A Review on High Performance Liquid Chromatography (HPLC)
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Gas Chromatography-Mass Spectroscopy
Gas Chromatography-Mass Spectroscopy Introduction Gas chromatography-mass spectroscopy (GC-MS) is one of the so-called hyphenated analytical techniques. As the name implies, it is actually two techniques that are combined to form a single method of analyzing mixtures of chemicals. Gas chromatography separates the components of a mixture and mass spectroscopy characterizes each of the components individually. By combining the two techniques, an analytical chemist can both qualitatively and quantitatively evaluate a solution containing a number of chemicals. Gas Chromatography In general, chromatography is used to separate mixtures of chemicals into individual components. Once isolated, the components can be evaluated individually. In all chromatography, separation occurs when the sample mixture is introduced (injected) into a mobile phase. In liquid chromatography (LC), the mobile phase is a solvent. In gas chromatography (GC), the mobile phase is an inert gas such as helium. The mobile phase carries the sample mixture through what is referred to as a stationary phase. The stationary phase is usually a chemical that can selectively attract components in a sample mixture. The stationary phase is usually contained in a tube of some sort called a column. Columns can be glass or stainless steel of various dimensions. The mixture of compounds in the mobile phase interacts with the stationary phase. Each compound in the mixture interacts at a different rate. Those that interact the fastest will exit (elute from) the column first. Those that interact slowest will exit the column last. By changing characteristics of the mobile phase and the stationary phase, different mixtures of chemicals can be separated. -
22 Chromatography and Mass Spectrometer
MODULE Chromatography and Mass Spectrometer Biochemistry 22 Notes CHROMATOGRAPHY AND MASS SPECTROMETER 22.1 INTRODUCTION We know that the biochemistry or biological chemistry deals with the study of molecules present in organisms. These molecules are called as biomolecules and they form the basic unit of every cell. These include carbohydrates, proteins, lipids and nucleic acids. To study the biomolecules and to know their function, they have to be obtained in purified form. Purification of the biomolecules includes many physical and chemical methods. This topic gives about two of the commonly used methods namely, chromatography and mass spectrometry. These methods deals with purification and separation of biomolecules namely, protein and nucleic acids. OBJECTIVES After reading this lesson, you will be able to: z define the chromatography and mass spectrometry z describe the principle and important types of chromatographic methods z describe the principle and components of a mass spectrometer z enlist types of mass spectrometer z describe various uses of mass spectrometry 22.2 CHROMATOGRAPHY When we have a mixture of colored small beads, it is easily separated by visual examination. The same holds true for many chemical molecules. In 1903, 280 BIOCHEMISTRY Chromatography and Mass Spectrometer MODULE Mikhail, a botanist (person studies plants) described the separation of leaf Biochemistry pigments (different colors) in solution by using solid adsorbents. He named this method of separation called chromatography. It comes from two Greek words: chroma – colour graphein – to write/detect Modern separation methods are based on different types of chromatographic methods. The basic principle of any chromatography is due to presence of two Notes phases: z Mobile phase – substances to be separated are mixed with this fluid; it may be gas or liquid; it continues moves through the chromatographic instrument z Stationary phase – it does not move; it is packed inside a column; it is a porous matrix that helps in separation of substances present in sample. -
Coupling Gas Chromatography to Mass Spectrometry
