1288 PRINCIPLES OF PHYSICAL CHEMISTRY sutrality, an equal num 7 Jan equal umber of Na* ions woul! pass from right to left acros the merabrane (ste the ‘equilibrium save shown below). ie ae tor oe ee a ° : piri ae ee The sma pres oa i measurement would tis articles on Sein eo On artes present on the Ie sige, Le, on te pateia se of te eas mbes A emit, the cemizal pat ime the sane on hse AL . H+ RT Guach = u* + RT 8 (ocd, @ hee ac fepreseas the activity of sodium toride i 5 ts lade ih of eiprete eabeaThs, web for ee eaprn (rach =a" aah = (Ya Na* (CT epee = (yah y (Cry, ey ‘of NaCl which is present on both sides of the membrane, (Gruc * (yu Qa Assuming that the mean ionic eau en y= de the Donna gue (ra)F(Na*),(CO), 83) acini execs i he conga wu e ee soe i seg esol ee (Net) (Cry (ate Na], (Cr, (2-8) (4-2) = ex : Cea deat roe of $2) <2 ene *=GIG +24) oo seo ie fon a. hat the misuse fx depends onc, he concentration of sa S58. stein of te pi Tis he xf of hence, Dehetre of he nme pron en PT tes Teh eet ete apc i ea at eet 8 to i of eee Ths, ogc, Ne Tenet er cas T= RT fh +2) + 4391-2) + (G2) we = UT (e,-ep428) 9) i Concentration of the sac is small 2s compas sos inn, ey the osmotic pressure woud be hen by HOR Nee), See ne } mee ioc page ae ety osmotic ese mesure’ wuld earn scale" HY ES fom If the concentration of ration of the salt cis large as compared 10 the concentration ¢, of the non- ® | 1205, MACROMOLECULES diffusible ion, then, aecording to Eq, 34, 2c is equ tc. Eq 35, therefore, takes the form T= 2RTey 05, “This equation does not involve the cosetraton of the chloride in. In other words, the effect of Donnan equilibrium oo osmotic presie i practically, eliminated by using a high concentration of earrate solution, This is precise) the condtion under which molar masses of such maéromolecules Should be determined from osmote pressure measurements. 23. Utraentritugation ‘The rate of sedimentation of suspended panicles under the action of gravity is very small However, it can be increased considerably by applying the ultracentiugaton tectoique devised by ‘Siauberg, A panicle sedimenting ina cexeifga field is subjected to an aceleration given by oi re cet angular veloity ofthe centrifuge (given by 2x% number of revolutions per secon) and Mee @ famace et the particle from the ais of rotation. The rate of sedimentation can, therefore, te inresed considerably by Snereasing the mrber of revolutions per second In some of te ulracentifuges, the number of revolution is as high s 1000 per seond. If he tie ina a distance of 10 em from he ais of rotation (.e., x=10 cm), the acecerato i the partatual etd would be ax = (Qn% 1000? x10 =. 394% 108 em per sec, This is nett 400,002 cr icer tan the acceleration die to aomal gravity which i only 98cm per set. The ae of {edimettation, therefore, canbe raised by 400,00 tnesthe normal rae. “ ‘Wien a polyiner sluts is plaed inthe ell of analtracenritue, the polymer molecules tend to diarcbute Themselves during rotation peredicalaly tothe axis of rotation throughout the cel in ‘RcGrdance with tel individ! molar asses. At constant force, the larger molecules move reply towards the periphery, rhe largest molecules being found nearest the periphery, at eau Strong beam of light pasting parallel othe ais of rotation atd perpendicular tothe etek to measure We reactive inde, ten the concentration ofthe polymer solaton in ay given ‘Revon can be measured. This enables determication of the velocity of the molecular movement eeunts tne periphery. a equilibrium, The viracentrifugation technique for the determination of (Boter mate of a polymer toy involve () the Sedimentation Velocity Method ox () the Sedimentation Foal Metod. While tbe fist metot yi the number-average molar mas, the scord roeod eds the mas-average ilar mas. a, The Sedimentation Velocity Method, Consider polymer solution being su in a ceatifuge tube sae the influence of very stong graviaonal field, The centrifuge force acting on the eee cmeetcle of mass m, at 2 dtance ftom the centre of rotation, is ma’r, where @ i the ‘tat loci of he ctr in radians pt svond. The parle i ls sbjed to» banyan free sae to Ge centifigl force 0 hatte result force acting on