EXPERIMENT 2. MEASUREMENT OF VOLUME

Transcription

1 #2. Measurement of Volume 19 EXPERIMENT 2. MEASUREMENT OF VOLUME For making accurate measurements in analytical procedures, next in importance to the balance is equipment for the measurement of volume. In this section burets, and pipets are discussed. The experimental procedures include gravimetric calibration of burets and pipets to provide checks on the accuracy of the markings as well as to aid in development of proper technique. The few minutes required to learn correct use of this equipment will in the long run save time and give better results. For example, a typical experienced person using incorrect technique takes about 10 s to read a buret and will have a standard deviation in the readings of 0.01 ml or more. A person of the same experience using correct technique takes about 6 s and will have a standard deviation in the readings of less than 0.01 ml. (A) Use of Burets Burets are designed to accurately deliver measurable, variable volumes of liquid, particularly for titrations. A 50 ml buret, the most common size, has 0.1 ml graduations along its length and can be read by interpolation to the nearest 0.01 ml. Parallax errors in reading are reduced by extension of every tenth graduation around the tube. For reproducible delivery, proper design of the tip is important. Any change affecting the orifice will also affect reproducibility; a buret with a chipped or fire-polished tip should not be employed for accurate work. Drainage errors are usually minimized if the tip is constricted so that the meniscus falls at a rate not exceeding 0.5 cm/s. The accuracy of the graduation marks on volumetric burets depends on the uniformity of the bore. The most convenient way to determine actual volumes is to weigh the amount of water delivered. Burets can be purchased already calibrated and accompanied by a calibration certificate. 1 However, burets are best calibrated by the user; the procedure is simple and also is an excellent means of learning to use burets correctly. Several characteristics of a buret should be recognized when the analyst chooses it to measure volumes. Because graduations on a 50 ml buret are 0.1 ml apart, volumes between the marks must be estimated. In this estimation the width of the lines must be taken into account. The thickness of a line on a 50 ml buret takes up about one fifth of the distance from one mark to the next and therefore is usually 1 Such certificates are of little value if the technique being used differs from that used in the calibration.

2 20 #2. Measurement of Volume equivalent to about 0.02 ml, as illustrated in Figure I-1. Generally it is preferable to read the bottom of the meniscus and to take as the value for a given line the point where the meniscus bottom just touches the top of the line. For most people this edge-to-edge technique gives consistent results. Figure I-1 shows readings of various meniscus positions with this technique. Parallax, another source of error in reading a buret, occurs if the eye is above or below the level of the meniscus. This error can be minimized by use of the encircling markings on the buret as guides to keep the eye level with the meniscus. Figure I-1. Enlarged sections of a 50 ml buret showing the meniscus at several positions; correct readings are given below each section. (For clarity the vertical scale is exaggerated over the horizontal.) The apparent position of the meniscus is significantly affected by the way it is illuminated. Lighting errors are minimized by use of a reading card consisting of a dull-black strip of paper on a white background. As in Figure I-2, the card is placed behind and against the buret so that the top of the black portion is almost flush with, or no more than a millimeter below, the bottom edge of the meniscus. For some solutions, such as permanganate and iodine, the bottom of the meniscus may be difficult to see; in such cases the top may have to be read.

3 #2. Measurement of Volume 21 Figure I-2. Reading card in correct position. To provide effective control of the delivery rate when titrating, operate the stopcock with the left hand around the barrel (if right-handed). The other hand is free to swirl the titration flask. Calibration of Burets Before using a buret, clean it thoroughly with soap and do not use a buret brush, which could scratch the interior surface. Rinse it well with distilled water; the buret is clean when water drains from the inside surface uniformly without the formation of droplets. Store the buret filled with distilled water and with the Teflon stopcock nut loosened slightly. Before use rinse the buret at least twice with titrant solution, tightening the Teflon stopcock nut only enough to prevent leakage of solution. After filling the buret with titrant, make certain no air bubbles are present in the tip. Bring the solution level to or slightly below the zero mark, remove the drop adhering to the tip, and wait a few seconds for solution above the meniscus to drain before taking the initial reading. Procedure Because water evaporates at an appreciable rate at room temperature complete the calibration procedure before starting the calculations. Before calibration ask the laboratory instructor to check your buret; if it is adequately clean, you may proceed to calibrate it. Before going to the balance, prepare a data page in your laboratory

4 22 #2. Measurement of Volume notebook with charts for two calibrations (see example in Table I-1). Also assemble the following equipment: clean buret, buret clamp, 50 ml conical flask with dry stopper, funnel, 400 ml beaker of distilled water, and buret reading card and white drybox glove. Approx. Interval (ml) Table I-1. Data and Calculations for Calibration of 50 ml Buret Buret Apparent True True Readings Volume Weight Weight Volume Correction (ml) (ml) (g) (g) (ml) (ml) Initial 0.03B 36.45OE Cumulative Correction (ml) A D 9.941F 9.97G C Using an electronic balance carry out the following procedure: 1. Place a thermometer in the beaker of distilled water. 2. Mount the buret in a buret clamp beside the balance. 3. Check the level and zero of the balance. 4. Read and record the temperature of the distilled water. 5. Fill the buret with distilled water, using the buret funnel. Run out water so that the meniscus is just below zero. 6. Ensure that no bubbles are trapped near the stopcock or in the tip. 7. Weigh the conical flask and dry stopper. Record the weight to the nearest milligram in the appropriate chart space in the notebook. 8. Read the initial buret volume and record it on the chart. 9. Drain approximately 10 ml of water into the conical flask. Touch off the final drop on the inner wall of the flask, below the farthest point of insertion of the stopper. Restopper the flask immediately. 10. Recheck the zero. Weigh the flask plus water and record the weight. 11. Read the buret and record the reading. 12. Repeat items 9 through 11 until 50 ml of water has been delivered and weighed. Keep the flask stoppered except when adding liquid. No more than 10 to 15 min should be needed to acquire a set of calibration data. 13. Empty the flask, wipe the neck, reweigh, refill the buret, and repeat the calibration, recording the data on the second chart. The letters after some entries refer to the arithmetic check of Eq. I-1, (next page)

