a ATE ~ gHEM SL! WAL TATE AND GUANTITATIVE CHEMISTRY LABORATORY | ACTINITY 06: SPECTROPIOTOMETRIC DETERMINATION OF AN Eau | 4 i PHOTO! IC DE iT LIBRIMM | CONSTANT AND METAL JON ANALYSIS alataPerce nuers 5 JL Favcisrwm CONSTANT DETERMINATION [As PREPARATION. OF STANDARD SHunons. To ESTASUCH A | CALIBRATION — CoRvE eeeetommmrnrsie. Tithe icoleionsi vin Table to Cheek: Hots puts cieewedl tect _ Ef Hees cons actommodate 25iml sp! _silvtion by meawring oot 1s ml _porhon of _purigied woter in a graduated cy)inder ancl transfering this Golume__te Cch of _qour_test fuber. Mark fhe level tha} shovld be occupied by 25- ml selufon in each Lf your te. Pipet 6) 1,2, 3, 4, and S-ml_ o 0-001M KEN inh scepante | labeled and clean test tubes ipa 10-0 mi of 0:2 M_Fe{No3)3 into each bbe and quand torvely dilte _& 25 mb [the merk you macle on tbe ) with OAM HN03. Shr or agittt each soluten — thomughly to ensure that equilibrium is es pblished . Satis TAGLE 4. Composinen OF TWE SET OF STANDARD FenCsat SoLuMeNS For PREPARING _CAIBRADON CURVE ee STawparo 0.2 Fe (Ns) Becain Ecrs Olm SovuTant Gaim 03) Cin orm HN03) —_ Hos BLawK (0.0 mL Oo mb Dilvte te 25me 4 Jo.0 mt Vo omt Dilute 75 me 1 jo.o mb 2 ml Dilvre fe 25 mi 3 10-5 mL 3 ml Dilute ty 2S me 4 {v-0 mL 4 ml Diivte fp 25 me 5 Jord mb sme Dilvke fe 25 me Tye chophotometer Set the wavelength scale 2% 447 nm. Rinse 9 covet thee mes with perhens the blank — selukon. side a ith hint -pree———— DATE _ “ara hondle the —cwvet on the cloudy cider. a. Ploce the cwvet) hee povrths filled with the blenk solvher, Lint the comple Compartment , adjuct as needed clependiing, on She _specho phohmeter being used, and close the cover. Set Yre__meter on the _spechughohmeter to fead zero absorbance, Remove the wet. Once the inshoment has been set, do not perform any addthars) adjustments for the temainder _| T op the experiment. 4. Empty the covet ancl vinse it three #mes with porkers | op _Solubon 1. Fill it approximately thee -pourths poll with Solution 1, Again, carepully Gry the Ovtride of the covet with | 7d 4 a clean lint-pree tssve paper. Remember to handle only the Vig of tre _ waver» Place the covet intr the Sample compartment J cluse Yee cover, Phen yead the absorbance at the given wavelength « Repeat Shep 4 gor__Selvtvns 2, 8 Qs. 5. Plot obsorbance , A (ordinate), vere [Fencs **] (abscises) por the six sofvtione en an appnprate Soptware | Drow the best ~pit line threvah the six pocts te esteblish the calibehon curve B. ABsorBANCE foe Me SET OF TST SALUDOS 1 In_clean test _fvbes, prepare the tect solukens jn Table Z._ Use _pipets pr tre__volumetne _mevsvements, Be ¢oreFu/ fy clean and __ nse the _pipets to avad centeenrn te Conteminoben of the reagents _prer + Me prepanton. See to it Hot you oe _vsing the _appmpriate Cancentahens of test Solon Components. *DATE TABLE £. COMPOSITION OF THE seT oF EGUILIGRIUM TEST soLUTDONS —— for ME peTeRmNanoN oF KC, Test 0.002 M_ Fe(N%3) 4 0-6627m Koon 0./M Hiog soLunon (in 0.1M_ Hw03) Cin 0.1 M Hw03) G 5 mt 1 me mb | i SB mt a ml 3 ml | 6 5 ml 2 mL 92 he 9 5 mt 4+ mt 4 mt Som 5 mL S. (7. Blank — the spectrophohmeter , Sir er ogitte each tert selotion Unk! equilibrium is Heche. Ringe the covet thtoughly Gemeee Maithvepérkens op the Fest pcaludiors and pill $F thee -fourthe pull Cleon # cm the outeicle op the Covet _| Be covkovs in handling the cuvets. Record! the absorbance| oe each test Solution. 