the particle i given by Résiant force = Ceaifugal free ~ Buoyant force = ma? —_mar G7) vitere my i the ris ofthe dslaed solvent, If p isthe density of the solvent and , the vole of Te cau parle hen mop, However, the measurement of ¥ is extremely iu. Hence is ees Ro define t quant, y the paral spec volune, a te increase in volume when one crams dy pore sie & doled ia Very large volume ofthe solvent. Then bviowy he aay od ll epesen! the increase in volume obtained on adding solute patil of mass m0 the solve In oter words, y= . Hence, Ea. 37 becomes anatr ~ motrip = mobr(t ~ ¥9) 8) ‘This resultant force, according to Newn's second law of motion wil cause the solute petite to wae However, the acceleration in this ease Will not last long since the medium exer @ clin force 02 it which is proportional the sedimentation velocity, df. Ths, Frictional force = frit Resultant force eo1286 PRINCIPLES OF PHYSICAL CHEMIsTAY where J is the frictional coefficient. The frictional force acts in the opposite direction to the resultant force. When the steady state i reached, we have from Eqs. 38 and 39, Said)» mor. ¥) : “o, Bg. 40 can be rearranged to give a quantity, $, defined 28 Sedimentation velociy/Ceatrifugal acceleration, dridt _ mA ~¥p) _ iv INIA he, Shia TEe 7 ‘The quantity $ is called the sedimentation coefficient; it isthe sedimentation rate for a unit cenriegal acceleration. For a given molecular species in a given solvent ata given tempera, § i a Constant. Mis expressed in the unt svedberg, named afer the Swedish chemist. Svedhrp (1884-1971) who did fundamental work on Prize in Chemisty in 1926, One svedberg= 071 For a spherical polymer gartcle of ratius.r, the frictional coefficient, f, is reed 10 the oefficieat of viscosity of the solvem, 7, by Stokes” equation, viz. f= 6m¢ : (82) Hence, from Eqs. 41 and 42, Fy = SNL. Seg a oie Toe : (a) that one can expect the frictional coefficient f determined from diffusion to be identical with the Getional coefficient f which is characteristic for sedimentation, The relationship between the rperimentally determined diffusion coefficient D andthe titional coefficient fis givea by Sikes. Einstein difasion equation, ve, i D= RIN) = ey lf weence fein 5) Ths, fom By. 48 and 4, Net ser We” Da 5p) * Ba Hy Which i the Svedberg equation for the determizaon of molar mass of a polymer. The sedimentation coefficient Protographic $ and the diffusion coefficient D must be corrected to late the same temperature, usually 20°C, and if both these ‘coefficients are concentration-dependent, their values ‘must be extrapolated to ero concentration, ‘The Experimenta Set-up. The ulioceatifuge is shown schematically in Fig. 1. The measuremeat of | AX sedimentation veleity in te ulttaceatrifuge leads tothe determination of the sedimentation coefficient. The dite = solution of the polymer contained ina ells inroduced Neat i ee fhe lraceifpe, The cel is Sed | FE 1- ering tr slineon ‘with two quartz windows to permit photographing ofthe NE SANE sc i vo As rotation in the centrifuge proceeds, the polymer motecules slowly sediment a ee eee ee ee eh Sewanee at enone ee ik con ee Sewanee Since manent eno te i pe ig iO ee os Soran mae Now, by definition, If the boundary is at a distance ry from the axis of the wltraceatrifige at time ty and at a Aisance foarte ais at tine, then negratin of Eq 4 yes a L fide Cs 7. oll) fests. +9 ain) “9, a ‘tn (in) 0) oc - “beanie he sno cet fo ge mmc independ fh ai vey 9 the rotor, the reason being that a increas, dh increases, 10,50 te Wo Jeoshe ute The selnenaionceficent for prota eae range af 10° Wox1T8 s. oo Example an sree ich operating 9 1. A ample of seco sou is placed aa i oe 9.08 tat orn pe), een ec af etn hw ‘lca bondry ve fa 30min a tne of 5c Rem te es of aon? Suton : Since fom 4 47,5 = (dani, Nase dis = Se? B:142900,000 © sspyaast 8 9 (257 ad 8 (65 ex) = 127x104 om ot 127x104 sme) 0X60 8) = 0229 cx. Asample of solution of bovine serum aBbunia is placed in au ultracetrifoge tic ae “speed r ‘from an initial p-value of sreiag ped of 104 ebay pan te tn eve oma ail a Sarco stale af 60cm ns etl of 57 i, eae the edict cet ove eran abun. Soltion : Subsimtig he gen da a Eq. 50, we have sorte” 5