5 #2. Measurement of Volume 23 Calculations Do not attempt to make calculations at the balance. Calculate the buret corrections, following the outline of Table I-1. Apply the arithmetic check on each calibration. If agreement between the duplicate runs is not within the designated acceptable limits, repeat the calibration. Check the calculations for the buret calibration by substituting the corresponding values from your table for the letters in the equation A - B + C = D - E + 5(G - F) If the arithmetic is correct, the two sides of the equation should agree within 0.01 ml. Duplicate calibration corrections should agree within 0.02 ml, and duplicate cumulative corrections within 0.04 ml at all volumes. If either of these conditions is not met, the technique for reading the buret probably needs improvement, and an additional calibration should be performed. Once the two calibrations agree within the tolerances specified, plot the average cumulative corrections against buret volume. The chart feature in the Excel application should be used or some other graphing program. A sample plot is shown in Figure I-3 (see next page). Two copies of the student s graph should be appended to your final report for this experiment. All subsequent buret readings at titration end point should be corrected by the use of this graph. The net correction for a titration volume is equal to the correction for the final reading minus the correction for the initial reading. This net correction is then added to the net reading. For instance, if the final reading is ml and the initial reading is 1.03 ml, the net correction interpolated from Figure I-3 is , or 0.02 ml. The correct titration volume is therefore = ml.

6 24 #2. Measurement of Volume Cum. Cor. (ml) Volume (ml) Figure I-3. Example of a calibration chart for a buret. Temp, C Density g/ml Temp, C Density g/ml Table I-2. Density of Water at Various Temperatures A more accurate table for the density of water at various temperatures will be posted in the D776 & D770 labs and available on the lab web site.

7 #2. Measurement of Volume 25 (B) Use of Pipets First clean the pipet so that water drains smoothly from the interior surface. Rinse the interior by drawing a portion of the liquid to be pipetted into the pipet with the aid of a suction bulb, then tilt and turn the pipet until all the inner surface has been wetted. Discard this portion of solution and repeat the operation twice. Then draw solution above the mark, wipe the tip and stem of the pipet carefully with a lintless tissue or a clean towel to remove external droplets, and allow the solution to drain until the bottom of the meniscus is even with the calibration mark. Keep the pipet vertical, with the tip in contact with a tilted beaker or flask wall, during this operation. Move the pipet to the receiving vessel and allow the solution to drain freely. During drainage, hold the pipet vertically, with the tip in contact with the inside wall of the container. Keep the tip in contact for about 5 s after free solution flow has stopped, and then remove it. Leave any remaining liquid in the tip; do not blow out this portion. A rubber suction bulb is recommended for all pipetting. Particularly, do not pipet poisonous or corrosive solutions by mouth. Rinse the pipet thoroughly after use. Avoid drawing liquid into the bulb. If the bulb is contaminated accidentally, thoroughly rinse and dry it before reuse. Calibration of Pipets Clean (with pipe cleaners if necessary) and calibrate the pipets provided, following the directions for pipet use given in the preceding section. Procedure Make sure your pipet is clean. Prepare a data page in your laboratory notebook for three calibrations (see Example I-1). Assemble the following equipment: clean 5, 10 and 20 or 25 ml pipet, pipetting bulb, 50-mL conical flask with dry stopper, 400-mL beaker of distilled water, and lintless tissue or clean towel. Example I-1. Trial #1 Trial #2 Trial #3 Temperature of water 26 C Weight of flask + water g Weight of flask g Weight of water delivered g True volume of pipet** ml Average **At 26 C, the density of water is (Table I-2). The true volume of g of water at 26 C is 9.975/ = ml.

8 26 #2. Measurement of Volume Use an electronic balance in this procedure: 1. Place a thermometer in the beaker of distilled water. 2. Check the level and zero of the balance. Ensure that the zero-adjustment knob, not the tare knob or the leveling feet, is used for the zeroing operation. 3. Weigh the conical flask and stopper, recording in your notebook the weight to the nearest milligram. 4. Read and record the temperature of the distilled water. Remove the thermometer. 5. Pipet a portion of the distilled water into the flask. Remove the final drop by touching the pipet tip to the inner wall of the flask below the farthest point of insertion of the stopper. Restopper the flask. 6. Recheck the balance zero. Weigh and record the weight of the flask and water. 7. Add a second portion, removing the stopper just before the addition. Restopper, recheck the zero of the balance, weigh, and record the weight. The three weighings for the set of two portions should not take any longer than 15 min. Calculations Leave the balance and calculate the delivered volumes as in Example I-1. If agreement between duplicates is not within ml, carry out another set of calibrations. Calibrate all the pipets provided 5 ml, 10mL and 20 or 25 ml. Use the average value of the duplicate calibrations for all subsequent measurements with the pipets. Unknown samples for Experiment #3 & #4 must be mixed thoroughly and dried.

