Sie sind auf Seite 1von 52

NATIONAL UNIVERSITY OF RWANDA FACULTY OF MEDICINE DEPARTMENT OF PHARMACY

LABORATORY PRACTICES OF CHEMISTRY

1. 2. 3. 4. 5.

SAFETY AND LABORATORY RULES COMMON LABORATORY MATERIALS AND THEIR USAGES PREPARATION OF SOLUTIONS TITRATIONS ORGANIC CHEMISTRY KAGISHA Vdaste, MSc

Academic year 2010

2 Chapter I: SAFETY AND LABORATORY RULES. I.1 Attitudes and Preparation a. Come to laboratory periods on time and mentally prepared by studying the experiment and planning your activities. This will help us to interact with each other. b. Be prepared physically; for example, don't try to do lab work on an empty stomach, or without sleep or when you have alcohol in your bloodstream. c. Write everything you do and see in your notebook so that you can trace your actions and make corrections if necessary. d. Wear sensible clothing, including shoes that are comfortable and permit rapid movement in case of emergency, and hair or hat that does not obstruct your view or dangle into the experiment. e. If you wear contact lenses, try to avoid wearing them in the lab. If you must wear contact lenses, your goggles must seal particularly well to your face. f. If you injure yourself, even slightly, report it to the safety-officer and/or your instructor, and seek first aid. If you experience eye irritation, flush your eyes at the nearest emergency eyewash station for 15 minutes (remove contacts) and seek medical attention immediately. g. If you have any existing physical conditions that might affect your performance, your health, or other peoples' health in the lab, please inform your instructor. This information will be kept confidential; examples might include pregnancy, medications, allergies, epilepsy, Special arrangements may be possible. I.2 Your Working Environment a GOGGLES for chemical splash protection are required at all times in labs or instrument rooms, i.e. all parts of the lab, even when your are not handling chemicals. The goggles will protect your eyes from most splashes and impacts. The goggles do not meet the standard if the air baffles are removed. Some people have trouble with their goggles fogging up. The best solution is to take a short break outside the lab to clean them. b. Rubber gloves are strongly recommended to protect you from absorption of chemicals through the skin. We also recommend a lab coat to protect your clothes and skin from your and your neighbor's spills. c. Keeping your bench space tidy will minimize breakage and spills of your valuable products. d. You are expected to clean up your own mess in community areas such as the IR room. e. Keep your glassware and other equipment cleaned up as you work. Having clean, dry glassware available at all times will save you much time in the long run. f. Be careful not to contaminate reagents with your spatulas or droppers. If you take too much of a reagent, give it to a needy neighbor - do no return it to the bottle. g. Do not wander off with the only bottle of a reagent that everyone needs; keep it in its assigned location. h. Be sure the aisles are passable.

Laboratory practices: Pharmacy department (Academic year 2010)

3 I.3. Glassware a. The most common laboratory injury is a cut occurring upon breakage of glass or porcelain. Most cuts can be prevented by careful work which prevents breakage. b. The safe procedure for inserting a glass tube or thermometer into a stopper with a hole is as follows: i. Be sure the tip of the tube is fire-polished. ii. Lubricate the glass with glycerol or water. iii. Be sure the hole in the stopper is large enough. iv. Grasp the glass about 1" (no farther) from the end and push and twist to insert it into the stopper. v. Be sure that the hand holding the stopper is not in line with the entering glass. vi. As the glass begins to slide into the rubber, move the hand holding the glass back a little, always keeping it no more than 1" from the rubber. vii. Most accidents occur because the glass snaps above the stopper from a force sideways (torque). Keeping your hand close to the stopper will help prevent your exerting a force sideways on the glass. viii. The above considerations apply also to attaching rubber hoses to condensers. The condenser should be in your hand (not clamped to an apparatus) and gripped close to the lubricated connector being inserted into the hose. c. Never use a thermometer as a stirrer! Always support a thermometer in a beaker or flask with a clamp. If a mercury thermometer breaks, immediately contact the laboratory instructor and restrict access to the area of contamination until cleanup can be arranged. d. Round-bottomed flasks will not stand upright by themselves and if rested on the counter will roll. They must be supported on a cork ring, in (not on) a beaker, or in a clamp. e. When glassware is assembled, care should be taken to use the minimum number of clamps needed for support, making sure: i. The clamp is attached to a vertical support bar. ii. No torque is applied by the clamp. iii. Top-heavy apparatus is prevented from rotating and tipping. iv. Hanging pieces are clamped - grease will not hold them against the force of gravity! f. Do not use a glass stopper to seal a hot container or you may never get it out again. Cork is recommended for organic solvents since rubber dissolves in organic solvents and viceversa. g. Graduated cylinders are metastable and tip easily with the touch of a sleeve. h. Report breakage of glassware to your instructor for disposal instructions. i. Think before cleaning equipment - it makes little sense to scrub a graduated cylinder that contained ether or a water-insoluble material with soap and water. I.4 Safety Equipment a. Fire Extinguishers for smothering fires. Departments policy regarding response to fires restricts the use of fire extinguishers to persons who are properly trained. Small fires may be extinguished by covering with a book or larger container. b. Fire Blanket for smothering fires.

Laboratory practices: Pharmacy department (Academic year 2010)

4 c. Safety Shower for rinsing chemicals off the body. d. Eye Wash Fountain for rinsing chemicals from the eyes. e. First Aid Kit - Note: even minor injuries must be reported to your instructor. f. At least two exits. g. Dustpan and broom for removing broken glass. I.5 Reagent a. Assume that a particular reagent is hazardous unless you know for sure it is not. b. Never taste a chemical unless specifically directed to do so. c. If you are instructed to smell a chemical, point the vessel away from your face and carefully fan the vapors toward your face with your hand and sniff gently. d. Dilute concentrated acids and bases by pouring the reagent into water (room temperature or lower) while stirring constantly. Never pour water into concentrated acids; the heat of solution will cause the water to boil and the acid to splatter. e. Avoid rubbing your eyes unless you know your hands are clean Use the fume hoods. Any experiment involving the use of or production of poisonous or irritating gases must be performed in a hood. f. Read the label. Read the label carefully, read it twice, before taking anything from a bottle. Many chemicals have similar names, such as sodium sulfate and sodium sulfite. Using the wrong reagent can spoil an experiment or can cause a serious accident g. Be aware of your lab neighbors' activities; you may be a victim of their mistakes. If you observe improper techniques or unsafe practices: h. Advise your neighbor. And advise your instructor if necessary. I.6 Toxic Hazards a. The materials used in the organic lab are the safest we can find consistent with your need to develop skills in working with hazardous materials in your career in science. b. Since you are wearing eye protection, the opportunity for liquids or solids to enter the eye is small. Chemicals in the eye should be immediately flushed with copious amounts of water using the eyewash fountain. c. To prevent inhalation of organic and inorganic vapors, do your experiments in the fume hood or under the minihoods on the bench. d. If your need to determine the odor of any material, waft it gently toward your nose with your hand - don't stick your nose in the container and inhale. e. Organic compounds can be absorbed through the skin, so be careful about spilling things. Wear rubber gloves to prevent contact with your skin, but treat the gloves as if they were bare skin, keeping them scrupulously clean. You might set aside a pen for laboratory work to minimize the possibility of contamination from your gloves via your pen to your hands and face. Obviously, chewing a pen or pencil that has been used in the lab would unwise. f. Organic vapors also can be absorbed into food or tobacco which you may ingest later. Moreover, any drinks brought into the lab could have things spilled into them. No food or drinks in the laboratory, not even stuffed in your backpack. If you do not have a locker to keep food in, please remove the food, drink and cigarettes to the hallway, or ask your instructor for a safe place to keep it. Smoking is not allowed in

Laboratory practices: Pharmacy department (Academic year 2010)

5 State buildings, as the nicotine and other contents of the smoke are well-known health hazards (look up the LD50 of nicotine if you are skeptical). g. If you spill a liquid on the bench, immediately soak it up with paper towels and, if it is volatile, transfer the towels to the hood. Inform your instructor as to the nature of the spill in case further action is warranted. h. If concentrated acid is spilled, add sodium carbonate or bicarbonate, solution or solid. If concentrated base is spilled, add dilute and/or weak acid (e.g. acetic). Indicator solution or paper will be available in the lab. If your skin (or clothing) comes in contact with the spill, immediately flush the skin or clothing with water for 15 minutes. i. Should you spill bromine solution anywhere, treat the spill immediately with sodium thiosulfate solution. j. Bottles of the reagents mentioned in g) and h) are available on the small counter above your bench. I.7 Heat Hazards a. Most organic compounds are flammable and may catch fire even in the absence of flame at high temperatures. b. Flames are rarely allowed in the organic laboratory. If flames are permitted by your instructor, plan your experiments so that you never leave your flame unattended. c. If you light a flame, you are responsible for the consequences, so check with your instructor for a safe location. d. If you use a bunsen burner, be sure to tie back your hair and be careful that hair or clothing are kept clear of the flame. e. If there is a flame in the neighborhood, do not pour flammables; organic vapors are usually denser than air and will flow along the bench without alerting you by their odors. f. Make sure you know the location of the nearest fire extinguisher and the nearest exit. g. Reactions that are exothermic or are being heated must be monitored; do not leave them without having someone watch. h. Never, never, never heat a closed system! Pressure will build up and cause the glass to fail, sending projectiles of glass in all directions. Do not depend on small leaks a substantial air exit must be provided. I.8 If There is a Fire a. In the lab where you are working. i. Shout "fire" to alert your neighbors and instructor if you discover it. ii. A small fire in a test tube or other container can usually be extinguished by covering the container with a watch glass or book. If the fire cannot be extinguished by one extinguisher or by sand or water, you will be instructed to evacuate, following the procedure in b). iii. One terrible possibility is that someone's clothing is set on fire. If the person runs, the flame will be increased by increasing the supply of oxygen. It must be smothered. Wrap the person in a lab coat, fire blanket, or whatever is handy to exclude oxygen. b. Elsewhere in the building (fire alarm sounds): i. Extinguish any flames and turn off electrical equipment. ii. Close any open windows and internal doors near you.

