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Procedure - Wear Gloves! A. Preparation of Standards and Calibration Cerve Volumetric pipets will be used to transfer the appriptae ves of the chemicale 5000 m vihmenic duke following the table below, Once de chemicals we accurately restened to the volumetric flaks, they will be dilunod so the lor with denied was Once these are diluted to the lise, cap and mix well by veing the flok MOTERY ETAVE Standard Number Mank 1 2 3 4 Volume of 0.0500 mg/ml. Fe standard tml) 6.00 1.00 /200 400 5.00 Volume of IM acetate (ml) 1.00 1.00 1.00 1.00 1.00 HO 1.00 100 -100 1.00 100 1.36 Valume of A3% phanhniline (ml.) 11.00 10.00 10:00 10.00 Create the calibration curve using Esed or anoder computer grphical program to create the best-fit line and how the equation of the 2 sobenit as part of the lab seport Do os forget in include Total Volene (ml) 500 5000 500 500 500 1000 These solutions develop for 45 maus, so they can to orase the color Degr coming de 45 minites hydroxylamine and phenahme se mand. The water begin timing. The desonized we can bed later so die them should be saved und Part B is complete se will befoor seting Then the standandi made in Pan A and the solutions from Part B will both be red soge Sving the tractions in Pt C be send they have been given that time standand iron, anupcitati acetabey have to be siled in ceder he ne Aho, these solutions analysis program the de graph Pine the graph and dats point Part B. Finding Unknown Iron The subetance being tested for the amount of mon peceat is actually wheeld Before being used by the speemophometer that must be doved and used to se new solutions Obina dean. dry 500 ml. velomenic flack and accurately weigh about 0.1 g f the unknown soll directly into the volumetric flask. Carefully add 5 drops of 6 Malfuric acid to help dove the sold then duteuing deionized water to the line on the flask. Cap and invest ou down standard solution that will be used to create the remaining solaces in this part of the lab. The unknown standard soloon shoold be colorless and therefore will never be sed in the spectanphotometer well Volonic pipets will be used to transfer the appropriate volumes of the chemicals no 500 ml. volemic flaiks following the table below to make the unknown slices that will be seated in the spectrophomeler Oper the chemicals ate acomtily tradem evolumetric flasks, they will
be dihnell to the line with dewind wairs Doce the by verting the flasks many times Unlooww. Dilate Solution Number Volume of Unhamn Fe standard solution (L) 1.00 100 1:00 Volume of IM acetate (ml) 100 100 100 20 Fint check that the Volme Speciniabotonacter instructions de IHOD 1.00 The three identical solutions mit derskop dat u en 45 from Part A. Begin cooning the 45 mi hydraylamine and a-pheandmine semised legiooming Test bods the standards and known 100 1.00 Pan C. Operating the Spectrophotometer Bring all the solutions from both Part A and some of dexperte they have waited 45 minutes from when they we cated. The amo for each group to get their ones. Do not sheme bere prog samples they must finish All samples mit be non sogether a There are two types of spectrophotometen in the lsh You between the One nedd is now and the aber w well, but there are different autruction depending From here on out I will call the newer modele de "Ne the Old spectrophotomer Select below the comme Valume octus send the lab after nough of these ins Now the strument is ready to use, Bele ning with the blank soli canned in Pur A. The purpose of the Ha one of our compound when there is ame presetit, por dem. In the lank shere should be us Feata mus the blank, take one ut, which look like a small sest tabe du solution is a witt beaker. Fill the coat wit the Mask sekon begini minning their these inmutent dearly till the diffen Bods types work zipidly phometer you dusose. and the ones from 1968 depending t Sound on the dal no sop of the Now she iatzumeneedi length ment. If is not set to the correct weegh, 55m The cas the dial so be set to display values in the set Abstance. Neste de digital scores has deploys the railings there is dot and four weeds Transmittance, Abocah, Conce button until the dia Fat Push the Mode ter the aumento desest to the balances before ing added to the blk To pedom with the bank solition Dump on, the cerWipe and
