Paper Chromatography - Used to seperate mixtures and give information to identify substances - itinvloves a stationary phase and a mobile phase - Seperation depends on the distribution of substances between these phases - to carry out: place substance on the line near bottom of the papers, place it in the solvent and observe how far the substance travels. - The paper is the stationary phase —because it does not move - The Solvent is the mobile phase —because it moves. nk- A pure substance will produce a single spot in all solvents - The compunds in a mixture may seperate into different spots depending on the solvent. - An impure substance will show up more than one spot - The position of the spot may change but a single spot means its a pure substance - Different solvents have different distribution. ° ° ° ° es ==, PORE SUBSTANCES: 8 3 3 i Why do we draw the base line with a pencil? If we draw the base line with a pen, the pen ink would move up the paper.Using paper chromotography to identify an unknown substance Rf - Retention factor 1. Measure the distance moved by the substance from the baseline 2. Measure the distance moved by the solvent from the base line Distance travelled by 3. Use the formula: solvent Rf= Distance moved by the substance/Distance moved by the solvent 4, Different compunds have different Rf values in different solvents, which can be used to identify the compund. Always measure from the center - of the spotLocating Agents In medicine, Proteins, which are made of complicated molecules sucha as amino acids are colourless. Colourless substances are sprayed with a liquid which reveals them to make them visible. These liquids are called LOCATING AGENTS. A locating agent is a chemical which reacts with the substance to produce a visibly coloured product.Testing the purity of Substances 1. Melting point (Solids) - A pure substance has a fixed and exact m.p => If impure substances are present the temperature of m.p will range —> If impurities are present, it will lower the temperature example: Pure steoric acid - melts exactly at 70 degree C with impurities - melts between 68 to 70 degree C 2. Boiling point (Liquids) - A pure substance will have a fixed and exact b.p. ~> Presence of impurities will raise the b.p and cause the liquid to boil in a range of temperatures. 3. Chromatography ~ A pure substance shows only one spot on a chromatogram. —> used to test purity —> used to identify complicated chemicals like drugs or food additives which cannot easily be dissolved or melted. ~> used to identify synthetic dyes in food is harmless or not -> used to identify chemicals like pesticides or herbicides present on green vegetables.Methods of Purification 1. Dissolving, Filtration, Evaporation and Condensation. - This technique is only suitable for seprating mixtures of solids which behaves differently in a particular solvent (Liquid): 1. Solid is soluble (Dissolves) 2. Solid is insoluble ( not dissolve) example: Sand and Water —> Filtration : Sand is removed by filtration as it collects as the RESIDUE in the filter funnel.example: Salt and water solution Separating Mixtures: Evaporation - Salt solution can be evaporated to leave pure 0) meer crystals in the evaporating dish Mixture Evoporating dish - Slower the evaporation, Larger the crystals. (so end woter) 7 -Once the water has evaporated, The + Bunsen burner concentrated salt solution (semi solid) is left | to cool down to form salt crystals —This is called crystallization.Simple Distillation - Simple distillation is used to seperate a pure liquid from a solid, if we want the liquid (Opposite to evaporation/crystallization) How it works: 1. we boil the solution, steam is given off (evaporation) 2. it is cooled and condensed in a condenser 2 consisting of a jacket of cold water, where the coldest water enters from the bottom and wy SES circulating out through the top, letting no vapour t Fim] escape, 3. The water collected this way is very pure, as all distillate the impurities are left behind in the flask.Fractional Distillation Fractional distillation is used to seperate two liquids which dissolve in one another. They are said to be MISCIBLE liquids because they mix completely together. How it works (Water and Ethanol): = Seperation relies on the difference in boiling points of the two liquids. water - 100 degree C; Ethanol - 78 degree C 11 when flask is heated, both of the liquids will start evaporating, but the one with the lower b.p will evaporate faster. 2. When the vapour hits the fractionating colournn, it will condense and drip down 3. After repeated evaporation and condensation, Around 80 Degrees pure ethanol will be collected. And around 100 degrees water can be collected in a seperate container. > Fractionating column used is normally packed with glass beads or other unreactive substances—> It provides larger surface area for condensation, i pres | 7 / J 4 ‘Adapter condenser Water inlet Fractionating “sc q Receiver fas Distillation, Mixture ¢ sve ie vgs —