Laboratory practices: Pharmacy department (Academic year 2010)

6 iii. Walk quickly through the nearest exit to the hallway and leave the building by the nearest stairwell. iv. The last person leaving the room, usually your instructor, will close the hall door. v Know the ways to put out a fire. a) If it is open fire, such as a large chemical spill on a lab bench, the correct extinguisher should be used as follows: Pull the pin. Point the extinguisher (of dry) or hose (if CO2) at the base of the fire. Squeeze the handle while moving the extinguisher back and forth. NOTE: Be careful not to spread the fire by getting the nozzle of the extinguisher too close--the material being emitted is under pressure. I.9 Laboratory Electrical Equipment a. During your s studies you will use a variety of instruments to analyze your samples. As with all electrical equipment, a certain amount of care is needed to prevent fire, shock and damage to the equipment. Be careful not to bring water, especially on your hands, into contact with connected electrical equipment. b. The hot plates you are provided are powerful and seldom need to be set higher than c. Much of the heating in organic chemistry is done with electrical heating mantles; these must be plugged into a variable transformer, not directly into the outlet or they will overheat and may cause a fire. d. Never transfer anything into a flask that is sitting in a heating mantle; use a cork ring, beaker or clamp to hold the flask during transfers. Organics spilled in a mantle will catch on fire when the electricity is connected, acids or bases will corrode the wires, and water will cause a short circuit. e. Never pour into a container on an electronic balance - they often have the wiring and knife edges under the pan are thus easily damaged. f. Turn off electrical equipment immediately after you have finished unless your instructor has stated otherwise (e.g. the gas chromatographs must be left on for an hour to stabilize). g. Report frayed cords, or non-functional equipment to your instructor. Do not put it back in the cupboard or you will be stuck with it again next time. h. No samples are allowed on top of any instruments. I.10 Pressure Hazards a. Never heat a closed system. b. When using a separatory funnel, vent frequently and remove the stopper immediately upon setting it upright for separation. c. Compressed gas cylinders must be strapped to the bench above their center of gravity when the protective caps are off. Pressure regulators are generally not interchangeable between gases for safety reasons. Gas cylinders should be free of regulators and protected by their cap before moving.

Laboratory practices: Pharmacy department (Academic year 2010)

7 I.11 Waste Disposal a. In order to minimize damage to the environment, and in compliance with State chemical wastes must be separated into categories and carefully labeled as to their contents. Please read and follow the labels on the waste bottles to ensure that your chemical wastes are treated safely and appropriately. You will find containers for: i. General Organic Waste (flammable) ii. Halogenated Hydrocarbons (non-flammable) iii. Chromic Acid Solutions (these have been phased out) iv. Lead v. Silver vi. Other Heavy Metals vii. Waste from specific experiments in some cases. viii. Acids ix. Bases x. In some experiments, acids and bases will be neutralized to a pH of 6 - 10 (State law) as part of the experiment and flushed down the drain with lots of water. Your instructor will give you instructions in particular cases. Indicator solution or paper will be available in the lab. xi. Broken thermometers create the special problem of spilled mercury (a toxic heavy metal). Report such accidents immediately to your instructor; usually any mercury which cannot be collected is reacted with sulfur or absorbed with a special kit before disposal as heavy metal waste. xii. Broken glass or porcelain is swept up into a dust pan and disposed of in a special container for broken glass. Please don't use your fingers.

I.12 Summary 1. Approved safety goggles must be worn at all times. 2. No food, drinks or smoking are allowed. 3. Shoes must be worn. No bare feet or thong sandals are allowed. 4. No lab-work is permitted when you are alone in the lab, specially with any solvents or chemicals. 5. No open flames are allowed except as directed by the instructor. 6. Inform yourself about the location of fire extinguishers, safety equipment, emergency showers and eye-wash, and the emergency telephone numbers and the nearest exit. 7. No unauthorized experiments may be performed. 8. Do not use broken or cracked glassware. Check glassware before using it. 9. Never taste or smell chemicals. Consult chemical-safety handbook before using any unknown chemical (which is to be used for the first time). 10. Avoid contact of chemicals with skin. The use of rubber gloves is recommended. have lab coates on you all the time 11. Dispose of chemical waste as directed by chemical-safety handbook or instructor. 12. Clean your work area and put away all equipment and glassware before leaving. make sure equipment is put away in the correct locker- your personal locker or the common locker. 13. Put paper trash and broken glass in trash containers. 14. Keep instrument room clean and free of paper. Laboratory practices: Pharmacy department (Academic year 2010)

CHAPTER II: COMMON LABORATORY MATERIALS AND THEIR USAGE Distilling flasks: The figure (a) is the ordinary distilling flask.The sizes vary between 25 and 5000ml. (b) is the so-called Claisen flask, a distilling flask with two necks; the thermometer is placed in the neck currying the side arm. Sizes vary between 25 and 2000ml. It is of particular value in distillation where foaming or bumping occurs and is widely employed in distillations under diminished pressure. (c) is identical with (b) except that it is provided with a second long and indented neck. It is sometimes termed a Claisen flask with fractionating side arm. In(d) the side arm outlet extends a short distance into the long neck of the flask , thus preventing any vapour which has been in contact with cork or rubber stoppers from condensing and flowing down sides the arm. It is usually employed for those liquids which attack cork or rubbers stoppers.

2.1 CONDENSER The various types in common use are shown in the figure bellow. Type(a) is a typically Liebig condenser,which consists of an inner glass tube surrounded by a glass jacket trough which water is circulated. The inner jacket is fitted into the outer jacket by means of rubber stoppers; rubber tubing used formerly, is less durable and is not recommended. (b) is an all-glass Liebig condenser of similar design to (a); the jacket is sealed to the condenser tube . Two convenient sizes of suitable for general use have jacket of 20 and 40cm length. In the Pyrex glass West condenser greater efficiency of cooling is obtained by having a lightwalled inner tube and a heavy-walled outer tube with a minimum space between them. (c) is the inner tube of Liebig condenser. It is used as an air.condenser when the boiling point of the liquid is above 140-1500 (d) is an all-glass Allihn condenser. The condensing tube is made with a series of bulbs; this increases the condensing surface and lessens the resistance to the passage of vapours when the condenser is employed for refluxing. (e) is a typical double surface condenser( Davies type). It is far efficient more than any of the preceding types and the jacket is usually shorter

Laboratory practices: Pharmacy department (Academic year 2010)

9 (f) is an efficient spiral condenser of Friedrich type. The hot vapours can be introduced either at the side or the bottom, thus permitting the use of the condenser either for condensing vapours from another reaction vessel or for ordinary reflux purposes. (g) is coil condenser provided with an internal glasss coil or spiral. Ina modification there is both an internal spiral as well as an outer cooling jacket. (h) is a Dewar type of refluxing condenser. It is usually charged with a freezing mixture, e.g., Dry ice mixed with alcohol or acetone.

Laboratory practices: Pharmacy department (Academic year 2010)

10

2.2 VARIOUS KIND OF FUNNEL

Type (a) an ordinary filtration funnel having a 600 angle Type (b) a wide-stemmed funnel is used when transferring powders. Type (c), (d)and (e) are known as reparatory funnels;

Laboratory practices: Pharmacy department (Academic year 2010)

11 .2.1 FUNNEL SUITABLE FOR FILTRATION

The Buchner funnel (a) is used in conjunction with a filter flask into which is fitted by means of rubber stopper. The use of either a flat annular rubber ring or a cone to provide a seal between flask and funnel as shown in (c) or (d) respectively is often more convenient. The Hirsch funnel (b) has sloping sides and is designed to deal with a smaller amount of precipitate than the Buchner funnel. The funnel in the assembly (d) is a substitute for the Hirsh funnel. It consists of an ordinary glass funnel fitted with a witt plate (e) which is a perforated porcelain plate 1-4 cm of diameter, upon the filter paper can rest. The slit sieve funnel (f) is constructed entirely of glass (Jena or Pyrex) and therefore posses obvious advantages over opaque (porcelain) Buchner or Hirsch funnel. Similar advantages are apparent with the sintered glass funnel (g), which is available in a number of porosities coarse, medium and fine) 2.3 APPARATUS FOR MEASURING MASS Mass is measured using weighing balances. There are different types of weighing balances such as beam balances and electrical balances

Laboratory practices: Pharmacy department (Academic year 2010)

12

2.4 APPARATUS FOR MEASURING TEMPERATURE Temperature is measured using thermometers. There are different types of thermometers. The figure below shows the common type used in chemistry laboratories 2.5 APPARATUS FOR MEASURING TIME The apparatus for measuring time are usually watches and clocks. For accuracy during experiments in the laboratory, stop watches and stop clocks are used. Some of the common types are shown in the figure bellow

Laboratory practices: Pharmacy department (Academic year 2010)

13 2.6 OTHER COMMONLY USED APPARATUS ARE REPRESENTED BELOW DIAGRAMMATICALLY IN THE FIGURE:

Laboratory practices: Pharmacy department (Academic year 2010)

14
Name of Apparatus 1 Separating Funnel 2 Round bottomed flasks 3 Flat-bottomed flask 4 Gas jar 5 Deflagratiing 6 Trough 7 Thistle 8 Desicccator 9 Boiling -tube 10 Test-tube 11 Test-tube rack 12 Teat pipette(dropper) 13 Funnel 14 Wash battle 15 conical flask 16 test-Tube holder 17 Pipe-clay triangle 18 Evaporiting dish 19 Spatula 20 Tongs 21 Wire gauze Use Seprating immiscible liquids Used when heating liquid substances because heat is suplied uniformly Used for general laboratory experiences Used for gas collection Used for holding burning substances Used foe holding somer amount of water for some experiments Used for delivering liquid substances Used for drying sustances or keeping substances free from moisture Used when heating liquid or solid substances Used for general laboratory experiments Used for holding boiling tubes and test-tubes Used for delivering liquid substances drop-wise Used for delivering liquids carefully into vessels Used to hold water for rinsing of vessels Used for general laboratory experiments Used for holding hot test-tubes Used for supporting crucibles during heating Used when evaporating liquids Used for scooping solid substances from containers Used for holding metallic or non-meatllic substances Used for storing chemicals Used when glas is being heated or burned. Causes even distribution of heat when heating substances in beakers or flasks Used for storing chemicals Used for supporting and holding pieces of apparatus during experiments Used when heating solid substances that require strong heating Used for supporting beakers and flaski in which liquids are being heated Pestle is used for crushing substances while the mortar holds the substances being crushed

22 Reagents bottles 23 Clamp and stand 24 Crucible 25 Tripod stand 26 Pestle and mortar

Laboratory practices: Pharmacy department (Academic year 2010)

15 CHAPTER III: PREPARATION OF SOLUTIONS 3.1 BASIC CONCEPTS 3.1.1 EXPRESSIONS OF CONCETRATION In chemistry, a solution is a homogeneous mixture composed of two or more substances. In such a mixture, a solute is dissolved in another substance, known as a solvent. An aqueous solution is a solution in which the solvent is water. Concentration is the measure of how much of a given substance (solute) there is mixed with another substance (solvent water). This can apply to any sort of chemical mixture, but most frequently the concept is limited to homogeneous solutions, where it refers to the amount of solute in a substance. For scientific or technical applications, concentration is expressed in a quantitative way. There are a number of different ways to quantitatively express concentration; the most common are listed below. They are based on mass or volume or both. Depending on what they are based on it is not always trivial to convert one measure to the other, because knowledge of the density might be needed to do so. At times this information may not be available, particularly if the temperature varies. Mass can be determined at a precision of ~ 0.1 mg on a routine basis with an analytical balance. Both solids and liquids are easily quantified by weighing. Some units of concentration -particularly the most popular one (molarity)- require to express the mass of substance in the moles. The mole (symbol: mol) is the SI basic unit that measures an amount of substance. A mole is the amount of substance of a system which contains as many elementary entities as there are atoms in 0.012 kilogram (or 12 g) of carbon-12 (12C), where the carbon12 atoms are unbound, at rest and in their ground state. The number of atoms in 0.012 kg of 12C is known as the Avogadro constant (NA), and is determined empirically. The currently accepted value is NA = 6.02214179(30)1023 mol-1. When the mole is used to specify the amount of a substance, the kind of elementary entities (particles) in the substance must be identified. The particles can be atoms, molecules, ions, etc. The atomic mass of a chemical element is the mass of NA atoms (=1 mol) of it. The atomic masses of the elements are included in the periodic table of the elements.

