Title Page
Contents
GENERAL ABSTRACT 11
CHAPTER 1. Development and Validation of a Sensitive Ultrahigh-Performance Liquid Chromatography-Tandem Mass Spectrometry Method for Quantitative Analysis of Bambermycin in Livestock and Aquatic Products: Implications for Food Safety Control and Regulatory Enforcement 13
1.1. ABSTRACT 14
1.2. INTRODUCTION 15
1.3. MATERIALS AND METHODS 18
1.3.1. Chemicals and reagents, and samples 18
1.3.2. Standard preparation 18
1.3.3. Sample preparation 19
1.3.4. UHPLC/MS/MS analysis 20
1.3.5. Method performance 23
1.3.6. Statistical analyses 23
1.4. RESULTS AND DISCUSSION 25
1.4.1. Optimization of UHPLC/MS/MS conditions 25
1.4.2. Optimization of sample preparation 29
1.4.3. Method performance 33
1.4.4. Method application 38
1.5. CONCLUSIONS 44
CHAPTER 2. Rapid Method for the Determination of Non-Steroidal Anti-Inflammatory Drugs and Corticosteroids Residues in Food Animal Tissues and Fishury Tissues 45
2.1. ABSTRACT 46
2.2. INTRODUCTION 47
2.3. MATERIALS AND METHODS 49
2.3.1. Chemicals and reagents 49
2.3.2. Standard preparation 49
2.3.3. Sample preparation 50
2.3.4. UHPLC/MS/MS analysis 51
2.3.5. Method validation 55
2.4. RESULTS AND DISCUSSION 57
2.4.1. Optimization of sample extraction 57
2.4.2. Optimization of UHPLC-MS/MS conditions 61
2.4.3. Method performance 61
2.4.4. Method application 73
2.5. CONCLUSIONS 92
REFERENCES 93
APPENDIX 103
ABSTRACT (in Korean) 104
Chapter 1. Development and validation of a sensitive UHPLC‒MS/MS method for quantitative analysis of bambermycin in livestock and aquatic products: implications for food safety control and regulatory enforcement 6
Table 1. UHPLC‒MS/MS conditions for bambermycin. 22
Table 2. Maximum residue limit (MRL) of bambermycin in eight food products. 24
Table 3. Method performance for bambermycin analysis in spiked beef, pork, chicken, milk, egg, eel, flatfish, and shrimp samples. 39
Table 4. Recovery of bambermycin beef, pork, chicken, milk, egg, eel, flatfish, and shrimp (between laboratories) 42
Chapter 2. Rapid method for the determination of non-steroidal anti-inflammatory drugs and corticosteroids residues in food animal tissues and fishury tissues 6
Table 1. The MRM parameters in UHPLC-MS/MS analysis anti-inflammatory 53
Table 2. MRL and marker residue criteria defined by the Korea Ministry of Food and Drug Safety 56
Table 3. Comparison of extraction methods used for beef, pork, and chicken samples (n=3). 59
Table 4. The correlation coefficients (R²) for linearity were calculated for beef, pork, chicken, milk, eggs, flatfish, eel, and shrimp samples. 62
Table 5. LODs and LOQs in 18 anti-inflammatory 64
Table 6. The matrix effects of the analytes. 65
Table 7. Method performance of 18 analytes in spiked beef, pork, chicken, milk, eggs, flatfish, eel, and shrimp samples. 67
Chapter 1. Development and validation of a sensitive UHPLC‒MS/MS method for quantitative analysis of bambermycin in livestock and aquatic products: implications for food safety control and regulatory enforcement 7
Figure 1. UHPLC-MS/MS chromatograms at a concentration of 10 µg/kg, captured at the same magnification, showcasing the performance of four distinct column types: (A) Kinetex EVO C18, (B) Luna C18, (C) Gemini NX C18, and (D) Kinetex C18 27
Figure 2. Comparative analysis of chromatography using different mobile phases at the same concentration (10 µg/kg): (A) 0.1% formic acid in ACN/0.1% formic acid, (B) 0.1% acetic acid in ACN/distilled water, (C) ACN/0.005 mol/L ammonium acetate, (D) 0.005 mol/L ammonium... 28
Figure 3. Evaluation of cleanup techniques for beef, pork, chicken, and flatfish samples (n=3). Statistical analysis was performed using two-way ANOVA (IBM SPSS Statistics, v29,... 31
Figure 4. Comparison of adding 2 g of NaCl to milk (n=6) (a) area (b) S/N 32
Figure 5. UHPLC‒MS/MS chromatograms of (A) blank samples of beef, pork, chicken, milk, egg, flatfish, eel, and shrimp matrices and (B) spiked samples with bambermycin at a concentration of ×1/2 MRL. 36
Chapter 2. Rapid method for the determination of non-steroidal anti-inflammatory drugs and corticosteroids residues in food animal tissues and fishury tissues 7
