Detection of pro‐inflammatory cytokines in healthy canine tears using Canine Cytokine SpikeMix™ mass spectrometry via multiple reaction monitoring

Abstract

AbstractPurposeTo evaluate the efficacy of the Canine Cytokine SpikeMix™ and MRM‐MS for detecting pro‐inflammatory cytokines in canine tears from healthy research Beagles.MethodsA complete ophthalmic examination was performed on 15 healthy research Beagles to verify no ophthalmic diseases. Tears were collected OU by placing a Weck‐Cel® cellulose spear in the ventral conjunctival fornix for 1 min. The Weck‐Cel® spear was placed in a 2.0 mL tube with a centrifuge filter forcing tears to flow through the filter into the bottom of the tube. The tears were analyzed using the Canine Cytokine SpikeMix™ and MRM‐MS. Descriptive data from this study was reported as the normalized total peak area (nTPA) and median (range) using data imported from the online MRM‐MS Skyline program.ResultsThe level of 16 pro‐inflammatory cytokines was successfully detected in all 15 dogs. The four cytokines with the highest median amounts in the samples were IL‐2 = 0.1243 (0.019–6.7289), IL‐6 = 0.964 (0.0036–16.9365), TNFα = 0.1644 (0.0096–0.7138), and CSF‐2 = 0.4022 (0.1475–2.6208).ConclusionsThis study revealed that 16 pro‐inflammatory cytokines in canine tears from healthy dogs can be detected with Canine Cytokine SpikeMix™ and MRM‐MS.