7. EXPERIMENTAL DETERMINATION OF CONSTANT AND COMPOSITION OF A COMPLEX 7.1 Spectrophotometric Method [5] This method is based on Beer’s law, which says that A =e XI x ¢, where A is the optical density or absorbance, € (epsilon) is the mole extinction coefficient, | is the length of the cell and ¢ is the concentration. Consider the formation of a complex given by the following reaction: (ML*} IM"yE] M*" +L ML" K; The complex [ML*"] exhibits a characteristic absorption pattern depending on its composition. The wavelength at which the complex absorbs most is termed as Amax. The value of the formation constant Kran be obtained by measuring the absorbance (4) of the sample solution at the Bmax of the complex. ‘According to the standard procedure, a known quantity of metal ion and ligand are equilibrated. Under equilibrium conditions, both metal and ligand can exist in the solution, either in the free state or complexed form. Thus, the total metal concentration [Cy] and total ligand concentration [Ci] can be given as the following: [Cu] = [M*"] + [ML*"] and [C, ]=(U+[Mt™ [Cu] = [M*"] + [ML*] and {C,] = [L] + [ML™] [M™ = [Cy] — [ML**] and (L] = [C,] - [ML*’] The term ‘c’ in the Beer's equation can be replaced by [ML*"] in this case, for the formation const: Thus, Beer's equation can be shown as [ML*"] = Hence, A A [M*"] = — = and [L] = Sa [M9] = (Gu) — Sand fb] = [a] The values from these two equations enable the solution of the equation .7.1 Job's Method Analogous to the spectrophometric method, the Job’s method also involves the measurement of absorbance of the complex at its Ay... This method involves the preparation of several sets with constant volume and variable concentrations of the complexes. Due to this, this method is also known as the method of continuous variation. Using this method, it is posible to determine the metal-ligand ratio (composition) of a complex [1]. The standard procedure involves preparation of 10 sets of solutions of a com- plex. Different volumes of metal and ligand solutions are equilibrated in these sets in such a way that the total volume of every set remains the same. An illustration of such a set preparation is shown in Table 3: Table 3 Preparation of sets for a Job's method experiment Set No. 1 2 3 4 5 6 7 8 9 WW M™" (al) 0 i 2 3 4) 5 [6] 7 8] 9 L (mi) wo fo}ls|7}o};5]4a)3]2]4 Tol m) | 10 |10 | 1 J 10 fro fio fio | ao | a0 Jao In each of the sets, the sum (C) of total concentmtion of the metal ion (Cy) and total concentration of the ligand (C,) remains constant (ie. C= Cyt G remains constant). The graph of the mole fraction of ligand (if, = C,/C) against absor- bance (A) measured at the characteris The mole fraction of ligand comesponding to the maximum absorbance can be identified by extrapolating the legs of the curve. From Beer's equation, the absorbance is directly proportional to the concentration of the complex. Hence, this point corresponds to the composition with the Janax Will appear as shown Figure 3.maximum concentration ot the complex. Absorbance Mole faction of igend > jure 3A plot of mole fraction of jand —+ absorbance measured at a character Fora complex ML,, ‘1’ represents the number of ligands per metal ion and can be mathematically equated as C,/Cyy. For example, if the complex is fepresenced Ola forinla MIL, the value Ua’ i 1, anid dei aid thar the metal ligand ratio in this case is 1:1. In such a case, the concentration of the complex will be at a maximum in set number six, which contains an equal volume of metal and ligand solutions. The mole fraction of ligand corresponding to this set can give the value of ‘y’ using the following equation: nf 1= mh For set number 6, mf, = C,/C = 5/10 = 0.5, Thus, nf 05 "T= 1-03— Even though Job’s method is a user-friendly method, it is necesary to make sure before using this method that the metal and ligand do not make more than one composition, Under certain circumstances, it becomes very difficult to create several sets with a constant volume. The application of Job’s method is not possible in such cases.7.2 Mole Ratio Method This method is a similar, yet simpler, version of the Job’s method. The difference between the two methods is that in the molar ratio method, the total analytical concentration of metal/ligand is kept constant instead of the sum of ligand and metal concentration. The absorbance is measured at a celength, where the complex absorbs strongly but the ligands and metal ion do not. A plot between absorbance versus ligand to the metal con- centration ratio (C,/Cy) is prepared as shown in Figure 4. Extrapolating the straight line portions until they cross, the value of R = (C,/Cy) gives the ratio of ligand to the metal ion in the complex. Hence, a formula of the complex ML,, can be easily determined. Limiting Logarithmic Method Theory ‘The limiting logarithmic method consists in measuring the optical densities of solutions of metal (M) ion and ligand (C) in which