Figure 1. Comparison of extraction methods used for milk, eggs, flatfish, and shrimp samples (n=3). 60
Figure 2. UHPLC-MS/MS chromatograms of flunixin (milk: in the following order: Blank samples, standard, and spiked samples (1/2 x MRL) in (A) Beef, (B) Pork, (C) Chicken, (D) Milk, (E) Eggs, (F) Flatfish, (G) Eel, and (H) Shrimp. 74
Figure 3. UHPLC-MS/MS chromatograms of meloxicam in the following order: Blank samples, standard, and spiked samples (1/2 x MRL) in (A) Beef, (B) Pork, (C) Chicken, (D) Milk, (E) Eggs, (F) Flatfish, (G) Eel, and (H) Shrimp. 75
Figure 4. UHPLC-MS/MS chromatograms of 4-Methylamino antipyrine in the following order: Blank samples, standard, and spiked samples (1/2 x MRL) in (A) Beef, (B) Pork, (C) Chicken, (D) Milk, (E) Eggs, (F) Flatfish, (G) Eel, and (H) Shrimp. 76
Figure 5. UHPLC-MS/MS chromatograms of aminopyrine in the following order: Blank samples, standard, and spiked samples (1/2 x MRL) in (A) Beef, (B) Pork, (C) Chicken, (D) Milk, (E) Eggs, (F) Flatfish, (G) Eel, and (H) Shrimp. 77
Figure 6. UHPLC-MS/MS chromatograms of antipyrine in the following order: Blank samples, standard, and spiked samples (1/2 x MRL) in (A) Beef, (B) Pork, (C) Chicken, (D) Milk, (E) Eggs, (F) Flatfish, (G) Eel, and (H) Shrimp. 78
Figure 7. UHPLC-MS/MS chromatograms of phenacetin in the following order: Blank samples, standard, and spiked samples (1/2 x MRL) in (A) Beef, (B) Pork, (C) Chicken, (D) Milk, (E) Eggs, (F) Flatfish, (G) Eel, and (H) Shrimp. 79
Figure 8. UHPLC-MS/MS chromatograms of acetanilide in the following order: Blank samples, standard, and spiked samples (1/2 x MRL) in (A) Beef, (B) Pork, (C) Chicken, (D) Milk, (E) Eggs, (F) Flatfish, (G) Eel, and (H) Shrimp. 80
Figure 9. UHPLC-MS/MS chromatograms of ketoprofen in the following order: Blank samples, standard, and spiked samples (1/2 x MRL) in (A) Beef, (B) Pork, (C) Chicken, (D) Milk, (E) Eggs, (F) Flatfish, (G) Eel, and (H) Shrimp. 81
Figure 10. UHPLC-MS/MS chromatograms of diclofenac in the following order: Blank samples, standard, and spiked samples (1/2 x MRL) in (A) Beef, (B) Pork, (C) Chicken, (D) Milk, (E) Eggs, (F) Flatfish, (G) Eel, and (H) Shrimp. 82
Figure 11. UHPLC-MS/MS chromatograms of carprofen in the following order: Blank samples, standard, and spiked samples (1/2 x MRL) in (A) Beef, (B) Pork, (C) Chicken, (D) Milk, (E) Eggs, (F) Flatfish, (G) Eel, and (H) Shrimp. 83
Figure 12. UHPLC-MS/MS chromatograms of firocoxib in the following order: Blank samples, standard, and spiked samples (1/2 x MRL) in (A) Beef, (B) Pork, (C) Chicken, (D) Milk, (E) Eggs, (F) Flatfish, (G) Eel, and (H) Shrimp. 84
Figure 13. UHPLC-MS/MS chromatograms of prednisolone in the following order: Blank samples, standard, and spiked samples (1/2 x MRL) in (A) Beef, (B) Pork, (C) Chicken, (D) Milk, (E) Eggs, (F) Flatfish, (G) Eel, and (H) Shrimp. 85
Figure 14. UHPLC-MS/MS chromatograms of dexamethasone in the following order: Blank samples, standard, and spiked samples (1/2 x MRL) in (A) Beef, (B) Pork, (C) Chicken, (D) Milk, (E) Eggs, (F) Flatfish, (G) Eel, and (H) Shrimp. 86
Figure 15. UHPLC-MS/MS chromatograms of triamcinolone in the following order: Blank samples, standard, and spiked samples (1/2 x MRL) in (A) Beef, (B) Pork, (C) Chicken, (D) Milk, (E) Eggs, (F) Flatfish, (G) Eel, and (H) Shrimp. 87
Figure 16. UHPLC-MS/MS chromatograms of flumethasone in the following order: Blank samples, standard, and spiked samples (1/2 x MRL) in (A) Beef, (B) Pork, (C) Chicken, (D) Milk, (E) Eggs, (F) Flatfish, (G) Eel, and (H) Shrimp. 88
Figure 17. UHPLC-MS/MS chromatograms of tolfenamic acid in the following order: Blank samples, standard, and spiked samples (1/2 x MRL) in (A) Beef, (B) Pork, (C) Chicken, (D) Milk, (E) Eggs, (F) Flatfish, (G) Eel, and (H) Shrimp. 89
Figure 18. UHPLC-MS/MS chromatograms of phenylbutazone in the following order: Blank samples, standard, and spiked samples (1/2 x MRL) in (A) Beef, (B) Pork, (C) Chicken, (D) Milk, (E) Eggs, (F) Flatfish, (G) Eel, and (H) Shrimp. 90
Figure 19. UHPLC-MS/MS chromatograms of anthranilic acid in the following order: Blank samples, standard, and spiked samples (1/2 x MRL) in (A) Beef, (B) Pork, (C) Chicken, (D) Milk, (E) Eggs, (F) Flatfish, (G) Eel, and (H) Shrimp. 91