P Date C/25/22 Serticize Questions: 1. Would your absorbance values be de ameif our cet w 200 on cal d 100 min why not No. THE WIDER CUVET WILL MALE LIGHT WIL BEWERD ON THE SPECTROPHOTO THE LIGHTARAN SANTHERA NA ZACH CUVET AND GROUSE 2 If solusion of copper was found to have extinction cont shoorbaance that should be found it a 002500 M 3. Why mait you always fine the caret up the same way when placing is the spectrophotometr What errons would occur if you didn't do this? TO ENSURE ACCURACY OF WOW MOT LIGHTWAVE MERGUREVENTS COULD BE SECTED BY & LONGTHENED SHORTH DISTANCE OF Splet O Flow LIGHT DMT 7 4. Determine a procedure to accurately create 15x10 mg Fe/ml, whinnering 00500 mg Fe/ml, standard solution and only having access 2000 100, 200, 300, 400, 500 and 1000 ml. vohanne pipen than one solution by dilution to accomplish this i ml. volmestic flasks and Hint you may have to make there out (sales PRINC 0.0500
086 ance Old an "L 1- UP Procedure Wear Gloves! MERGE COLORATION SAT AP A. Preparation of Standards and Calibration Curve Volumetric pipets will be used to transfer the appropmate vokames of the chemicals into 500 ml. volumetric flasks following the table below. Once the chemicals are accurately transferred to the volumetric flasks, they will be diluted to the line with deionized water. Once the solutions are dilated to the line, cap and mix well by inverting the flasks many times Standard Number Blank 1 2 3 4 5 Volume of 0.0500 mg/ml Fe standard (L) 0:00 1.00 200 3.00 400 5.00 Volume of IM ammonium acetate (ml.) 1:00 1.00 1.00 1.00 1.00 100 Volume of 1016 lydroxylamine HCI (ml) 1.00 1.00 1.00 1.00 1.00 1.00 Volume of 0.30% phenanthroline (mL) 10.00 10.00 10.00 10:00 10.00 10.00 Total Volume (ml) 50.0 50.0 50.0 50.0 50.0 50.0 These solutions develop for 45 minutes, so they cannot be tested until they have been given that time to crease the color Begin counting the 45 minutes as soon as the standard inon, ammonium aceste hydroxylamine and o-perianthenlase are mixed. The water does not have to be added in order to begin timang. The desonized water can be added later to dilute them to the lice. Also, these solutions should be saved unnd Part B is complete as well before testing Then the standards made in Pan A and the solutions from Part I will both be tested together following the instructions in Part C Create the calibration curve using Escel er soother computer graphical analysis program. Use the program to create the best-fit line and show the expansion of the line on the graph Print the graph and submit as part of the lab report. Do not forget to include the black as a data point! Part B. Finding Unknown Iron % The substance being tested for the amount of iron present is actually a white sold. Before being tested by the spectrophotometer this must be dissolved and used to create new solutions, Obtain a clean, dry 500 ml volumetne flack and accursely weigh about 01 g of the inkoown sold directly to the volumetne flask. Carefislly add 5 drops of 6 M sulfuric acid to help dissolve the solid then done using deionized water to the line on the flaik Cap and invert to mix well. This is caur unknown standsed solution that will be used to create the remaining icons in this part of the lab. The known standard solution should be colorless and there will neret be tested in the spectrophotometer Volantric pipets will be used to transfer the appropriate vohames of the chemicals into 500 ml. volesenic flasks following the bible below to make the unknown roluons that will be tooed in the specamphotometer Cace the chemicals are accurately transfereed in the volumetric flasks, they will