Laboratory practices: Pharmacy department (Academic year 2010)

16 The molar mass of a molecule is the mass of NA such molecules. The molar mass of a molecule is equal to the sum of the atomic masses of its constituting atoms. For example, table salt is sodium chloride (NaCl). The atomic mass of sodium, given in the periodic table is 22.990 g/mol, and of chlorine is 35.453 g/mol. The molar mass of NaCl is, therefore: M(NaCl) = 22.990 + 35.453 = 58.443 g/mol. The molar mass M of a molecule, multiplied by the number of moles n, is equal to the total mass m (g) of the molecules: m = nM [1] In contrast to mass, a substance's volume is variable depending on ambient temperature and pressure. The volume of a liquid is usually determined by calibrated glassware such as burettes and volumetric flasks. For very small volumes precision syringes are available.

Volumetric flask Volumetric flasks (Fig.1; V=1l, 250 ml, 100ml, 5 ml, etc) are calibrated at a standard state temperature and pressure (25oC, 101.325 kPa). (The measurement of mass does not require such restrictions.) The use of graduated beakers and cylinders is not recommended as their indication of volume is mostly for decorative rather than quantitative purposes. The volume of solids, particularly of powders, is often difficult to measure, which is why mass is the more usual measure.

Laboratory practices: Pharmacy department (Academic year 2010)

17

3.1.2 CONCENTRATION: Molar concentration (Molarity) denotes the number of moles of a given substance per litre of solution.

C=

n V

Following the SI system of units, the National Institute of Standards and Technology, the United States authority on measurement, considers the term molarity and the unit symbol M to be obsolete, and suggests instead the 'amount-of-substance concentration' (c) with units mol/m3 or other units used alongside the SI such as mol/l. This recommendation has not been generally implemented in academia yet. When discussing the molarity of minute concentrations, such as in much pharmacological research, molarity is sometimes expressed in milimolars (mmol/l) or micromolars (1 millionth of a molar). 3.2 TO MAKE SOLUTION OF GIVEN MOLARITY AND VOLUME 1. Find the FW of the solute, usually from label. 2. Determine the molarity desired. 3. Determine the volume desired. 4. Determine how much solute is necessary by using the formula. 5. Weigh out the amount of solute. 6. Dissolve the solute in less than the desired final volume of solvent. Place the solution in a volumetric flask or graduated cylinder. Add solvent until exactly the required volume is reached, Bring To Volume, BTV EXAMPLE How much solute is required to make 300 mL of 0.8 M CaCl2? ANSWER FORMULA FW X molarity x volume = g solute needed 111.0 g) (0.8 mole) (0.3 L) = 26.64 g Laboratory practices: Pharmacy department (Academic year 2010)

18 mole L

Molal concentration, (Molality) denotes the number of moles of a given substance (n) per kilogram of solvent (ms)(not solution). The SI unit for molality is mol/kg. The determination of molality only requires a good balance, because the masses of both solvent and solute can be obtained by weighing. Using a balance is often more precise than working with volumetric flasks burettes and pipettes. Another advantage of molality is that it does not change with the temperature as it deals with the mass of solvent, rather than the volume of solution. Volume typically increases with increase in temperature resulting in decrease in molarity. Molality of a solution is always constant irrespective of the physical conditions like temperature and pressure. In a dilute aqueous solution near room temperature and standard atmospheric pressure, the molarity and molality will be very similar in value. This is because 1000 g of water roughly corresponds to a volume of 1 l at these conditions (supposing the density of water

Laboratory practices: Pharmacy department (Academic year 2010)

19 1 g/ml), and because the solution is dilute, the addition of the solute makes a negligible impact on the volume of the solution. However, in all other conditions, this is usually not the case. The mole fraction ?i, (also called molar fraction) denotes the number of moles of solute as a proportion of the total number of moles in a solution.
Xi = ni

ni
i

Mole fractions are dimensionless quantities. For instance: 2 mole of solute A is dissolved in 6 nA 2 moles of solvent B. Mole fractions: XA = = = 0.25 n A + nB 2 + 6 This measure is used very frequently in the construction of phase diagrams. It has a number of advantages: the measure is not temperature dependent, does not require knowledge of the densities of the phase(s) involved, a mixture of known mole fraction can be prepared by weighing off the appropriate masses of the constituents. As both mole fractions and molality are only based on the masses of the components it is easy to convert between these measures. Other, common used expressions of concentration are: v Mass percentage (denotes the mass of a substance in a mixture as a percentage of the mass of the entire mixture); v v Mass -volume percentage, Volume-volume percentage.

3.3 WEIGHT/VOLUME % Grams of solute 100 mL total solution EXAMPLE1 20 g of NaCl in 100 mL of total solution = 20% (w/v) solution. EXAMPLE2 How would you prepare 500 mL of a 5 % (w/v) solution of NaCl? ANSWER By equation 1. Total volume required is 500 mL 2. 5% = 0.05

Laboratory practices: Pharmacy department (Academic year 2010)

20 3. (0.05) (500 mL) = 25 4. 5. 6. 25 is the amount of solute required in grams. Weigh out 25 g of NaCl. Dissolve it in less than 500 mL of water. In a graduated cylinder or volumetric flask, bring the solution to 500 mL.

TWO OTHER FORMS OF % v/v mL solute 100 mL solution w/w g solute 100 g solution 3.4 WEIGHT/WEIGHT How would you make 500 g of a 5% solution of NaCl by weight (w/w)? ANSWER 1. Percent strength is 5% w/w, total weight desired is 500g. 2. 5% = 5g/100g 3. 5g X 500 g = 25 g = NaCl needed 100 g 4. 500 g 25 g = 475 g = amount of solvent needed Laboratory practices: Pharmacy department (Academic year 2010)

21 Dissolve 25 g of NaCl in 475 g of water PARTS Parts may have any units but must be the same for all components of the mixture. EXAMPLE: A solution is 3:2:1 ethylene:chloroform:isoamyl Might combine: 3 liters ethylene 2 liters chloroform 1 liter isoamyl alcohol PPM AND PPB v ppm: The number of parts of solute per 1 million parts of total solution. v ppb: The number of parts of solute per billion parts of solution. PPM EXAMPLE: 5 ppm chlorine = 5 g of chlorine in 1 million g of solution, or 5 mg chlorine in 1 million mg of solution, or 5 pounds of chlorine in 1 million pounds of solution 3.5 PPM TO MICROGRAMS/mL For any solute: 1 ppm in water = 1 microgram mL Each star represents 1 mg of dioxin. What is the concentration of dioxin in tube expressed as ppm (parts per million)? ____________ What is the total amount of dioxin in beaker? ___________ alcohol

Laboratory practices: Pharmacy department (Academic year 2010)

22

Each star represents 1 mg of dioxin. What is the total amount of dioxin in tube? 25 mg What is the concentration of dioxin in tube expressed as ppm? ____________ 1 ppm in water = 1 g mL 25 mg/500 mL = 0.05 mg/mL = 50 g/mL So the concentration is 50 ppm 3.6 MAKING DILUTIONS In common chemical practice, one frequently needs to prepare a new solution by dilution of the stock solution. This can be done following the mixing rule C0V0 ?? C1V1 where V0 is a volume of the stock solution at concentration C0, and V1 is a required volume of a final solution with its final concentration C1. The following example shows how to prepare 50 ml (V1) of 0.1 mol/l (C1) solution by dilution of the stock solution at concentration C0=1 mol/l: According to the mixing rule, we need the volumeV0 of a stock solution with concentration C0 C0V0 =?O C1V1 C1V1 V0 = C0
V0 = 50mlx 0.1mo / l = 5ml 1mol / l

Laboratory practices: Pharmacy department (Academic year 2010)

23 Therefore, we pour a small amount of the stock solution into the beaker and transfer 5 ml of it into 50 ml volumetric flask using the pipet. 0.1 mol/l solution is then obtained by filling the flask with distilled water up to the 50 ml mark. 3.6 STOCK SOLUTION We define a stock solution as a concentrate, that is, a solution to be diluted to some lower concentration for actual use. We may use just the stock solution or use it as a component in a more complex solution. We refer to the solution that we end up using as a working solution. If you are comfortable making dilutions then you can appreciate the many advantages of working with stock solutions. Although it is never absolutely necessary to use a stock solution, it is often impractical not to use them. Stock solutions can save a lot of time, conserve materials, reduce needed storage space, and improve the accuracy with which we prepare solutions and reagents. Here are several illustrated types of applications using stock solutions. How do I make a 10 millimolar solution of HCl ? It is easily you only need concentrate HCl and then dilute it into certain volume for desired molarity. Concentrate HCl with percentage of 38% HCl has molarity about 12.39 M, assume that you will make 10 millimolar (mM) HCl solution with 1 L in volume thus how many mL of concentrate HCl do you need? Just simple calculate using the dilution formula: V1xM1 = V2xM2 10 mM = 0.01 M thus 1 L x 0.01 M = V2 x 12.39 V2 = 8.07 L V2 = 0.807 mL so you need to take 0.0801 mL of concentrated HCl with graduated pipet and then put it into 1 L volumetric flask and finaly add distilled water until it reach 1 L. You can use other HCl with another molarity that prepared in your laboratory, for the example if there is 6M HCl, use this HCl and then calculated the solution needs using the same formula above.

