filtrates & residues estoy INNES O. SCHRECK Univ of Norte Celrado ‘realy C0 30630 Determining Iron Content in Foods by Spectrophotometry Paul E. Adams Beall High School, E. Main Street, Frostburg, MD 21832 Secondary school chemistry students ean have a satisfy ing introduction to colorimetry by obtaining reasonably close results for the determination of iron in various foods. ‘The classic reaction between the iron(IIP) ion and the thiocyanate ion to yield a reddish-brown complex ion, works quite well for this activity The various ions that in- terfere with the formation of this eomplex are silver, cop- per, nickel, cobalt, titanium, uranium, molybdenum, mer- cury, zine, cadmium, bismuth, phosphate, arsenate. Muoride, oxalate, tartrate, and sulfate (7), Most of these ions either are absent or in low concentrations in food. ‘ChemCom (2) has a visual comparison activity related to the iron content in food, and this procedure ean be used as fan extension, This experiment works very well in other chemistry courses as an experience in spectrophotometric ‘analysis and its concomitant skill, It is precisely in this, latter context that it is useful At high concentrations of thiocyanate ion, the following equilibrium is pushed far to the right Fe (aq) +6 SCN” (ag) = [FeSCN}gI* aa) ‘The food sample is heated until it is reduced to a fine ash, Hydrochloric acid is added to dissolve the iron in the residue and to prevent the hydrolysis of the iron( IID) ion. Aer filtering, thiocyanate solution is added and the ab- sorbanee is determined with a spectrophotometer. Using a previously prepared iran(II1) complex concentration-ab- sorbanece graph, the result can be obtained with the help of interpolation o* extrapolation. ‘The solution made when the iron(11D) complex is mixed with the thiocyanate ion is not stable under normal tem: perature and light conditions. The third dilution (de- scribed below) lost 5% of its absorbanee within 10 min and 10% ofits absorbance in 30 min, Spectrophotometrie read- ings should be made without delay: Equipment and Supplies List + spectrophotometer-meter or digital readout (digital readout IS preferred). Colorimetric probes with associated software are available for attachment onto computars IBM, Mac, Ap ple Ib, These will gather the data and graph results, «+ erucble or evaporating dish + pipets or burets 40,0010 M FeCl, stock solution (at least 11 mL/group) dis- solve 0.270 g of FeCle6H,0 in about 500 mL of distilled Water. Acdify with 5 mi of concentrated HCI. Dilute to 1 L ‘with distilled water. Mix wel. This is more than suiiont. A Tange quantity must be made due to the mass of solute re ‘quired for such a low concentration, + L5 MKSCN (at least 35-50 mLigroupl. For 10 groups, dis solve 729 4 of KSCN in distilled water to make 500 mL. of solution, Mi wel + 2.09 HCI at leat 40-70 mL/group) For 10 groupe, add 170 mL. of concentrated HC to 500 mL of distilled water. Mix, Dilute to L with cisiled water Mix Procedure Part 1. Preparation and Absorbance ‘of the Standard Solutions 1. Starting with a 0.0010 M FeCl, stock solution, prepare 10 mL of each of the following. Use a pipet or buret for the FeCl, volume measurements. + Caution: Never pipet by mouth, Always use suction ball forsimilardeviee for dravving up fluids with a pipot. Use 2.0 M HCI to dilute the stock solution, not distilled water. Use a pipet or buret for the HCI volume measure ‘ments, Place each solution into a clean dry test tube with ‘4 minimum capacity of 20 mL. Do not mix with a stirring rod because that could contaminate the solutions. Mix by swirling the solutions in their test tubes. If time is a factor, these dilutions will have to be prepared for the student in, advance, ‘9, 0.000050 M iron) ion solution is made by diluting 0.50 mL-of the ion(TID chloride stack solution with 9.50 mf-of| 2.0 M hydrochloric acid 0.00010 Miron II jon