Title page

ACKNOWLEDGEMENTS

Contents

ABBREVIATIONS 22

ABSTRACT 24

Ⅰ. Introduction 28

Ⅱ. Materials and methods 36

1. Materials 36

1.1. Plant materials 36

1.2. Chemicals 36

1.3. Instrumental analysis 37

2. Screening for antioxidant activities of Ulmus davidiana 38

2.1. Extraction and solvent fractionation of Ulmus davidiana 38

2.2. Proximate analyses of Ulmus davidiana extracts 40

2.3. Antioxidant activity assays 41

2.3.1. Measurement of total phenolics and flavonoids 41

2.3.2. DPPH radical scavenging assay 41

2.3.3. Nitric oxide scavenging assay 42

2.3.4. Superoxide anion radical scavenging assay 43

2.3.5. ABTS radical scavenging assay 43

2.3.6. Reducing power assay 44

2.3.7. Superoxide dismutase (SOD) assay 44

2.3.8. Isolation of mitochondria 45

2.3.9. Determination of lipid peroxidation in liver homogenates 45

2.3.10. Calculation of half maximal inhibitory concentration (IC50(이미지참조)) 46

3. Isolation and identification of antioxidant compound 46

3.1. Isolation procedures of Ulmus davidiana 46

3.2. Identification of antioxidant compound by HPLC 48

3.3. Structural analysis of antioxidant compound 50

3.3.1. UV spectrophotometry 50

3.3.2. FT-IR spectrometry 50

3.3.3. LC/ESI-MS analysis 51

3.3.4. Nuclear magnetic resonance (NMR) spectroscopy 52

4. Effects of Ulmus davidiana on quality and antioxidant activity of Yackwa during storage 55

4.1. Preparation of Yackwa 55

4.2. Proximate analyses of Ulmus davidiana on Yackwa 56

4.3. Effects of Ulmus davidiana on physicochemical and sensory characteristics of Yackwa during storage 56

4.3.1. Water activity of Yackwa 56

4.3.2. Expansion ratio of Yackwa 56

4.3.3. Color measurement of Yackwa 57

4.3.4. Texture analysis of Yackwa 57

4.3.5. Sensory characteristics of Yackwa 57

4.4. Effects of Ulmus davidiana on oxidation stabilities of Yackwa during storage 58

4.4.1. Extraction of oil from stored Yackwa 58

4.4.2. Measurement of acid value (AV) 59

4.4.3. Measurement of peroxide value (POV) 59

4.4.4. Measurement of TBA value 60

4.5. Effects of Ulmus davidiana on antioxidant activities of Yackwa during storage 61

4.5.1. Preparation of Yackwa extract 61

4.5.2. Antioxidant activities of Ulmus davidiana on Yackwa 62

5. Statistical analysis 62

Ⅲ. Results and discussion 63

1. Evaluation of antioxidant activities of Ulmus davidiana extracts 63

1.1. Proximate analyses of Ulmus davidiana extracts 63

1.2. Antioxidant activities of Ulmus davidiana extracts 65

1.2.1. Total phenolic and flavonoid contents of Ulmus davidiana extracts 65

1.2.2. DPPH radical scavenging activity of Ulmus davidiana extracts 68

1.2.3. Nitric oxide scavenging activity of Ulmus davidiana extracts 72

1.2.4. Superoxide anion radical scavenging activity of Ulmus davidiana extracts 76

1.2.5. ABTS radical scavenging activity of Ulmus davidiana extracts 79

1.2.6. Reducing power of Ulmus davidiana extracts 82

1.2.7. Superoxide dismutase (SOD) activity of Ulmus davidiana extracts 84

1.2.8. Lipid peroxidation in liver homogenates of Ulmus davidiana extracts 88

1.2.9. Correlation between the total phenolic contents and antioxidant activities of Ulmus davidiana extracts 91

2. Antioxidant activities of Ulmus davidiana fractions 94

2.1. Total phenolic and flavonoid contents of Ulmus davidiana fractions 94

2.2. DPPH radical scavenging activity of Ulmus davidiana fractions 98

2.3. Nitric oxide scavenging activity of Ulmus davidiana fractions 103

2.4. Superoxide anion radical scavenging activity of Ulmus davidiana fractions 106

2.5. ABTS radical scavenging activity of Ulmus davidiana fractions 109

2.6. Reducing power of Ulmus davidiana fractions 114

2.7. Superoxide dismutase (SOD) activity of Ulmus davidiana fractions 116

2.8. Lipid peroxidation in liver homogenates of Ulmus davidiana fractions 119

2.9. Correlation between the total phenolic contents and antioxidant activities of Ulmus davidiana fractions 122

3. Isolation and identification of antioxidant compound 126