9 #3. Acid - Base Titration 27 EXPERIMENT 3. ACID-BASE TITRATIONS: DETERMINATION OF CARBONATE BY TITRATION WITH HYDROCHLORIC ACID BACKGROUND Carbonate Equilibria In this experiment a solution of hydrochloric acid is prepared, standardized against pure sodium carbonate, and used to determine the percentage of carbonate in a sample. An aqueous solution of hydrochloric acid is almost completely dissociated into hydrated protons and chloride ions. Therefore, in a titration with hydrochloric acid the active titrant species is the hydrated proton. This species is often written H3O+, although the actual form in solution is more correctly (H2O)nH+. For convenience we designate it simply H+. Carbonate in aqueous solution acts as a base; that is, it is able to accept a proton to form bicarbonate ion. CO H+ <==========> HCO - 3 (1) Bicarbonate is able to combine with another proton to form carbonic acid: HCO H+ <==========> H2CO3 (2) Equilibrium expressions for the dissociation of bicarbonate and carbonic acid may be written [H+] [CO 2-3 ] K2 = [HCO - (3) 3 ] and K1 = [H+] [HCO - 3 ] [H2CO3] (4)

10 28 #3. Acid - Base Titration where K1 and K2 are the first and second acid dissociation constants for H2CO3; the experimentally determined values are K1 = 3.5 x 10-7 and K2 = 5 x When successive protonation reactions such as (1) and (2) occur, the extent to which the first reaction proceeds before the second begins depends on the difference between the two acid dissociation constants. By combination of Equations (3) and (4) with those for charge and mass balance, [H+] can be calculated for any ratio of hydrochloric acid to initial carbonate concentration, that is, at any point on a titration curve of carbonate with hydrochloric acid. Because complete and rigorous solution is time consuming, here only procedures for calculating the ph at several convenient points in a titration of 0.1 M sodium carbonate with 0.1 M hydrochloric acid (Figure 1) are covered briefly. An analytical textbook should be consulted for a more detailed discussion of this topic. ph at Point A in Figure 1. At point A no acid has been added, and only sodium carbonate is present in solution. The ph is determined by the extent of carbonate reaction with water to give HCO - 3 and OH - 1 : CO H2O <==========> HCO OH- (5) Here water acts as an acid, providing a proton to carbonate ion, the base. The equilibrium constant for this reaction may be written Kb = [HCO - 3 ] [OH -] [CO 2-3 ] (6) 1 Reactions of ions of a solute with water often are called hydrolysis reactions. They are more properly considered, however, as simply another example of a Bronsted acid-base reaction in which water acts as an acid or a base.

11 #3. Acid - Base Titration 29 Figure 1. Curve for the titration of carbonate with hydrochloric acid. Multiplying the right side of Equation (6) by [H+]/[H+], we see that Kb is equal to Kw/K2, where Kw is the dissociation constant for water. Kw = [H+] [OH-] = at 25 C (7) and K2 is the second dissociation constant for carbonic acid [Equation 3]. If the initial concentration of carbonate and the values of Kw and K2 are known, [OH-] can be calculated from Kw K2 = [HCO - 3 ] [OH -] [CO 2-3 ] (8) Assume that the equilibrium for Equation (5) lies far to the left, so that the carbonate ion concentration is still essentially 0.1 M. Since bicarbonate and hydroxide are formed in equimolar amounts, [HCO - 3 ] = [OH-] (9) Substitution of numerical values and Equation (9) in Equation (8) gives x = [OH -]2 0.1 (10)

12 30 #3. Acid - Base Titration and [OH-] = 4.5 x 10-3 M (11) From Equation (7) [H+] = x 10-3 = 2.2 x M (12) so the ph is In our use of Equation (6) we assume that the reaction HCO H 2O <=========> H2CO3 + OH- (13) does not occur to an appreciable extent; that it does not can be verified by substituting the value for [H+] found in Equation (12) in Equation (4) and calculating [H2CO3]. If [H2CO3] is found to be greater than 5% of the total carbonate concentration, the [H+] calculated from Equations (6) and (7) will be appreciably in error. In this case the expression should be solved either exactly, by including all species (which is tedious), or by successive approximations. Calculation shows that [H2CO3] at Point A is negligibly small, so our assumption is valid. The additional assumption that [CO 2-3 ] is essentially 0.1 M also is confirmed because Equations (9) and (11) show that [HCO - 3 ] is less than 5% of [CO 2-3 ]. Note from this discussion that K w = Kb, or K2 Kw = K2Kb. Thus, if Ka for an acid HA is known, Kb for the corresponding base A- can be calculated in aqueous solutions. An acid HA and base A- are called a conjugate acid-base pair; HA is the conjugate acid of A- and A- the conjugate base of HA.

13 #3. Acid - Base Titration 31 ph at Point B. At Point B in Figure 1, 0.5 mole of hydrochloric acid has been added for each mole of carbonate. The solution now contains an equimolar mixture of carbonate and bicarbonate. We can calculate the ph at this point by rearranging Equation (3) to [H+] = [HCO - 3 ] K 2 [CO 2-3 ] (14) Since the bicarbonate and carbonate concentrations are equal, the hydrogen ion concentration is equal to K2, and the ph is Accurate calculations of concentrations of species during titrations must include the effect of dilution by the titrant, but thus far those caused by the addition of hydrochloric acid have not been considered. To correct calculations of concentrations of the major components for dilution, multiply each calculated concentration by the factor V/(V + v), where V is the volume of the original solution and v is the volume of hydrochloric acid added at any point. Although in the present example the effect is slight, in many systems the correction is significant. ph at Point C. The first equivalence point (C in Figure 1) is reached when 1 mole of hydrochloric acid per mole of carbonate has been added. This solution contains only sodium bicarbonate; [H+] is calculated by and the ph is 8.4. [H+] = K1K2 = (3.5 x 10-7) (5 x 10-11) = 4.2 x 10-9 M (15) ph at Point D. Protonation of half the bicarbonate gives an equimolar solution of bicarbonate and carbonic acid (Point D). This is again a buffer system, this time involving the first dissociation constant of carbonic acid. The calculation is handled in the same way as for Point B, with K1 used in place of K2, to yield a ph of 6.5.