the concentrations of the (Wo species are varied in turn at a constant total concentration. It is assumed that Beer's law is applied, ie. the optical density at the absorption peak ‘is proportional to the concentration of the complex. For the reaction mM+IL > Maly ‘The stability constant, K is given by [ML] iM iLy ‘On taking logarithms, we get logtMaLd ‘The valves of m, I and K may be found by pl sets of solutions versus composition. @ One with constant metal and variable ligand concentra- sion, and Gi) The other with constant ligand and variable metal / |= mlog(M] + HNogiL] + log K =) the logarithms of the optical densities of two concentration (Fig, 1260) ‘The slope of the curve in case () yields “I while in case of (i) “m” is obtained. From these values of “I” and “m” and equation (3), values of K and the composition of the complex can be calculated. Limitations (The method is onty suitable at low conecntrations, Gi) The value of sability consiant obtained by the method is __ higher than that given in the literature. Log (M) oF Log (L) = w “Wy Log Optcal Density,higher than that given in the literature, Log (M) oF Log (L) > 3. Molar Rarlo Method : This method is vary simple as com- ppared to the continuous variation method. The diflerence between two ‘methods is that in the molar ratio method, the total analytical concentra- FIG: 1280. Unig logattnic tion of metal (or ligand) is held constant rather than the sum of ligand B and metal concentration. Optical density is measured at such a wavelenght of light at which complex absorbs strongly whereas the ligands and metal ion do not. ‘Then draw a graph between absorbance, ic., optical density versus Ha 1 ae meat comznenrs (=) xtrapolate the straight line Hence, formula of the complex MX can be easily determined. 4, Matching Absorbance Method : This method is quite useful for the determination of stability constants. This method involves the Fig, 12.61. Nola ratio method. following steps : @ The absorbance of two solutions having different metal to ligand ratios are measured at a wavelength where only the complex absorbs strongly, and the ligand and metal ion do not. (i) Then dilute the solution of higher absorbance to such an extent until the absorbance of the ‘wo solutions becomes the same. (il) Let the concentration of metal and ligand in solution (1) be Cus and Cx, respectively. For ‘the solution (2) Cxq and Cz are the corresponding quantities. (ix) For: Tcompler, we have (Mi) = Ci (MX), C= Gi —IMxh, [Ma] = Cia — MX 0%] = Co—(MX}2 ‘When the avssorbance of the two solutions becomes equal then (MX],={MXlz= Cs where Cs is texas ator onvewtro (W) The itabity constant K can be calculated foym the HAbility constant expression which con thins Cx cnt Kos antnown by oliminating Cs. Tag x= 2M). IM) Benge ~ IMal0d Ce cs CGB * GHG) Ga) (CaKu)= (Cu) ° ae © (Gai Cu) (Coa + Cad Tht, the stability constant can be easily determined (5) Bjerrum Method : Bjerrum determined formation consiants of metal complexes and chelate compcunde in aqueous solution. Bjerrum descoed the slepwise formation of see of metal complexes of the type ML, MLz.... . ML, which are defined by equations (6) to (8). Mu) © MeL Mi ‘(M)(L} meats ih f = Mba) ~® Mis Lp MLL Kes gheyOf the type ML, ML; ..... ML, which are defined by equations (6) to (8), +i— {ML} (6) Merb MEK Iti = tM) o ML+L— Mla = nity \ (ML) -@®) Migs +L Mig Ks tty Equation (6) can be written at (O4L = Ks (04) (41 ®) ‘Equation (7) can be written as (MLa} = Ky (MLL) (i) ‘Substituting the value of [ML] from equation (9) in (10), [Mia] = KK: (LP [M) ly, equation (8) can be written as (MLa] = Ki, KaKae.-KefLI" [MI] (12) Bjerrum introduced a function 7, which is defined as the average number of ligand molecules bound er mole of metal. t may be expressed mathematically as : ge PMLI+2 IML] + 3IMLa) + + 0M} a3), (M] + [ML] + [ML] + [ML3} +... + [MLa} Substituting eqs (9) (11) and (12) in (13) we get jee —KulMML] + 2K Ko(M [L}"+ nk Ko Kp .. + KelMIILD” 0M) + KuIMJIL] 4K, KIMI? + .#K; Ke Ks KM) (LI ‘Canceling “M" throughout, we get y= MMIII + 2K Ko IL) + Ks Ko Ks «. KIL)" ~ Ad 14+ KifL] + Ki KalL}?+ + Ki Ka Ky + Kall] Equation (14) isthe so called Bjerrum formation function. Ifthe concestation of unbond ligand (A) can be calculated experimentally, can be calculated form the equation zeb=i 0) Mi where Ly and M, denote the total concentration of ligand and metal respectively. ‘Solution of equation (14) for the known values of n and corresponding (L) values yields values of the formation constant, ey, Ki, Kay KioeK Let vs illustrate the Bjerrum method by calculating the formation constants for the complex between (CuSO, and 5-sulphosalicylic acid (FL). Procedure “Titrate 100 mi of each solution e., CuSO, and 5- sulphosalicylic acid with standard NaOH solution ‘at 25°C , noting the pH values after addition of the titrant base NaOH . Plot the pH versus ml of base ‘curves and note the point of inflexion ofthe curve