes equations to calculate chlorine dosage based on gallons of NaOCl solution and water produced. Tables are provided to determine NaOCl solution strength based on ounces of 12.5% NaOCl added to water and to calculate chemical feed rate based on desired chlorine residual, water flow rate, and NaOCl solution strength. Examples are included to demonstrate using the equations and tables to calculate values like chlorine dosage, NaOCl solution percentage, and chemical feed pump rate.
The document describes procedures for operating and calibrating a UV-Visible
spectrophotometer. It includes steps for turning on the instrument and software, cleaning
cuvettes, auto-zeroing with a blank sample, running samples to obtain spectra and absorbance
maxima. Experiments are presented to study the effect of concentration on absorbance using
paracetamol solutions, determine the concentration of an unknown using standard solutions, and
use a calibration curve method to find the concentration of an unknown sample. The document
provides objectives, requirements, theoretical background and procedures for various experiments
conducted using a UV-Vis spectrophotometer.
Particles in the Biotech Product Life Cycle: Analysis, Identification and Con...SGS
This presentation looks at the different technologies available for detection of particles generated during the drug development lifecycle and their control using a formulation approach for particles generated as a result of agitation and freeze/thaw, events commonly observed during sample shipment and temperature excursions.
This document summarizes the quantification of human C-reactive protein (CRP) using mass spectrometry. CRP is a biomarker for inflammation, atherosclerosis, and infection. Samples were digested with trypsin and specific CRP peptides were quantified using multiple reaction monitoring mass spectrometry. Standard curves for two peptides showed excellent accuracy and precision between 93.6-104.4% and 96.8-102.4% respectively. Quality control samples across four lots demonstrated good accuracy from 87.5-104.3% and precision below 13.31%. Representative chromatograms of quality control samples at different concentrations are also shown.
With increasing pressure of a higher sample throughput and fewer chemists, purification labs in medicinal chemistry groups need to be more productive now than ever before.
This presentation will describe a technique that allows the analyst to obtain a higher purity and better resolution using information from the preliminary analytical screening of these samples prior to purification.
This presentation discusses the history of size exclusion chromatography from gel permeation to modern day columns, and introduces the next evolution in polymer chromatography, the ACQUITY Advanced Polymer Chromatography system.
Three anti-cancer drugs were extracted from plasma samples using a 96-well plate and analyzed using liquid chromatography mass spectrometry. The analysis identified all drug compounds in under 1.4 minutes with high accuracy and sensitivity. This high-speed analysis method using 96-well plates is compatible with high-throughput anti-cancer drug research workflows.
This document discusses polymeric nanoparticles for encapsulation and controlled release of bioactive compounds. Section 1 discusses polysaccharide-based nanocomplexes of chitosan and alginic acid for co-encapsulation and sustained release of 5-fluorouracil and temozolomide. The nanoparticles had sizes around 100-200 nm, encapsulation efficiencies between 20-80%, and provided sustained release over one week. Section 2 explores chitosan grafted with low molecular weight polylactic acid for protein encapsulation and reducing initial burst release. The grafted polylactic acid prolonged release in acidic media and reduced initial burst. Section 3 examines amphiphilic nanoparticles using chitosan grafted with polyl
Practical Implementation of the New Elemental Impurities Guidelines May 2015SGS
The International Conference on Harmonization (ICH) released its Q3D Guideline for Elemental Impurities in December 2014, initiating reviews and changes in quality testing programs in bio/pharmaceutical companies around the world. In advance of the implementation dates, companies need to assess the risks of potential elemental impurities in their process and materials streams.
In this presentation, experts will review the requirements of elemental impurities guidelines from ICH, the European Pharmacopeia, and United States Pharmacopeia, outline practical recommendations to address implementation challenges, and discuss key considerations for analytical testing programs.
Pierre-Charles Bierly provides examples of quality control tools and methods he has developed for his work, including spreadsheets, graphs, and programs. These include quality control charts to monitor precision, mass and charge balance calculations to check analysis completeness, concentration calculation programs, and alkalinity and solubility calculation programs. The tools are intended to help establish quality control programs and solve various process problems.
Stress testing and physico-chemical characterization of monoclonal antibodies...Quality Assistance s.a.