3. Fam fhe calibrton cume use the recorded absorbance yalve + each test colvhen tw determine the equilibrivny —moler __ cencentotion ap FeNCs*"- C. CHcuiaros OF Ke Table 3. Set op Stondard Solutors te eshbiish a sbndardizahon cune. Table 9. Absorbance aa fre set oF test selvdons, Toble 5. Calculation op ke _l_metay Jon ANANSIS A. preraranon oF sTCK AND sTAHDARD SOLUNMONS 1. Reger to Toble GC for grmup!s assigned met) ionDATE _ | TABLEG. METAL Ion ASSIGNMENT PER Grour WITH voLUmME OF ComPLERINE AGENT Pe STMck So.vNON pre pARATnON) Geovps meTAL roms Complenswe AGENT as see 3 Gu lomt cf concentrated Mx 4,5 and 6 Fe>* Pome 6.4 WM KSCN 2. Pepare 00 ml of 9 0./0M stock sebtion . To clo thic, you will need fr Enow the formula of the Cy** or Fe # jon- conining solt provided in the rerqent area jh order f_colcvlate the mass of salt por its _peparton» Measvte the mass of the salt. 2. Tnnsper the s2lt_ hh a f00-mk yelumehe flack , halp- tiled with 0.01M NH3 , add the apprpnrte complexing agent [sec Table), ard then ai lte tr the mark aith 001M _HNG, 4. Prepar _phesolvhon in Table 7. vsing clean, l2beled test tubes, Check Hart gour issved fest thee can gecommedate 25 mb of __seluen by messun'ng ovt Z5- mt port of punpred voter in 0 gaduated cylinder "end tanegeming this volume _t “exch %F your Whos. Mork te level thot shold be occupied by 25-ml | selubon (7 erch oF your be. Use serlogye Pipets * dispence the wlumes o stock selon prepsreel in Sep 3. TPbLE 7. VOLUMES OF STOCK SOLUTION AND DLLENT Fyre STANDARD SouuTon Ss PREPARATION STAWOARD SowTen STOCK SELUDON DILUENT (hloM meta tow) Co.Dim HIN03)) Blank Omt Duvre HE 25-ml mare ' ym Dior Me 2E-mt__mark |e 5 mt ouvre mT We 25 “Ml mark 3 [o mt Nive my WE 25 - mt mark 4 15 mb owe TD He 25 - ml magDATE Eee ZEIUP ES DIT pion SoS ara mae | deveRMNATION OF A may | \. Apter the Specho photometer hog been fiened_ ie tee atleast Jo_minvtes set the wavelength 4» its minimum (~35¢nm) ond set the 26m en the _spect»phobmeter , Rinse twice a cuvet with the blank — sclur Fill the cuvet at least three-fourths pil) with the blani< seloton and dey the outside op the cet with clean -lnt- pree tissue paper, temoving _zingerprints 2nd moisture . Ploce the cuvet inh the sample Compartment and adu st such that the meter should toad ze dbeorbence, 2. Set the wavelength op the spechephotometer to the shortest wavelength setting , Place fhe Slvtien S in @ Covet | folowing the blank seluben procedure. Measure snd record the wavelength 2nd sbsarbance- Scan the wavelength range the spectrephhmeter (+350 nm ~~ 750nm while recording absorbance values every ~50nm. - Remember t set the absorbance 3+ rere with } the blank solukon at each Chosen wavelength. 