14 32 #3. Acid - Base Titration ph at Point E. At the second equivalence point (E) the ph is determined by the extent of dissociation of carbonic acid, the principal species present, and [H+] is calculated from Equation (4): K1 = 3.5 x 10-7 = [H+] [HCO - 3 ] (0.1) [50/( )] = [H +] (16) Therefore, [H+] = 1.07 x 10-4 M = (17) the ph is 3.97, or rounding to 2 significant figures, 4.0. Detection of the Equivalence Point Either the first or second equivalence point (C or E in Figure 1) can be used for carbonate analysis. In neither case is the ph change large in the region of the equivalence point. An uncertainty of 0.1 ph unit at either end point results in an uncertainty of about 1% in the amount of hydrochloric acid required. The error can be reduced if the titration is carried to a preselected indicator color. When a solution is titrated to the second equivalence point, a better approach is to take advantage of the dissociation of carbonic acid into a solution of carbon dioxide in water. H2CO3 <=========> H2O + CO2 (g) (18) Shaking or boiling a solution of carbonic acid causes the equilibrium to be driven to the right through loss of carbon dioxide. If a carbonate or bicarbonate solution is titrated to just before the equivalence point at ph 4 and then shaken or boiled, 2 the ph will rise to about 8 as the concentration of carbonic acid drops (dotted line in Figure 2). The ph is no longer controlled by dissociation of a relatively large concentration of carbonic acid but by a small concentration of bicarbonate. When the titrations continued, the ph goes down sharply because the amount of carbonic acid formed is small and the buffering effect negligible (dashed line in Figure 2). 2 In mammals the CO 2 produced through biological oxidation is carried by the blood to the lungs, where it is exchanged for oxygen. Part of the CO 2 is present in the blood as H 2 CO 3. Since the time available in the lungs for exchange is short, the dissociation of H 2 CO 3 to CO 2 and H 2 O is accelerated by the enzyme carbonic acid anhydrase, a zinc-containing protein of high molecular weight. Thus nature need not resort to either boiling or shaking.

15 #3. Acid - Base Titration 33 Standard Solutions Some standard solutions can be prepared directly by weighing or measuring carefully a definite quantity of a pure substance, dissolving it in a suitable solvent, and diluting it to a known volume. None of the strong acids, however, is convenient to handle and measure accurately in concentrated form. Therefore a solution of approximately the desired molarity is prepared, and the exact value is determined by standardization against a primary-standard base. Figure 2. Effect of removal of carbon dioxide on ph change the second equivalence point in a titration of carbonate with hydrochloric acid. Band indicates region of change of indicator color. Primary standards are stable, nonhygroscopic substances that react quantitatively and are easy to purify and handle. A high equivalent weight is advantageous because weighing errors are minimized. Among the excellent primary standards available are potassium acid phthalate, benzoic acid, oxalic acid dihydrate, and sulfamic acid for standardizing bases and sodium oxalate, tris(hydroxymethyl)aminomethane, 4-amino pyridine, and sodium carbonate for standardizing acids. Pure anhydrous sodium carbonate, besides having all the properties of a suitable primary-standard base, has the added advantage in this experiment of being the same compound as the substance determined. This tends to compensate for determinate errors in end-point selection.

16 34 #3. Acid - Base Titration PROCEDURE Reagent List: Unknown Sample - must be mixed thoroughly and dried HCl concentrated - approx. 12M sodium carbonate (Na2CO3) - must be dried Bromocresol Green - indicator Put a little less than 1 liter of distilled water into a clean 1-liter bottle. Calculate the volume of 12 M HCl ( conc. ) required to prepare 1 liter of 0.2 M HCl, and measure this quantity into a small graduated cylinder. Transfer it to the bottle and mix thoroughly. Label. Standardization of HCl with Primary-Standard Na2CO3 Dry 1.5 to 2.0 g of pure Na2CO3 in a glass weighing bottle or vial at 150 to 160 C for at least 2 hours. 3,4 Check to see if the standard reagent was previously dried for the class. Allow to cool, in a desiccator if necessary, and then weigh by difference (to the nearest 0.1 mg) three or four 0.35 to 0.45 g portions of the dry material into clean 250-ml conical (Erlenmeyer) flasks. Add about 50 ml of distilled water to each and swirl gently to dissolve the salt. Add 4 drops of bromocresol green indicator and titrate with the HCl solution to an intermediate green color. At this point stop the titration and boil the solution gently for a minute or two, taking care that no solution is lost during the process. Cool the solution to room temperature, wash the flask walls with distilled water from a wash bottle, and then continue the titration to the first appearance of yellow. Just before the end point the titrant is best added in fractions of a drop. 5 Record the buret reading and add to it the buret calibration correction. 3 Na 2 CO 3 tends to absorb H 2 O from the air to form Na 2 CO 3.H 2 O, and CO 2 to form NaHCO 3. At least several hours of drying at 140 C is necessary to remove all H 2 O and CO 2. 4 Use a pencil or felt marking pen to label the container with the name or sample number of the contents and with your locker number. The container may be placed inside a small glass beaker, and a watch glass, raised with several bent portions of glass rod, placed on top for protection. Avoid leaving chemicals or equipment in the drying oven longer than necessary, this not only causes crowding, but increases the chance of equipment being broken or samples contaminated by spilled chemicals. 5 To deliver amounts less than 1 drop from a buret, first let a droplet form on the tip, and then touch the tip momentarily to the inside wall of the flask. Rinse the wall with a small amount of distilled water from a wash bottle to ensure that the titrant is washed into the solution. Do not rinse the tip of the buret.