Accelerated stress studies of biotech products can be carried out to study the main degradation pathways of the recombinant proteins to evaluate their stability, or to identify the different peaks / entities observed while running stability indicating separative methods such as ion exchange or size exclusion chromatography.
by Dr Géry Van Vyncht, R&D Director
For more informations : www.quality-assistance.com
This document provides information on dosage guidelines and administration instructions for several vasoactive medications including adrenaline, noradrenaline, dobutamine, dopamine, nitroglycerine, nitroprusside, heparin, fentanyl, and midazolam. It includes details on drug compatibility, dilution, concentration, dosage ranges, stability times, and infusion rate tables based on patient weight. Proper light protection during preparation and a maximum 24 hour infusion time are noted for several of the drugs.
Similar to Optimization parameters in Countercurrent Chromatography (20)
How to prepare and share your Raw NMR data along with your publication. Sharing your NMR data will enhance the transparency and reproducibility of structure elucidation while improving the dereplication process. This is a guidance document proposed by the CENAPT team. See also the corresponding website at : https://cenapt.pharm.uic.edu/resource/.
This document combined the microscopic analysis, DNA barcoding results, and phytochemical fingerprints for the botanical identification of the following commercial plant materials: Epimedium sagittatum (leaf powder), Marrubium vulgare (crushed aerial parts), Pausinystalia johimbe (bark powder), Senna alexandrina (leaf powder), Trigonellum foenum-graecum ( seed powder), and Trifolium pratense (crushed aerial parts).
Presentation from Dr. Guy Harris at the annual meeting of the American Society of Pharmacognosy (ASP) in Lexington Kentucky (2018) during the CENAPT/Gilson Workshop on Countercurrent Chromatography.
Workshop presented by Gregoire Audo from Gilson Purification at the annual meeting of the American Society of Pharmacognosy (ASP 2018) in Lexington, Kentucky.
Slides presented at the annual meeting of the American Society of Pharmacognosy, Lexington Kentucky (2018). Compile basic information of the principles of countercurrent separation, choice of solvent systems, and determination of partition coefficient.
Prepared by Drs. Charlotte Simmler, Brent Friesen, Guido Pauli from the Center for Natural Product Technologies (CENAPT)
You will find here all the elements presented by the CENAPT team ( Drs. Guido Pauli and Charlotte Simmler) and pertaining to the NMR workshop at the American Society of Pharmacognosy (ASP 2017, Portland Oregon).
These slides summarize the different steps related to the implementation of quantitative NMR for purity analysis.
This document lists and provides links to various commercially available continuous centrifugal precipitation chromatography (CCS) instruments. It mentions the Pharmatech CCC-1000, Kromaton FCPC-B-D, and CCBiotech bench-top centrifugal precipitation chromatograph models. The technology was invented at NIH and licensed to CCBiotech. It also lists instruments from Armen, Dynamic Extractions, Tauto, AECS, Everseiko. CCS is a process that can continuously fractionate high molecular weight molecules in a salt or solvent gradient applied through a membrane.
More from Center for Natural Product Technologies (9)
Authoring a personal GPT for your research and practice: How we created the Q...Leonel Morgado
Thematic analysis in qualitative research is a time-consuming and systematic task, typically done using teams. Team members must ground their activities on common understandings of the major concepts underlying the thematic analysis, and define criteria for its development. However, conceptual misunderstandings, equivocations, and lack of adherence to criteria are challenges to the quality and speed of this process. Given the distributed and uncertain nature of this process, we wondered if the tasks in thematic analysis could be supported by readily available artificial intelligence chatbots. Our early efforts point to potential benefits: not just saving time in the coding process but better adherence to criteria and grounding, by increasing triangulation between humans and artificial intelligence. This tutorial will provide a description and demonstration of the process we followed, as two academic researchers, to develop a custom ChatGPT to assist with qualitative coding in the thematic data analysis process of immersive learning accounts in a survey of the academic literature: QUAL-E Immersive Learning Thematic Analysis Helper. In the hands-on time, participants will try out QUAL-E and develop their ideas for their own qualitative coding ChatGPT. Participants that have the paid ChatGPT Plus subscription can create a draft of their assistants. The organizers will provide course materials and slide deck that participants will be able to utilize to continue development of their custom GPT. The paid subscription to ChatGPT Plus is not required to participate in this workshop, just for trying out personal GPTs during it.
PPT on Direct Seeded Rice presented at the three-day 'Training and Validation Workshop on Modules of Climate Smart Agriculture (CSA) Technologies in South Asia' workshop on April 22, 2024.