Coupling Gas Chromatography to Mass Spectrometry Introduction The suite of gas chromatographic detectors includes (roughly in order from most common to the least): the flame ionization detector (FID), thermal conductivity detector (TCD or hot wire detector), electron capture detector (ECD), photoionization detector (PID), flame photometric detector (FPD), thermionic detector, and a few more unusual or VERY expensive choices like the atomic emission detector (AED) and the ozone- or fluorine-induce chemiluminescence detectors. All of these except the AED produce an electrical signal that varies with the amount of analyte exiting the chromatographic column. The AED does that AND yields the emission spectrum of selected elements in the analytes as well. Another GC detector that is also very expensive but very powerful is a scaled down version of the mass spectrometer. When coupled to a GC the detection system itself is often referred to as the mass selective detector or more simply the mass detector. This powerful analytical technique belongs to the class of hyphenated analytical instrumentation (since each part had a different beginning and can exist independently) and is called gas chromatograhy/mass spectrometry (GC/MS). Placed at the end of a capillary column in a manner similar to the other GC detectors, the mass detector is more complicated than, for instance, the FID because of the mass spectrometer's complex requirements for the process of creation, separation, and detection of gas phase ions. A capillary column is required in the chromatograph because the entire MS process must be carried out at very low pressures (~10-5 torr) and in order to meet this requirement a vacuum is maintained via constant pumping using a vacuum pump. -
Lecture Content
EMT 518: METHODS IN ENVIROMENTAL ANALYSIS III (2 UNITS) Lecturer: Professor O. Bamgbose SYNOPSIS Electro-analytical method: Potentiometry, Reference electrode – Calomel, Ag/Agcl, indicator electrodes – 1st, 2nd and 3rd order, Metal Electrodes, membrane electrodes – glass electrode, types of liquid junction potential, solid state electrode, potentiometric titration, end point location in potentiometric titration –visual, plot of E/V, plot of derivative curves 1st and 2nd electrogravimetry, fixed potential, constant current, constant cathode potential coulometry: constant current coulometry, coulometric titration. Voltammetry: classical polarography, Description of dropping mercury electrode, condition for polarographic determination, qualitative and quantitative analysis conductance methods: description of limiting ionic conductance, conductance cell, conductomertic titration. Thermal methods: Thermogravimetry, differential thermal analysis (DTA) LECTURE CONTENT POTENTIOMETRY Is a measurement of a given chemical species in an equilibrium system by the use of an electrode, while potentiometric titration is the technique that is used for following the changes in the concentration of chemical species as function of added titrant using an electrode. In both cases a cell is needed and a cell consists of the following: (1) Reference electrode (2) Liquid junction (3) Analyte solution (4) indicator electrode. It is also possible to have a cell without liquid junction. REFERENCE ELECTRODE. In carrying out a potentiometric determination the half cell potential of one electrode must be known which should be constant, reproducible and completely insensitive to the reference electrode and must be fully polarised throughout the duration of the measurement i.e the potential of the reference electrode does not change through the whole measurement. A classical example of reference electrode is the calomel electrode. -
Development of a New Analytical Method for Determination of Veterinary Drug Oxyclozanide by Electrochemical Sensor and Its Application to Pharmaceutical Formulation
chemosensors Article Development of a New Analytical Method for Determination of Veterinary Drug Oxyclozanide by Electrochemical Sensor and Its Application to Pharmaceutical Formulation Ersin Demir 1 and Hulya Silah 2,* 1 Department of Analytical Chemistry, Faculty of Pharmacy, Afyonkarahisar University of Health Sciences, Afyonkarahisar 03200, Turkey; [email protected] 2 Department of Chemistry, Faculty of Art & Science, Bilecik ¸SeyhEdebali University, Bilecik 11210, Turkey * Correspondence: [email protected]; Tel.: +90-228-214-1484 Received: 25 February 2020; Accepted: 26 March 2020; Published: 30 March 2020 Abstract: A novel highly selective, sensitive and simple analytical technique was recommended for the investigation of anthelmintic veterinary drug oxyclozanide based on square wave anodic stripping voltammetry (SWASV) by using a carbon paste electrode (CPE). According to the cyclic voltammetric data, the oxidation and electron transfer processes of oxyclozanide were found as irreversible and adsorption-controlled, respectively. The voltammetric anodic peak response was characterized with respect to pH, accumulation potential, accumulation time, frequency and pulse amplitude, etc. Under these optimized experimental conditions, the anodic peak density of oxyclozanide was linear to