17 #3. Acid - Base Titration 35 Calculate the molarity of the HCl solution. The procedure outlined in the discussion of calculations below may be used as a guide. Relative deviations of 1 individual values from the average should not exceed about 2 parts per Determination of Carbonate in a Sample Mix the sample VERY THOROUGHLY and then dry it in a weighing bottle or small beaker for at least 2 hours at 150 to 160 C. Weigh into clean 200-ml conical flasks, to the nearest 0.1 mg, 0.35 to 0.45 g samples and titrate as in the standardization procedure. Calculate and report the percentage of Na2CO3 in the sample. Use the Q test as the criterion for rejection of suspect experimental data. Either the median or the average may be reported. When the median is chosen the median value for the molarity of the HCl should be used in the calculations rather than the average value. CALCULATIONS The percentage of Na2CO3 in a sample can be calculated in two steps: (1) the determination of the molarity of the HCl titrant from the standardization titrations and (2) the calculation of the percentage of Na2CO3 from titrations of the sample. 1. Molarity of HCl. In titrations of Na2CO3 with HCl to the ph 4 end point, 2 moles of HCl are added for each mole of Na2CO3: 2HCl + Na2CO3 <=========> H2CO3 + 2NaCl (19) The HCl molarity is obtained from the following relations: MHCl = = moles HCl liter = moles Na2CO3 x 2 (ml HCl/1000) (wt of Na2CO3) x 2 (mol wt Na2CO3) (ml HCl/1000) (20)

18 36 #3. Acid - Base Titration The factor 2 required because each mole of Na2CO3 reacts quantitatively with 2 moles of HCl. 2. Percentage of Na2CO3 in Sample. The percentage of Na2CO3 in the sample is calculated as follows: %Na2CO3 = wt of Na 2CO3 in sample g sample x 100 = (moles Na 2CO3)(mol. wt. Na2CO3) wt of sample x 100 (21) = (ml HCl) (molarity HCl) (mol wt Na 2CO3) 1000 x 2 x wt. of sample x 100 Remember: Poor results are often caused by errors in calculation rather than by faulty laboratory technique. Check all calculations and significant figures before reporting results. Ensure you have reported the correct unknown number.

19 #4. Precipitation Titration 37 EXPERIMENT 4. PRECIPITATION TITRATIONS: ARGENTIMETRIC DETERMINATION OF CHLORIDE BACKGROUND In this experiment the reaction between silver ion and chloride is used to illustrate volumetric precipitation analysis, that is titration in which the reaction product is an insoluble substance. Reaction (1) is rapid and the stoichiometry is accurate - so accurate, in fact, that this reaction was used in the early determination of several atomic weights. Ag+ + Cl- <==========> AgCl (s) (1) In this experiment the method of end-point detection is the Fajans method, which uses the formation of a colored adsorbed layer on the silver chloride precipitate. The titration illustrates two important techniques - the direct preparation of a standard solution and the taking of aliquots. The operations of pipetting, using a buret, and simple quantitative transfer of solutions are emphasized. The taking of aliquots frequently saves time and is especially useful when the sample size is small. A disadvantage in the use of aliquots is that errors in the initial weighing and dilution steps are not revealed by replicate titrations. Silver nitrate is an excellent primary standard. It may be dried at 100 C for 1 to 2 h to remove adsorbed surface water, if desired, although drying is normally unnecessary. The solid tends to discolor somewhat upon heating, apparently because of reduction of silver(i) to the metal by traces of organic material. Solutions of silver nitrate are stable although slow photoreduction occurs if protection from prolonged exposure to bright sunlight is not provided. The Fajan's Method The end point in many precipitation titrations can be detected through the use of an adsorption indicator. Adsorption indicator methods were first developed by Fajans. Dichlorofluorescein, a weak organic acid, may be used for the titration of chloride with silver(i). The dichlorofluoresceinate anion tends to be adsorbed on the surface of precipitated silver chloride, but not so strongly as chloride. In the

20 38 #4. Precipitation Titration first stages of a titration, chloride ions in solution are adsorbed preferentially on the surface of the precipitate. As the titration proceeds, the chloride ion concentration decreases until it approaches a small value at the equivalence point. After the equivalence point a slight excess of the silver ion is present, and the attraction between adsorbed silver ion and dichlorofluoresceinate anion causes the latter to be adsorbed onto the surface of the precipitate. This adsorbed layer is pink, and the appearance of this pink color on the precipitate is taken to be the end point. Because the color change takes place on the surface of the precipitate, it is sharper if the surface area is large. Certain compounds, such as dextrin, keep precipitates of silver halides from coagulating and provide the needed surface area. In acidic solutions dichlorofluoresceinate is not a satisfactory indicator because, being the anion of a weak acid, it forms protonated species that will not function as an indicator. Therefore buffer is often added to control the ph during the titration. Preparation for Experimental Work Before starting laboratory work, read the experimental procedure carefully and set up a summary data page in your laboratory notebook. Enter your data directly onto this page as you collect it. PROCEDURE Reagent List: Unknown sample - must be mixed thoroughly and dried Silver nitrate Potassium chloride 0.5M Acetic acid - 0.5M Na acetate - supplied Dextrin - small vials dichlorofluoresein - 0.1% soln. - indicator