Mending Clothing to Support Sustainable Fashion_CIMaR 2024.pdfSelcen Ozturkcan
Ozturkcan, S., Berndt, A., & Angelakis, A. (2024). Mending clothing to support sustainable fashion. Presented at the 31st Annual Conference by the Consortium for International Marketing Research (CIMaR), 10-13 Jun 2024, University of Gävle, Sweden.
The cost of acquiring information by natural selectionCarl Bergstrom
This is a short talk that I gave at the Banff International Research Station workshop on Modeling and Theory in Population Biology. The idea is to try to understand how the burden of natural selection relates to the amount of information that selection puts into the genome.
It's based on the first part of this research paper:
The cost of information acquisition by natural selection
Ryan Seamus McGee, Olivia Kosterlitz, Artem Kaznatcheev, Benjamin Kerr, Carl T. Bergstrom
bioRxiv 2022.07.02.498577; doi: https://doi.org/10.1101/2022.07.02.498577
ESR spectroscopy in liquid food and beverages.pptxPRIYANKA PATEL
With increasing population, people need to rely on packaged food stuffs. Packaging of food materials requires the preservation of food. There are various methods for the treatment of food to preserve them and irradiation treatment of food is one of them. It is the most common and the most harmless method for the food preservation as it does not alter the necessary micronutrients of food materials. Although irradiated food doesn’t cause any harm to the human health but still the quality assessment of food is required to provide consumers with necessary information about the food. ESR spectroscopy is the most sophisticated way to investigate the quality of the food and the free radicals induced during the processing of the food. ESR spin trapping technique is useful for the detection of highly unstable radicals in the food. The antioxidant capability of liquid food and beverages in mainly performed by spin trapping technique.
Current Ms word generated power point presentation covers major details about the micronuclei test. It's significance and assays to conduct it. It is used to detect the micronuclei formation inside the cells of nearly every multicellular organism. It's formation takes place during chromosomal sepration at metaphase.
Describing and Interpreting an Immersive Learning Case with the Immersion Cub...Leonel Morgado
Current descriptions of immersive learning cases are often difficult or impossible to compare. This is due to a myriad of different options on what details to include, which aspects are relevant, and on the descriptive approaches employed. Also, these aspects often combine very specific details with more general guidelines or indicate intents and rationales without clarifying their implementation. In this paper we provide a method to describe immersive learning cases that is structured to enable comparisons, yet flexible enough to allow researchers and practitioners to decide which aspects to include. This method leverages a taxonomy that classifies educational aspects at three levels (uses, practices, and strategies) and then utilizes two frameworks, the Immersive Learning Brain and the Immersion Cube, to enable a structured description and interpretation of immersive learning cases. The method is then demonstrated on a published immersive learning case on training for wind turbine maintenance using virtual reality. Applying the method results in a structured artifact, the Immersive Learning Case Sheet, that tags the case with its proximal uses, practices, and strategies, and refines the free text case description to ensure that matching details are included. This contribution is thus a case description method in support of future comparative research of immersive learning cases. We then discuss how the resulting description and interpretation can be leveraged to change immersion learning cases, by enriching them (considering low-effort changes or additions) or innovating (exploring more challenging avenues of transformation). The method holds significant promise to support better-grounded research in immersive learning.
When I was asked to give a companion lecture in support of ‘The Philosophy of Science’ (https://shorturl.at/4pUXz) I decided not to walk through the detail of the many methodologies in order of use. Instead, I chose to employ a long standing, and ongoing, scientific development as an exemplar. And so, I chose the ever evolving story of Thermodynamics as a scientific investigation at its best.
Conducted over a period of >200 years, Thermodynamics R&D, and application, benefitted from the highest levels of professionalism, collaboration, and technical thoroughness. New layers of application, methodology, and practice were made possible by the progressive advance of technology. In turn, this has seen measurement and modelling accuracy continually improved at a micro and macro level.
Perhaps most importantly, Thermodynamics rapidly became a primary tool in the advance of applied science/engineering/technology, spanning micro-tech, to aerospace and cosmology. I can think of no better a story to illustrate the breadth of scientific methodologies and applications at their best.
6. Column Loading
0
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16000
0 50000 100000 150000 200000 250000
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mg loaded
mg loaded & volume
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0 2000 4000 6000 8000
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mg loaded & volume
Friesen, J. B.; McAlpine, J. B.; Chen, S.-N.; Pauli, G. F.,
Countercurrent Separation of Natural Products: An Update. Journal of
Natural Products 2015, 78, 1765-1796.