corresponding to the mixed solution (Fig. 12.62). Read off the horizontal distance between the curves. Thus find values of F and plot the formation curve of ‘W against —log [L*, the values of the latter having been found at various pH from the relation :“d ‘Ml of 0.1 NaOH Soin, —— Logit? —> 12.42. Petentometie thaton of 5 auphosalioc - (HL) In the presence of CuSO. oe Fig. 12.63. The formation curve. [HL soe ~ [CuL"]~2 [Cul] 4 wr, OL, ~ 19 (Ka Ku) ~ (Kun) ‘Ku and Ku are the second and tird dissociation constants of HLs which are 3.23 x 107 and 1.81x respectively. The yelcs of Ki ed I ound fom te fomaton cure (ig, 1263) ot 70S and a1 ae Ki=22K10" and K2=6.3 x 1 5. Polarographic Method : This method is based on the fact that the reduction potentials of metal ions are usually shifted to the more negative values asa result of complex formation. Consider the reaction: ' 7 ML, + ne + He > MHg+mL which may be imagined to occur in two steps :® 08) Mia > M + mL mo) & ol M 4 ne + Hg > MHg+ mL a where k, k; and kz correspond to respective formation consianis of the above reaction ; M is the metal, [the ligands, ML. is the complex compound and isthe number of electrons involved in the reduction. ingane derived from the above reactions a relation connecting the potential at the dropping mercury cathode, E, the half wave potertial Eos, the current i and the limiting current i, (the maximum of the polarographic wave)vic, Be Ha OOD ng - 00) Pt of & agin loge) wil of couse beasts lie fo a reversible potential, the slope of the line being 0.0S9In where mis the number of electrons transfered in the reduction. By coinsidering equations (17), (18) and (19) Lingane found that at 25°C sss vat -m an 221 eas where Eis the standard penal ofthe meal amalgam electrode, the ratio kyvkz depends upon experimental conditions and electrode charac- teristics, m isthe number of co-ordinated ligands, (Lis the ligand con- centration. Hence, te slope of line obtained by ploting Eas against log toot 2.059 lr from which m can be found asthe value fm is known, Eos (Eas. Volts Fig. 12.64. Polarographi study of te cic ne Gin Sanat Fai om ener (17) is found from the equation: onset Bs Fos en logiLy 8091 soak where Eos, is the half wave potential in the absence of ligands. The plot of the eft hand side of equation (21) against L yields straight ine, the intercept of which on te ents axis vs 2.0591 fog k and hence k can be determined. I is also clear that the slope of the Tine see st 0 that m can be calculated, + If the metal ion forms a relatively insoluble salt whose solubility product is kao ‘ud a compen which fe raat tre sable he stability constant of the complex may be determined by measuring the increased solubility of the salt caused by the presence of the ligand. Consider the reaction of silver acetate with acetate ion. If a sodium acetate solution of known high concentration is equilibrated ot constant temperature with an excess of silver acetate, the equilibrium given in eq. (1 is sats AgOAc gt Ag’ + OAC Ky a [As {0AcT Agt+20Ac™ g2 Ag(OAc)i p, = fAM@AcH -@ [agOac? fier equilibration, the excess solid silver acetate is removed by filtration and the solution is analysed for total silver. Then, the following relationship holdsCaq*= (Ag"] + [Ag(OAC2)1 @ ‘where Cag* is the silver solution concentration. From the solubility product expression, we get. TAg*] = Kep/{OAc™ J, and this then becomes [A8'1=Ky/Crsore ~@ because of the large exess of sodium acetate, Substituting equations (2) and (3) in the formation constant ‘expression which gives Br= (Cag Ky/Creore/l(Ky/Craoae Crsoie)') 4) Each of these quantities is known, and the overall stability constant 2 may be calculated, Electromotive Force Method : A meial in contact with a solution of its ions gives a potential, E, which is govemed by equation (1) E=E’ RT /n Poin (M™)/(M] solid) If two different cells ofthis type are connected, the voltage isthe difference of the two cell potentials and can be measured with high precision. If two cells are identical ée., both have same metal and the same metal ion concen‘ration, the measured voltage will bo zero. Ifa ligand be added 10 cell the complex equilibrium will cause © change in the free metal ion M™+L ge ML @ concentration and a net voltage will be observed. The observed voltage is_given by eq. (3) or (4). Buy = Ei-Ey (RT /m F) Inf", /T 2a ~ (EM aM eid Eete = —(RT/n F) In ((M™7],/(M™"]2) ‘where T is the absolute temperature, Ris the gas constant ; F is the Feraday (96500 coulomb); and n is the number of electrons transferred, in this case (M° —»M™) equal to 2, Thus, the observed voltage is direcly reiated to the free motal jon conceavations in the clls. In practice successive portions of ligands are added to the metal ion solution in one cell, and the corresponding voltages measured. From the known ‘otal concentration of the metal ion, ligand, and uncomplexed metal ion, the stability constant may be evaluated.