Laboratory practices: Pharmacy department (Academic year 2010)

24 CHAPTER IV: TITRATION 4.1: INTRODUCTION Titration is an analytical method that can be used to determine the amount of an unknown present in a sample. Titration is the process of adding a reagent solution of known concentration (the titrant) to an unknown until all of the substance of interest is used up. In order for a titration to be useful for analysis, a number of different conditions need to be met. Among these are: a reaction with known ratios of reactants and products; stable reagents for the analysis; and a method for determining when the reaction is complete. 4.2: ACID-BASE TITRATION 4.2.1: TITRATION OF ACETIC ACID Objective: Determine the concentration of acetic acid by titration with a standard sodium hydroxide solution. Background: Review acid/base titrations in your textbook. A weak acid is a compound that partially ionizes in aqueous solution producing hydronium (H3O+) ions. The general equation for the dissociation of any weak acid can be written as: HA (aq) + H2O (l) ? A- (aq) + H3O+ (aq) (1) ? The addition of a strong base results in a neutralization reaction in which hydroxide ions (OH-) react with hydronium to produce water: H3O+ (aq) + OH- (aq) ? ? 2 H2O (l) (2) As hydronium is consumed in the neutralization reaction, the equilibrium in equation 1 is shifted to the right according to Le Chateliers Principle. The overall reaction for the neutralization process can be written as the sum of equations (1) and (2): HA (aq) + OH- (aq) ?-?A- (aq) + H2O (l) (3) The concentration of the unknown solution can be determined by measuring the volume of titrant added to reach the equivalence point. The equivalence point occurs when all of the acid has been neutralized by the base. At the equivalence point: MAVA = MBVB (4) Where MA is the molarity of the acid, MB is the molarity of the base and VA and VB are the volumes of the acid and base respectively. The equivalence point for the titration will be determined by using an indicator that changes color at the equivalence point. Phenolphthalein is pink in bases and clear in acids. When the titration mixture changes from clear to pink, the equivalence point has been reached. Procedure: Safety goggles must be worn at all times in the laboratory. 1. Obtain about 35 mL of the acetic acid solution in a clean dry Erlenmeyer flask. Also obtain about 70 mL of the sodium hydroxide solution in another clean dry Erlenmeyer flask (be sure to record the concentration). 2. Rinse a 50 mL buret with two separate 5 mL portions of sodium hydroxide. 3. Fill the buret with the NaOH, noting the starting volume. 4. Rinse a 25 mL pipette with a small portion of the acid solution.

Laboratory practices: Pharmacy department (Academic year 2010)

25 5. Pipette a 25.00 mL acid sample into a clean dry 150 mL beaker. Add three drops of phenolphthalein to the beaker. 6. Slowly add one to two mL of NaOH to the acid, stir. Continue adding NaOH in one to two mL increments and record the pH. When the pink color flashes through the entire sample, add NaOH drop wise. 7. When the solution remains pink you have reached the equivalence point, record the volume of NaOH. 8. Record the pH of your solution at the equivalence point. 9. Repeat with two additional trials. QUESTIONS 1) Write the balanced net ionic reaction that occurs between acetic acid and sodium hydroxide. 2) The pOH of a solution is 3.78; determine pH, [H3O+], and [OH-]. Is the solution acidic, basic, or neutral? 3) Explain why the pH at the equivalence point is not 7.00. 4) Write a balanced net ionic reaction for the titration of nitric acid with potassium hydroxide 5) What are the Arrhenius definitions of an acid and a base? 6) What are the Brnsted-Lowry definitions of an acid and a base? 7) Plot the curve pH=f(VNaOH) 4.2.2: ASPIRIN TITRATION Introduction: Purpose: In this experiment you will run a titration to determine the amount of aspirin (acetyl salicylic acid) present in an aspirin tablet. Aspirin is an acid. The active ingredient is acetyl salicylic acid. Different strengths of aspirin are based on the amount of active ingredients that they contain. Titration is a way to determine how much acid is in a solution by adding just enough base of a known concentration to neutralize the acid. In a neutralization, the number of moles of acid, H+, are combined with an equal number of moles of base, OH-. In the titration you will be performing, you will dispense base into a known amount of acid solution to find the unknown concentration. If you wanted to know the concentration of an unknown base, you could titrate the base with an acid in the same manner. The aspirin will be titrated against a solution of base, 0.100 M NaOH. Base will be dispensed from a burette into a beaker containing the dissolved (in ethanol) acid and phenolphthalein indicator, which will show a faint pink color in basic solutions. Reagents Materials: 0.100 M NaOH Burette Ethyl alcohol Mortar and pestle Aspirin tablets Phenolphthalein indicator 150 mL flask

Laboratory practices: Pharmacy department (Academic year 2010)

26

Procedure: 1. Grind a tablet into a fine powder by using a mortar and pestle. 2. Tare a piece of weighing paper on the balance. Carefully transfer as much powdered sample to a piece of paper and then determine the mass. 3. Place the powdered sample in a 150mL beaker. 4. Add a 10.0 mL portion of ethyl alcohol to the beaker and stir. 5. Add 25.0mL of water to the beaker. 6. Put 3 drops of the phenolphthalein indicator in your flask. Put a magnetic stir bar in your flask and place the flask on the center of the stir plate. 7. The burette is filled with 0.100M NaOH. Make sure there are no bubbles apparent in the burette. Record the initial volume on the burette. 8. Begin titrating, Add the NaOH in 1.0mL increments, making note of when the color change occurs. Continue adding base 5.0 mL past the equivalence point (the equivalence is approximately when the solution turned pink from the phenolphthalein). 9. Repeat steps 1-8 for the remaining tablets. Amount of Active Ingredient in Product Tested v v Calculate the moles of base used to neutralize the acid for each aspirin. Acetyl salicylic acid (C9H8O4) is not a strong acid, which means that for every mole that dissolves, not an entire mole of H+ dissociates from the acid. Nevertheless, what hydrogen ions that did dissociate were completely neutralized by the hydroxide added from the base. How many moles of H+ were neutralized? For simplicitys sake, we are going to assume that acetyl salicylic acid is a strong acid, and, therefore, the initial moles of H+ equals the initial moles of acid. Since we are comparing aspirin to aspirin, we will be able to obtain a relative comparison of the amount of acid in each aspirin. Calculate the mass of the acid for each aspirin based on the number of moles that reacted with base. Check the label on the bottles and determine if your calculation in #3 is valid.

4.2.3: TITRATION OF VINEGAR Chemists and laboratory technicians are often called upon to make analyses of the constituents of food products. Food and drug manufacturers are required to make public the ingredients in their products and often the percent of some or all of the components in a product. Vinegar is used in many food products. Sauces, relishes, and pickles all contain large quantities of vinegar. Vinegar is used in food preparation as a source of acidity (tartness) and flavor. Vinegar is produced by bacterial action upon fermented juices, wine or beer. The ethanol (CH3CH2OH) in the juice or wine is oxidized to acetic acid (CH3COOH) and minor amounts of other organic compounds which give the characteristic flavor of each type of vinegar. Acetic acid is the chemical in vinegar, which makes vinegar an important food additive. The pickling industry requires vinegar of a certain percent acetic acid in order to assure pickles of Laboratory practices: Pharmacy department (Academic year 2010)

27 good quality. Vinegar with a higher or lower concentration of acetic acid will make worthless pickles. As purchased from the store, most vinegar contains 4-5% acetic acid by weight. This means that in a 100g (approximately 100mL) sample of vinegar, there is 4-5g of acetic acid. Laboratory analysis of vinegar is done by a technique known as acid-base titration. In this technique, the acid (here acetic acid) in the solution is reacted with a base (here sodium hydroxide) to give a neutral salt and water.
O NaOH + OH ONa sodium acetate water O + H
2O

sodium hydroxide

acetic acid

When exactly all of the acetic acid is reacted or consumed by the sodium hydroxide, the solution changes from acid to neutral. We can observe this change by using an indicator which will change color at our desired pH. If we know how much sodium hydroxide we added to neutralize the acetic acid, then we will know the amount of acetic acid in our sample. This is because one mole of sodium hydroxide reacts with one mole of acetic acid. We use an indicator to tell us when the titration is complete. Indicators are organic compounds that change color when there is a sudden change in the pH of the solution. The end point of the titration is when a sudden change in the pH of the solution occurs. Therefore, we can tell the completion of the titration when we observe a change in the color of our solution to which a few drops of indicator have been added. The best way to monitor such a change in the pH is to use the indicator phenolphthalein which changes color from colorless to pink at pH 8.0-9.0. By performing a titration we can calculate the concentration of acetic acid in the vinegar. To do so we must know the volume of the acetic acid (5.00mL), the molarity of the base (the molarity will be approximately 0.200M, record the exact molarity from the label), and the volume of the base used to reach the end point of the titration. The volume of base will be read from the buret which is filled with the 0.200M sodium hydroxide. At the beginning of the titration the buret is filled to its maximum capacity. The base is then added slowly (dropwise) from the buret to the vinegar in an Erlenmeyer flask. Continuous swirling ensures proper mixing. The titration is stopped when the indicator shows a permanent pink coloration. The buret is read again. The volume of the base added is the difference between the initial volume (50.00mL) and the volume left in the buret at the end of the titration. Equipment/Materials: pH meter NaOH solution droppers buret buffer pH 7 & 10 vinegar stirrer and stir bar buret clamp

Laboratory practices: Pharmacy department (Academic year 2010)

28 beaker (100, 250 mL) funnel graph paper Procedure: There will be two brands of vinegar in the lab. Work in pairs. Each member of the pair will titrate one of the two vinegar samples 1. Rinse a 50mL burette with about 5mL of the 0.200M sodium hydroxide solution. Discard the rinse solution in the labeled waste container. After rinsing, fill the burette with the 0.200M sodium hydroxide solution. Use a clean dry funnel to fill the burette most of the way. Tilt the burette at a 45 degree angle, and slowly turn the stopcock to allow the solution to fill the tip (perform this operation over a beaker). If you do not succeed the first time, repeat the operation until the liquid in the burette forms a continuous column from top to bottom. Clamp the burette onto a ring stand using a burette clamp. Bring the memiscus exactly to the 50.00mL mark using a dropper. Record the exact concentration of the sodium hydroxide on your report. 2. Use a 5mL volumetric pipette and pipette filler to add 5.00mL of vinegar to a 125mL Erlenmeyer flask. Allow the vinegar to drain completely from the pipette by holding so that its tip touches the wall of the flask. Record the volume of vinegar and the initial burette reading on your report. 3. Add 3 to 5 drops of phenolphthalein to the flask with the vinegar. Also add about 10 mL of deionized water to the flask using your graduated cylinder to measure the water. The water is added to dilute the natural color that some commercial vinegars have. In this way, the natural color of the vinegar will not interfere with the color change of the indicator. 4. While holding the neck of the Erlenmeyer flask in your left hand, and swirling it, open the stopcock of the burette slightly with your right hand and allow the dropwise addition of the base to the flask. 5. Using your pH meter, record pH continuously per 1 mL of sodium hydroxide added. 6. At the point where the base hits the vinegar solution the color may turn temporarily pink but this color will disappear upon mixing the solution by swirling. Continue the titration until a faint permanent pink color appears. Stop the titration. Record the level of the sodium hydroxide in the burette on your report. Be careful not to add too much base, an error called "over titration". If the indicator in your flask turns deep pink or purple, you have over titrated and will need to repeat the entire titration with a new sample of vinegar. 7. Repeat this procedure (steps 1-4) two more times with fresh 5.00 mL sample of vinegar each time. Record your data from this second run on your report sheet. 8. Calculate the molarity of the vinegar for all three titrations. pipet 10mL analytical balance

Laboratory practices: Pharmacy department (Academic year 2010)

29 9. Average the three molarities. Calculate the percent concentration of acetic acid in your vinegar sample. Compare your results with the vinegar composition listed on the label of the vinegar you titrated. 10. Obtain the data for the alternate brand of vinegar (the brand that you did not titrate) from your lab partner. Titration of Vinegar - Pre-Laboratory 1. Write the balanced equation for the neutralization reaction and calculate the number of moles of vinegar neutralized. 2. Why was the buret rinsed with NaOH? 3. Calculate the molar mass of acetic acid. (HC2H3O2) 4. Calculate the number of grams of acetic acid in the vinegar 5. What is the Molarity of an acid, if it takes 25.27 mL of 0.198 M NaOH to titrate 5.00 mL of the acid. 6. Calculate the percent acetic acid in the vinegar from question 2 7. Graph the pH (y) vs volume NaOH (x).