7. Model Compounds:
HO
H
H H
H
O
O
OH
OH
O
O O
HO
H H
H
CH3
OH
OH O
OHO
OH
N
O
OH
O
OH
O
HO
O
O
OH
O OHO
O
H
HO
H
HO
H
H
OHH
O
OH
OH
O
OH
O
O
OH OH
OH
O
O
OH
OH
HO
O
O
H
HO
H
HO
H
H
OHH
O
OH
OH
N
N
N
N
O
O
N
H
O
OH
NH2
N
N
OH
S
O
O
O
SO
O
O
S
O
O
O
3Na
N
H
N
O
OH
H
H
O
O
O
O
O
O
O
O
OH
OH
OH
OOH
HO
The GUESSmix
Friesen J.B, Pauli G.F. Journal of Liquid
Chromatography and Related Technologies, 28:
2777-2806, 2005
b
O
Q
r
R
U
F
Y
C
I
E
MZ
V
G
T X
H
D
N
A
8. GUESSmix Used to Evaluate SSs
The distribution coefficient (K) is a constant for a particular substance
in a particular solvent system.
Independent of: - column volume
- instrument model
- normal or reverse-phase mode
- run time
- rotation speed
- flow rate
G Mix H/tBME/ACN/Water 4:6:5:5 10/12/06
-0.1
0.8
0 25 50 75 100 125 150 175 200 225 250 275 300 325 350 375
mL & mn
A
280nm
230nm
K-Based Chromatography
9. CCC Chromatogram
0
0 20 40 60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500 520 540 560 580 600 620 640 660 680 700
A254
mL
GUESSmix in ChMWat 10:7:3 12/14/12, 35 degrees N Phase
Compounds were identified using TLC and CCC peaks
Q HD
FUE
C
V
r
11. Emphasize the importance of K
Using GUESSmix to explore solvent system families.
Friesen, J.B. Pauli, G.F. Analytical Chemistry 79: 2320-2324 (2007)
Symmetry
Midline
0
0 0.25 0.5 0.75 1 1.25 1.5 1.75 2 2.29 2.67 3.2 4 5.33 8 16 ¥K
A
M
Q
V
U
F
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Z E
16. Fig. 3. APCI-MS (pos. mode) molecular weight profile
chromatogram generated by sequential injections of even
numbered fractions of the HPCCC preparative separations of S.
terebinthifolius berries on the Midi and selected ion traces of
peaks 1–3. Each signal contained the complete APCI-MS profile
information of compounds present in a single assay tube (distance
between two monitoring signals is equivalent to 50.0 mL – Midi I
to III and 80 mL – Midi IV).
J Chromatogr A. 2015 Apr 10;1389:39-48. doi: 10.1016/j.chroma.2015.02.005.
Schinus terebinthifolius scale-up countercurrent chromatography (Part I): High
performance countercurrent chromatography fractionation of triterpene acids with off-
line detection using atmospheric pressure chemical ionization mass spectrometry. Vieira
MN, Costa Fd, Leitão GG, Garrard I, Hewitson P, Ignatova S, Winterhalter P, Jerz G
Column Loading
n-heptane/ethyl acetate/methanol/water (6:1:6:1)
17. Figure 4. HSCCC chromatogram of crude flavonol glycosides from Ginkgo biloba
leaves at flow rate 1.2 mL/min. n-hexane/butanol/ethyl acetate/methanol/0.5%
acetic acid (2:1:7:2:8)
J. Sep. Sci. 2007, 30, 2153 – 2159
Qiang Zhang, Li-Juan Chen, Hao-Yu Ye, Lei Gao, Wenli Hou, Minghai Tang, Guangli Yang, Zhenhua Zhong,
Yuan Yuan, Aihua Peng, Isolation and purification of ginkgo flavonol glycosides from Ginkgo biloba leaves
by high-speed counter-current chromatography
Column Loading
18. (a) HSCCC chromatogram when injection volume was 100 mg for MD-R.
(b) HSCCC chromatogram when injection volume was 600 mg for MD-R.
HEMWat 10:2:5:7, 800 rpm, lower phase mobile 2 mL/mi. 280 nm TBE300
J Chromatogr B 2016 Feb 1;1011:99-107. doi: 10.1016/j.jchromb.2015.12.051.
Separation and preparation of 6-gingerol from molecular distillation residue of Yunnan ginger rhizomes by high-speed counter-current
chromatography and the antioxidant activity of ginger oils in vitro. Gan Z, Liang Z, Chen X, Wen X, Wang Y, Li M, Ni Y.