oxyclozanide concentrations in the range from 0.058 to 4.00 mg/L. The described electrochemical method was successfully carried out for the oxyclozanide in pharmaceutical formulation and tap water with mean percentage recovery of 101.5 % and 102.2 %, respectively. The results of pharmaceutical formulation studies were statistically compared to the high-performance liquid chromatographic method. Keywords: carbon paste electrode; determination; oxyclozanide; pharmaceutical formulation; voltammetry 1. Introduction Pharmaceuticals are necessary for human and animal health. They are an important class of emerging contaminants in the environment because, after usage, these compounds and their metabolites are widespread in the environment by several mechanisms. -
Four Channel Liquid Chromatography/Electrochemistry
Four Channel Liquid Chromatography/Electrochemistry Bruce Peary Solomon, Ph.D. The new epsilon family of electrochemical detectors from BAS can Hong Long, Ph.D. Yongxin Zhu, Ph.D. control up to four working electrodes simultaneously. There are several Chandrani Gunaratna, Ph.D. advantages to using multiple detector electrodes. By using four different Lou Coury, Ph.D.* applied potentials with electrodes placed in a parallel arrangement, a Bioanalytical Systems, Inc. hydrodynamic voltammogram can be generated quickly through Corporate R&D Laboratories 2701 Kent Avenue acquisition of four data points for every analyte injection. This speeds West Lafayette, IN method development time. In addition, co-eluting compounds in complex 47906-1382 mixtures can be resolved on the basis of their observed half-wave * corresponding author potentials by using the same arrangement of electrodes, also in parallel. This article presents a few examples of four-electrode experiments performed with epsilon detectors in the BAS R&D labs during the past few months, using both radial-flow and cross-flow thin-layer configurations. The epsilon Platform the past fifteen years, our contract These instruments are fully network- research division, BAS Analytics, able and will be upgradable over the BAS developed and introduced the has provided analytical data of the Internet. New techniques and fea- first commercial electrochemical de- highest quality to the world’s leading tures may initially be ordered àla tector for liquid chromatography pharmaceutical companies using carte, or added at any time when the over twenty-five years ago. With this state-of-the-art products from BAS, need arises. In the coming months, issue of Current Separations,BAS as well as other leading vendors. -
Analytical Applications of Nuclear Techniques
ANALYTICAL APPLICATIONS OF NUCLEAR TECHNIQUES ANALYTICAL APPLICATIONS OF NUCLEAR TECHNIQUES The following States are Members of the International Atomic Energy Agency: AFGHANISTAN GUATEMALA PERU ALBANIA HAITI PHILIPPINES ALGERIA HOLY SEE POLAND ANGOLA HONDURAS PORTUGAL ARGENTINA HUNGARY QATAR ARMENIA ICELAND REPUBLIC OF MOLDOVA AUSTRALIA INDIA ROMANIA AUSTRIA INDONESIA AZERBAIJAN IRAN, ISLAMIC REPUBLIC OF RUSSIAN FEDERATION BANGLADESH IRAQ SAUDI ARABIA BELARUS IRELAND SENEGAL BELGIUM ISRAEL SERBIA AND MONTENEGRO BENIN ITALY SEYCHELLES BOLIVIA JAMAICA SIERRA LEONE BOSNIA AND HERZEGOVINA JAPAN SINGAPORE BOTSWANA JORDAN SLOVAKIA BRAZIL KAZAKHSTAN SLOVENIA BULGARIA KENYA SOUTH AFRICA BURKINA FASO KOREA, REPUBLIC OF SPAIN CAMEROON KUWAIT CANADA KYRGYZSTAN SRI LANKA CENTRAL AFRICAN LATVIA SUDAN REPUBLIC LEBANON SWEDEN CHILE LIBERIA SWITZERLAND CHINA LIBYAN ARAB JAMAHIRIYA SYRIAN ARAB REPUBLIC COLOMBIA LIECHTENSTEIN TAJIKISTAN COSTA RICA LITHUANIA THAILAND CÔTE D’IVOIRE LUXEMBOURG THE FORMER YUGOSLAV CROATIA MADAGASCAR REPUBLIC OF MACEDONIA CUBA MALAYSIA TUNISIA CYPRUS MALI TURKEY CZECH REPUBLIC MALTA DEMOCRATIC REPUBLIC MARSHALL ISLANDS UGANDA OF THE CONGO MAURITIUS UKRAINE DENMARK MEXICO UNITED ARAB EMIRATES DOMINICAN REPUBLIC MONACO UNITED KINGDOM OF ECUADOR MONGOLIA GREAT BRITAIN AND EGYPT MOROCCO NORTHERN IRELAND EL SALVADOR MYANMAR UNITED REPUBLIC ERITREA NAMIBIA OF TANZANIA ESTONIA NETHERLANDS UNITED STATES OF AMERICA ETHIOPIA NEW ZEALAND URUGUAY FINLAND NICARAGUA UZBEKISTAN FRANCE NIGER GABON NIGERIA VENEZUELA GEORGIA NORWAY VIETNAM GERMANY PAKISTAN YEMEN GHANA PANAMA ZAMBIA GREECE PARAGUAY ZIMBABWE The Agency’s Statute was approved on 23 October 1956 by the Conference on the Statute of the IAEA held at United Nations Headquarters, New York; it entered into force on 29 July 1957. The Headquarters of the Agency are situated in Vienna. Its principal objective is “to accelerate and enlarge the contribution of atomic energy to peace, health and prosperity throughout the world’’. -
