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Chromatography, Gel: Chromatography on non-ionic gels without regard to the mechanism of solute discrimination.
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Quantification of the Immunosuppressant Tacrolimus on Dried Blood Spots Using LC-MS/MS

1iC42 Clinical Research and Development, University of Colorado, Anschutz Medical Campus, 2Division of Clinical Pharmacology, Cincinnati Children's Hospital Medical Center, 3Food and Drug Administration (FDA), Center of Drug Evaluation Research - Office of Generic Drugs, 4Transplant Clinical Research, University of Cincinnati

JoVE 52424


 Medicine

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Quantitative Detection of Trace Explosive Vapors by Programmed Temperature Desorption Gas Chromatography-Electron Capture Detector

1Chemical Sensing & Fuel Technology, Chemistry Division, U.S. Naval Research Laboratory, 2NOVA Research, Inc., 3Bio/Analytical Chemistry, Chemistry Division, U.S. Naval Research Laboratory, 4Navy Technology Center for Safety and Survivability, Chemistry Division, U.S. Naval Research Laboratory

JoVE 51938


 Chemistry

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Column Chromatography

JoVE 10217

Source: Laboratory of Dr. Jimmy Franco - Merrimack College

Column chromatography is one of the most useful techniques for purifying compounds. This technique utilizes a stationary phase, which is packed in a column, and a mobile phase that passes through the column. This technique exploits the differences in polarity between compounds, allowing the molecules to be facilely separated.1 The two most common stationary phases for column chromatography are silica gel (SiO2) and alumina (Al2O3), with the most commonly used mobile phases being organic solvents.2 The solvent(s) chosen for the mobile phase are dependent on the polarity of the molecules being purified. Typically more polar compounds require more polar solvents in order to facilitate the passage of the molecules through the stationary phase. Once the purification process has been completed the solvent can be removed from the collected fractions using a rotary evaporator to yield the isolated material.


 Organic Chemistry

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Ion-Exchange Chromatography

JoVE 10269

Source: Laboratory of Dr. B. Jill Venton - University of Virginia

Ion-exchange chromatography is a type of chromatography that separates analytes based on charge. A column is used that is filled with a charged stationary phase on a solid support, called an ion-exchange resin. Strong cation-exchange chromatography preferentially separates out cations by using a negatively-charged resin while strong anion-exchange chromatography preferentially selects out anions by using a positively-charged resin. This type of chromatography is popular for sample preparation, for example in the cleanup of proteins or nucleic acid samples. Ion-exchange chromatography is a two-step process. In the first step, the sample is loaded onto the column in a loading buffer. The binding of the charged sample to the column resin is based on ionic interactions of the resin to attract the sample of the opposite charge. Thus, charged samples of opposite polarity to the resin are strongly bound. Other molecules that are not charged or are of the opposite charge are not bound and are washed through the column. The second step is to elute the analyte that is bound to the resin. This is accomplished with a salt gradient, where the amount of salt in the buffer is slowly increased. Fractions are collected at the end of the column as


 Analytical Chemistry

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A Convenient Method for Extraction and Analysis with High-Pressure Liquid Chromatography of Catecholamine Neurotransmitters and Their Metabolites

1School of Public Health of Southeast University, Laboratory of Environment and Biosafety Research Institute of Southeast University in Suzhou, 2Key Laboratory of Child Development and Learning Science (Ministry of Education), School of Biological Science & Medical Engineering, Southeast University, 3School of Public Health, Tianjin Medical University, 4British Columbia Academy, Nanjing Foreign Language School

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JoVE 56445


 JoVE In-Press

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Deep Proteome Profiling by Isobaric Labeling, Extensive Liquid Chromatography, Mass Spectrometry, and Software-assisted Quantification

1St. Jude Proteomics Facility, St. Jude Children's Research Hospital, 2Departments of Structural Biology and Developmental Neurobiology, St. Jude Children's Research Hospital, 3Heilongjiang University of Chinese Medicine

JoVE 56474


 Biochemistry

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Construction of Models for Nondestructive Prediction of Ingredient Contents in Blueberries by Near-infrared Spectroscopy Based on HPLC Measurements

1United Graduate School of Agricultural Science, Tokyo University of Agriculture and Technology, 2Faculty of Agriculture, Tokyo University of Agriculture and Technology, 3Institute of Agriculture, Tokyo University of Agriculture and Technology

JoVE 53981


 Chemistry

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Internal Standards

JoVE 10225

Source: Laboratory of Dr. B. Jill Venton - University of Virginia

The goal of many chemical analyses is a quantitative analysis, where the amount of a substance in a sample is determined. In order to accurately calculate the concentration of an unknown from a sample, careful sample preparation is key. Every time a sample is handled or transferred, some of the sample can be lost. There are strategies however, for minimizing sample loss. There are also strategies for coping with sample loss and still making accurate measurements of concentration. To minimize sample loss, the ideal is to minimize the number of sample handling and transfer steps. For example, massing a solid sample directly into a flask that a solution will be made in reduces a transfer step. If it's necessary to transfer from one flask to another and a dilution is being made, then triple rinsing the glassware helps ensure all the sample is transferred. Other strategies are more specific to the sample. For example, samples that adsorb to glass, such as proteins, might better be handled in polypropylene disposable tubes. The tubes are not hydrophilic, so if a small amount of sample is to be pipetted in water, it is best to have already added the water to the tube, so the sample can be pipetted directly into the solve


 Analytical Chemistry

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Isolation and Characterization Of Chimeric Human Fc-expressing Proteins Using Protein A Membrane Adsorbers And A Streamlined Workflow

1Department of Chemical and Biomolecular Engineering, Ohio University, 2Biomedical Engineering Program, Russ College of Engineering and Technology, Ohio University, 3Department of Dermatology, Brigham and Women's Hospital, Harvard Medical School

JoVE 51023


 Bioengineering

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Characterization, Quantification and Compound-specific Isotopic Analysis of Pyrogenic Carbon Using Benzene Polycarboxylic Acids (BPCA)

1Department of Geography, University of Zurich, 2Department of Earth and Ocean Sciences, University of South Carolina, 3Department of Earth Sciences, ETH Zurich, 4Laboratory of Ion Beam Physics, ETH Zurich, 5Department of Geological Sciences, Stockholm University

JoVE 53922


 Chemistry

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