以毛豆為原料萃取煙醯胺單核苷酸之研究

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姓名

曾馨儀(Hsin-I Tseng) 查詢紙本館藏

畢業系所

化學工程與材料工程學系

論文名稱

以毛豆為原料萃取煙醯胺單核苷酸之研究
(Extraction of Nicotinamide Mononucleotide from Edamame)

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摘要(中)

保健食品原料來源主要由動物、植物或微生物而來，而本實驗選擇植物來源做為原料。萃取有許多方法如蒸餾法、微波輔助萃取法、超臨界流體萃取等。純化方法也有許多種，如層析法、離子交換法、超臨界流體技術亦能作為純化方法。
煙醯胺單核苷酸(NMN)是衍生自核糖及煙醯胺的一種核苷酸，NMN是由體內的B群維生素製成的，是在所有生物體中天然存在的一種分子，能防止肥胖、抗衰老、保護神經、防止心血管疾病等優點。
本實驗使用煙醯胺單核苷酸含量最高之食物－毛豆作為原料，透過天然食物萃取出煙醯胺單核苷酸並純化，可作為新興保健食品。選用丙酮及水作為萃取煙醯胺單核苷酸之溶劑，並使用真空烘箱將毛豆萃取液做濃縮。結果顯示，利用水作為溶劑較易溶出煙醯胺單核苷酸。使用穿透式電子顯微鏡(TEM)觀察到煙醯胺單核苷酸大小約為50-70奈米。利用紫外光-可見光分光光譜儀(UV-Vis)顯示出煙醯胺單核苷酸之吸收波峰在290 nm。最後用薄層分析法(TLC)將純煙醯胺單核苷酸及毛豆萃取液做比較，確認毛豆萃取液中成功萃取出煙醯胺單核苷酸。

摘要(英)

The raw material of health food is mainly from animal, plant or microorganism, and this experiment chose plant source as the raw material. There are many methods of extraction such as distillation, microwave-assisted extraction, supercritical fluid extraction, etc. There are also many purification methods, such as chromatography, ion exchange, supercritical fluid technology can also be used as purification methods.
　　Nicotinamide mononucleotide (NMN) is a nucleotide derived from ribose and nicotinamide. NMN is made from the body′s B-complex vitamins, a naturally occurring molecule in all living organisms, and can prevent obesity, anti-aging, neuroprotection, cardiovascular disease, and other benefits.
In this study, the food with the highest content of nicotinamide mononucleotide, edamame, was used as the raw material. Extraction and purification of nicotinamide mononucleotides from natural food can be used as a new health food. Acetone and water were used as solvents to extract nicotinamide mononucleotides, and the extract was concentrated in a vacuum oven. The results show that the nicotinamide mononucleotides were more easily extracted by using water as the solvent than acetone. The size of nicotinamide mononucleotide was observed to be 50-70 nm by using transmission electron microscope. The absorption peak of nicotinamide mononucleotide was shown to be around 290 nm by UV-Vis spectroscopy. Finally, the pure nicotinamide mononucleotide was compared with edamame extract by thin layer analysis (TLC), and it was confirmed that nicotinamide mononucleotide was successfully extracted from edamame extract.

關鍵字(中)

★ 毛豆
★ 煙胺醯單核苷酸
★ 萃取

關鍵字(英)