Column Loading
19. Phytochem Anal. 2015 Nov-Dec;26(6):444-53. doi: 10.1002/pca.2579. Rapid Separation of Three Proanthocyanidin Dimers from Iris lactea Pall.
var. Chinensis (Fisch.) Koidz by High-Speed Counter-Current Chromatography With Continuous Sample Load and Double-Pump Balancing Mode.
Lv H, Yuan Z, Wang X, Wang Z, Suo Y, Wang H.
Figure 5. HSCCC chromatograms of the EPS at different load masses. HSCCC conditions: solvent system: EBuWat (9:1:10);
revolution speed: 900 rpm; separation temperature: 30 °C; flow rate: 2.2mL/min; detection wavelength: 280 nm; sample size:
(A) 50mg, (B) 100mg, (C) 150mg, (D) 200mg of the EPS in 5mL of the upper phase and 5 mL of the lower phase.
Column Loading
30. Sf and Flow Rate
Fig. 3. Stationary (upper) phase retention ratio in percentage of the column volume plotted versus the square root of the lower
mobile phase flow rate. CCC column volumes and rotor rotations: Mini 1.6mm I.D. 20.8mL and 1800 rpm; Mini 0.8mm I.D.
19.5mL and 2100 rpm; SFCC 1-coil 54mL and 800 rpm; SFCC 3-coil 156mL and 800 rpm. HepEMWat 2:3:2:3, head to tail
flowing direction. The regression equations give A intercepts and B slopes (Eq. (5)). The R2 regression coefficient is listed
below its equation.
Berthod2009_JCA_1216_4169_SmallVolume
34. Figure 3. HSCCC chromatogram of crude flavonol glycosides from Ginkgo biloba leaves. HSCCC conditions: column volume: 40 mL;
solvent system: hexane/butanol/ethyl acetate/methanol–0.5% acetic acid (1:0.5:3.5:1:4); stationary phase: lower; 1600 rpm; detection
wavelength: 254 nm; temperature: 25 oC; sample concentration: 20 mg/mL; Sf phase at 1.0, 1.2, and 1.5 mL/min flow rates: 78.1, 60.1,
and 45.4%, respectively.
J. Sep. Sci. 2007, 30, 2153 – 2159
Qiang Zhang, Li-Juan Chen, Hao-Yu Ye, Lei Gao, Wenli Hou, Minghai Tang, Guangli Yang, Zhenhua Zhong,
Yuan Yuan, Aihua Peng, Isolation and purification of ginkgo flavonol glycosides from Ginkgo biloba leaves
by high-speed counter-current chromatography
Flow Rate
35. Phytochem Anal. 2015 Nov-Dec;26(6):444-53. doi: 10.1002/pca.2579. Rapid Separation of Three Proanthocyanidin Dimers from Iris lactea Pall.
var. Chinensis (Fisch.) Koidz by High-Speed Counter-Current Chromatography With Continuous Sample Load and Double-Pump Balancing Mode.
Lv H, Yuan Z, Wang X, Wang Z, Suo Y, Wang H.
Figure 4. HSCCC chromatograms of the EPS at different flow rates. HSCCC conditions: EBuWat (9:1:10): 900 rpm: 30
°C;: 50mg of the EPS in 5mL of the upper phase and 5mL of the lower phase; detection 280nm; flow rate: (A)
1.2mL/min, (B) 1.5mL/min, (C) 1.8mL/min, (D) 2.2mL/min.
Flow Rate Gradients
37. Fig. 3. HSCCC chromatograms of the EFS. Peak 1: procyanidin B3, Peak 2: procyanidin B1, Peak 3: catechin, and Peak 4:
procyanidin B7. HSCCC conditions: solvent system: HEMWat (0.75:12.5:1:12.5, v/v/v/v); stationary phase: upper phase: 900
rpm; 30 C; 300 mg of the EFS: 280 nm; flow rate: 0–100 min, 1.5 mL/min, ∼100 min, (A) 1.5 mL/min, (B) 2.0 mL/min, (C) 2.5
mL/min, (D) 3.0 mL/min. Rs of 1.5 mL/min:67.19%; 2.0 mL/min:65.94%; 2.5 mL/min:64.06%; 3.0 mL/min:62.50%.