solution ix made by dilating 1.00 mL. ‘ofthe iron! II) chloride stock solution with 9.00 mL, of 20 Mhydrochlorie seid «0.00015 M iron II) fon solution is made by diluting 1.50 ml. ‘of the iron TI) chloride stock solution with 8.50 ml of 2.0 Mhydrochlorie acid 4, 0.00020 AF iron ITT fon solution is made by diluting 2.00 mL. ‘ofthe iron I) chloride stock soltion with 8.00 ml-of 20 M hydrochloric seid «0.00025 M iron TI) jon solution is made by diluting 2.50 mL. ‘ofthe iron Il) chloride tock soliton with 7.50 mL of 2.0 M hydrochloric seid 2. Add 5.0 mL. of 1.5 M KSCN to each of the prepared solutions. Mix each solution well. This will dilute 10 mL of solution to 15 mL. and cause its concentration to decrease to two-thirds its original molarity. This two-thirds concen- tration is the actual concentration that is being read in the spectrophotometer. '3. Read the absorbance of each solution on a spectro- photometer set at 480 mu wavelength (1). This step should not be delayed for an extended period because the color does fade significantly within 15-80 min. Part 2, Determination ofthe Iron Content in a Food Sample 1. Obtain the mass of a clean, dry crucible (or evaporat- ing dish if the food sample is going to be too large). Add about 5 g of a finely chopped sample of the food to be tested ‘and determine its mass to three decimal places. 2, Heat the sample over a Bunsen burner until it has been reduced to an ash. ‘The heating process could cause some smells and smoke, so use a well-ventilated room. A fume hood is desirable. No splashing should occur 3. After the sample has cooled, transfer the contents to a small beaker if the crucible or evaporating dish does not, Volume 72 Number7 July 1995 649have at least 10-15 mL capacity. With a buret or pipet, add 10.00 mL of 2.0 M HCl to dissolve the iron(IID present in the ash. Stir gently for about five minutes Filter the mixture. Collect the fil- trate 4. Mix 5.00 ml. of the filtrate with 5.00 mL of 1.5 M KSCN. Mix well. Because the solution concentration is halved by add- ing the 5.00 mL of KSCN, the concentration of the filtrate will be twice what is determined by absorbance. 5. Obtain the absorbance of this solution soon after mixing. ‘The color does change signifi= cantly within 15-30 min, 6. If the laboratory drain is connected to a municipal waste 0.0 treatment plant and ifloeal regu- ° lations permit, the solutions or- dinarily can’ be disposed of through dilution 20 ABSORBANCE os 0s 1 18 2 Concentration of [Fe(SCN),,]*; (M x 10°) ‘Absoroanco versus conceptraton often thiceyanat fon Note: the actual concentration, for example at Data and Calculations Part 1 1. Construct a data table of standard comparison infor- ‘mation: (sample data consisting of average values from a class are provided) ‘Standard Comparison Information " [Feild trom step 1 [Fett in complex absorbance of ion from step 2 comple ion o 0 o ‘9.000050 .ooacaa Oa ‘.00010 .000087, 079 ‘00018, o.00010 wiz 9.00020 0.00013 aa 900025 ____0.00017 18 2. Using the data above, plot molar concentration of the solution (M) on the x-axis and absorbance A) on the y-axis, ‘Draw a straight line representing the best fit for the points, fn the graph. This graph (see figure) will serve as a means of converting absorbance to molar eoncentration when eal: culations from the food source are done. Pant2 1. Construct a data table like the following: (sample data provided) ‘Type of fod: broccoli Mass of food: 4.540 Absorbance of selution prepared from the food fash: 042 2. Using the graph from Part 1, determine the concentra tion of the solution in Part 2 step 4. For absorbance of 0.42, the molar concentration of the solution would be about 3.2 « 10° M 3, Determine the molar concentration of iron(II) in the filtrate obtained in Part 2 step 3. Remember that the con- centration of the filtrate will be twiee the concentration of the solution due to the doubling of the volume when the KSCN was added, (22% 10° Myx 410M, 650 Journal of Chemical Education 41,18 0.00010 0.0 10" M, 4, Determine the total moles of iron in solution in Part 2, step 8. Determine the mass (in milligrams) of that many. ‘moles of iron, Bocause there were 10 mL of filtrate: ‘Molar concentration infiltrate «0.01000 L.