3.1. Isolation of antioxidant compound 126

3.2. Identification of antioxidant compound 133

3.3. Antioxidant activities of compound 141

4. Effects of Ulmus davidiana on quality and antioxidant activity of Yackwa during storage 147

4.1. Proximate analyses of Yackwa 147

4.2. Effects of Ulmus davidiana on physicochemical and sensory characteristics of Yackwa during storage 149

4.2.1. Water activity of Yackwa 149

4.2.2. Expansion ratio of Yackwa 152

4.2.3. Color measurement of Yackwa 154

4.2.4. Texture analysis of Yackwa 158

4.2.5. Sensory characteristics of Yackwa 160

4.3. Effects of Ulmus davidiana on oxidation stabilities of Yackwa during storage 165

4.3.1. Acid value (AV) of Yackwa 165

4.3.2. Peroxide value (POV) of Yackwa 169

4.3.3. TBA value of Yackwa 173

4.4. Effects of Ulmus davidiana on antioxidant activities of Yackwa during storage 176

4.4.1. DPPH radical scavenging activity of Yackwa 176

4.4.2. Nitric oxide scavenging activity of Yackwa 178

4.4.3. Superoxide anion radical scavenging activity of Yackwa 181

4.4.4. ABTS radical scavenging activity of Yackwa 184

4.4.5. Reducing power of Yackwa 189

4.4.6. Superoxide dismutase (SOD) activity of Yackwa 192

4.4.7. Correlation between preparation treatments, oxidation stabilities, and antioxidant activities of Yackwa 195

Ⅳ. Conclusion 198

REFERENCES 204

List of Appendix 19

Appendix 1. Gallic acid standard curve. 234

Appendix 2. Rutin standard curve. 235

Appendix 3. Various and their levels for control composite design for Ulmus davidiana on Yackwa. 236

Appendix 4. Score sheet for sensory characteristics 237

Appendix 5. DPPH radical scavenging activity of extracts from Ulmus davidiana root and bark 238

Appendix 6. Nitric oxide scavenging activity of extracts from Ulmus davidiana root and bark 239

Appendix 7. Superoxide anion radical scavenging activity of extracts from Ulmus davidiana root and bark 240

Appendix 8. Reducing power of extracts from Ulmus davidiana root and bark 241

Appendix 9. Superoxide dismutase (SOD) activity of extracts from Ulmus davidiana root and bark 242

Appendix 10. Inhibitory activity of lipid peroxidation of extracts from Ulmus davidiana root and bark 243

Appendix 11. DPPH radical scavenging activity of Ulmus davidiana fractions 244

Appendix 12. Nitric oxide scavenging activity of Ulmus davidiana fractions 245

Appendix 13. Superoxide anion radical scavenging activity of Ulmus davidiana fractions 246

Appendix 14. Reducing power activity of Ulmus davidiana fractions 247

Appendix 15. Superoxide dismutase (SOD) activity of Ulmus davidiana fractions 248

Appendix 16. Inhibitory activity on lipid peroxidation of Ulmus davidiana fractions 249

Appendix 17. Change in acid value (AV) of Yackwa with different concentrations of Ulmus davidiana extract during storage 250

Appendix 18. Change in peroxide value (POV) of Yackwa with different concentrations of Ulmus davidiana extract during storage 251

Appendix 19. Change in TBA value of Yackwa with different concentrations of Ulmus davidiana extract during storage 252

Appendix 20. Change in DPPH radical scavenging activity of Yackwa with different concentrations of Ulmus davidiana extract during storage 253

Appendix 21. Change in nitric oxide scavenging activity of Yackwa with different concentrations of Ulmus davidiana extract during storage 254

Appendix 22. Change in superoxide anion radical scavenging activity of Yackwa with different concentrations of Ulmus davidiana extract during storage by using PMS-NADH method 255

Appendix 23. Change in ABTS radical scavenging activity of Yackwa with different concentrations of Ulmus davidiana extract during storage using TEAC value 256

Appendix 24. Change in ABTS radical scavenging activity of Yackwa with different concentrations of Ulmus davidiana extract during storage 257

Appendix 25. Change in reducing power of Yackwa with different concentrations of Ulmus davidiana extract during storage 258