2. Plot the clef Ibsorb»ynce # (ordinate) versvs Wwavelengeh Dd (abscissa) with an appropriate Sophuare ty determine Jax [the wavelength where maximum _ dbear phen occurs far your met! on). C. PloMNG THE cmIBRAPON CYRVE 1. Set the spechaphoh meter at [mx. Record the absurbance salukon 5 at Amayx, Measure the absorbance for the other _stendard _solukune in Table 7 staring with the mos} dilute —_[Yhe_blank) . \p_ at lest four op the shn dard —_|_sdokuns ae nok i _th@_Dosorbance_enge oF. 6 to 74 Prepare _additivnal shndord _Sdlottons. 64DATE Plot abcorbance A (ordlivrate) versus molar cententrahin __(pesciss2) for tre six _selubons in Tobie 7 (or the additens) strrderd — sotons you prepared ). Dow the pest shright line thrugh the clot» ponds h eshblich the calibrebcn Cove. Table &. ABSORBANCE OF STANDARD SoLUDONS D- DETERMINATION GF UNKNOWN METAL [0nN ConcENTRANOM [- Combine 5-mi__porkene ip any fue Shndard selubens in Table 7. Take mote o¢ phe storting concentmhers of the samples phat you are mixing , Mix fhorghly . Prepoe tue adel Hone! S?~p les 2. meacvie te abcorbrnce ao your three samples in the same Wenner 96 the dhsorbance velyes were mersered . Ye Stondord tolubens. Read the c@bbmhen cuve 4 determine the molar __Cancentak'on op the meh! im in yor samples. 3 E xpress the “oncentahon of your mebl ion in ynts op __ mess Jude (ie. percent, pats ger _milion ate TABLE 4. Yyty For UNENIWN MmeTAC (en CON CENTRATOM EGuiPmesT USES - meter Covette _(Gises_2 Plastic) Sermlogic Pipette. Pasteve igette, Gaduatee NinersDATE BRIEF DSKCRIPNON ANO oKET oF LT. EQUILIBRIUM CONSTANT VETEXMINATION ve + | | = pur exact meacurements of each meagent in every fest tube. > tranifer each volubon in a onvute filing (t 8/4 te the hp after rising with falank soln partons Sx ‘Solution 5 3] = pl te ver and pace ah ene in Ine gethnptmeter | ‘olution? | Fetateralicainiine teastucnt ado fhameinektcam ence! iol ‘solution 4 4] arsvroance ty vert alter ing thon read: Reytal for she wank ][ |] remnining srubens : I. metal ton tmatytis = mieasure salt required fs thre trick solution wing an onalydcal belaree then trnifer ha vilumeme flak = Add the corresponding complening agen’ and dilute tre mark wlth HNO, SOLUTION F zxlo-¢ mel SCN 9 xlo-sm p.02s & Sel'n . Samm 3 3 x[0-% mol SON” a 1,2 x(0-# M oe 0.075 ¢ sol’n | Savnow f 4/07 mol SCN™ 1.6xlo-4 4 0.025 gal'n sat 5 Sx | Peete ool fe Seno 5 a «mors OF sey, /NIDAL = solunen aw lt keen, leekSew 0.002 mo) KSCN Irmol SCY” 7.x /0-% Too m KSC Ie KSeN fmol KCN mol SCV sou nen 7 .. ImL Kscw . [kL Kscn 0-002 mol KSCN | Ime! $60" y~¢ 000m. KCN IL ksen Tool KsCN~ mol SCN SOLUTION § a ai KeON TL Kscn , 6.00% mo) Keen [mal SCN” 6 x/0-® Tom. KSCR KSC dmel_ KSC Seen 4 S| _Ksen 002 mol Kicn__/mel SC" 8 xD 7000 mt_KSCN Ie Keen Irol sew- MON Sen wDATE Sorvnen 10 Smt KSeN Iu keep 9.002 mut sen (mmol SCN} x Jo~F ————— 1000 mt IKScn IL kecw — jrrol kgcn mol SCN | | TABLE 8B. CompoTanons Gu en TF utnee (0.96) = CC Ee vk