Journal of Liquid Chromatography & Related Technologies, 38: 1486–1493, 2015
DOI: 10.1080/10826076.2015.1063506 Separation and Purification of Four Flavan-3-ols From Iris
Lactea Pall. var. Chinensis (Fisch.) Koidz by High-Speed Counter-Current Chromatography with Flow-Rate Gradient
HUANHUAN LV, JIAN OUYANG, XIAOYAN WANG, XIAOFENG MA,2YOURUI SUO, and HONGLUN WANG
Flow Rate Gradients
38. Flow Rate & rpm
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p
m
flow mL/min
flow rate & rpm
Friesen, J. B.; McAlpine, J. B.; Chen, S.-N.; Pauli, G. F.,
Countercurrent Separation of Natural Products: An Update.
Journal of Natural Products 2015, 78, 1765-1796.
39. Flow Rate & Volume
0
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flow rate & volume
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v
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u
m
e
flow mL/min
flow rate & volume
Friesen, J. B.; McAlpine, J. B.; Chen, S.-N.; Pauli, G. F.,
Countercurrent Separation of Natural Products: An Update.
Journal of Natural Products 2015, 78, 1765-1796.
44. 0
200
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1000
1200
1400
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0 500 1000 1500 2000
v
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m
L
rpm
rpm and volume
rpm & Volume
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rpm
rpm and volume
Friesen, J. B.; McAlpine, J. B.; Chen, S.-N.; Pauli, G. F.,
Countercurrent Separation of Natural Products: An Update.
Journal of Natural Products 2015, 78, 1765-1796.
55. no temperature regulation
0
0.00 0.25 0.50 0.75 1.00 1.25 1.50 1.75 2.00 2.25 2.50 2.75 3.00 3.25 3.50 3.75 4.00
A254
25 C
0
0.00 0.25 0.50 0.75 1.00 1.25 1.50 1.75 2.00 2.25 2.50 2.75 3.00 3.25 3.50 3.75 4.00
A254
30 C
0
0.00 0.25 0.50 0.75 1.00 1.25 1.50 1.75 2.00 2.25 2.50 2.75 3.00 3.25 3.50 3.75 4.00
Sf = 0.54
Sf = 0.53
Sf = 0.53
2.29 2.67 3.2 4.00 5.33 8.00 16
8
2.29 2.67 3.2 4.00 5.33 8.00 16
8
2.29 2.67 3.2 4.00 5.33 8.00 16
8
Friesen JB, Pauli GF GUESSmix-guided optimization of elution–extrusion counter-current separations. Journal of Chromatography A 1216: 4225-4231 (2009)
Temperature
56. Temperature
Temperature influences both
K values and resolution.
Generally, K decreases while Temperature
increases.
Generally, Temperature influences
resolution because compounds respond
differently to Temperature changes.
Sf as a function of Temperature
0.4
0.5
0.6
0 10 20 30 40degrees C
Sf
Friesen JB, Pauli GF GUESSmix-guided optimization of elution–extrusion counter-current separations. Journal of Chromatography A 1216: 4225-4231 (2009)
57. Temperature
Generally, K tends toward unity while
Temperature increases.
K Value as a Function of Temperature
0
2
4
6
8
10
1 2 3 4 5 6 7 8
temperature
K
F
U
V
Q
M
N
E
5 10 15 20 25 30 35room
Friesen JB, Pauli GF GUESSmix-guided optimization of elution–extrusion counter-current separations. Journal of Chromatography A 1216: 4225-4231 (2009)
59. Parameter Type Parameter Experimental report
Essential Important Optional
Operational Flow rate E
Rpm E
Solvent system solvent and volume ratios E
Mobile phase identity E
Flow direction (head-to-tail, tail-to-head) E
Stationary phase volume ratio (Sf) E
Switch volume (Vex) of elution extrusion if used E
Column equilibration and sample injection method I
Temperature I
Pressure variation during experiment O
Gravitational field generated by rotation O
Solvent system phase composition O
SS interfacial tension O
SS density difference of phases O
Viscosity of each phase O
pH of aqueous phase O
Sample Loading mass of sample E
Loading volume E
Recovery mass of individual compounds I
Enrichment I
Composition of active fractions and analytical method I
Purity of target analytes and determination method I
Partition coefficient (K) of target analytes I
Percent recovery of target analytes O
Pauli GF, Pro S, Friesen B Countercurrent Separation of Natural Products Journal of Natural Products 71: 1489-1508 (2008)
dx.doi.org/10.1021/np800144q
Reporting Operational Parameters