Abstract
Oxytocin (OT) is a mammalian neuropeptide with various functions in regulating birth, lactation, parenting, and social recognition. The study of OT became of increasing interest for the petcare industry due to its role in animal behavior and socialization. In the present study, a simple, sensitive, and accurate liquid chromatography-mass spectrometry (LC-MS) method for quantifying OT in dog saliva was developed and validated. OT and its deuterated internal standard (OT-d5) were detected with multiple reaction monitoring (MRM) in a positive ion mode using an AB Sciex 6500+ QTRAP mass spectrometer. Chromatographic separation was achieved by using an ACE Excel C18 column and a gradient elution at a flow rate of 0.5 mL/min over a 5 min run. The mobile phases consisted of 0.1% (v/v) acetic acid in water and 0.1% (v/v) acetic acid in acetonitrile. After development and optimization, the performance of the method was validated to prove its reliability. Calibration curves were linear over the range of 50–20,000 pg/mL and recovery of OT was above 87.8%. The validated method was successfully applied to evaluate OT concentrations in multiple batches of dog saliva samples.
Highlights
Oxytocin (OT) is a mammalian neuropeptide containing nine amino acids and has various regulatory functions in multiple conditions such as birth, lactation, parenting, social recognition, and relationship bonding [1,2,3,4]
Formic acid (FA), AA, and trifluoroacetic acid (TFA) are commonly used as mobile phase additives for compounds that were detected in the positive ion mode
OT spiked in 50% aqueous ACN with different levels of FA, AA, and TFA were directly infused into MS system to evaluate the responses of OT
Summary
Oxytocin (OT) is a mammalian neuropeptide containing nine amino acids and has various regulatory functions in multiple conditions such as birth, lactation, parenting, social recognition, and relationship bonding [1,2,3,4]. Besides these functions of endogenous OT, previous studies showed that social affiliations were promoted after intranasal administration of OT in humans [5,6]. Multiple categories of analytes can be measured in saliva, including enzymes, electrolytes, metabolites, and proteins [13]. Certain analytes, such as cortisol and c-reactive protein, have already been used in dog studies for evaluation of stress and inflammatory lesions [14,15]
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