Caz _(oto) Co) he te OP as Gorvnen 4 (0 )0) - AES a Cr = (6. )C C2= 4x fo A ts 7 Suomen 2 (0-1)(5) = Co 0s) ae Lunt, Cp = 002 M som 3 (0-1¢) (ie) = C2 (25) Gey Cs) C2 = 6109 M Zs Sivnor 4 Cote) (I) = C2 fs) cy = (0-10) (1) Cz = 0-06 25) & suomon s (910) (%) = C2 (sy (2:12) (2) joa = 0.08M Zs TABLE 4. ComPutanons x LINEAR = REGRESSION LAUATION ; ye lt bx ~ 1.7014 Re = 0.9733 SCONCENTRANDN OF MeTAL tow FROM CALIBRATION CURVE (mel de) Wa 42,3 C_ (2o-Is}- 1-704) cz 01058 Tt. 8 | —— Gr MeTAL ON Flom CALIBRATION Ovave x Tim 0.01658 zy = 0.04074 ; — 1 0.016 $8 3 3 6016 X= 0.0058 +sDate Ls ConcenTRAMeN UF METAL iON ( rl git att, 3 | 0.01656 mol, 55854 0.925943 9], E [mol fea "AVERAGE CONCENTRATON OF meTAL [ON (mass [vocome) 7 = = TRAC S6.45493 x= 2.171974 } Tu 20. 92S 993 2 TRIAL > 0-425993 = 01426993 TABLE 5. Computations + UFENCS*TD exunisaium FROM CALIBRATION CURVE tinor Linear Regression Equation : y > Sous. Ux 4 0025S Solution Y (A = 0-812) Solution 1 (A= 1-066) 0.512 - 0.0785 E dst - bones Sou. & Sous -y Solution ¢ (1-447 + A) Solution 4 (A? 1-844) 1u1- O°02s¢ 2 SS BOB zal 1-QWY + 0.0¢cS Z SOUS. ‘ S048. y i Solution 10 LA 2-14) eB ISU = 00S aru se S049 & + MOLES FeNcs* At couriegum Solution (© Iu 1.08355 5 07% mol Fenes* ssc-xi0.% Fences 1000 wit. 1: ve Solution 7 1 2,04318 x (074 mol Fence” aig-xo’® fencsty_| 1000 wl te Solution ¥ ie ie 20S 808 x 1OvY mol FEES 9 cc 0g 102% wel FN 1000 wnt. te Solution 4 -4OSSU x (0° yo) FEN we {000 mt —__Slution 10 ae eel a me x AE KAN yp wre FENCE Gantt x (07! me Fen 100 wt itDATE “fre Veaunspawm ANO (SCN "Jeowiignum _Ssuhon | fe®® 4 SN = Fence’? (re**] nit oar E ee = EE Swab 3 ais = s jo mt am EE a {con Diniiat a xt 1088S « 10°4 Sere nner [fer] = veiw" - 0kass x4 s ¢.aU4s xe Mm 1 mL {sen} £2 x04 y.onacs xy + etter eT " qu Moxe Ss ‘soiution 7 r : fe2* + SCN = Fencs*® (603 ital 1 1x"? 4x04 ° _ (0.00rm) (2 mt) ces cs eS to mt Eo xwd-x — Axford-x 2.04518 x0 xe g0us18 xto74 teal) 073 - 9 rosie xi = 7-4Su82 XIN [sm]: Hx york = c04ag ee N= 1 qsue2 x7" Solution $ “fet SCNT Fence Cscnt=T initial 1 xio"* axed be p+ le norm) [2 me 4 le MOL x x fe wh seal E inpt-x xp -x _ 2-48%08 x10 . | X4 trtort~ 208808 x 1074 fi [febt] + Into“ - -9SROE xjo-N + TANNA xiot om a (sory? oxford = z-esnon x04» 4 ANIA x0 om Glution 4 a fet ye sense Fees? Coon" Tintin i | iio Sx ior4 ° 1.002 m) (4 mL) , “yy Ce > x WW mt F E iso t« B x10" 4-uossa x10 Ke F:WOSSG x 10°F {Fe} = 1xpo-> - 3.60SS x0Y * @-3Amy xl“ M “J Gx ped = $.4oss@ x10% = Heaquid ye Slution 10 + SCNT (sen) initiq (©t00 M)(¢ wt)eras a VxtoS - Gawi7 xR = gTSRS xd HM “Aged ne? 4 22417 to + 577883 x 10-4 [+ moves OF Fe3* gexcren AND MOLES OF CCN” Reacten | Sinet FENR ys Fereacred * SN vented Solution wal Fe" reageeg 2 1:08 SES X10°" yo} |_mol SNvencked = 1-0E3ES ajo“ mo! Solution 7 mol Fe reacted > 20USIE X 10°" mol mol SCN reacted = 2-04318 x10 “8 aml Solution & wml Fe™* reacted > 2-880 x10 “& yo! mol Sen reared * 2-8CR0¢ x10~4 nel Solution 4 mmol Fe reqareg > 9 W°SS© x10“ mol mn) SON" reacked > 7: WOSSe x07 mol