Laboratory practices: Pharmacy department (Academic year 2010)

30 Titration of Vinegar - Data and Laboratory Report Name______________________________ Lab Day_____________ 1st run: buret reading initial: _____________________ buret reading final: ______________________ VB: ML NaOH pH ___________________________________ ML NaOH pH

Laboratory practices: Pharmacy department (Academic year 2010)

31 2nd run: buret reading initial: _____________________ buret reading final: ______________________ VB: ___________________________________

ML NaOH

pH

ML NaOH

pH

Laboratory practices: Pharmacy department (Academic year 2010)

32

3rd run: buret reading initial: _____________________ buret reading final: ______________________ VB: ___________________________________

ML NaOH

pH

ML NaOH

PH

Laboratory practices: Pharmacy department (Academic year 2010)

33 Brand: ______________________ 1st run MB VB VA MA % 2nd run 3rd run Manufacturers claim for % acidity:

1. What is the average % acetic acid in each sample? How do these results compare with label values (compare by using the percent difference)? 2. If your acetic acid sample contained a small amount of an alkaline impurity, would you expect the molarity of this sample to be the same, greater or less than a sample that contains the same amount of acetic acid, but no alkaline impurity? 3. If you over titrated your vinegar sample (the color was dark pink), would the calculated molarity of the acetic acid be the same, larger or smaller as the solution if it was not over titrated? 4.3: COMPLEXOMETRIC TITRATION 4.3.1: TITRATION OF CALCIUM AND MAGNESIUM IN WATER PURPOSE The purpose of this experiment is to determine the hardness of water by measuring the concentrations of calcium and magnesium in water samples by complexometric titration. Determining water hardness by EDTA All natural waters contain dissolved cations and anions. Water dissolves many ions as it flows through minerals. Although water hardness is defined as the quantity of cations with a +2 or +3 charge, calcium ion and magnesium ion are the most common of such ions in natural water. The formation of solid calcium carbonate is an endothermic process. Thus, when water containing both carbonate and calcium ions are heated, calcium carbonate can precipitate out onto the walls of pipes, boilers, and household items such as tea pots. This can shorten the life-time of some of these items.

Laboratory practices: Pharmacy department (Academic year 2010)

34 In addition, an insoluble scum develops when hard water comes into contact with soap. Both calcium and magnesium ions are responsible for this precipitate. This scum can be very difficult to clean. However, there is some evidence that hard water has beneficial health effects. Selenium, for example, may help prevent cancer. Soft water drinking supplies have been associated with an increased heart attack risk. The quantity of hardness ions will be determined by titration. EDTA, a weak acid, will be used as the titrant. In its ionized form, it is able to form soluble complexes with calcium and magnesium cations. The indicator added to the sample is Eriochrome Black T. Initially, the indicator will form a complex with the cations. When complexed it is red in color. As the EDTA is added dropwise to the sample, it replaces the Erio T and forms more stable complexes with calcium and magnesium. When the indicator is released by the metal ions, it has a distinct blue color. Therefore, the endpoint of the titration is marked by the color change from red to blue. Procedure v v v v v v Pipet 25-ml of the water sample into an Erlenmeyer flask and dilute to a total volume of approximately 50 ml. Add at least one ml of pH 10 buffer solution (1/2 of a Beral pipet) to the sample. The pH should be 10. To check pH, standardize pH meter. Place the magnetic stirrer in the beaker and turn on the stirrer slowly; making sure that the bar does not hit the electrode. Add a few drops Eriochrome Black T indicator to the Erlenmeyer. Fill the burette with EDTA solution 0.01M. Record the initial burette reading. Immediately begin to titrate the sample two drops at a time. Be careful to titrate slowly near the endpoint, as the color will take about 5 seconds to develop. Thus, add the last few drops at 3-5 second intervals. The endpoint color is blue. Record the initial and final burette reading to the nearest 0.1 mL.

CALCULATIONS v Hardness is expressed as parts per million (mg per liter) of equivalent CaCO3. For example, if the titration required 5 ml EDTA, the calculation would be:

Calculate the hardness of your sample in ppm of calcium carbonate

Compare the hardness of your sample with the founding of others

Laboratory practices: Pharmacy department (Academic year 2010)

35 4.3.2: DETERMINATIONS OF ALUMINUM AND MAGNESIUM IONS IN DRUG TABLET (ANTACIDS) 4.3.2.1 INTRODUCTION Antacids are useful to relieve acid indigestion, upset and sour stomach, or heartburn. Antacids may be divided into two classes: (a) chemical antacids work by chemical neutralization of gastric acid, for example sodium bicarbonate; and (b) adsorptive antacids act by adsorbing the acid, including aluminum and magnesium salts, and calcium carbonate. The former category shows the most rapid action, but may cause "acid rebound," a condition in which the gastric acid returns in greater concentration after the drug effect has stopped. The latter category is less prone to the rebound effect. Antacids containing aluminum ion prescribed with a low-phosphate diet can treat hyperphosphatemia or prevent formations of phosphate urinary and kidney stones. Antacids containing magnesium hydroxide and oxide with a larger dosage than normally required can produce a laxative effect. Antacids with aluminum and magnesium hydroxides, or aluminum hydroxide alone effectively prevent significant stress ulcer bleeding in postoperative patients or those with severe burns. Antacids including calcium ion use as diet supplements to prevent osteoporosis, but constipation and renal stone formation side effects may develop. According to the information in the Medline Plus of the National Library of Medicine (see the More Readings) related to antacid production in the U.S., there are 80 common brands. Antacids with aluminum used as the active ingredient includes alumina, aluminum hydroxide, and basic aluminum carbonate. Antacids with magnesium ion usually contain magnesia, and magnesium trisilicate, magnesium carbonate, magnesium alginate, magnesium hydroxide, and magaldrate, Antacid with calcium ion ordinarily has calcium carbonate. Among these brands, major productions consist of aluminum and magnesium ions. 4.3.2.2 ANALYTICAL METHODS In this experiment, the aluminum ion and magnesium ion contents in commercial antacid will be determined. First, the antacid tablets are weighted, dissolved in dilute nitric acid with heating, and then diluted with deionized water to a fixed volume. Analytical methods are designed with three protocols based on complexometric direct and back titrations along with proper indicators in appropriate buffer solutions. EDTA, ethylenediaminetetraacetic acid, (in quantitative analysis usually using disodium dihydrogen ethylenediaminetetraacetate, Na2EDTA) can form strong 1:1 complexes with most metal ions. It is most widely used as a chelating agent in analytical chemistry. Almost all metallic ions with two valences or more may be quantitatively analyzed using complexometric direct or back titration with EDTA. Complexometric back titration generally performs when the metallic ions form a stable complex with EDTA in a slow reaction or when a metal ion blocks an indicator. The blocked indicator cannot release metallic ions, thus no color change will be observable at the endpoint of complexometric direct titration. Both conditions exist in the case of aluminum ion, thus the ion is best determined by complexometric back titration along with heating to enhance the complexation of Al-EDTA.

Laboratory practices: Pharmacy department (Academic year 2010)

36 4.3.2.3 TOTAL ALUMINUM AND MAGNESIUM PROTOCOL The total aluminum and magnesium ion contents in antacids can be determined using complexometric back titration. In this determination, the sample solution is kept in a pH 10.0 buffer solution followed by adding an excess known amount of EDTA. Heating the solution prior to adding the indicator Eriochrome Black T or Calmagite is necessary to avoid aluminum ion blocking the indicator and to facilitate the complexation of the two metallic ions with EDTA. In the presence of unchelated EDTA, Eriochrome Black T or Calmagite remains pure blue color due to its free form. The amount of unchelated EDTA can be then determined using the back titration with standardized zinc solution. At the endpoint, a change to purple color is observable. The formation of Zn-Eriochrome or Zn-Calmagite complex is responsible for this color change. The endpoint detection should avoid unnecessary change to deep wine red color. Avoiding titrating with excess zinc solution will result in good accuracy. Table 1 is a summary used to illustrate the sequence and schematic quantities of metallic ions during performing the total aluminum and magnesium protocol. 4.3.2.4 ALONE ALUMINUM PROTOCOL The amount of aluminum ion alone in antacids can also be analyzed using complexometric back titration. In this analysis, the sample solution is controlled in a pH 5.0 buffer solution followed by adding an excess known amount of EDTA. At this low pH, the Al-EDTA complex can form, whereas the Mg-EDTA complexation is completely inhibited. This protocol is similar to total aluminum and magnesium protocol. To prevent aluminum ion blocking the indicator and to facilitate the Al-EDTA complexation, heating the mixture prior to adding the indicator xylenol orange is imperative. Xylenol orange in its free form is lemon yellow color. The amount of unchelated EDTA can be then determined using complexometric back titration with standardized zinc solution. At the endpoint, a change to light orange-red color is observable. This change is due to the formation of Zn-xylenol orange complex. Avoiding persisting deep red color at the endpoint will obtain an accurate result. Table 2 is a summary used to illustrate the sequence and schematic quantities of metallic ions during performing the aluminum alone protocol. 4.3.2.5 ALONE MAGNESIUM PROTOCOL The quantity of the magnesium ion alone in antacids can be determined using complexometric direct titration with EDTA. The sample solution is kept in a pH 10.0 buffer solution followed by adding a large amount of triethanolamine to completely generate Al-triethanolamine complexation and to entirely mask the formation of Al-EDTA complex. No heating is required in this protocol. Swirling can help to speed up the formation of Al-triethanolamine complex. The indicator Eriochrome Black T or Calmagite changes from blue to wine red color due to Mg-Eriochrome or Mg-Calmagite complexation. The endpoint detection is at which the color turns back to pure blue. This color change is due to whole liberation of Mg ion from the Mg-Eriochrome or MgCalmagite complex. Table 3 is a summary used to illustrate the sequence and schematic quantities of metallic ions during performing the magnesium alone protocol.

