Antibiotics are widely used in veterinary medicine for prevention of disease in food producing animals. Excessive or unconscious usage of antibiotics might result residue in food such as milk, meat, honey and others. These residues can cause toxicity, allergic reactions. In additions low antibiotics residues in food products, consumed for long time and lead to the spread of drug-resistant bacteria. On the other hand, milk goes through production, processing and circulation until reaching consumers. In order to ensure consumers’ safety and to prevent any harmful health effects, WHO and EU established the MRL for antibiotics. LC-MS/MS has become the method of choice for analysis antibiotics residues due to do its high sensitivity, selectivity, robustness and cost effective.