21 #4. Precipitation Titration 39 Preparation of 0.05 M AgNO3 Weigh accurately on an analytical balance a clean dry 50 ml beaker. Transfer it to a top loading balance, and weigh into it enough AgNO3 to prepare 500 ml of 0.05 M solution.1 Reweigh the beaker plus compound on the analytical balance. Dissolve the AgNO3 in a small portion of chloride free distilled water.2 Transfer this solution quantitatively to a clean 500 ml volumetric flask with the aid of a funnel, and dilute to volume with distilled water. Mix well and store protected from light. Check the concentration of this solution by titration of aliquots of a standard solution of potassium chloride, as directed in the following procedure. Preparation of the Sample Solution Weigh to the nearest 0.1 mg in a 50 ml beaker, ~1 g of unknown sample, and dissolve it in a small volume of distilled water. Transfer the solution quantitatively to a 100 ml volumetric flask, dilute to volume, and mix well. Pipet 10 ml aliquots of this solution into several 200 or 250 ml conical flasks, using a calibrated pipet. Add about 10 ml of distilled water to each flask. Preparation of 0.15 M Potassium Chloride Weigh to the nearest 0.1 mg in a 50 ml beaker, 3 to 3.5 g of primary standard potassium chloride, and dissolve it in a small volume of distilled water. Transfer the solution quantitatively to a 250 ml volumetric flask, dilute to volume with demineralized water, and mix well. Pipet 10 ml aliquots of the standard potassium chloride solution into a set of 200 or 250 ml conical flasks, using a calibrated pipet. Add about 10 ml of distilled water to each flask. Titrate the standards and the samples with silver nitrate solution according to the Fajan's procedure. It is recommended that titrations of aliquots of standard and unknown samples be carried out in alternation. Fajan's Titration Procedure To each of the aliquots of potassium chloride standard and unknown sample add about 1 ml of 0.5 M acetic acid M sodium acetate buffer and 0.1 g of dextrin. For the first aliquot of standard or sample, add 5 drops of 0.1 % dichlorofluorescein solution and immediately titrate until the precipitate becomes pale pink. For subsequent aliquots first titrate with AgNO3 solution to within 1 ml of the end point, and then add 5 drops of 0.1 % dichlorofluorescein solution.3 Immediately continue to titrate until the precipitate becomes pale pink, swirling the flask continuously.4 No blank correction is necessary.

22 40 #4. Precipitation Titration Empty all solutions and a first rinse only into the container marked "Silver Waste". Calculate the percentage of chloride in the sample using (a) the molarity of the silver nitrate based on the titrations of the standard potassium chloride aliquots and (b) the molarity based on the weight of silver nitrate taken. CALCULATION EXAMPLES The weight % of Cl in the unknown sample can be calculated using the following equations: wt Cl Wt.% Cl = wt Samplein aliquot x 100 Since one mole of Ag+ reacts with one mole of Cl, wt Cl = (ml AgNO 3)(M AgNO3)(35.45 g/mol) 1000 ml/l % wt Cl = (ml AgNO 3)(M AgNO3)(35.45 g/mol) (1000 ml/l)(wt sample in aliquot) (100) The weight of sample in the aliquot taken for each titration is given by; (initial wt sample) calibration vol of 10 ml pipet 100 ml The molarity of the AgNO3 based on the weight taken is calculated from the moles AgNO3 and volume of solution: mol AgNO3 = g AgNO3 g AgNO3/mol = wt AgNO L AgNO3 = ml/(1000 ml/l) M AgNO3 = g AgNO3 ( g/mol)(0.500 L)

23 #4. Precipitation Titration 41 The molarity of the AgNO3 solution based on the titration of aliquots of standard KCl is obtained from; M AgNO3 = (M KCl) (ml KCl) (ml AgNO3) The molarity of the KCl solution is based on the weight of the salt taken and can be calculated in the same way as that for the AgNO3 solution M KCl = (g KCl) (g KCl/mol)(0.250 L) Notes 1 AgNO3 is corrosive and should not be handled near an analytical balance. 2 Chloride free distilled water is available in the distilled water taps located on the back bench. 3 Dichlorofluorescein promotes photoreduction of silver in the AgCl precipitate. The resulting finally divided dark silver obscures the end point. Addition of the indicator near the equivalence point reduces this difficulty. If the approximate location of the end point is unknown indicator must be added early in the first titration, and the approximate end point volume determined. To lessen photodecomposition, which may cause error even when the indicator is added late in the titration, avoid direct sunlight on the titration flask. 4 If a gradual indicator change makes end point selection uncertain, record the buret reading. Then add another drop and record any change; repeat this procedure. Select as the end point the reading corresponding to the drop that causes the greatest change.