Laboratory practices: Pharmacy department (Academic year 2010)

37 4.3.2.6 REAGENTS PREPARATION 1. M EDTA solution: Weigh approximately 1.861 g of reagent grade disodium dihydrogen ethylenediaminetetraacetate dihydrate, Na2H2EDTA2H2O (molecular weight = 372.25) to precisely 0.0001 g. Quantitatively transfer it into a 500 mL of volumetric flask, add a half full of deionized water, swirl to enhance its dissolution, and then dilute to the calibration mark. Stopper the flask and mix it well by inverting and shaking repeatedly. (provided) 2. 0.01 M Standardized Zinc solution: Weigh out reagent grade zinc sulfate heptahydrate, ZnSO47H2O (molecular weight = 287.53) approximately 1.438 g to precisely 0.0001 g. Transfer it quantitatively into a 500 mL volumetric flask, add a half full of deionized water with swirling to dissolve it, and then dilute to the calibration mark. Stopper the flask and mix it thoroughly by inverting and shaking constantly. If a 1.4377 g of the zinc salt is used, the exact molarity of the solution should be 0.01000 M. 3. Acetate-acetic acid buffer solution: Dissolve about 20.0 g of sodium acetate trihydrate, CH3COONa` 3H2O, in a 1000-mL beaker containing about 1000 mL of deionized water. Slowly add 6 M hydrochloric acid with stirring throughout until a pH 5.0 0.1 of the solution is detected by a pH meter. (provided) 4. Bicarbonate-carbonate buffer solution: Dissolve about 30 g of sodium carbonate, Na2CO3, in a 1000-mL beaker containing about 1000 mL of deionized water. Slowly add 6-M hydrochloric acid with stirring throughout until a pH 10.0 0.1 of the solution is detected by a pH meter. (provided) 5. Xylenol orange indicator: Dissolve 0.10 g of the acid or sodium salt form of xylenol orange in 50 mL of absolute ethanol. The prepared indicator with lemon yellow color is suitable for analyzing solutions at pH = 5.0. The indicator prepared from the acid form is indefinitely stable, whereas that from the salt form may use only for several months. (provided) 6. Eriochrome Black T indicator: Dissolve 0.25 g of Eriochrome Black T with 50 mL of absolute ethanol. The blue indicator is suitable for analyzing solutions at pH = 10.0. If the indicator appears purple in color, add dropwise a pH 10 buffer solution until the color changes back to blue color. (provided) 4.3.2.7 SAMPLE PREPARATION (PROVIDED) 1. Obtain an antacid tablet from your TA and record its brand name, active ingredient(s) and the declared quantity of each component. 2. Weigh the tablet precisely to the nearest 0.0001 g. Grind it with a clean and dried mortar and pestle to make powder as fine as possible. Place the powder onto a weighing paper on a tare balance and precisely weigh about 0.7 g of it. Transfer the powder quantitatively to a clean 250-mL of Erlenmeyer flask. Then add about 25-mL of demonized water and 5-mL of conc. hydrochloric acid to the flask. 3. Boil the mixture for about 20 minutes on a hot plate. Place a stem funnel into the flask mouth so that the vapor can condense quickly back to water. This simultaneously helps to wash down any powder that sticks onto the flask wall. If any powder remains on the wall, wash it down with a small amount of demonized water and continue heating.

Laboratory practices: Pharmacy department (Academic year 2010)

38 4. Remove the flask from the hot plate and allow it to cool to near room temperature or rapidly in a water bath. Filter the mixture using gravity filtration into a 250-mL volumetric flask. Rinse the flask and the solid on the filter paper with about 10-mL of deionized water twice to make sure that all metallic ions are transferred into the volumetric flask. 5. Dilute the solution to the calibration mark with demonized water. Stopper the flask and mix the solution well by inverting and shaking it repeatedly. Label this solution The antacid sample solution. 4.3.2.8 EXPERIMENTAL PROCEDURE Part A: Standardization of the EDTA Solution 1. Prepare the 0.01M standardized zinc solution according to the Reagents Preparation Procedure. 2. Pipet a 10.00 mL aliquot of 0.01 M standardized zinc solution into a 125 mL Erlenmeyer flask followed by adding about 20 mL of the bicarbonate-carbonate buffer solution (pH 10.0 0.1). 3. Add 3 drops of Eriochrome Black T indicator and mix it well. The solution should appear wine red in color. 4. Direct-titrate the solution with 0.01M EDTA solution until the color changes to pure blue at the endpoint. 5. Repeat the titration twice. 6. Calculate the exact concentration of the EDTA solution. Part B: Determination of Total Aluminum and Magnesium Contents 1. Pipet a 10.00 mL aliquot of the antacid sample solution into a 125 mL Erlenmeyer flask followed by adding about 20 mL of the bicarbonate-carbonate buffer solution (pH 10.0 0.1). Transfer quantitatively a 30.00 mL aliquot of 0.01 M EDTA solution to the flask using a burette. 2. Boil gently the mixture for 5 min. on a hot plate to speed up the formation of Al-EDTA complex. Add 3 drops of Eriochrome Black T indicator and mix it well. The solution should appear pure blue in color. If the EDTA is not enough to chelate all of the metallic ions completely, the solution should be wine red in color at this moment. In this case, put an additional 5.00 mL or more aliquot of the EDTA solution to this wine red solution. Boil again until the color changesto pure blue. 3. Back-titrate the solution with standardized zinc solution until the color changes to purpleblue at the endpoint (no wine red color should persist). 4. Repeat the titration twice. 5. Calculate the total millimoles of aluminum and magnesium ions in the antacid sample solution and in the tablet.

Laboratory practices: Pharmacy department (Academic year 2010)

39 Part C: Determination of Alone Aluminum Content 1. Pipet a 10.00 mL aliquot of the antacid sample solution to a 125 mL of Erlenmeyer flask. Add about 20 mL of the acetate-acetic acid buffer solution (pH 5.0 0.1) to mask the formation of Mg-EDTA complex. Transfer quantitatively a 20.00 mL aliquot of 0.01 M EDTA solution to the lask using a burette. 2. Boil it gently on a hot plate for 5 min. to speed up the formation of Al-EDTA complex. Add 10 drops of xylenol orange indicator and mix well. The solution should appear lemon yellow in color at this moment. If the EDTA is not enough to chelate aluminum ion, the solution should be deep red color. In this case, put an additional 5.00 mL or more aliquot of the EDTA solution to this deep red solution. Boil again until the color changes to lemon yellow. 3. Back-titrate the solution with a standardized zinc solution until the color changes to light orangeyellow at the endpoint (no deep red color should remain). If the light orangeyellow color shortly turns back to lemon yellow, continuously titrate the solution until a light orange-yellow color persists for more than 3 minutes. Note: The turning back slowly to lemon yellow color results from the complex formation of EDTA with zinc ion at a low pH solution, which is thermodynamically more stable and kinetically slower than the Zn-indicator complexation. Thus, in this protocol slow titration will give good results. 4 Repeat the titration twice. 5 Compute the millimoles and weights of aluminum ion in the antacid sample solution and in the tablet. 4.3.2.9 DATA TREATMENT Part A: Standardization of the EDTA Solution 1. Calculate the exact concentration of EDTA solution. Part B: Determination of Total Aluminum and Magnesium Contents 2. Calculate the total concentration of aluminum and magnesium of the sample. Part C: Determination of Alone Aluminum Content 3. Calculate the concentration of aluminum of the sample. 4. Base on part B and C, Calculate the concentration of magnesium of the sample. 4.3.2.10 QUESTIONS: 1. From your calculation, compare your own results with declared concentration for the sample you analyzed. Explain if there are significant differences. 2. If back-titration is fast in the Part C for the determination of aluminum content alone, what would be the effect on the reading of the standardized zinc solution used and the calculated concentration of aluminum content in the sample? Explain. What would be the effect on the calculated active ingredient values in an antacid for each g? Explain: (i) An insufficient time of boiling the solution in the Part B. (ii) Adding indicator prior to heating in the Part C

Laboratory practices: Pharmacy department (Academic year 2010)

40 4.3.2.10 HAZARDS Disodium dihydrogen ethylenediaminetetraacetate dihydrate, zinc sulfate heptahydrate, potassium acetate, and sodium carbonate may cause irritation to skin, eyes and the respiratory tract. Nitirc acid is corrosive and damageable to the skin, eyes, and respiratory tract. Triethanolamine causes skin irritation and severe eye irritation. Ethanol is highly flammable liquid; there should be no open flames in the laboratory. Avoid breathing the vapors. Use with adequate ventilation. Xylenol orange, Eriochrome Black T and calmagite may cause irritation. Avoid contact with the eyes and skin. Magnesium hydroxide, aluminum hydroxide and hydrotalcite are the active ingredients in commercial antacids, thus there is no hazard in normal handling. Solutions will spatter if heated too strongly. To avoid this, heat solutions slowly and allow only gentle boiling. Placing a funnel into the flask mouth will prevent spatter. Immediately flush the affected skin with plenty of water for at least 15 minutes. Protective gloves and goggles must be worn at all times. . 4.4: REDOX TITRATION 4.4.1: MANGANIMETRIC TITRATION INTRODUCTION Iron ablets are prescribed for anaemia. The iron in commercial iron tablets is in the form of Fe2+. This can be oxidised to Fe3+ by the manganate (VII) (permanganate) ion. This is the reaction which will form the basis of our titration. MnO4- + 8 H+ + 5 e-? Mn2+ + 4 H2O Fe2+? Fe3+ + e5 Fe2+ + MnO4- + 8 H+ ? 5 Fe3+ + Mn2+ + 4 H2O v The purple MnO4- ion becomes colourless when it reacts with the Fe2+ ions. The endpoint of the titration (that is, when all the Fe2+ ions have been used up) is that point when the purple colour of MnO4- no longer disappears after addition to the iron solution. v The iron tablet also contains chalk powder, sucrose, and other minor ingredients. In order to prepare this tablet for analysis you will need to grind it and transfer it to a volumetric flask. I recommend using 2 tablets and in addition, the tablets should be dissolved in H2SO4. APPARATUS beakers, 250 cm3 weighing bottles burettes, 50 cm3 measuring cylinder, 25 cm3 pipettes, 25 cm3 thermometer volumetric flasks, 250 cm3 Bunsen burner, tripod & gauze Conical flasks, 250 cm3 REAGENTS potassium manganate (VII) solution sulphuric acid iron tablets distilled water