★ edamame
★ nicotinamide mononucleotide
★ extraction

論文目次

摘要……………………………………………………………………………………………………………………………….i
Abstract………………………………….………………………….………………………………………….……ii
致謝………………………………………………………………………………………………………………………..……iii
Table of contents………………………………………………………………………………………….…iv
List of Tables………………...………………………………………………………………………………vii
List of Figures..………………………………………..…………...………………..……..viii
Chapter 1 Introduction …………………………………………………...…………….………1
1.1 Research Background ………………………………………………………...………...1
1.1.1 Market Introduction………………………………………………..………....……..2
1.1.2 Nicotinamide Mononucleotide (NMN) Introduction…………4
1.1.3 Nicotinamide Mononucleotide (NMN) Synthesis Mechanism………………………………………………………………………………………………………………………………6
1.2 Research Motivation…………………………………………………………...…...……7
1.3 Research Objectives……………………………………………………...……………...8
Chapter 2 Literature Review…………………………………………………………......…..…9
2.1 Synthesis of Nicotinamide Mononucleotide……………………………………9
2.1.1 Chemical Synthesis of Nicotinamide Mononucleotide………9
2.1.2 Microbial Preparation of Nicotinamide Mononucleotide…………………………………………………………………………………………………………10
2.1.3 Application of Nicotinamide Mononucleotide……………………11
2.2 Natural Extract Preparation Technology………………………………..13
2.2.1 Extraction Technology……………………………………………………….…….13
2.2.2 Purification Technology……………………………………………………..……..17
Chapter 3 Experimental………………………………………………………………..…20
3.1 Materials……………………………………………………………………………..…20
3.2 Preparation of Nicotinamide Mononucleotide Solution Extracted from Edamame…….………………………………………………………………………………21
3.2.1 Edamame Processing………………………………………………………………22
3.2.2 Add Solvent and Mesh Edamame…………………………………………….……22
3.2.3 Centrifugal…………………………………………………………………....……23
3.2.4 Vacuum Oven……………………………………………………….………..……23
3.3 Characteristic Analysis of Nicotinamide Mononucleotide.…………………………………………………………………………………………………………24
3.3.1 Optical Microscope (OM)……………………………………………….…………24
3.3.2 Scanning Electron Microscope (SEM) and Energy-Dispersive X-ray Spectroscopy(EDS) Composition Analysis….25
3.3.3 Transmission Electron Microscope (TEM)…………………………………27
3.3.4 Fourier-Transform Infrared Spectroscopy (FT-IR)………28
3.3.5 Ultraviolet-Visible Spectroscopy (UV-Vis)………………….…30
3.3.6 Thin Layer Chromatography (TLC)…………………………………………….…31
Chapter 4 Discussion and Results……………………………………………………………..34
4.1 Pure NMN analysis……………………………………………………………………..34
4.1.1 Observe the Morphology of Pure NMN by Optical Microscope (OM)………………………………………………………………………………………………………………34
4.1.2 Observe the Morphology of Pure NMN by CFE-SEM………………34
4.1.3 Observe the Morphology of Pure NMN by TEM……………….36
4.1.4 The UV-Vis Spectral Analysis of Pure NMN……………………………38
4.1.5 Analysis of FT-IR Functional Group of Pure NMN Powder……………………………………………………………………………………………………………………………….…39
4.1.6 Pure NMN Powder was Dissolved in Acetone and Observed its Morphology by OM…. ……………………………………………………………………………..41
4.1.7 UV-Vis Spectroscopy Analysis of Pure NMN with Acetone as Solvent……………………………………………………………………………………………………………………42
4.2 Analysis of Extracting NMN from Edamame with Acetone as Solvent………………………………………………………………………………………………………………………43
4.2.1 Synthetic Image of NMN Solution Extracted from Edamame with Acetone as Solvent…………………….……………………………………………...………43
4.2.2 The Surface Morphology of the NMN Solution Extracted from Edamame with Acetone as the Solvent was Observed by OM…………….……………………………………………………………………………………………………………………………….…44
4.2.3 The UV-Vis Spectroscopy Analysis of the NMN Solution Extracted from Edamame with Acetone as the Solvent……………45
4.3 Analysis of NMN Extracted from Edamame with Water as the Solvent……………………………………………………………………46
4.3.1 Synthetic Diagram of NMN Solution Extracted from Edamame with Water as the Solvent.…………………………………46
4.3.2 Observation of the OM Surface Morphology of the NMN Solution Extracted from Edamame with Water as the Solvent………….…………………………………………………………………………………….48
4.3.3 Observation of the SEM Surface Morphology of the NMN Solution Extracted from Edamame with Water as the Solvent…………………………………………………………………………………………48
4.3.4 UV-Vis Spectroscopy Analysis of NMN Solution Extracted from Edamame with Water as the Solvent……………….…50
4.3.5 FT-IR Functional Groups Analysis of NMN Solution Extracted from Edamame with Water as the Solvent…………...51
4.4 Thin Layer Chromatography (TLC)…………………………………………….………52
4.4.1 Silica Gel Thin Layer Chromatography Aluminum Plates ……………………………………………………………………………………………………………………53
4.4.2 Thin Layer Chromatography Cellulose F Plates…………...58
4.5 Ultrasonic extraction………………………………………………………….….61
4.6 CO2 supercritical fluid extraction……………………………………63
4.7 Stability test………………………………………………………………………….…67
Chapter 5 Conclusions…………………………………………………………………...….69
References……………….……………………………………………………………..……71
Appendix……………………………………………………………………………………...77

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指導教授

陳郁文(Yu-Wen Chen)

審核日期

2021-7-7

推文