Identification of bradykinin, substance P, and neurokinin A in human dental pulp.

Methodology was developed that uses reverse phase, high performance liquid chromatography (HPLC) to identify and quantify bradykinin, substance P, and neurokinin A contained in dental pulp tissue. Pulp tissue was prepared and homogenized from teeth frozen in liquid nitrogen, Known amounts of three substances found in inflamed tissue were added to the homogenized tissue and also to bovine serum albumin (BSA) (positive control), and supernatants were analyzed using HPLC. Other pulp tissue was prepared and analyzed without the addition of the substances. Recovery from the pulp and BSA with added substances was similar, with bradykinin recovered maximally. In pulp tissue without additions, all three substances were recovered. Thus HPLC appears to be a viable alternative to other methods for identification of these substances and allows for their quantification.