ofthe effect of the beat of dissolution ofthe solute into the solvent and ofthe different nest capacity ofthe solution as compared tothe solvent. A char giving {he relation between the enthalpy ofthe solution and the conceiration ofthe ‘solution calle an enthalpy concentration diagram. Figure 5.14sthe enthalpy ‘oocentration diagram of NaQH-H,0 system. On the diagram, various is0- therms are ploted. Such diagrams a useful in the evaluation of (the heat required fo be added vo or removed from the solution whenbeatng or soning of the solution i satied out nd b) the temperature of the resent inter when to different solutions having Jiferea cosceataions ahd temperatures att rmixadtopetherby te lse method. ‘The fllowing example wlllhelp clarity tbe applications. ‘Eizmple Sav find he ampere of 25% NOOH whan, prcpued by Alting {46% NeOH yea 298.18 K 25°C) with wate at 108 K (35°C). Al percentages re by ‘elation Oa be char (Fig. 5.14), place Arepesemting 46% NaOH tye a1 298.13 K (GSC) Pace pint B representing pure wate (05 ND) at 308 K (38°C). Jain the poise and. The ie line AB intersects the veri ais 29% concentration oan Isotherm of 329.5 K (65°C) Ans tet hg an 2 8BeessaaEE oan Te aa OO Ow eon mas (ERB Eenatpy-coneernvonon Diagram for NaOH-H,O System"Methods in ENZYMOLOGY Aspopular of these procedures utilizes @ simple, commercis Dyenometer, This is a ves of fted volume containing a ground-glas int into which is fitted a stopper containing a small eapillary hole to allow the escape of sir and overfiow of liquid. The solution of interest is placed into the open and the stopper is then inserted care- fully and slowly. I the liquid level is sufciently high, the insertion ofthe topper causes air to be displaced and some liguid rises through the ‘topper and overflows through the small hoe. In this way the pyenometer can be filled routinely with a roproducible volume. The pyonometer i fiat weighed on an analytical balance while empty and dry. Then the solution of protein ix added, the stopper seated carefully, the external walls wiped clean of the overflowing liquid, and the weighing is repeated. ‘The filing of the pyenomseter should be performed ina constant-tempers- ture bath, and itis generally advisable that this temperature be greater than that of the balance room én which the weighings are made. It ie important that the liquid level in the capillary of the stopper be at the very top; any alight overlow car be removed by gentle and rapid wiping of the top of the stopper with a piece of filter paper. Ifthe temperature ofthe balance room is too high, expansion ofthe liquid inthe pyenometer will cause los of iquid and erroneous results unless a pycnometer with & ‘cover cap is used, These, like the simpler pycnometers, are available ‘commercially from chemical supply houses. With a Hitde practice the ‘weight of @ fixed volume of solution is readily determined. After the sample is recovered, the pyenometer is rinsed and then filled with a solue tion at another protein concentration or with the solvent, and the weigh- ing operation is repeated. Finally the pyenometer is filed with water and this weight determined. It is advisable to check the weight of the ‘empty pyenometer between the measurements of individual solutions both because the accurate weight of the empty pyenometer is ocesary and because this furnishes a check on the cleanlines of the pyenometer. ‘Thus the weights ofa fixed volume of diferent protein solutions, solvent, and water are determined, Sinee the temperature is known and the density of water at that temperature is readily available, the weight of the contents of water in the filled pyenometer is used to determine the volume of the penometer. ‘This volume is then used with the weights of the diferent solutions to determine the densities of the correponding Solutions, ARteriatively, the ratio of the density of any specific solution is readily determined by the ratio of the weights of the