An automated differential thermal and potentiometric titration apparatus for binding studies

A differential pH-thermal titration apparatus is described which can detect pH differences with a sensitivity of +/- 0.0001 pH units and a thermal sensitivity of +/- 0.00002 degree C at a time constant of 0.1 s. With a reaction which yields 1 kcal mol-1, the current system can detect concentrations as low as 4 X 10(-6) M or, in a 2 ml volume, a total amount of 40 nmol. With a time constant of 0.1 s, the sensitivity is 20 +/- 4 micro degrees C. The experimental protocol is specified by a microprocessor and three modes of operation are possible: titration at constant rate of reagent addition, titration at variable rates of addition so that the contents of both cells are at either constant pH or at a constant temperature and variable rate when a rate of change is specified. Experimental data are collected in files, corrected for heat loss, initial baseline drift, and changes in volume. The final corrected data from the standardized run of 0.01338 M HCl in 0.2 M KCl at 25 degrees C calibrate the pH scale and yield the calorimetric conversion constants and pKw which are calculated and stored for subsequent corrections for the titration of an unknown acid or the measurement of binding constants and heats.