Solution 10 mol Fe>* reacked = 4° 22417 y jo “& me] “aml Sen" reacted = 4.22417 x 1p “* mo) + MOLES oF fFe>* ar couLiBRiUM Solution | g teys x 1074 mol Fes “4 cote {ie Solution 7 md 1-98U81_x (074 mol Fert a 1c Slution & ’ S| 7-ydjar ¥ 10-4 mpl Fes at are} te —_| Stating 4 Me bn oto gg ae tH tor ol FE. att ar tot mol-Fe 2X —$§ a ¢-1158% x 10% mol_Fe 3 tc “CONTINUATION DF TABLE © COMPUTATIONS AT PAGE 7 wGenens oF THE Resuits loratiod Curva My CredncP a] wl Absorfante Gall lost . ! foobar | [o.oaous INEAR REGRESS | NI = F043. bx [4 g.0 ss nd = 0.9466 Tre grOph above shows the plot fur the different cConcentahons of Focns in soutions Gnd their cespective abiorbances, My graph generally depicts an increasing trera berleen the FOONT™ concentrohons and the abworbanu Therefore, we can infer that as the concentration of Fens increases, the absorbance, als inureasts. The, curt) is significant for the calaulahon of thy molar concehtahin of thy analy ot tquilbvivm, alelehgt vt. Mesotbahve| ofc lurgn [F 4 a [ae [oe ae Pe «fo Thegrmph above, shows the plot far tht wavelength that the spluin wos introduced fe and ine absorbance of tre stluhon at that wavtltngth: According to the graph, the highest abvorboance recorded wass when the warolength iy Jet at 350m. Terefore, tre folutions Amax is at dS0nm,orf of Metal ior] uf Absofodncb | Golurion|— Fo\whon |6) °. oe | 03 | oly | ops | ope | ol] « Y= 4el26 im The graph above snows the plot for tne different concentrmhons of Fe® in the sol Uhon and their aigorbanes of they Amax, which is gOnm. However, some of the data ig deerma unreliacl, becawt tv gah incks lineanhy. Tri problem can bts vowed by interferences present in tre sdkahon or the equipment wed scm ag Sormtthy in tht cuvette. 1 remedy thir problem, sume of rhe data may be omitted. Moiat cohceptratioh of Metailior «ef xbsorbahc | (Blank) Sojuripn 7 fo}urjon [2 | s 1S yle te. Bx FI. fov R24 0.A143 — There an fwd _gynphs mode by the eiperinented al Sawn above. one ir shown wing all fhe results for the blank Soluhon tp doluhion $. me ther gerrh snows the an rine wlok Soluntn up Jowhth 2 only To snury tht accarake onlonlani mn of the anctntahon of thy Metal ion wing the calibtahow curve, fhe *perimenters omitted nswlts far Seluhons 3 fo $. Mis amithon iS also hy Dane thal the Re of Or darn fri witmn thy, vonge 0:40 fo 04.0, wit With indicaks that the dan gathered is liable os | a wDATE | @RROR ANacrsis oO | + PossiBLE CONTAMINATION Tre grup Ws unable keep | aecount of how the other _grvps had prepored och sample because _Yhit expen'ment is Camied out in Clecters. Glacswares or equipment trot was conmineted my hove been vhlized in some cpses. + JMPROPER LaBoratoRy TECHNIQLES Since everyone was Unpemiliar with this 2pprach aleng mith Specipic equipment , given that the expenment was } t 7 done in Clostecs , if is ossible that occarsons/) Pp. 2 ei Seme__shdents pailed + follow the correct _procecturec E Washing and Aning wvettes- i ii 3) Revsing dis pesable plaske cuvettes with scatches 1s