24 42 #5. Determination of Trace Iron EXPERIMENT 5. SPECTROPHOTOMETRIC DETERMINATION OF TRACE IRON AS A 1,10-PHENANTHROLINE COMPLEX BACKGROUND 1,10-phenanthroline (phen) forms an intensely red complex with iron(ii) that may be exploited to determine iron concentrations in the range of parts per million. The reaction is 3 phen + Fe2+ <==============> Fe(phen)3 2+ The reagent has the structure shown below. Other reagents which are used for the determination of iron are shown also. N N C N N C N N N N N N 1,10-Phenanthroline Bipyridine 2,4,6,-Tripyridyl- s- triazine The molar absorptivity (a in Beer's law) of the complex is 8650 L/mol-cm at 522 nm, the wavelength of maximum absorption. The complex forms rapidly is stable over the ph range 3 to 9, and may be used to determine iron(ii) concentrations in the range of 0.5 to 8 ppm. Iron(III), if present, must be reduced to iron(ii) to produce the colored species. A suitable reagent for this purpose is hydroxylamine hydrochloride, NH2OH.HCl.

25 #5. Determination of Trace Iron 43 The concentration of iron in the sample could be calculated from Beer's law if the molar absorptivity and solution thickness were known. It is preferable, however, to prepare one or more standards and compare absorbance readings of the sample and standard solutions, since this technique minimizes the effects of instrument and solution variation. Although spectrophotometric methods are normally accurate to about 1%, higher accuracy and precision can be obtained with more sophisticated instruments. In most cases an accuracy of 1% at the level of milligrams per liter is sufficient. The standard in this experiment, FeSO4 (NH4)2SO4 6H2O, though not a primary standard, is available with a purity greater than 99%, which is adequate. PROCEDURE Reagent List: Unknown Sample DO NOT DRY ferrous ammonium sulfate sulfuric acid (H2SO4) - conc. 18 M hydroxylamine hydrochloride - 10% solution needed 0- phenanthroline 0.25% soln. sodium acetate - 10% solution needed cobalt chloride HCl conc. 12M Due to the availability of Spectronic 20 spectrophotometers, this experiment demands that you share an instrument with a partner. One set of standard solutions per group is acceptable. Report your partner s name to the instructor. Collect an unknown sample which is contaminated with Fe from your laboratory instructor. Preparation of Standard Iron Solution Weigh to the nearest 0.1 mg, enough FeSO4 (NH4)2SO4 6H2O to prepare 250 ml of a solution M in iron. Transfer the salt to a 250-mL volumetric flask, dissolve it in distilled water, add 8 ml of 3 M H2SO4, dilute to volume with distilled water, and mix. Pipet 10 ml of this solution into 100-mL volumetric flask, add 4 ml of 3 M H2SO4, and dilute to volume. 1 1 The iron concentration of this standard solution should be known to within about 0.5%. Do not exceed 4 ml of H 2 SO 4, or the buffering capacity of the sodium acetate, to be added later, may be exceeded.

26 44 #5. Determination of Trace Iron ANALYTICAL PROCEDURE Preparation of Solutions for Measurement Obtain the total mass of your sample (mass maybe used as a check) and then place in a 500-mL volumetric flask. Dissolve in distilled water, add 20 ml of 3 M H2SO4 and then dilute to volume with distilled water. Pipet 5 ml portions of this stock solution into two 50-mL volumetric flasks. Standard Solutions Into a set of four 50-mL flasks, pipet 5, 10, 15, and 20 ml of the standard iron solution. 2 Prepare a blank by adding 0.4 ml (about 10 drops) of 3 M H2SO4 to another 50-mL volumetric flask. 3 Add to each of the seven flasks, in order, 1 ml of 10% hydroxylamine hydrochloride solution, 10 ml of 0.25% phenanthroline solution, and 4 ml of 10% sodium acetate solution, 4 mixing after each reagent is added. Dilute to volume. Measurement of Absorption Spectrum To obtain an absorption spectrum of the iron-phenanthroline complex, first measure and record % transmittance values of the most concentrated standard solution at 20-nm intervals over the spectral range of 400 to 640 nm. (The procedure is given in later pages). Convert the %T readings to absorbance, and plot absorbance (vertical axis) against wavelength (horizontal axis) in your laboratory notebook. From this graph select the wavelength to be used for measurement of the unknown solutions. Check the selected value with your laboratory instructor before proceeding. Measurement of Standards and Samples Using matched tubes, read the % transmittance (%T) of each solution several times at the wavelength of maximum absorbance. Set the %T reading to 100 with the blank solution between each group of six sample and standard readings. Note: All solutions may be discarded in the sink upon completion of the experimental work. 2 A 5-mL pipet should be used for this purpose. 3 A 100-mL volumetric flask may be used if convenient; in that case use 0.8 ml of 3 M H 2 SO 4. 4 The sodium acetate plus sulfuric acid gives an acetic acid-sodium acetate buffer in the ph region of about 4.5 to 5. At a ph much below 4 bipyridine exists predominantly in the protonated form, and complex formation with the iron may not be complete.

27 #5. Determination of Trace Iron 45 CALCULATIONS Prepare a calibration curve by plotting the absorbance (calculated from the % transmittance) of the standard solutions (vertical axis) against the concentration of iron(ii) in the unknown solutions and calculate the mg of iron in the original sample. Also calculate the mg of iron by the least squares procedure described below. Here is a example SIMILAR to the calculations you must perform: By use of the foregoing procedure standard absorbance values were obtained as follows: Std 1: 0.120, 0.125, 0.130; Std 2: 0.248, 0.255, 0.252; Std 3: 0.382, 0.385, 0.384; Std 4: 0.504, 0.502, The sample readings were 0.337, 0.335, for Sample 1, and 0.341, 0.336, and for Sample 2. If the weight of ferrous ammonium sulfate taken was g, how many milligrams of iron did the unknown contain? The Graphical Method First calculate the concentration of iron in each of the standard solutions: mg Fe in stock soln = (at. wt. Fe)(0.204)(1000 mg/g) (mol. wt. FeSO4.(NH4)2SO4.6H2O)(250 ml) = mg/ml in 250-mL Flask (0.116) (V10) (100.0 ml) = mg/ml in 100-mL flask (0.0116) (V10) (50.0 ml) = mg/ml = 2.32 µg/ml in Std 2 For Stds 1, 3, and 4 the corresponding values are 1.16, 3.48, and 4.65 µg/ml. Next plot a calibration curve using the average absorbance values, as shown in the accompanying figure. Finally, read the concentration of iron in the two unknown solutions from the graph (3.09 µg Fe/mL in this example). In this example the two sample solutions gave essentially identical absorbance readings.

