Development and analytic validation of an electron ionization gas chromatography/mass spectrometry (EI-GC/MS) method for the measurement of 3-bromotyrosine in canine serum

Sattasathuchana, Panpicha; Berghoff, Nora; Grützner, Niels; Thengchaisri, Naris; Rangachari, Venkat R; Suchodolski, Jan S; Steiner, Jörg M (2016). Development and analytic validation of an electron ionization gas chromatography/mass spectrometry (EI-GC/MS) method for the measurement of 3-bromotyrosine in canine serum. Veterinary clinical pathology, 45(3), pp. 515-523. Wiley-Blackwell 10.1111/vcp.12373

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BACKGROUND

The activation of eosinophils causes the release of eosinophil peroxidase and subsequent production of 3-bromotyrosine (3-BrY), a stable byproduct. In people, 3-BrY is used as a biomarker for eosinophil activation. The method for measuring 3-BrY concentrations in biologic samples from dogs has not previously been described.

OBJECTIVES

The objective of this study was to develop and analytically validate an electron ionization gas chromatography/mass spectrometry (EI-GC/MS) method for the measurement of 3-BrY in canine serum samples.

METHODS

Pooled canine serum samples were utilized to validate the assay. Serum samples from healthy control dogs (n = 41) were used to evaluate 3-BrY concentrations and establish a reference interval.

RESULTS

The analytic validation revealed that the limit of blank and limit of detection were 0.33 and 0.63 μmol/L, respectively. The coefficients of variation for precision and reproducibility for 3-BrY were < 13.9% and < 11.0%, respectively. The means ± SD of observed-to-expected ratios for linearity and accuracy were 109.6 ± 17.2% and 98.7 ± 11.3%, respectively. The reference interval was determined as ≤ 1.12 μmol/L (median [range]: ≤ 0.63 μmol/L [≤ 0.63-1.13]).

CONCLUSIONS

The EI-GC/MS assay described here for the measurement of 3-BrY in canine serum samples was precise, reproducible, linear, and accurate. Further studies are underway to determine the diagnostic utilities in canine patients with eosinophilic diseases.

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