Quantification of peramivir in dog plasma by liquid chromatography/tandem mass spectrometry employing precolumn derivatization

•The LLOQ of this method is better than previous published methods.•The esterification derivatization was used to increase the HPLC retention time and ionization efficiency in this article.•Peramivir is a novel anti-influenza drug which was shown to be safe and have well activity.

Peramivir is a novel influenza neuraminidase inhibitor used for anti-influenza. In this article, a novel method was developed to determine peramivir in dog plasma using a derivatization treatment step to increase the retention time and enhance the signal intensity. The sample preparation consisted of a protein precipitation extraction followed by derivatization with 10 M hydrochloric acid–methanol (10:90, v/v) and determined by liquid chromatography coupled with tandem mass spectrometry. The selected reaction monitoring mode of the positive ion was performed and the precursor to the product ion transitions of m/z 343 → 284 and m/z 299 → 152 were used to measure the derivative of peramivir and Ro 64-0802 (internal standard, an active metabolite of oseltamivir). The chromatographic separation was achieved using a ZORBAX RX-C8 (2.0 mm × 150 mm × 5 μm) analytical column with an isocratic mobile phase composed of acetonitrile–water–formic acid (30:70:0.1, v/v/v, 0.2 mL/min). The method was linear over a concentration range of 0.25–250 ng/mL. The average intra-day/inter-day precision values were 4.04–8.17% and 3.02–7.08%, respectively, while the average accuracy value was 93.99–106.48%. This method has been successfully applied to the preclinical dog research of peramivir following intragastric administration.