Appendix 26. Change in superoxide dismutase (SOD) activity of Yackwa with different concentrations of Ulmus davidiana extract during storage 259

ABSTRACT IN KOREAN 260

Table 1. Proximate composition of extracts from Ulmus davidiana root and bark 64

Table 2. Comparative total phenolic and flavonoid contents of extracts from Ulmus davidiana root and bark 66

Table 3. IC50(이미지참조) values for radical scavenging and antioxidant activities of extracts from Ulmus davidiana root and bark 71

Table 4. ABTS radical scavenging activity of extracts from Ulmus davidiana root and bark using TEAC value 81

Table 5. Correlation between the total phenolic contents and antioxidant activities of Ulmus davidiana Extracts 93

Table 6. Comparative total phenolic and flavonoid contents of Ulmus davidiana fractions 97

Table 7. IC50(이미지참조) values for radical scavenging and antioxidant activities of Ulmus davidiana fractions 101

Table 8. ABTS radical scavenging activity of Ulmus davidiana fractions using TEAC value 111

Table 9. Correlation between the total phenololic contents antioxidant activities of Ulmus davidiana fractions 125

Table 10. DPPH radical, superoxide anion radical, nitric oxide, and ABTS radical scavenging activities of compounds 1 from Ulmus davidiana as compared to other antioxidants 142

Table 11. Proximate composition of Yackwa 148

Table 12. Change in water activity of Yackwa with different concentrations of Ulmus davidiana extract during storage 150

Table 13. Change in expansion ratio of Yackwa with different concentrations of Ulmus davidiana extract during storage 153

Table 14. Change in L, a and b values of Yackwa with different concentrations of Ulmus davidiana extract during storage 155

Table 15. Change in hardness of Yackwa with different concentrations of Ulmus davidiana extract during storage 159

Table 16. Change in sensory characteristics of Yackwa with different concentrations of Ulmus davidiana extract during storage 161

Table 17. Correlation between preparation treatments, oxidation stabilities, and antioxidant activities of Yackwa 197

Figure 1. Extraction and fractionation of Ulmus davidiana. 39

Figure 2. Isolation procedure of antioxidant compound from Ulmus davidiana by column chromatography. 49

Figure 3. Isolated compound 1 (-)-Catechin-7-O-β-D-apiofuranoside. 54

Figure 4. DPPH radical scavenging activity of extracts from Ulmus davidiana root and bark (◆: Ulmus davidiana root extract; ■: Ulmus davidiana bark extract; ▲: L-ascorbic acid). 69

Figure 5. Nitric oxide scavenging activity of extracts from Ulmus davidiana root and bark (◆: Ulmus davidiana root extract; ■: Ulmus davidiana bark extract; ▲: L-ascorbic acid). 74

Figure 6. Superoxide anion radical scavenging activity of extracts from Ulmus davidiana root and bark by using PMS-NADH method (◆: Ulmus davidiana root extract; ■: Ulmus davidiana bark extract; ▲: Tannic acid). 77

Figure 7. Reducing power of extracts from Ulmus davidiana root and bark (◆: Ulmus davidiana root extract; ■: Ulmus davidiana bark extract; ▲: L-ascorbic acid). 83

Figure 8. Superoxide dismutase (SOD) activity of extracts from Ulmus davidiana root and bark (◆: Ulmus davidiana root extract; ■: Ulmus davidiana bark extract; ▲: L-ascorbic acid; ●: BHT). 86

Figure 9. Inhibitory activity on lipid peroxidation of extracts from Ulmus davidiana root and bark (◆: Ulmus davidiana root extract; ■: Ulmus davidiana bark extract; ▲: α-tocopherol). 89

Figure 10. DPPH radical scavenging activity of Ulmus davidiana fractions (◆: n-hexane; ■: dichloromethane; ▲: ethyl acetate; ×: n-butanol; +: water; ●: L-ascorbic acid). 99

Figure 11. Nitric oxide scavenging activity of Ulmus davidiana fractions (◆: n-hexane; ■: dichloromethane; ▲: ethyl acetate; ×: n-butanol; +: water; ●: L-ascorbic acid). 105

Figure 12. Superoxide anion radical scavenging activity of Ulmus davidiana fractions by using PMS-NADH method (◆: n-hexane; ■: dichloromethane; ▲: ethyl acetate; ×: n-butanol; +: water; ●: tannic acid). 108

Figure 13. Reducing power of Ulmus davidiana fractions (◆: n-hexane; ■: dichloromethane; ▲: ethyl acetate; ×: n-butanol; +: water; ●: L-ascorbic acid). 115