d be diluted to the line with deionized water. Once the solutions are diuted to the line, cap and mix well by inverting the flasks many times. Unknown Dilate Solution Number Volume of Unknown Fe standard solution (ml) 1:00 1.00 Volume of IM acetate (ml) 100 1:00 1.00 Volume of 10% hydroxylami HC (ml) 100 1.00 1:00 Volime of 0.39% penanthroline. (mL) 10:00 10.00 10.00 Toal Vabime (L) 41 30.0 50.0 50.0 The three identical solutions must develop their color over 45 minunes, jair like the irandand solutions from Pan A. Begin conting the 45 meson as the unknown, ammonium acetate, hydroxylamane and o-phenantheoline are mixed. The water does not have to be added in order to begin timing Test both the standards and unknown solumons at the same time uring the same spectrophotometer Part C. Operating the Spectrophotometer Being all the solutions from both Part A and B to one of the spectrophotometers around the lab after they have waited 45 minutes from when they were created. There are not enough of these instruments for each group in get their own. Do not alternate between groups Choce a group beguns minning their amples they must firesh All samples mest be run together at the same time on the same instrument There are two types of spectrophotometers in the lab You will be able to clearly tell the difference between the ran. One model in new and the other was manufactured in 1988 Both types work equally well, but there are different instructions depending on the rype of spectrophotometer you choose From here on out I will call the newer models the "New" spectropherometer and the ones from 1988 the "Old" spectrophotomett. Select below the correct set of instructions depending on yo instrument Seccaphomet instructions. Fint check that the wavelengths to 530 mm This can be found on the dial on top of the instrument. If it is not set the correct wavelength, rom the dial until it Now the instrument needs to be set to display values in the unit Absorbance. Next to the digital screen that daplays the readings there is a dot and four words. Tranmince, Absorbance, Concentration and Factor Push the Mode button until the dot is ex to Absorbance Now the merament u ready to use. Before ring your samples, the stroment must be blinked with the Idink solution created in Part & The purpose of the blank is to set the instrument to detect none of our compound when there is none present, ut like when we are the balances before using them. In the blank there should be mat no on standard was added to the blank. To pertom die blank, take one covet, which fook like a small set tube, and fill it with the blank solution. Das dos subcian in a ware besker. Fill the cover with the blank soktion again and pour into the waste beaker Repeat one more time Then fill the cuvet with the blank solution, take one KimWipe and
56. ce 2 Part B. Finding Unknown Iron % Mass of unknown UNEA Fem Yemery longtegle. lage Unknown Dilute Solution 1 UhmNH DÒNG Solion a Unknown Dilute Solution 3 (Show calls for the station of Uk 1 Te "= 100 mg Fe Average unrounded Average Standard Deviation (Sallation) Agr TALFF MIV. = M₂ V₂ Absorbance Concentration of Fe Untounded 0.309 w Po/mL. 0.344 Dilete Salt 1) 100 Percentage of Iron as the Unknown Sold Concentration of Unknown Pe Standard Solution made from solid Percentage of low in the Unknown Sold Unrounded Concentration of Unknown Fe Standard Solution made from sold unrounded lewat 100. 10. F/L LF/ml. mg/ml. mg/L. Fr/L
Name Questions: 1. Would your absorbance values be the same if our cuvet was 2.00 cm instead of 1.00 cms? Why of why not No. THE WIDER CUVET WILL CONTAIN MORE LIGHT ASSORAMIC SOLUTION THEREFORE MOLE LIGHT WILL BE LESORGED IN A 200 CM CUVET AND REDUCE THE RUNDING EN THE SPECTROPHOTOMETER, 2. If a solution of copper was found to have an extinction coefficient of 51.6 cm 'M', calculate the absorbance that should be found for a 0.02500 M solution using a 1.00 cm cuvet. 3. Why must you always line the cuvet up the same way when placing in the spectrophotometer? What errors would occur if you didn't do this? TO ENSURE recuency of MGRUNT LIGHTWAVE ME SUROVENT COULD BE SEENED BY A LENGTHENED/ SHORTIOVED DISTANCE OF SALE RECURENTAT Cutt Ruve LIGHT LIGHT 4. Determine a procedure to accurately create a 15x10 mg Fe/ml solution using a 0.0500 mg Fe/ml, standard solution and only having access to 100.0 ml. and 500 ml. volumetric flasks and 100, 200, 300, 400, 500 and 1000 ml volumetric pipets (Hine you may have to make more than one solution by dilution to accomplish this) 12345 10 8.0000018 100 mt (soal tecke 0.0500 wasten! NEP