= moles of iron (6-4 10° My x0.01000 L= 6.4 « 10°? moles roles of Fe 55.8 ghmol x 1000 mai mg Fe 1036 mg (6.4 107 mol) 55.8 g/mol « 1000 met 5. Determine the mass (in milligrams) of iron per gram of food used. Milligrams of Fein sample! grams of sample 0.038 mg 4.540 g = 7.9% 10°" gig Conclusion 1. From the Handbook of Chemistry and Physies (3) or ChemCom (2) (pp 535-537) or other sources (4), determine fan accepted value for the amount of iron in the analyzed food. Accepted values for the iron content ofa few foods are presented in the next section, 2. Determine the percent difference between your value and the value found in a reference book. What. are some reasons for the differences between the two values? (Reference value-experiment value) 100/reference value Values from different reference sources often are not the same. This helps the students understand that there are ‘many factors from the sample itself (such as growing con- ditions, soil,varioty of that vegetable, part ofthe vegetable analyzed, ete.) that might result in different analytical outcomes, 3. Discuss your body’s needs for iron, Research in ency- clopedias 5, 7) and health texts will provide ample sources, of information Student responses to this question usually center around the hemoglobin in the blood ar the blaod's oxygen ‘carrying role and that is satisfactory: However, the instruc- tor might wish to add that a 70 kg male has about 4-5 g of iron in his body and 60-70% of t (in the forms of hemozlo- bin, myoglobin, and heme enzymes) is related to the oxy-{gon carrying role. The remainder is present as iron bound, {o protein in compounds such as transferrin, ferritin, and hemosiderin (8) 4. For someone of your sex, age, and weight, what is the body's daily requirement for iron? (4-7) ‘Typical Values of Iron Requirements (3) Category Ane (yr) Weightfeg) ron mg) male 15-18 6 2 mao 19-28 m 10 female 158 55 15 female 19-24 58 15 5. According to your results, what mass of the vegetable analyzed would you have to consume to get your daily sup- ply needs? daily requirement of iron (mgvimg of iron per gram of food) Results Good results have been obtained from broccoli, peas, eau- liflower, spinach, beans, nuts, and evaporated tap water (evaporate 200 mL or more). ‘When using nuts, a good bit of smoke and soot are gen- crated. Ifsoot remains on the side of the erucible or evapo- rrating dish, the ashes should be transferred to another container before dissolving with HCI. Although raisins and bother dried fruit are a good souree of fron, the incineration, process might be too long to be worth the lab time. If the graphing is done on a computer or a graphing eal- calator, the axes might have to be switehed so that interpo- lation or extrapolation will proceed more smoothly. Comparisons of values obtained experimentally and the values obtained from the 7st edition of CRC (3) are shown, in the table. Two samples of tap water were evaporated to, dryness and the residue in the beaker was treated as an 8sh The results fom the two analyzed samples were quite Iron Supplied by Foods and Water Experimental Value CRC Value mg Felg ‘Sample mg Fela ‘vos a TO frozen green peas 12x10? 4.6107 almonds 34x10? 36x 10% cabbage 12x10 6x10 pinto beans 47 «10% (tied) 3.0% 10%(cooked) 200 mitap water 26x10 400 miL-tep water 21xto* Literature Cited 1 Ngee A atk of Quetta age Anas Wi New ak, 38, Ala ond Bok of Fat Mak finan; Phar Bak, 18, 1 Bogle ier pra tae e181. 1, * oy ltr tak Bacher: WB Swedes Co, Papin, Volume 72 Number7 July 1995 651