This document describes a procedure to use a spectrophotometer to determine the concentration of iron (Fe2+) in an unknown water sample. Standard iron solutions of known concentrations are prepared and their absorbance values measured to generate a calibration curve. The absorbance of the unknown sample is then measured and its concentration determined using the calibration curve. The purpose is to quantify the amount of iron in the unknown water sample.
This document describes a procedure to use a spectrophotometer to determine the concentration of iron (Fe2+) in an unknown water sample. Standard iron solutions of known concentrations are prepared and their absorbance values measured to generate a calibration curve. The absorbance of the unknown sample is then measured and its concentration determined using the calibration curve. The purpose is to quantify the amount of iron in the unknown water sample.
This document describes a procedure to use a spectrophotometer to determine the concentration of iron (Fe2+) in an unknown water sample. Standard iron solutions of known concentrations are prepared and their absorbance values measured to generate a calibration curve. The absorbance of the unknown sample is then measured and its concentration determined using the calibration curve. The purpose is to quantify the amount of iron in the unknown water sample.
INTRODUCTION Spectrophotometers measure the amount of light absorbed by a sample. All colored solutions absorb visible light and can, therefore, be analyzed ith a spectrophotometer. !his e"periment uses a spectrophotometer to determine the concentration of iron #II$, or Fe %& , ions in an un'non sample after analyzing the absorption values for a series of iron #II$ standards, solutions of 'non concentration. Although solutions containing iron #II$ ions are colorless, the addition of ortho(phenanthroline comple"es the iron #II$ ions, immediately forming an orange species. More iron #II$ ions in a sample result in a deeper orange color that ill absorb more light. PURPOSE !he purpose of this e"periment is to determine the concentration of iron #II$ in an un'non sample. MATERIALS )** ppm Fe &% solution +imipes *.&,- ortho(phenanthroline solution spectrophotometer graduated )* m. pipet computer ith .ogger /ro )* m. graduated cylinder distilled ater , 0 )** m. volumetric flas's cuvette un'non or collected ater sample SAFETY 1 Alays ear goggles and an apron in the lab. PROCEDURE Part I: Preparation of tan!ar! ). .abel , )** m. volumetric flas's2 * ppm #or blan'$, ).* ppm, &.* ppm, 3.* ppm, 4.* ppm. &. 5se pipets to deliver *.* m., ).* m., &.* m., 3.* m., and 4.* m. of the )** ppm Fe %& solution to the appropriate )** m. flas's. 6. Add , m. of *.&,- ortho(phenanthroline solution, measured in a graduated cylinder, to each flas'. Add enough distilled ater to each flas' to dilute to the )**.* m. mar'. !hese are the standard solutions. Part II: Settin" #p t$e Spe%trop$oto&eter ). Connect the spectrophotometer to the computer using the 5S7 cable. !he cuvette holder 8 light source should already be connected to the spectrophotometer. &. Start the .ogger /ro program. 6. If the 9:; < 7I= bac'ground does not appear, select the Connect Interface Spectrometer Scan for Spectrometer from the e"periment menu. 3. !o calibrate the Spectrometer, chose Calibrate Spectrometer from >"periment menu. !he calibration dialog bo" ill display the message2 ?@aitingA4* seconds for lamp to arm up.B !he minimum arm up time is one minute. NOTE: For best results, allow the spectrometer to warm up for at least fve minutes. Folloing the instructions in the dialog box to complete the calibration, use a rinsed cuvette flled about full with the 0 ppm Fe 2+ solution as instructed. Be sure to wipe the outside of the cuvette with a Kimwipe and chec to mae sure the non!frosted, clear sides are in the li"ht path. #he cuvette should be inserted all the wa$ throu"h the cell holder. %ou should feel that the cuvette is "entl$, but frml$, held in place so that $ou cannot twist the cuvette. &lic Finish &alibration and then clic 'K. Part III: Fin!in" t$e 'a(e)en"t$ of &a*i&#& a+or+an%e an! ettin" t$e %o))e%tion ettin" ). 9inse a cuvette tice ith about )(& m. of 4 ppm Fe &% solution. Cispose of the solution as directed by your instructor. &. Fill the treated cuvette about D full of 4 ppm Fe &% solution. Clean the cuvette ith a +imipe and place sample in the cuvette holder of the spectrometer. Clic' Clic' to end the data collection. 6. Add a title to your graph by double clic'ing on the top of the graph and adding title in the bo". /rint a copy of this graph for each member of your group. 3. ;ou can read the absorbance using the >"amine tool, by clic'ing on . !hen move the cursor along the spectrum. !he avelength and absorbance ill be displayed in the ne dialog bo" in the data indo. Cetermine the avelength of ma"imum absorbance. 5se this avelength throughout your e"periment. ,. Clic' on the ?Configure Spectrometer CataB Icon located on the right hand side of the toolbar. 5nder the ?Set Collection ModeB icon in the display clic' ?Abs vs. ConcentrationB. !he avelength of the ma"imum absorbance ill automatically be selected. ;ou may choose a different avelength by unclic'ing the avelength and clic'ing another avelength. Clic' ?:'B hen done to close the display. ;ou do not need to store the data.
PART IV: Contr#%tin" a Beer, La' -rap$ ). !o set up the graph to reflect ppm, under the data menu, select Column :ptions E Concentration. Change the units to ppm. &. Cispose of the solution in the cuvette. 9inse the cuvette tice ith about )(& m. of the ).* ppm Fe &% solution. Fill the cuvette D full of the ).* ppm Fe &% solution. @ipe and place the cuvette in the spectrophotometer. Clic' . :nce the absorbance stabilizes, clic' the +eep button. >nter the concentration of the solution and clic' :+. 6. C: F:! C.IC+ S!:/ 5F!I. A.. S!AFCA9CS GA=> 7>>F 95FH 9epeat the above step to obtain absorbance readings for each of the other 'non Fe &% solutions. Clic' +eep to save each absorbance reading. 3. After the final 'non Fe &% solution, clic' . ,. Clic' ?linear fitB to see the eIuation for the standard solutions. Part V: Preparation an! ana).i of t$e #n/no'n ). Fill a cuvette about D full ith your un'non, hich ill be provided by your instructor. &. 9ecord the absorbance of the un'non solution in the Cata !able. 6. Chose ?Interpolation Calculator #not Interpolate$ from the Analyze menu. A helper bo" ill appear, displaying the absorbance and concentration of the un'non. 9ecord the concentration of the un'non. Clic' ?:'B. 3. Move the bo"es for the linear fit and interpolation calculator so that they are easily readable and do not interfere ith the line. Add a title to the graph. /rint a copy of your graph for each member of your group. FamesJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJJ /eriodJJJJJJJJJ ClassJJJJJJJJJJJJJJJJJJJ CateJJJJJJJJJJJ DETERMINATION OF IRON IN WATER DATA TABLE Con%entration A+or+an%e *.* ppm #blan'$ ).* ppm &.* ppm 3.* ppm 4.* ppm 5n'non JJJJJ CALCULATIONS: Ca)%#)ate t$e %on%entration of iron in t$e #n/no'n o)#tion in +ot$ pp& an! M0 1UESTIONS 2to +e an'ere! in )a+ report3 ). @hy as ortho(phenanthroline added to the solutionsK &. @hat is the purpose of preparing and analyzing standard iron solutionsK 6. @hat other items could be analyzed using this methodK