Journal
ANALYTICAL BIOCHEMISTRY
Volume 360, Issue 1, Pages 47-55Publisher
ACADEMIC PRESS INC ELSEVIER SCIENCE
DOI: 10.1016/j.ab.2006.10.015
Keywords
ITC; calibration; dilution enthalpy; NaCl(aq); data analysis
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An isothermal titration calorimeter of the perfusion type (MicroCal model VP-ITC) is calibrated using the heat of dilution of NaG in water. The relative apparent molar enthalpy function (L-phi) for NaCl(aq) varies strongly and nonlinearly with concentration in the low-concentration region (< 0.2 M) that is sampled easily and extensively in a single program of injections of NaCl solution into water. This nonlinearity makes it possible to calibrate with respect to two quantities: the measured heat and the active cell volume. The heat factor is determined with typical standard error 0.003; its value in the current case is 0.987. The cell volume factor is 0.93 but is quite sensitive to possible systematic errors in the temperature and in the literature values for L-phi. Both correction factors are closely tied to the delivered volume from the injection syringe, which required a correction factor of 0.973, attributed to an instrumental gear ratio error. Temperature calibration of the instrument showed a small offset of 0.12 K at the temperature 25 degrees C of the experiments, but the error increased to more than 1 K at 46 degrees C. The experiments were not able to distinguish clearly between mixing algorithms that assume instantaneous mixing on injection and those that assume instantaneous injection followed by mixing; however, examination of these algorithms has revealed an error in a program widely used to analyze isothermal titration calorimetry data. (c) 2006 Elsevier Inc. All rights reserved.
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