another _pacter. Scratches en the cuvette will hove © similor _impsct in hot less light than anherpoted sill rtach the detectr, dve te Gcottering or refrection erpect. once more, this will fick the inthmert intr indicabing 2 higher abéorbrnce than is actually the cace. * Possigle soLunon PREPARANON ERROR Possible errs jin preparing the Strek Colutions may have occured Such ae im proper mining 6 reagents, indecurate divkion of _somples, opponescence ypormaten, ar msupy cient me in order por the _eachon 4 reach equil/onum before, rePding "the absorbance due to the limited tme in ferforming the ¢xpen'ments. ‘NON LINEAR? RELANeMSHIP PRETWEEN fpsonBANCE AND CONCENTRATION, Possible eners ante m the _expmmenhl lure and. inter, fo he. cwvettes which some have _gentohes . Scratches on the _Wvettes cwse_ li ht saatering, moking deviakine fen Navan hy.DATE a — | cemecism OF PROCEDURES +The mark made on the test bes dees rot aceuntely measure sme, Veliring micro pore 26 2 marker in indicabing the Volume measure ments oc evch sample oli bred covld _resvlt in inaccuracy in the readinge. No markings exists on erch he since test bbes are not designed ty provide precise volume _ measurements. f } - + The phcedures didnot spect Fy exactly how the experimenter « should “vege” and “carepully dey” the aucee This gave Conpysion + some and led te the Shucertte | hosing _veming techniques pen Arying and __ rinsing. 44 4 "Te precede did not inclide making replicetes Which wold ensure mere accunte ets + “fb account for aendem errors by calculating avenge values A Jw number tp replicates can recvl+ jn less reliable olata rnd greater sus cephbr lity te vanations. * Since the expen'men hon was __perpyrmed in Clusters tovblesheoring err Was dippicult. Procedvval enorc are also diggicultts _idenh’ since all op the ctor an only be @mpwed When everyone ie clone papery each task.HOTS AND DAEAONL cs 7 DATE Spectroweomyy Fee. AE He Miembtle hls aA Memement of lifer mcMoIn Metudeen retry one electomequehe. ovation. WF entails analyzing He tnt spectrum ol a femme eth, aoe, oF seattert ih ever 4b lawn nom abo MH mabe Up, Srudure | Ant chorocterietict (Harabook of Soretrouurpy , 4.) SpreInscomy encompasies Verio teerniqiea Ann Mee Aiea raefon oF 4ne elecromog mene SGM, em at Irgrarea (IR) , vhibiy, viiaicler (WV) | Ke-vey, OF germ rou reoiont. Te Interaction of electomuanehy rediaton sith tre emp, wide estes IM ty ateorgton oF emisiion % Protons, W dre qundamental idee leering sprotancrpy - A Sorell of spectescony , caltes aprcre prone meta) desica WAP tmuowving heh mud Vine kb dotorbea or trowmittes 0) 6 EmpIe Gt A jwdton of wiauelengtn Or prequenay Gooian et ot., 2020). The snout of @ chomeyen qprmoa tn a chemical Fwsctiom betes Om arate Yn m cinlcol Sams ane the evameeht PEC yor thot onalyte TE QuERRLeA sting Sractogrotemmeiny = Aerating fo the amount of ovate im dhe nical simp , ar amount of chromonin qrogsera 03 ne feochen % Pororione’ (echmann & Whey, £028). 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