Reference Materials for Microanalytical Nuclear Techniques
IAEA-TECDOC-1295 Reference materials for microanalytical nuclear techniques Final report of a co-ordinated research project 1994–1999 June 2002 The originators of this publication in the IAEA were: Industrial Applications and Chemistry Section and IAEA Laboratories, Seibersdorf International Atomic Energy Agency Wagramer Strasse 5 P.O. Box 100 A-1400 Vienna, Austria REFERENCE MATERIALS FOR MICROANALYTICAL NUCLEAR TECHNIQUES IAEA, VIENNA, 2002 IAEA-TECDOC-1295 ISSN 1011–4289 © IAEA, 2002 Printed by the IAEA in Austria June 2002 FOREWORD In 1994 the IAEA established a Co-ordinated Research Project (CRP) on Reference Materials for Microanalytical Nuclear Techniques as part of its efforts to promote and strengthen the use of nuclear analytical technologies in Member States with the specific aim of improving the quality of analysis in nuclear, environmental and biological materials. The objectives of this initiative were to: identify suitable biological reference materials which could serve the needs for quality control in micro-analytical techniques, evaluate existing CRMs for use in micro-analytical investigations, evaluate appropriate sample pre-treatment procedures for materials being used for analysis with micro-analytical techniques, identify analytical techniques which can be used for characterisation of homogeneity determination, and apply such techniques to the characterization of candidate reference materials for use with micro-analytical techniques. The CRP lasted for four years and seven laboratories and the IAEA’s Laboratories in Seibersdorf participated. A number of materials including the candidate reference materials IAEA 338 (lichen) and IAEA 413 (single cell algae, elevated level) were evaluated for the distribution of elements such as Cl, K, Ca, Cr, Mn, Fe, Zn, As, Br, Rb, Cd, Hg and Pb. -
Complexometric Titration and Atomic Emission Spectroscopy) That Allow Us to Quantify the Amount of Each Metal Present Individually
Title Quantitative Analysis of a Solution Containing Cobalt and Copper Name Manraj Gill (Lab partner: Tanner Adams) Abstract In this series of experiments, we determine the concentrations of two metal species, copper and cobalt, in a mixture of the two by initially using an ion exchange column to separate them from the mixture. We consequently use two different techniques (complexometric titration and atomic emission spectroscopy) that allow us to quantify the amount of each metal present individually. This step-by-step analysis not only goes through the basis for the experimental approach but also is a breakdown of the measurements that leads us to the final results: the Co:Cu ratio in the unknown mixture. Purpose In this series of three experiments (Parts I – III), we aim to initially separate a solution containing a mixture of Cobalt, Co (II), and Copper, Cu (II), ions in an unknown ratio and in unknown concentrations. The end goal of the entire experiment is to determine how much Co and Cu were originally present in the sample we obtained in Part I. But to do so, as explained in further detail in the ‘Theory and Methods’ section below, we initially separate the two ions (Part I) based on their unique chemical properties and then we attempt to quantify the separated ions through two different methods (Parts II, III). The overall purpose is an analysis of the efficiency of the separation and quantification approaches used to measure the constituent compounds in a mixture. This “efficiency” can be determined by comparing the obtained measurements to the known amounts that were used to make the solution. -
Development and Evaluation of a Calibration Free Exhaustive Coulometric Detection System for Remote Sensing
University of Louisville ThinkIR: The University of Louisville's Institutional Repository Electronic Theses and Dissertations 5-2014 Development and evaluation of a calibration free exhaustive coulometric detection system for remote sensing. Thomas James Roussel University of Louisville Follow this and additional works at: https://ir.library.louisville.edu/etd Part of the Mechanical Engineering Commons Recommended Citation Roussel, Thomas James, "Development and evaluation of a calibration free exhaustive coulometric detection system for remote sensing." (2014). Electronic Theses and Dissertations. Paper 1238. https://doi.org/10.18297/etd/1238 This Doctoral Dissertation is brought to you for free and open access by ThinkIR: The University of Louisville's Institutional Repository. It has been accepted