1. Analytical methods

2. Chromatography – general principles Mobile phase Stationary phase Compound for analysis - solute

3. Principles The sample to be analyzed is introduced in small volume to the stream of mobile phase. The solute’s motion through the column is slowed by specific chemical or physical interactions with the stationary phase as it traverses (passes through) the length of the column. The amount of retardation depends on the nature of the solute, stationary phase and mobile phase composition.

4. HPLC – High Performance Liquid Chromatography Mobile phase – liquid solvent Stationary phase - small silica beads (typical size = 5 micro metres) (1) Solvent reservoirs, (2) Solvent degasser, (3) Gradient valve, (4) Mixing vessel for delivery of the mobile phase, (5) High-pressure pump, (6) Switching valve in "inject position", (6') Switching valve in "load position", (7) Sample injection loop, (8) Pre-column, (9) Analytical column, (10) Detector (i.e. IR, UV), (11) Data acquisition, (12) Waste or fraction collector.

5. GLC or GC – Gas Chromatography

6. GC in more detail Stationary phase = 1.5 - 10 m in length and have an internal diameter of 2 - 4 mm Mobile phase – a gas - typical carrier gases include helium, nitrogen, argon, hydrogen and air. Which gas to use is usually determined by the detector being used Gas-liquid chromatography (GLC), or simply gas chromatography (GC), is a common type of chromatography used in organic chemistry for separating and analyzing compounds that can be vaporized without decomposition

8. Fluorescence spectroscopy Fluorescence spectroscopy aka fluorometry or spectrofluorometry, is a type of electromagnetic spectroscopy which analyzes fluorescence from a sample. It involves using a beam of light, usually ultraviolet light, that excites the electrons in molecules of certain compounds and causes them to emit light of a lower energy, typically, but not necessarily, visible light. A complementary technique is absorption spectroscopy.

10. Atomic absorption spectroscopy atomic absorption spectroscopy is a technique for determining the concentration of a particular metal element in a sample. The technique can be used to analyze the concentration of over 70 different metals in a solution.

12. IR or Infrared Spectroscopy Symmetrical and antisymmetrical stretching, scissoring, rocking, wagging and twisting Infrared spectroscopy (IR spectroscopy) is the subset of spectroscopy that deals with the infrared region of the electromagnetic spectrum. It covers a range of techniques, the most common being a form of absorption spectroscopy. As with all spectroscopic techniques, it can be used to identify compounds or investigate sample composition

14. BondCompound TypeFrequency range, cm -1 C-H Alkanes 2960-2850(s) stretch 1470-1350(v) scissoring and bending CH 3 Umbrella Deformation 1380(m-w) - Doublet - isopropyl, t- butyl C-HAlkenes 3080-3020(m) stretch 1000-675(s) bend C-H Aromatic Rings3100-3000(m) stretch Phenyl Ring Substitution Bands870-675(s) bend Phenyl Ring Substitution Overtones2000-1600(w) - fingerprint region C-HAlkynes 3333-3267(s) stretch 700-610(b) bend C=CAlkenes1680-1640(m,w)) stretch CºCAlkynes2260-2100(w,sh) stretch C=CAromatic Rings1600, 1500(w) stretch C-O AlcoholsAlcohols, Ethers, Carboxylic acids, EstersEthersCarboxylic acids Esters 1260-1000(s) stretch C=O AldehydesAldehydes, Ketones, Carboxylic acids, EstersKetonesCarboxylic acidsEsters 1760-1670(s) stretch O-H Monomeric -- Alcohols, Phenols3640-3160(s,br) stretch Hydrogen-bonded -- Alcohols, PhenolsAlcohols Phenols 3600-3200(b) stretch Carboxylic acids3000-2500(b) stretch N-HAmines 3500-3300(m) stretch 1650-1580 (m) bend C-NAmines1340-1020(m) stretch CºNNitriles2260-2220(v) stretch NO 2 Nitro Compounds 1660-1500(s) asymmetrical stretch 1390-1260(s) symmetrical stretch

16. NMR - Nuclear Magnetic Resonance