Introduction to Instrumental Analysis What are instrumental techniques

Introduction to Instrumental Analysis

What are instrumental techniques? Modern chemists use a range of instruments to analyse and identify substances. Most produce quantitative data, which requires expert interpretation. There are many different types of machine used for analysis, each producing a different type of information, such as: l whether a substance is pure or a mixture l the molecular mass of a compound l the types of bonds in a molecule l the arrangement of atoms in a molecule l the isotopes of different atoms in a substance. 2 of 37 © Boardworks Ltd 2009

Advantages of instrumental techniques Instrumental techniques are very powerful. They have many advantages over more old-fashioned chemical methods of analysis. This is because they: l do not usually damage the substance during testing l are much more sensitive than chemical techniques l can identify all the substances in a mixture l require only tiny amounts of a substance l are reliable and accurate l are quick. However the test results often require expert interpretation. 3 of 37 © Boardworks Ltd 2009

Using instrumental techniques A small amount of the substance under investigation is placed inside a machine, which then analyses its chemical contents. The scientist is then able to interpret and evaluate the results, and identify the elements and compounds in the substance. This could be forensic, health or environmental purposes. 4 of 37 © Boardworks Ltd 2009

Classification of instrumental analysis 1. Spectroscopy : which uses instruments generally known as a spectrometers. 2. chromatography: which uses instruments generally known as chromatographs 3. and electroanalytical chemistry: which uses electrodes dipped into the analyte solution. 5 of 37 © Boardworks Ltd 2009

General principle of analysis with electronic instrumentation • the standards and samples are provided to the instrument, the instrument measures the property, the electrical signal is generated, and the signal, or often the numerical value of the property itself, is displayed on a readout and subsequently related to concentration 6 of 37 © Boardworks Ltd 2009

instrument components In general, an instrument consists of four components: 1. sensor that converts a property of the solution into a weak electrical signal. 2. signal processor that amplifies or scales the signal and converts it to a useable form 3. readout device that displays the signal for the analyst to see. 4. power supply to provide the power to run these three components. 7 of 37 © Boardworks Ltd 2009

spectrochemical methods : all instrumental methods of analysis that involve the absorption or emission of light Spectroscopy: The science that deals with light and its absorption and emission by solutions and other material substances is called or spectrometry The broad term for the instrument used is spectrometer . 8 of 37 © Boardworks Ltd 2009

The Electromagnetic Spectrum 9 of 37 © Boardworks Ltd 2009

Absorption and Emission of Light 10 of 37 © Boardworks Ltd 2009

Instruments manufactured to measure ultraviolet or visible light absorption by a test sample have a light source from which a beam of light is formed. 11 of 37 © Boardworks Ltd 2009

What is spectroscopy? Spectroscopy is the process of investigating substances using electromagnetic radiation. There are many different spectroscopic techniques, each using a different frequency of electromagnetic radiation, including UV and visible light, infrared, radio waves and x-rays. Spectroscopy allows chemists to identify elements and investigate the detailed structure of compounds (including bonding and atom arrangement). 12 of 37 © Boardworks Ltd 2009

How do chemists use spectroscopy? Spectroscopic techniques are often the first thing a chemist might turn to in the analysis of a new chemical. When they find an interesting natural product, such as a molecule from tree bark which may have anti-cancer applications, they need to understand its structure. Spectroscopy will allow them to get a very detailed view of how its atoms fit together. When chemists carry out a reaction, they need to find out what they have made, and spectroscopy is the quickest and most reliable way of doing so. 13 of 37 © Boardworks Ltd 2009

Atomic absorption spectroscopy (ABS) is a technique that allows elements to be identified, and their concentration measured down to just a few parts per billion. ABS has many uses: l environmental chemistry – to analyse pollutant concentrations in air and water l medicine – to analyse concentrations of toxic chemicals in blood and urine l building – to check for impurities in concrete and steel l mining – to check how much metal is in an ore. 14 of 37 © Boardworks Ltd 2009

How does spectroscopy work? All spectroscopy uses the principle that electromagnetic radiation can be absorbed by atoms and molecules. Different parts of a molecule absorb different frequencies of radiation: Electromagnetic radiation Absorbed by Spectroscopic technique radio waves protons in nuclei nuclear magnetic resonance spectroscopy ultra violet and visible light electrons in atoms atomic absorption spectroscopy infrared electrons in bonds infrared spectroscopy 15 of 37 © Boardworks Ltd 2009

