Determination of Henry’s constant, the dissociation constant, and the buffer capacity of the bicarbonate system in ruminal fluid

Abstract Despite the clinical importance of ruminal acidosis, ruminal buffering continues to be poorly understood. In particular, the constants for the dissociation of H 2 CO 3 and the solubility of CO 2 (Henry's constant) have never been stringently determined for ruminal fluid. The pH was measured in parallel directly in the rumen and the reticulum in vivo, and in samples obtained via aspiration from 10 fistulated cows on hay- or concentrate-based diets. The equilibrium constants of the bicarbonate system were measured at 38°C both using the Astrup technique and a newly developed method with titration at 2 levels of partial pressure of CO 2 (pCO 2 ; 4.75 and 94.98kPa), yielding mean values of 0.234±0.005mmol∙L −1 ∙kPa −1 and 6.11±0.02 for Henry's constant and the dissociation constant, respectively (n/n=31/10). Both reticular pH and the pH of samples measured after removal were more alkalic than those measured in vivo in the rumen (by ΔpH=0.87±0.04 and 0.26±0.04). The amount of acid or base required to shift the pH of ruminal samples to 6.4 or 5.8 (base excess) differed between the 2 feeding groups. Experimental results are compared with the mathematical predictions of an open 2-buffer Henderson–Hasselbalch equilibrium model. Because pCO 2 has pronounced effects on ruminal pH and can decrease rapidly in samples removed from the rumen, introduction of a generally accepted protocol for determining the acid-base status of ruminal fluid with standard levels of pCO 2 and measurement of base excess in addition to pH should be considered.