Figure 14. Superoxide dismutase (SOD) activity of Ulmus davidiana fractions (◆: n-hexane; ■: dichloromethane; ▲: ethyl acetate; ×: n-butanol; +: water; ●: L-ascorbic acid, ○: BHT ). 118

Figure 15. Inhibitory activity on lipid peroxidation of Ulmus davidiana fractions (◆: n-hexane; ■: dichloromethane; ▲: ethyl acetate; ×: n-butanol; +: water; ●: α-tocopherol). 121

Figure 16. DPPH radical scavenging activity of the first subfraction of Ulmus davidiana. 128

Figure 17. DPPH radical scavenging activity of the second subfraction of Ulmus davidiana. 129

Figure 18. DPPH radical scavenging activity of the third subfraction of Ulmus davidiana. 130

Figure 19. DPPH radical scavenging activity of the fourth subfraction of Ulmus davidiana. 131

Figure 20. HPLC spectrum of 4th(이미지참조) subfraction No. 3 of ethyl acetate fraction from Ulmus davidiana. 132

Figure 21. UV spectrum of antioxidant compound 1. 137

Figure 22. FT-IR spectrum of antioxidant compound 1. 138

Figure 23. LC/ESI-MS spectrum of antioxidant compound 1 : (a) SIM profiles for m/z 423.0 and 867.0; (b) LC/ESI-MS spectrum of the ion at m/z 423.0 appearing at retention time 2.41 min. 139

Figure 24. ¹H-NMR spectrum of antioxidant compound 1 in CDCl₃. 140

Figure 25. 13(이미지참조)C-NMR spectrum of antioxidant compound 1 in CDCl₃. 140

Figure 26. Change in acid value (AV) of Yackwa with different concentrations of Ulmus davidiana extract during storage (+: control; ■: 0.05% extract; ▲: 0.1% extract; ×: 0.2% extract; ◆: L-ascorbic acid; ●: BHT). 168

Figure 27. Change in peroxide value (POV) of Yackwa with different concentrations of Ulmus davidiana extract during storage (+: control; ■: 0.05% extract; ▲: 0.1% extract; ×: 0.2% extract; ◆: L-ascorbic acid; ●: BHT). 170

Figure 28. Change in TBA value of Yackwa with different concentrations of Ulmus davidiana extract during storage (+: control; ■: 0.05% extract; ▲: 0.1% extract; ×: 0.2% extract; ◆: L-ascorbic acid; ●: BHT). 175

Figure 29. Change in DPPH radical scavenging activity of Yackwa with different concentrations of Ulmus davidiana extract during storage (+: control; ■: 0.05% extract; ▲: 0.1% extract; ×: 0.2% extract; ◆: L-ascorbic acid; ●: BHT). 177

Figure 30. Change in nitric oxide scavenging activity of Yackwa with different concentrations of Ulmus davidiana extract during storage (+: control; ■: 0.05% extract; ▲: 0.1% extract; ×: 0.2% extract; ◆: L-ascorbic acid; ●: BHT). 180

Figure 31. Change in superoxide anion radical scavenging activity of Yackwa with different concentrations of Ulmus davidiana extract during storage by using PMS-NADH method (+: control; ■: 0.05% extract; ▲: 0.1% extract; ×: 0.2% extract; ◆: L-ascorbic acid; ●: BHT). 183

Figure 32. Change in ABTS radical scavenging activity of Yackwa with different concentrations of Ulmus davidiana extract during storage using TEAC value (+: control; ■: 0.05% extract; ▲: 0.1% extract; ×: 0.2% extract; ◆: L-ascorbic acid; ●: BHT). 187

Figure 33. Change in ABTS radical scavenging activity of Yackwa with different concentrations of Ulmus davidiana extract during storage (+: control; ■: 0.05% extract; ▲: 0.1% extract; ×: 0.2% extract; ◆: L-ascorbic acid; ●: BHT). 188

Figure 34. Change in reducing power of Yackwa with different concentrations of Ulmus davidiana extract during storage (+: control; ■: 0.05% extract; ▲: 0.1% extract; ×: 0.2% extract; ◆: L-ascorbic acid; ●: BHT). 191

Figure 35. Change in superoxide dismutase (SOD) activity of Yackwa with different concentrations of Ulmus davidiana extract during storage (+: control; ■: 0.05% extract; ▲: 0.1% extract; ×: 0.2% extract; ◆: L-ascorbic acid; ●: BHT). 194