臺灣博碩士論文加值系統

高尿酸血症（Hyperuricemia , HUA）是指血液中尿酸水平表現異常高的症狀。抑制黃嘌呤氧化酶（Xanthine Oxidase , XO）是預防HUA的有前途策略。
本文研究探索玉米副產品萃取物（corn by-products, CBP）和黃豌豆水解產物（yellow field pea , YFP）對抗高尿酸血症的潛力，並各自分析體外XO抑制活性實驗。 最後CBP根據體外XO抑制活性實驗數據的條件，通過高效液相色譜法(high performance liquid chromatography, HPLC)以定性和定量鑑定CBP萃取物的物質。 YFP通過HPLC收集蛋白質水解產物的餾分並且通過nanoUHPLC-ESI-Q-TOF質譜法鑑定序列。

玉米副產品乙醇萃取物（EBP）表現出最高的粗萃取率（14.51 ± 0.88％）和總黃酮含量（3.15 ± 0.06 mg CE eq / g EBP）。EBP樣品對XO抑制為IC50：0.49 ± 0.02 mg / mL並確定含有桑色素、柚皮素、槲皮素、山奈酚和芹菜素，其含量為34.28、30.58、29.49、33.18、20.32 mg/g extract, DP of EBP。 YFP的鹼性蛋白酶水解產物（YFPAH）表現出最高水解率（89.02 ± 1.83％）和蛋白質含量（97.15 ± 0.98％ BSA eq / g），YFPAH樣品對XO抑制為（IC50：0.42 ± 0.003 mg / mL），其鑑定的序列顯示為Ser-Arg-Gly-Gln-Ile-Glu-Glu-Leu（SRGQIEEL）。

玉米副產品萃取物和黃豌豆水解產物都被發現具有優異的XO抑制活性。所獲得的結果表明CBP和YFP可作為潛在天然生物活性抗高尿酸血症的來源之一。

Hyperuricemia (HUA) is a symptom characterized by the high level of uric acid in the blood. The inhibition of Xanthine Oxidase (XO) is a promising strategy for preventing HUA. This study has explored the potential both of them for anti-hyperuricemia, of an extract from corn by-products (CBP) and hydrolysates from yellow field pea (YFP). Both of them were analyzed the sample contents and in vitro XO inhibition activity experiments. CBP identified the conditions of the in vitro XO inhibition activity experiments and data by high-performance liquid chromatography (HPLC) to the qualitative and quantitative identification of extracts. YFP collected peaks of proteins hydrolysate by HPLC and identified sequence by nanoUHPLC-ESI-Q-TOF mass spectrometry.

The ethanolic extract of corn byproducts (EBP) demonstrated the highest dry crude extract rate (14.51 ± 0.88 %) and total flavonoids content (3.15 ± 0.06 mg CE eq /g of EBP). The most active plants examined where the sample of EBP (IC50: 0.49 ±0.02 mg/mL) and identified content morin, naringenin, quercetin, kaempferol and apigenin (34.28, 30.58, 29.49, 33.18, 20.32 mg/g extract, DP of EBP). The alcalase hydrolysate (YFPAH) by YFP demonstrated the highest dry crude extract rate (89.02 ± 1.83%) and protein content (97.15 ± 0.98% BSA eq /g of YFPAH. The sample of YFPAH by YFP (IC50: 0.42 ± 0.003 mg/mL) and identified sequence was shown as Ser- Arg- Gly- Gln- Ile- Glu- Glu- Leu (SRGQIEEL). All of them were found to have excellent XO inhibitory activity. Results obtained indicated that CBP and YFP can be used potentially as a readily anti-hyperuricemia of the natural bioactive source.

Master Thesis I
Recommendation of professor II
Verification From the Oral Examination Committee III
CMU Graduate Student Academic Ethics Statement IV
Acknowledgement V
Chinese abstract VII
Abstract VIII
Forward X
Table of contact XI
List of figures XIV
List of tables XIV
List of supplementary materials XV
I. Commercial protease XV
II. The drugs of experimental XVI
III. The commercial proteases of experimental XVII
IV. Experimental equipment XVIII
List of abbreviations XIX
Chapter I - Introduction 1
1.1 Research gaps/Justifications for the proposed study 2
1.2 Hypothesis of the proposed study 3
1.2.1 Corn by-product (CBP) 3
1.2.2 Yellow field pea (YFP) 4
1.3 Objective of the proposed study 5
1.4 Experimental design 6
1.5 Experimental structure 7
1.6 Significance of the proposed research 8
Chapter II - Literature review of hyperuricemia 9
2.1 xanthine oxidase (XO) 9
2.1.1 Structure 9
2.1.2 Biochemistry 10
2.1.3 Involvement in pathological conditions 10
2.2 Hyperuricemia (HUA) 12
2.2.1 Uric acid (UA) 12
2.2.2 Definition 13
2.2.3 Epidemiology Frequency 14
2.2.4 Hyperuricemia Risks 15
2.2.5 Mortality & Morbidity 16
2.2.6 Factors 17
2.2.7 Hyperuricemia symptoms 20
2.2.8 Treatment - Asymptomatic hyperuricemia 23
2.2.9 Treatment -Symptomatic hyperuricemia 25
Chapter III - Manuscript One 28
Inhibitions of xanthine oxidase activities by corn by-products 28
3.1 Abstract 28
3.2 Literature review of extracts with XO inhibitors 29
3.3 Materials and methods 35
3.3.1 Experimental materials 35
3.3.2 Preparation of corn by-products (CBP) 35
3.3.3 The crude extracts of CBP 36
3.3.4 Determination of total flavonoid content 36
3.3.5 Xanthine oxidase inhibitory activity of CBP extracts 36
3.3.6 Qualitative and quantitative identification by HPLC 37
3.3.7. Statistical analyses 39
3.4 Results 40
3.4.1 Raw materials of corn by-products 40
3.4.2 The crude extracts of corn by-product (CBP) 41
3.4.3 Determination of total flavonoid content 43
3.4.4 Xanthine oxidase inhibitory activity of CBP extracts 47
3.4.5 Qualitative and quantitative identification by HPLC 51
3.5 Discussion 56
Chapter IV - Manuscript Two 62
Inhibitions of xanthine oxidase activities by yellow field pea 62
4.1 Abstract 62
4.2 Literature review of peptides with XO inhibitors 64
4.3 Materials and methods 70
4.3.1 Experimental materials 70
4.3.2 Preparation of yellow field pea (YFP) 71
4.3.3 Determination of protein content (%) 72
4.3.4 XO inhibitory activity of yellow field pea 73
4.3.5 Ultra-filtration of YFP 74
4.3.6 Analyzed by HPLC 74
4.3.7 Sequence identification analysis by Q - TOF 76
4.3.8 Statistical analyses 77
4.4 Results 78
4.4.1 The hydrolysate of yellow field pea 78
4.4.2 Analysis of protein content by YFP 80
4.4.3 XO inhibitory activity of YFP hydrolysate 84
4.4.4 XO inhibition by Ultra-filtration of YFPAH 88
4.4.5 XO inhibition by HPLC fractions of <1 kDa permeate 90
4.4.6 Peptide identification 93
4.5 Discussion 96
4.6 Conclusions 99
Chapter V - General Discussions and Conclusions 100
5.1 General discussions 100
5.2 General conclusions 103
5.3 Novelty of the thesis findings 106
5.4 Limitations of this thesis work 107
5.5 Anticipated future direction of this study 108
References 109

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