研究目的
To assess the potential application of the conventional 2D and the 3D fluorescence spectroscopy to detect and discriminate three antibiotics at different concentrations in milk, and establish their potential to predict antibiotic concentrations from the spectral peak heights.
研究成果
The study demonstrated great potential of fluorescence spectroscopy in the multi-dimensional mode coupled with chemometric tools to rapidly and sensitively detect penicillin G, sulfadiazine and tetracycline above their maximum residue limits in milk. Moreover, the milk samples were further discriminated according to the type of antibiotic they contained. Antibiotic concentration in milk could be predicted from the fluorescence spectral data of various fluorophores with great accuracy and selectivity. Therefore this technique could be used for inline monitoring of antibiotics in milk processing plants.
研究不足
The study focused on only three antibiotics (penicillin G, sulfadiazine, and tetracycline) in milk. Further studies are required to evaluate its performance towards detection of these antibiotics and many other types in the raw and processed fluid milk of different composition.
1：Experimental Design and Method Selection:
Multi-dimensional fluorescence spectroscopy (3D and 2D) coupled with chemometric tools were tested for potential application to detect, discriminate and quantify penicillin G (PG), sulfadiazine (SF) and tetracycline (TC) in milk by direct measurement.
2：Sample Selection and Data Sources:
Milk samples were prepared according to the following antibiotic concentrations: Penicillin G: 0, 1, 2, 3, 4, 5, 6 μg/l; Tetracycline: 0, 25, 50, 100, 150, 200, 250 μg/l; Sulfadiazine: 0, 25, 50, 100, 150, 200, 250 μg/l.
3：List of Experimental Equipment and Materials:
F-4500 FL Spectrophotometer (Hitachi High Technology, Tokyo, Japan), Penicillin G, sulfadiazine and tetracycline were obtained from Aladdin Reagent (Shanghai) Co, Ltd. Skimmed milk powder was purchased from WALMART supermarket (Changchun, China).
4：Experimental Procedures and Operational Workflow:
Fluorescence spectra were recorded using F-4500 FL Spectrophotometer with the incidence angle of excitation of 90°. Excitation and emission slits were both set at 5 nm. The emission spectra of Maillard reaction products (MRP) & riboflavin (RF) (400–600 nm), and tryptophan (300–400 nm) were recorded three times for each of the two replicates at room temperature.
5：0°. Excitation and emission slits were both set at 5 nm. The emission spectra of Maillard reaction products (MRP) & riboflavin (RF) (400–600 nm), and tryptophan (300–400 nm) were recorded three times for each of the two replicates at room temperature. Data Analysis Methods:
5. Data Analysis Methods: Principal component analysis (PCA) was applied separately to the spectral regions corresponding to the 3-dimensional data, MRP/RF, vitamin A and tryptophan to obtain similarity maps and spectral patterns. Linear regression analysis was applied to determine any correlations between antibiotic concentration and the peak height in various fluorescence spectral data.
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