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.