Laboratory practices: Pharmacy department (Academic year 2010)

41

PROCEDURE. v Accurately note the mass of one iron tablet. v Crush these tablets to a fine powder in a mortar with a pestle. v Rinse all of the crushed tablets into a conical flask and add about 25 cm3 of dilute sulphuric acid (1 M H2SO4 ). Add more water (the final solution should have a final volume of 100ml) and / or warm if you experience difficulty in getting the tablet to dissolve completely. v Titrate 10.00 mL samples of your tablets solution with 0.005 M potassium permanganate. Remember that you have reached the endpoint of the titration when the purple colour of the permanganate just begins to stay - this indicates that all of the Fe2+ ions have reacted. Do at least three determinations although you may have to do more to ensure that your values are within 0.3 mL of each other. v Repeat the titration accurately in the chosen manner. QUESTIONS 1. Calculate the mass of iron in one tablet. 2. Calculate the percentage mass of iron in a tablet 3. Compare your values with other members of the group and find average values. The following steps will help you in your calculations 1. Determine the number of moles of MnO4- used to react with 10.0 mL Fe2+ solution (you know the volume and concentration). 2. . Calculate the number of moles of Fe2+ that reacted (use the reaction equation) 3. . Calculate the number of moles that would have reacted in 50.0 mL of Fe2+ solution (since this contains 1 tablet). 4. The number of moles of Fe2+ is the same as the number of moles of FeSO4. Calculate the mass of FeSO4 in 1 tablet. 4.4.2: IODOMETRY Iodometry is a method of volumetric chemical analysis, a titration where the appearance or disappearance of elementary iodine indicates the end point. It involves the following reaction: I2 (aq) + 2e2I- (aq) 1. Titration with reducing agent Iodine, I2, is sparingly soluble in water but dissolves in potassium iodide solution, KI (aq), in which it forms a complex ion, KI3, potassium iodide. This solution of Iodine and potassium iodide is known as Lugol's solution or Lugol's iodine. First made in 1829, it was named after the French physician J.G.A. Lugol. I2 (aq) + 2II3- (aq)

Laboratory practices: Pharmacy department (Academic year 2010)

42 PROCEDURE v Put in burette an aqueous solution of sodium thiosulphate Na2S2O3 whose the concentration is 0.1M v Put in conical flask 20ml of aqueous solution of lugol (2g I2+6gKI) v Titrate the iodine solution by Na2S2O3 until the solution becomes pale yellow v Add some drops of starch ( the solution becomes blue) v Continue the titration until the blue color despairs. QUESTIONS Write down the reaction for this titration if the redox potential of involved couples are: I2/I-, E0=0,54V 2S2O4 /S2O32- E0=0.09V 2. TITRATION WITH OXIDIZING AGENT In acidic solutions, Hydrogen peroxide (H2O2) reacts as an oxidizing agent and oxidizes Iodide Ito I2. The elementary iodine formed is then reduced to iodide I- by thiosulfate. Knowing the quantity of thiosulfate used in the titration, we can then deduce both the quantity of Iodine and the quantity of Hydrogen peroxide in the solution. We have to consider 2 Half-reactions: 1. The reduction of Hydrogen peroxide (H2O2) to H2O; Standard potential: E= 1.77V 2. Oxidation of Iodide I- to Iodine I2; Standard potential: E= 0.53V This reaction is speeded up by the addition of a catalyst known as Ammonium molybdate. By the end, we have to consider the reaction between Iodine and thiosulfate. Procedure v Transfer 10ml of aqueous solution of hydrogen peroxide by mean of pipette, 10ml of 5% solution of potassium iodide,15ml of sulfuric acid 1M , 10ml of aqueous solution of ammonium moybdate o.1%. v And titrate with sodium thiosulphate solution of concentration of 0.1M. v Add 3 more drops and write the final volume of Na2S2O3 used. Questions a) Write down the reactions involved in this titration b) From these equations deduce the concentration of hydrogen peroxide. Preparation of starch solution v Weight about 5mg of soluble starch and knead carefully with cold water v Transfer the found dough into 100ml of boiling water v Boil the solution about 2minutes ( until the solution becomes transparent) v Give the time to starch to settle at the bottom of recipient v Use the liquid part as indicator 4.5: PRECIPITATION TITRATIONS A precipitation titration involves the formation of precipitates during the course of a titration. The titrant reacts with the analyte forming an insoluble material and the titration continues till the very last amount of analyte is consumed. The first drop of titrant in excess will react with an indicator resulting in a color change and announcing the termination of the titration. Laboratory practices: Pharmacy department (Academic year 2010)

43

IV.1 ARGENTOMETRIC TITRATIONS The most widely applicable precipitation titrations involve the use of silver nitrate with chlorides, bromides, iodides, and thiocyanate. Since silver is always there, precipitation titrations are referred to as Argentometric titrations. A. MOHR METHOD This method utilizes chromate as an indicator. Chromate forms a precipitate with Ag+ but this precipitate has a greater solubility (Ksp=1.1.10-12) than that of AgCl (Ksp=10-10), for example. Therefore, AgCl is formed first and after all Cl- is consumed, the first drop of Ag+ in excess will react with the chromate indicator giving a reddish precipitate. - Before the EP, Ag+ forms a precipitate with Cl- : Ag+ + ClAgCl -At the EP, Chromate forms a precipitate with Ag+: 2 Ag+ + CrO42Ag2CrO4 In this method, neutral medium should be used since, In alkaline solutions (pH>10.5), silver will react with the hydroxide ions: 2 Ag+ 2 HOAg2O +H2O In acidic solutions (pH< 6.5), chromate will be converted to dichromate: Ag2CrO4 + 2 H+ Cr2O72- + Ag+ + H2O Blank solution preparation v In the beaker of 250ml ,put about 40ml of distilled water and 10 drops of K2CrO4 5? v Add the solution of Ag NO3(0.05M) drop by drop until the solution becomes brownish. v Keep the solution to locate easily the EP during titration. Chloride titration v By means of pipette, introduce 10ml of sodium chloride in a beaker of 250ml and add 10drop of indicator(K2CrO4 5?| ) v Titrate by a standard solution of AgNO3(0.05M). During titration agitate continuously to have a homogenous solution. v Determine the concentration of NaCl. This procedure is valid for the titration of all halogen B. VOLHARD METHOD The reactant used in this method is SCN- in form of KSCN or NH4SCN. This method utilizes Fe3+ (iron III) as an indicator. Fe3+ is in form of FeCl3 or NH4Fe(SO4) 2.12 H2O.(Ferric ammonium sulfate). Direct titration

Laboratory practices: Pharmacy department (Academic year 2010)

44 Titration of Ag+ Before the EP, Ag+ forms a precipitate with SCN- : Ag+ + SCNAgSCN -At the EP, a red color due to the addition of a drop of SCN- is obtained. The following equation describes what happens at EP. Fe3++ 6SCN[Fe (SCN) 6] 3Indirect titration or Back Titration Back titration is an analytical chemistry technique which allows finding the unknown concentration of a reactant by reacting it with an excess volume of another reactant of known concentration. The resulting mixture is then titrated back, taking into account the molarity of the excess which was added. For this specific demonstration, the concentration of an analyte (chloride for example) is determined by reacting it with a known number of moles of excess reagent (Ag NO3 silver nitrate). The excess reagent is then titrated with SCN- .The indicator used is Fe3+. At the EP, a red color resulting in the formation of [Fe (SCN) 6] 3- is obtained. PROCEDURE v Take 25 ml of analyte solution and put them in a beaker v Add 2ml of concentrated HNO3 and a known volume of Ag NO3 0.1M( this volume must be in excess and during this practice 40ml will be used ) v Add again 1ml of NH4Fe(SO4)2 12H2O and agitate about one minute to coagulate the precipitate v Titrate the excess of Ag+ in the mixture with KSCN 0.1M until the red color appears (this color can be observed easily when the precipitate has been deposited) v Determine the concentration of halide by taking into consideration the excess of AgNO3 . 4.6: TITRAGE POTENTIOMETRIQUE INTRODUCTION Les mthodes potentiomtriques en chimie analytiques sont bases sur la relation entre le potentiel dune pile lectrochimique et les concentrations des espces chimiques prsentes en solution. Lapplication de la potentiomtrie met en jeu une pile lectrochimique compose dune lectrode de rfrence qui maintient un potentiel constant et dune lectrode indicatrice qui rpond la composition de lchantillon. Un pont salin vite le mlange de la solution de rfrence avec celle de lchantillon. Pendant le titrage, la solution standard (solution titrante) est ajoute par fraction ; on mesure et on enregistre le potentiel de la pile aprs chaque addition de ractif. Au tour du point dquivalence qui est repr par le changement important de E, le ractif titrant est ajout en trs petite quantit.

Laboratory practices: Pharmacy department (Academic year 2010)

45 NB : La mthode de dosage potentiomtrique sadapte toute sorte de dosage de raction chimique. Elle est indique pour les solutions colore ou opaque qui ne permettent pas lutilisation des indicateurs ordinaires ETALONNAGE DE LELECTRODE DE VERRE Le pH mtre (lectrode de verre) est une lectrode membrane slective spcifique aux ions H3O+ .A force ionique fixe la ddp est reli au pH et donc la concentration des ions H3O+ par la relation E= E0 -2.3RT/F pH Cette formule est une variation linaire de E en fonction du pH avec la pente gale au coefficient de Nernst 2.3RT/F de 58mV 200C. Ltalonnage analytique avec des solutions de pH connu (normalement des solutions tampons) correspond bien ce comportement dit nernstien c..d. idal. La reprsentation graphique E= f( pH) est labore lors de ltalonnage analytique et la droite obtenue est appele la droite dtalonnage. Etant donn que la sensibilit de llectrode peut varier au cours du temps (sur quelques jours), vous devez talonner votre pH-mtre . Plus vous talonnez rgulirement, plus vous obtenez des mesures prcises en accord avec lquation de NERNST (rponse nernstienne) Procdure Chaque marque de pH-mtre son protocole dtalonnage. Mais il y a un principe presque commun Principe de ltalonnage deux points v On rgle les potentiomtres lorsque llectrode est plonge dans une solution talon de pH = 7 (milieu de lchelle des pH dans leau). v On rgle les potentiomtres lorsque llectrode est plonge dans une solution talon de pH = 4 NB : Si la manipulation couvre la fois le milieu acide, neutre et basique on emploi la fois les tampons 4,7, et 10 MODE DEMPLOI DE LELECTRODE DE VERRE v Avant de se servir de llectrode : sortir llectrode de la solution de KCl. Rincer dlicatement llectrode avec de leau distille. Absorber dlicatement la goutte deau qui ruisselle avec un papier absorbant. v Plonger llectrode dans la solution tudier. Veiller ce que le barreau magntique tourne bien au dessous de llectrode et quil ne risque pas de heurter celle-ci. v Aprs lexprience : sortir llectrode de la solution tudie. Rincer la dlicatement avec de leau distille. Absorber dlicatement la goutte deau qui ruisselle avec un papier absorbant. Plonger llectrode dans la solution de conservation si on doit rutiliser llectrode. v En fin de TP : rincer llectrode avec de leau distille. Absorber dlicatement la goutte deau. Remettre llectrode dans la solution de KCl concentre. Veillez ce que la bulbe de llectrode plonge bien dans cette solution. QUESTIONS 1. Tracer le graphe E=f (pH) 2. Calculer la pente de ce graphe 3. Pourquoi est-il ncessaire dtalonner un appareil comme lectrode de verre avant son utilisation.