How does IR spectroscopy work? 16 of 37 © Boardworks Ltd 2009

MRI and NMR Magnetic resonance imaging (MRI) scans are often used in hospitals to provide images of bone and tissue. The images are made by investigating the nuclei of atoms with radiowaves. Nuclear magnetic resonance spectroscopy (NMR) is another name for MRI. Why do you think doctors choose to use the term MRI instead of NMR? 17 of 37 © Boardworks Ltd 2009

Paper chromatography is used to separate mixtures, especially dyes or pigments. chromatogram Dots of single dyes are placed alongside a dot of the unknown mixture. The solvent is drawn up the paper by capillary action. As the solvent moves up the paper, the pattern of the single dyes can be compared to that of the mixture. Which dyes does the mixture contain? 18 of 37 © Boardworks Ltd 2009

Which ink? 19 of 37 © Boardworks Ltd 2009

Rf values 20 of 37 © Boardworks Ltd 2009

Thin layer chromatography All chromatography involves a stationary phase and a mobile phase. In thin layer chromatography (TLC) the stationary phase is a layer of silica gel fixed onto a glass plate The mobile phase is a solvent which travels up the plate, silica gel carrying the substances. In paper chromatography, what are the stationary and mobile phases? 21 of 37 © Boardworks Ltd 2009

How does TLC work? TLC uses the same principals as paper chromatography. Capillary action still draws the solvent up the matrix; however while the molecules in paper chromatography are separated based on mass, in TLC, separation often depends upon solubility or charge, due to the interaction of solute and matrix. A dry sample is placed in the silica gel matrix. As the solvent front moves up the gel, it dissolves the sample and carries it up the matrix with it. Some of the particles in the sample stick more strongly to the silica gel than others, so they lag behind the solvent. Eventually the different substances in the sample separate out, with similar molecules travelling a similar distance. 22 of 37 © Boardworks Ltd 2009

TLC or paper chromatography Thin layer chromatography has a number of advantages over paper chromatography. 1. The glass plate is rigid, not flexible like paper, so it is easy to control. 2. After separation, the substances in the mixture can be recovered. The silica gel holding the separated substance is scraped off the glass plate and added to a solvent. The substance will dissolve and the silica gel can easily be removed by filtration. The glass plates can be re-coated with silica gel and used over and over again. 23 of 37 © Boardworks Ltd 2009

UV and locating agents Many substances are white or colourless, and so aren’t visible on a TLC plate. One way of making colourless substances show up is to use UV light. This usually works well for organic compounds. An alternative method is to use a chemical locating agent – a chemical that reacts with the substance to form a coloured compound. For example, when ninhydrin is exposed to an organic compound it stains it purple-brown. 24 of 37 © Boardworks Ltd 2009

Gas chromatography (GC) is used widely in many analytical laboratories, including forensic police labs, synthetic chemical labs, and drugs testing labs. In paper and thin layer chromatography, the mobile phase is a liquid. However, as the name implies, the mobile phase in GC is an inert gas. The stationary phase is usually a long thin tube of silica gel. 25 of 37 © Boardworks Ltd 2009

How does gas chromatography work? Like all forms of chromatography, GC uses a stationary phase to impede the movement of a mobile test substance. Different substances are attracted to the matrix by different amounts, and therefore journey along it at different speeds. In GC, the sample is injected into the machine, where it is vaporized. It is then washed over the matrix by an inert gas. Some substances will be more attracted to the matrix than others. These will take much longer to reach the detector. The detector measures the abundance of a substance at a given time, and this data is plotted on a graph. 26 of 37 © Boardworks Ltd 2009

Using gas chromatography 27 of 37 © Boardworks Ltd 2009

Testing for banned substances using GC All kinds of athletes are banned from taking performanceenhancing drugs – including racehorses! High-level competition horses are regularly tested for banned substances, such as painkillers that help them run through injury, or steroids that reduce inflammation. Urine samples are collected from the horses at events, and then sent to labs to be tested by gas chromatography. 28 of 37 © Boardworks Ltd 2009

Testing for banned substances 29 of 37 © Boardworks Ltd 2009

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