for inclusion in Electronic Theses and Dissertations by an authorized administrator of ThinkIR: The University of Louisville's Institutional Repository. This title appears here courtesy of the author, who has retained all other copyrights. For more information, please contact [email protected]. DEVELOPMENT AND EVALUATION OF A CALIBRATION FREE EXHAUSTIVE COULOMETRIC DETECTION SYSTEM FOR REMOTE SENSING by Thomas James Roussel, Jr. B.A., University of New Orleans, 1993 B.S., Louisiana Tech University, 1997 M.S., Louisiana Tech University, 2001 A Dissertation Submitted to the Faculty of the J. B. Speed School of Engineering of the University of Louisville in Partial Fulfillment of the Requirements for the Degree of Doctor of Philosophy Department of Mechanical Engineering University of Louisville Louisville, Kentucky May 2014 Copyright 2014 by Thomas James Roussel, Jr. All rights reserved DEVELOPMENT AND EVALUATION OF A CALIBRATION FREE EXHAUSTIVE COULOMETRIC DETECTION SYSTEM FOR REMOTE SENSING By Thomas James Roussel, Jr. -
Separation Science - Chromatography Unit Thomas Wenzel Department of Chemistry Bates College, Lewiston ME 04240 [email protected]
Separation Science - Chromatography Unit Thomas Wenzel Department of Chemistry Bates College, Lewiston ME 04240 [email protected] LIQUID-LIQUID EXTRACTION Before examining chromatographic separations, it is useful to consider the separation process in a liquid-liquid extraction. Certain features of this process closely parallel aspects of chromatographic separations. The basic procedure for performing a liquid-liquid extraction is to take two immiscible phases, one of which is usually water and the other of which is usually an organic solvent. The two phases are put into a device called a separatory funnel, and compounds in the system will distribute between the two phases. There are two terms used for describing this distribution, one of which is called the distribution coefficient (DC), the other of which is called the partition coefficient (DM). The distribution coefficient is the ratio of the concentration of solute in the organic phase over the concentration of solute in the aqueous phase (the V-terms are the volume of the phases). This is essentially an equilibration process whereby we start with the solute in the aqueous phase and allow it to distribute into the organic phase. soluteaq = soluteorg [solute]org molorg/Vorg molorg x Vaq DC = --------------- = ------------------ = ----------------- [solute]aq molaq/Vaq molaq x Vorg The distribution coefficient represents the equilibrium constant for this process. If our goal is to extract a solute from the aqueous phase into the organic phase, there is one potential problem with using the distribution coefficient as a measure of how well you have accomplished this goal. The problem relates to the relative volumes of the phases. -
HPLC Vs. UPLC
HPLC vs. UPLC WHAT IS HPLC? A: HPLC is High Performance Liquid Chromatography, which is formerly referred to as high-pressure liquid chromatography. HPLC is one of the most dominant and widely used technologies used in analytical labora- tories for the last 30 years. HPLC system can handle pressure up to 6000 psi. WHAT IS UPLC? A: UPLC is Ultra Performance Liquid Chromatography. UPLC became the modern standard HPLC platform due to its power for increasing sample throughput, chromatographic efficiency, sensitivity and decreasing run time. UPLC system can handle pressure up to 15,000 psi. WHAT’S THE DIFFERENCE BETWEEN UPLC AND HPLC? A: Both HPLC and UPLC are both liquid chromatography techniques used to separate the different com- ponents found in mixtures. However, UPLC operates at higher pressures (15,000 psi) and allows for lower particle sizes in columns, while HPLC operates at lower pressures (max <6000 psi). UPLC improves analyte resolution and sensitivity, lower solvent consumption, and shorten run times. THE ADVANTAGE OF UPLC: UPLC can shorten run time to decrease TAT, provides “ better resolution to eliminate the potential impact from complicated matrix (See below Figure), and has higher sensitivity for low components analysis. HPLC - broader, wider peaks with some overlap at UPLC - sharper peaks with clear separation and peak the baseline level. specificity – clear lift-off and touch-down to peak. 1945 Fremont Drive dyadlabs.com Phone+1.801.973.8824 Salt Lake City, UT 84104 USA [email protected]@dyadlabs.com 1 HPLC vs. UPLC WHAT’S THE APPLICATION OF UPLC? A: The main purposes for using UPLC are for identifying and quan- tifying the individual components of the complicated samples (e.g.