Author ORCID Identifier

Year of Publication


Degree Name

Master of Science (MS)

Document Type

Master's Thesis


Agriculture, Food and Environment


Veterinary Science

First Advisor

Dr. Scott D. Stanley


Bone resorptive diseases affect humans and horses, alike. Examples of these diseases include osteoporosis, Paget’s disease, and osteogenesis imperfecta in humans, and navicular disease in the horse (Mitchell et al., 2019; Suva et al., 2021). Bisphosphonates are used to treat these diseases, and may remain in the bone for several years after administration, demonstrating the need for newer drug testing methods.

The purpose of the discovery phase of this study was to use blood samples of in-training horses which had been given an initial administration of the bisphosphonate tiludronate and identify protein biomarkers that changed in response to this class of drugs. In the targeted phase, the objective was to validate the previously identified protein biomarkers that can be added into a biomarker library. This library is meant to be referenced when drug testing racehorses and can lengthen the detection window of prohibited or restricted substances.

Fractionation, digestion, and de-salting were performed on sera samples from 19 equine subjects in-training. These samples were injected in an Orbitrap™ Exploris 480d coupled with an Ultimate 3000 RSLCnanoj. Label-free quantitation of proteins was performed in both phases. Four key protein biomarkers were identified in the discovery phase and validated in the targeted phase.

Digital Object Identifier (DOI)