**1. Introduction**

Kanamycin is widely used in the treatment of animal infections, added as growth promoters or feed additives for preventive therapy [1]. The antibacterial mechanism of kanamycin is that it can irreversibly bind to the bacterial ribosomal 30S subunit and inhibit its protein synthesis [2]. Because of its potential ototoxicity and nephrotoxicity [3–6], the indiscriminate use of kanamycin will enhance bacterial resistance and cause kanamycin-residue accumulation in animal-derived food, which threatens human health. Therefore, the European Union has promulgated regulations on the maximum residue limits (MRLs) of kanamycin in different food matrices (100 μg/kg for muscle, 100 μg/kg for egg, 600 μg/kg for liver, 2500 μg/kg for kidney, 150 μg/kg for milk and 100 μg/kg for chicken meat) [7].

Kanamycin was isolated in 1957 [8]. It is a mixture of several closely related compounds, such as main constituent kanamycin A (>95%), as well as minor constituents kanamycin B, C, and D (<5%). The major components are shown in Figure 1 [9]. In addition, degradation products such as 2-deoxystreptamine and paromamine can also be present [10]. Kanamycin A and C are isomers, whereas kanamycin B and D have different functional groups [9].

**Figure 1.** Structure of kanamycin A, B, C, and D and amikacin.

#### **2. The Pre-Treatment Methods of Food Sample**

The key point of detecting kanamycin is to remove the impurities or extract kanamycin from matrices. The usual techniques for extraction and cleanup of kanamycin from matrices include protein precipitation, liquid-liquid extraction (LLE), and solid-phase extraction (SPE) [11]. Based on these techniques, pre-treatment methods for kanamycin detection in food samples are summarized as follows.