28 46 #5. Determination of Trace Iron Absorbance µg Fe/mL Figure IV-1. Calibration curve for the iron(ii) phenanthroline complex, based on data given in example. From the concentration of iron in the unknown solution the total iron in the original sample is calculated as follows: For Unknown Solution 1; (3.09 µg/ml)(50.0 ml) = 154 µg in the 50-mL flask. Since this amount of iron came from a 10-mL aliquot of a 100-mL sample, the total iron in the sample is 154 µg ml V10 = 1540 µg, or 1.54 mg Report the total weight in milligrams of iron in your sample.

29 #5. Determination of Trace Iron 47 THE SPECTRONIC 20 SPECTROPHOTOMETER. OPERATING PROCEDURE & PROCEDURE FOR MATCHING SAMPLE CELLS The Spectronic 20 Operating Instructions for Bausch and Lomb Spectronic Turn on the instrument by rotating the amplifier control clockwise. (Knob - front - lower left) 2. Set the wavelength control (knob - top panel right side) to the desired wavelength. With the amplifier control, set the meter needle to zero on the percent-transmittance scale. The setting corresponds to infinity on the absorbance (optical density on older instruments) scale. 3. Dry the outside of the matched sample tubes with a lintless towel or tissue. Insert a tube containing a blank 1 into the sample compartment. Position the tube in the instrument with the aid of the index mark. Close the cover of the compartment. 4. Rotate the light control (knob - front - lower right) until the meter reads 100 on the percent-transmittance scale (zero on the absorbance scale). 5. Remove the blank tube and recheck the zero reading. Replace the blank tube with one containing a standard or sample. Read the absorbance directly and record it. The cover of the sample compartment should be closed for all readings. Variations of ±1% in the readings are normal. 6. If readings are to be taken at another wavelength, remove the sample tube and insert the blank tube. Turn the light control counterclockwise before changing the wavelength setting; otherwise, increased sensitivity of the photodetector at the new wavelength may result in a signal of sufficient magnitude to damage the meter. Set the new wavelength. 7. To record multiple wavelength readings or to record a spectrum, repeat steps 4, 5, and 6 until readings at all desired wavelengths have been taken. Readjust the light control whenever the wavelength setting is changed. The zero 1 A blank solution is one that contains all the reagents for an analysis but not the material under test.

30 48 #5. Determination of Trace Iron percent-transmittance setting (dark current) also should be checked periodically and readjusted as needed with the amplifier control. Readings taken at 10- to 20-nm intervals are sufficient to outline an absorption spectrum except at absorption peaks or shoulders, where additional points should be recorded to characterize the curve more completely. Near the ends of the spectral range of the instrument (below about 350 and above 650 nm) a 100% transmittance reading may be impossible to obtain with the blank. SAMPLE-CELL MATCHING Sample Cells (Cuvettes) in Spectrophotometry Several different types of sample cells are used in spectrophotometry. Less expensive instruments are designed to use test tubes for liquid samples. To ensure that the solution path length is the same for sample and standard solutions, a matched set of tubes is needed; the use of a single tube for all measurements, although possible, is inconvenient. Tubes are matched by placing a solution of intermediate absorbance in each and comparing absorbance readings. One tube is picked arbitrarily as a reference, and others are selected that give the same reading within 1%. Tubes should be matched in a separate operation before any spectrophotometric experiments are begun. Follow the procedure outlined below.

31 #5. Determination of Trace Iron 49 PROCEDURE Selection of Matched Tubes for Bausch and Lomb Spectronic 20 Spectrophotometer Obtain a supply of 13- by 100-mm test tubes that are clean, dry, and free of scratches. You will need at least 3 test tubes which are matched. Half-fill each tube with a solution of approx. 2% CoCl2.6H2O. The solution contains about 2 g of CoCl2.6H2O in 100 ml of 0.3 M HCl. 2 Take the tubes to a Spectronic 20. Set the wavelength to 510 nm on the spectrophotometer. (See operating instructions for Bausch and Lomb Spectronic 20 at the beginning of this section.) With the amplifier control, set the instrument to read zero. Place a vertical index mark near the top of one of the tubes, and insert the tube into the sample compartment. Adjust the light control so that the meter reads 90% transmittance. Using this tube to check periodically the 90% reading, insert the other tubes and record their transmittance. If the percent transmittance is within 1% of the initial tube, place an index mark so that the tube can be inserted in the same position every time. If it is not within 1%, rotate the tube to see whether it can be brought into range. In future measurements insert each tube in the same position relative to its index mark. Once you have obtained at least 3 tubes that match proceed with the section Measurement of Absorption Spectrum. To compensate for variations between instruments, use the same one for both tube matching and experimental work. Note: The 100 ml of 2% CoCl2.6H2O is more than enough for several groups to share. 2 This solution is recommended because it is stable, has a broad absorption band at about the center of the visible region, and transmits about 50% in a 1-cm cell.