Since amantadine, rimantadine, arbidol, laninamvir, oseltamivir, peramivir, and zanamivir may be used as antiviral agents to treat avian influenza, we herein developed a simultaneous assay using LC-MS/MS. This method was applied to chicken products (including yakitori (grilled chicken), fried chicken, chicken steak, and boiled eggs) as well as chicken tissues (muscle, fat, the liver, gizzards, and heart) and eggs.Samples were extracted with methanol-water (9 : 1), purified by a tandem column with an InertSep® MAX cartridge (upper part) and InertSep® MCX cartridge (lower part), and then measured by LC-MS/MS. The sample matrix had no effect on the identification of compounds. Chromatographic separation was performed on a ZIC-HILIC column using a mobile phase of 1% acetic acid solution and 1% acetic acid solution in acetonitrile, resulting in complete separation and other obstructive peaks from the sample matrices. An external solvent calibration curve was used for quantification.The application of the method to 6 samples of chicken tissues and eggs achieved good results of between 77.9 and 97.5% for trueness and between 1.7 and 9.2% for concurrent accuracy. The method was also applied to 9 samples of processed products, including grilled chicken and fried chicken, and achieved good results with true percentages ranging between 72.6 and 99.2% and concurrent accuracies between 3.0 and 11.2%. Therefore, the developed method may also be applied to processed products.The limit of quantification (LOQ) of the developed method was 0.01 mg/kg.The method was then applied to 42 types of commercial processed products, including yakitori, fried chicken, steamed chicken, chicken steak, and boiled eggs, and no antiviral agents were detected.Collectively, the present results confirmed that the method developed herein is applicable to not only chicken tissues, but also their processed products.
Keywords: Highly Pathogenic Avian Influenza; LC-MS/MS; antiviral agent; chicken product; chicken tissue.