Laboratory practices: Pharmacy department (Academic year 2010)

46 DOSAGE POTENTIOMETRIQUE DUN POLYACIDE Lacide phosphorique est un triacide faible dont les pKa dans leau des trois couples acidobasiques valent respectivement 25oc : - couple acide phosphorique/anion dihydrognophosphate : H3PO4 /H2PO4- :2.15 - Couple anion dihydrogenophosphate /anion (mono)hydrogenophosphate : H2PO4-/HPO42-:7.2 - couple anion (mono)hydrogenophosphate/anion phosphate: HPO42-/PO43-:12.10 Les valeurs de pKa1 et de pKa2de lacide orthophosphorique sont calculs en se servant des valeurs de pH au dpart, au premier et au deuxime point quivalent. pH au dpart : pH= (Eo-E)F /2.3RT ou pH=1/2pKa1-1/2log Ca Au premier point quivalent : pH=1/2pKa1+1/2pKa2 Au deuxime quivalent : pH : pH=1/2 pKa2+1/2pKa3 Mode opratoire - Etalonner llectrode de verre utiliser au moyen des solutions tampons -Transvaser 30ml de lacide orthophosphate 0.05 M dans bcher de 150 ml, puis ajouter quelques gouttes de mthylorange (1er point quivalent) - Titrer au moyen dune solution de NaOH 0.1M - Aprs avoir atteint le premier point quivalent, ajouter quelques gouttes de phnolphtaline - Continuer le titrage en utilisant la mme solution de NaOH jusquau virage de la solution (2me point quivalent) - Rpter lexprience trois fois NB : Ajouter chaque fois 1ml et noter le potentiel obtenu Dpasser le 2me point quivalent de 10ml au moins Questions 1. Montrer clairement comment vous avez procd pour prparer les diffrentes solutions utilises. 2. Donner les concentrations des espces prsentes en solution au premier et au deuxime point quivalent. 3. Tracer la courbe de titrage (la courbe drive). 4. Calculer les valeurs de pKa1 et pKa2 de lacide orthophosphorique. CHAPTER V: ELECTROCHEMISTRY ELECTROCHEMICAL CELL Introduction The device used to study reactions electrically is called an electrochemical cell. Such a cell consists of two electrodes, or metallic conductors, dipping into an electrolyte, an ionic conductor, which may be a solution, a liquid, or a solid. An electrode and its electrolyte comprise an electrode compartment. If the two electrolytes are different, then two compartments may be joined by a salt bridge, which is an electrolyte solution that completes the electrical circuit by permitting ions to move between the compartments and so enables the cell to function (see figure bellow).

Laboratory practices: Pharmacy department (Academic year 2010)

47 Electrons

V
_
Voltmeter

ANODE

Salt bridge

CATHODE

Cations and anions M M Mn+ Mm+

Oxidation

Reduction

Reaction at electrodes In electrochemical cell, the oxidation half-reaction takes place in one compartment and the reduction half reaction takes place in the other compartment. As the reaction proceeds, the electrons released in oxidation half reaction: In one compartment travel through the external circuit, and enter the cell through the other electrode, where they bring about the reduction: Electrical neutrality is preserved in the electrolytes by the flow of cations and anions in the opposite directions through the salt bridge as shown in the figure above. MATERIALS 2 beakers of 500ml Blade or rod of copper Blade or rod of zinc Digital voltmeter with high internal resistance Two external conductors in copper U-shape tube SALT BRIDGE PREPARATION v REAGENTS CuSO4 ZnSO4 Agar-agar KNO3 Distilled water

In the conical flask heat 100ml of a solution obtained by mixing 30g KNO3 ,3g of agaragar and distilled water. This mixture should be covered and gently heated, with stirring,

Laboratory practices: Pharmacy department (Academic year 2010)

48 on a hotplate until the agar is completely dissolved (solution becomes clear) and bubbles are just beginning to form. Dip one end of the U-shaped tube into the hot agar mixture. Capillary action will draw the agar into the tube, filling it completely. Monitor this process to prevent the formation of bubbles within the capillary. Agar filled tubes should be immediately placed in fresh saturated solution of KNO3 to prevent shrinkage of the agar during cooling.

SALT BRIDGE STORAGE Usable bridges should be placed in fresh KNO3 solution for long-term storage. Bridges may be covered and safely stored in the refrigerator for up to 90 days. Salt bridges can be discarded after each use or reused several times. Procedure v Plunge the blade of Zn into the left beaker which contains 400ml of ZnSO4 solution v Plunge the blade of Cu into the right beaker which contains also 400ml of CuSO4 solution v Link these two blades to voltmeter poles by means of external conductors. See if anything changes. v Connect these two solutions in the beakers by salt bridge prepared. v If the voltmeter shows negative value, reverse the poles Questions a. b. c. d. Sketch the prepared electrochemical cell Write down the reaction at electrodes and the overall reaction Based on this scheme explain how this electrochemical cell works This electrochemical cell is joined using the salt bridge containing KNO3. What is the purpose of the salt bridge? e. They are spectator ions in this process .They are SO42-,K+ and NO3-. On the diagram show the direction in which these ions travel f. In which direction do Cu2+ ions travel? In which direction do Zn2+ ions travel? v

Laboratory practices: Pharmacy department (Academic year 2010)

49 CHAPTER VI: ORGANIC CHEMISTRY 6.1: PREPARATION OF BUTYL ACETATE CH3-CO2H + CH3-(CH2)2-CH2OH H+ CH3-CO2-(CH2)3-CH3

Mix together 37g (46 ml, 0.5 mol) of butan-1-ol and 60g (60 ml, 1 mol) of glacial acid in a 250- or 500- ml round-bottomed flask, and add cautiously 1 ml of concentrated sulphuric acid (use a small measuring cylinder or a calibrated dropper pipette). Attach a reflux condenser and reflux the mixture for 3-6 hours (1). Pour the mixture into about 250 ml of water in a separatory funnel; remove the upper layer of crude ester and wash it again with about 100 ml of water, followed by about 25 ml of satured sodium hydrogen carbonate solution and 50 ml of water. Dry the crude ester with 5-6 g of anhydrous sodium sulphate. Filter through a small funnel containing a fluted filter-paper and distil on wire gauze or from an air bath. Collect the pure butyl acetate at 124-125 C. The yield is 40 g (69 %). Note. (1) A slightly better yield of ester can be obtained by increasing the quantity of acetic acid to 90120 g and refluxing for 12-18 hours.

Laboratory practices: Pharmacy department (Academic year 2010)

50 V.2 : SYNTHESE DE LACIDE 2-ACETOXYBENZOIQUE actylsalicylique) ET DU SALICYLATE DE METHYLE. A. Gnralits Formules :


O C OH O O O C O CH3 Salicylate de mthyle OH C CH3 Acide actylsalicylique

(aspirine

ou

acide

Obtenus par: - Actylation (estrification par lacide actique) de lacide 2- hydroxybenzoque (ou acide ortho-hydroxybenzoque ou encore acide salicylique)
O C OH

OH

O C O CH3

- Le salicylate de mthyle salicylique et le mthanol. Raction destrification


O R C OH
+

OH

rsulte de la formation dun ester entre lacide

O R' OH R C O R'
+

H2O

Formes actives de lacide: O R C Cl - Chlorure dacide:

Laboratory practices: Pharmacy department (Academic year 2010)

51
R C O O C O
O O C CH 3
+

- ou anhydride dacide:

OH CH 3 C O O C O O CH 3
+

H + / H 2O H 3O +

CH 3COOH

COO H

COO H
Aspirine (Ac. actylsalicylique)

COO H + OH CH 3 OH

O C OCH 3 + OH Salicylate de mthyle H 2O

B. Matriel - Produits -Mode opratoire 1. Matriel Berlin de 600 cm3 Erlenmeyer de 100 cm3 Thermomtre Plaque chauffante Papier filtre Filtre de Bchner Pipette gradue Pipette Pasteur Verre de montre Spatule Tubes essai 3. Mode opratoire a) Prparation de laspirine Peser avec prcision sur un verre de montre 1g dacide salicylique. Transfrer le produit pes dans un petit erlenmeyer. Ajouter 3 cm3 danhydride actique et, immdiatement aprs, cinq gouttes dacide phosphorique 85 %. Agiter lerlenmeyer pour homogniser, chauffer le mlange ractionnel au bain-marie 50 60 C pendant 15 minutes (sous la hotte). Agiter doucement le mlange avec une tige de verre. Laisser refroidir le mlange en agitant occasionnellement. Ajouter prudemment 15 cm3 deau distille afin de dcomposer lanhydride en excs. Chauffer ensuite la solution jusqu ce quelle soit limpide et laisser refroidir. Les cristaux daspirine vont se former. Baguette en verre 2. Produits ou ractifs Acide salicylique Anhydride actique Mthanol Acide sulfurique et acide phosphorique 85 % concentrs Acide actique Alcool thylique Solution de FeCl3

Laboratory practices: Pharmacy department (Academic year 2010)

52 Quand le mlange ractionnel a atteint la temprature de la pice, agiter afin de mettre les cristaux en suspension, filtrer sur Bchner et laver les cristaux avec de petites quantits deau distille. Filtrer, laver les cristaux et les scher soigneusement. Peser laspirine obtenue et dterminer le rendement de la raction. Lacide 2- actoxybenzoque se dcompose quand il est chauff et ne possde par consquent pas de point de fusion bien dfini. (Le domaine de la temprature de dcomposition vs de 129 C 133 C) b) Prparation du salicylate de mthyle v v v v v v Peser 1g dacide salicylique sur un verre de montre. Transfrer le produit dans un tube essai. Ajouter 5 cm3 de mthanol et 3 gouttes de H2SO4 concentr (prudence). Plonger le tube dans un bain-marie et chauffer ensuite rapidement le bain jusqu 80 C. Maintenir cette temprature pendant une dizaine de minutes. Constater lodeur caractristique du liquide qui se trouve dans le tube essai en fin de raction. A quel produit cela vous fait-il penser ?

Laboratory practices: Pharmacy department (Academic year 2010)