A determination method for clenbuterol residue in pork based on optimal particle size gold colloid using SERS

Clenbuterol is often used as a feed additive to increase the percentage of lean meat in livestock. Meat containing clenbuterol can cause many illnesses and even death for people. In this paper, the particle growth method was used to prepare gold colloids of different sizes, and the enhanced effectiveness of gold colloids of different sizes on clenbuterol in pork was investigated. The results showed that the gold colloid with the best enhanced effectiveness for clenbuterol had a particle size of approximately 90 nm. Second, a sample collection component was designed to detect clenbuterol from bottom to top, solving the problem of poor reproducibility of Surface-enhanced Raman scattering (SERS) detection caused by different droplet sizes and shapes. Then, the influence of different volumes of samples and concentrations of aggregating compounds on the enhanced effectiveness was optimized. The results showed that, based on the sample collection components designed in this article, 5 µL of enhanced substrate, 7.5 µL of clenbuterol and 3 µL of 1 mol/L mixed detection of NaCl solution had the best enhanced performance. Finally, 88 pork samples (0.5, 1, 1.5,…, 10, 12, 14 µg/g) with different concentrations were divided into correction sets and prediction sets in a ratio of 3:1. Unary linear regression models were established between the concentration of clenbuterol residue in the pork and the intensity of the bands at 390, 648, 1259, 1472, and 1601 cm^-1. The results showed that the unary linear regression models at 390, 648, and 1259 cm^-1 had lower root mean square errors than those at 1472 and 1601 cm^-1. The intensity of the three bands and the concentration of clenbuterol residue in the pork were selected to establish a multiple linear regression model, and the concentration of clenbuterol residue in the pork was predicted. The results showed that the determination coefficients (R²) of the correction set and the prediction set were 0.99 and 0.99, respectively. The root mean square errors (RMSE) of the correction set and the prediction set were 0.169 and 0.184, respectively. The detection limit of clenbuterol in pork by this method is 42 ng/g, which can realize the crude screening of pork containing clenbuterol in the market.