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Improved Taste and Flavor by Optimized Roasting and Extraction of Vietnamese Robusta Coffee and Cosmeceutical Efficacies for Anti-aging and Skin Whitening

초록/요약

In this study, we assessed changes in the flavors and tastes of coffee extracted from Vietnamese Robusta with different roasting times and temperatures, as well as different extraction temperatures. In addition, Vietnamese Robusta extracts were in vitro tested for cosmeceutical properties -- anti-aging and skin whitening. Vietnamese Robusta green beans were roasted for different times ranging from 5 to 20 minutes and at different temperatures ranging from 100 to 250 degrees. The roasted coffee was then extracted at five different temperatures ranging from 90 to 120 degrees. The coffee flavor was evaluated in terms of 5 key odorants: guaiacol; 4-ethylguaiacol; 2-ethyl-3,5-dimethylpyrazine; 2,3-diethyl-5-methylpyrazine, and 2-furfuryl. The taste of coffee was evaluated in terms of 6 main compounds that confer bitterness and sourness: caffeine, trigonelline, citric acid, acetic acid, formic acid, and chlorogenic acid. According to the contents of key odorants and taste compounds, the optimized roasting and extracting conditions were identified: 200 g of VN Robusta green beans, roasting at 230oC for 18 minutes, and then extracted with Vibiemme Espresso Machine at 110oC. For the in vitro anti-aging tests, Vietnamese Robusta green beans were roasted at six different levels and extracted by hot water. After measuring the radical scavenging activity, the good roasting condition was chosen for in vitro test on human fibroblast cells. First, MTT assay was conducted to identify the toxicity of the coffee extract. After that, proteome analysis using 2-D was conducted to observe the changes of proteins that are involved in skin aging process. Deoiled VN Robusta extract at roasting level 4 (200 g, 2300C, 15 min) has stronger anti-oxidative activity with SC50 value for DPPH scavenging is 33.2 mg/L. Deoiled VN Robusta extract inhibits the degradation of elastin and dermal extracellular matrix with the down-regulation of 4 proteins including CELA2B, CELA3A, CELA3B and MMP3. Also the two proteins, which are reported to increase expression in senescent fibroblast, were down-regulated in NF treated by deoiled VN Robusta extract. Deoiled VN Robusta extract significantly up-regulated the natural anti-oxidant enzymes of human fibroblast cells including GPX2, GCLM, PRDX4, GSTA1 and NQO1. This extract as well as destroyed or inhibited free radicals which resulted in down-regulation of NF-kB1 and TNF-alpha. For the in vitro skin whitening tests, Vietnamese Robusta green beans were roasted at six different levels and extracted oil by supercritical carbon dioxide. After measuring the radical scavenging activity and tyrosinase inhibition activity of coffee oils, the good roasting condition was chosen for in vitro test on B16F1 mouse melanoma cells. First, MTT assay was conducted to identify the toxicity of the coffee oil. After that, melanin content was measured to evaluate the inhibition of Vietnamese Robusta oil on melanin production. Finally, proteome analysis using 2-D was conducted to observe the changes of proteins that are involved in melanogenesis. VN Robusta oil at roasting level 4 (200 g, 2300C, 15 min) has stronger melanogenesis inhibition activity with SC50 value for DPPH scavenging is 400 mg/L and IC50 value for Tyrosinase Inhibiton is 17.73 mg/L. Melanin content of melanoma cells treated with 10 mg/L VN Robusta oil decrease about 25%. VN Robusta oil inhibits the early period of melanogenesis which is proved by the down-regulation of 8 identified proteins including α-MSH, MC1R, ACTHR, MITF, PKC-β, SCF, TYR and TRP-2. These results indicate that Robusta in general and Vietnamese Robusta in particular is not only the important source for coffee drink but also is a potential material for cosmetic industry with good skin whitening activity and strong anti-aging effect.

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목차

TABLE OF CONTENTS

Acknowledgement i
Abstract iii
Table of Contents iv
List of Figures viii
List of Tables x

PART 1. Introduction 1
1.Over view of the world coffee and Vietnamese Robusta 1
2.Health benefits of coffee 1
3.The skin aging mechanism 3
4.The melanomagenesis of skin 5

PART 2. Improved Taste and Flavor of Vietnamese Robusta by Roasting and Extraction 7
1.Object 7
2.Materials and methods 7
2.1.Materials 7
2.2.Roast coffee beans 8
2.3.Espresso extract 8
2.4.Collect headspace 8
2.5.Gas chromatography (GC) analysis 8
2.6.High Performance Liquid Chromatography (HPLC) analysis 10
3.Results and discussion 10
3.1.Changes of taste, flavor contents in Vietnamese Robusta versus roasting temperature and time 10
3.2.Optimized roasting and extracting conditions for Vietnamese Robusta Espresso taste and flavor 23
4.Conclusion 31

PART 3. The Anti-aging Effect of Vietnamese Robusta Extract 32
1.Object 32
2.Materials 32
2.1. Materials and methods 32
2.2.The DPPH radical scavenging Assay 33
2.3.MTT Assay 33
2.4.Protein Extraction 34
2.5.Proteome analysis 34
3.Results and discussion 35
3.1.Anti-oxidative activity of Vietnamese Robusta extract 35
3.2.Toxicity of Vietnamese Robusta extracts on human fibroblast 37
3.3.Effect of Vietnamese Robusta extracts on cellular senescence related proteins of human fibroblast 38
3.4.Effect of Vietnamese Robusta extracts on anti-oxidant related proteins of human fibroblast 42
4.Conclusion 46

PART 4. The Skin Whitening Effect of Vietnamese Robusta Oil Extracted by The Supercritical Fluid 47
1.Object 47
2.Materials and Methods 47
2.1.Materials 47
2.2.The DPPH radical scavenging Assay 48
2.3.Tyrosinase Inhibition Assay 48
2.4.MTT Assay 48
2.5.Melanin Content Assay 48
2.6.Protein Extraction 49
2.7.Proteome analysis 49
3.Results and discussion 49
3.1.Anti-oxidative activity of Vietnamese Robusta Oils 49
3.2.Tyrosinase Inhibition Activity of Vietnamese Robusta Oils 52
3.3.Toxicity of Vietnamese Robusta Oil on melanoma cells 54
3.4.Effect of Vietnamese Robusta Oil on Melanin Production of melanoma cells 55
3.5.Effect of Vietnamese Robusta Oil on melanogenesis related proteins of melanoma cells 56
4.Conclusion 59

PART 5. Conclusion 60

REFERENCES 61

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