THE EFFECT OF PRETREATMENTS ON THE CURCUMIN CONTENTS AND ANTIOXIDANT ACTIVITIES OF FRESH AND SOLAR TUNNEL DRIED CURCUMA (Curcuma xanthorrhiza Roxb.

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1 THE EFFECT OF PRETREATMENTS ON THE CURCUMIN CONTENTS AND ANTIOXIDANT ACTIVITIES OF FRESH AND SOLAR TUNNEL DRIED CURCUMA (Curcuma xanthorrhiza Roxb.) PENGARUH PERLAKUAN AWAL TERHADAP SENYAWA KURKUMIN DAN AKTIVITAS ANTIOKSIDAN PADA TEMULAWAK (Curcuma xanthorrhiza Roxb.) SEGAR DAN YANG DIKERINGKAN DENGAN MENGGUNAKAN SOLAR TUNNEL BACHELOR THESIS Submitted to The Faculty of Agricultural Technology in partial fulfillment of the requirements for obtaining the Bachelor Degree By: RIANA NATALIA SETIYAWAN NIM : DEPARTMENT OF FOOD TECHNOLOGY FACULTY OF AGRICULTURAL TECHNOLOGY SOEGIJAPRANATA CATHOLIC UNIVERSITY SEMARANG 2017

2 THE EFFECT OF PRETREATMENTS ON THE CURCUMIN CONTENTS AND ANTIOXIDANT ACTIVITIES OF FRESH AND SOLAR TUNNEL DRIED CURCUMA (Curcuma xanthorrhiza Roxb.) PENGARUH PERLAKUAN AWAL TERHADAP SENYAWA KURKUMIN DAN AKTIVITAS ANTIOKSIDAN PADA TEMULAWAK (Curcuma xanthorrhiza Roxb.) SEGAR DAN YANG DIKERINGKAN DENGAN MENGGUNAKAN SOLAR TUNNEL By: RIANA NATALIA SETIYAWAN NIM : This thesis has been approved and defended in front of the examination committee on 24 th January 2017 Semarang, 24 th January 2017 Faculty of Agricultural Technology Soegijapranata Catholic University Supervisor Dean, Dr. V. Kristina Ananingsih, ST, MSc Dr. V. Kristina Ananingsih, ST, MSc Co-Supervisor Dr. R. Probo Y. Nugrahedi STP, MSc

3 THE AUTHENTICITY OF A THESIS STATEMENT I hereby declare that the thesis entitled THE EFFECT OF PRETREATMENTS ON THE CURCUMIN CONTENTS AND ANTIOXIDANT ACTIVITIES OF FRESH AND SOLAR TUNNEL DRIED CURCUMA (Curcuma xanthorrhiza Roxb.) contains no work that ever proposed to acquire a bachelorship title in a University, and along to my knowledge, there is no work ever written or published by others, except the ones uses as references in this thesis and mentioned in the list of references. If it is proven in the future that partially or whole thesis is the result of plagiarism, therefore I will be willing to be revoked with all the consequences in accordance with the law and regulations applied at Soegijapranata Catholic University and/or valid law and regulations. Semarang, 24 th January 2017 Riana Natalia Setiyawan

4 SUMMARY Curcuma (Curcuma xanthorrhiza Roxb.) is Zingiberaceace family rhizome. The yellow pigment that gives a yellow color in curcuma is known as bioactive compound namely curcuminoid. Curcuminoid comprises curcumin, demethoxycurcumin, and bisdemethoxycurcumin. Curcumin compound in curcuma is the main active of all three curcuminoids. Curcumin has antioxidant activity that can be used as radical scavenging, anti-inflamation, anti-tumor, anti-allergy, and anti-dementia. Based on the benefits, curcuma is developed industrially. However, curcuma is easily spoiled and incorrect handling makes nutrients and bioactive compounds are degraded. Dried curcuma can be the solution. Dried curcuma is obtained through the drying process that can prolong the shelf life, but the process can lead to the loss of some nutrients and bioactive compounds contained in the product. The objective of this research is to determine the effect of pretreatments (citric acid soaking and steam blanching) on the curcumin contents and antioxidant activities (%inhibition) of fresh and solar tunnel dried curcuma (Curcuma xanthorriza Roxb.). There were pretreatments before drying process, namely steam blanching for 3, 5, 10 minutes, soaking in the 0.5% and 1% citric acid solution for 10, 20, 30 minutes. Curcuma was dried using Solar Tunnel Dryer until the water content reached below 7%. After that, curcuma was extracted with methanol for 30 minutes. The curcumin content was analyzed by using High Performance Liquid Chromathography (HPLC) at 425 nm and spectrophotometer at 421 nm. The antioxidant activity was analyzed by DPPH method using spectrophotometer at 517 nm. The results show that soaking curcuma with citric acid solution and steam blanching could accelerate the drying process and increased the antioxidant activity (%inhibition). Curcuma soaked in the 1% citric acid solution for 30 minutes is the best pretreatment to accelerate the drying process and reduced the probability of antioxidant activity lost (% inhibition is 92.25±2.41%) and curcumin content is ± ppm for HPLC method and is ± ppm for spectrophotometric method. i

5 RINGKASAN Temulawak (Curcuma xanthorrhiza Roxb.) merupakan rimpang dari keluarga Zingiberaceace. Temulawak mengandung pigment bewarna kuning yang diketahui mengandung senyawa bioaktif yang disebut kurkuminoid. Kurkuminoid terdiri atas kurkumin, demetoksikurkumin, dan bisdemetoksikurkumin. Senyawa kurkumin mempunyai aktivitas antioksidan yang dapat digunakan sebagai penangkal radikal bebas, anti peradangan, anti tumor, anti alergi, dan anti penuaan. Berdasarkan kelebihan yang dimiliki, temulawak mulai dikembangkan di bidang industri. Namun, temulawak mudah rusak dan penanganan yang kurang tepat dapat mengakibatkan nutrisi dan senyawa aktif terdegradasi. Temulawak kering dapat menjadi salah satu solusinya. Temulawak kering diperoleh melalui proses pengeringan yang dapat memperpanjang umur simpan, tetapi prosesnya dapat membuat kehilangan beberapa nutrisi dan senyawa aktif yang terkandung di dalamnya. Tujuan dari penelitian ini adalah untuk mengetahui pengaruh perlakuan awal( perendaman asam sitrat dan steam blanching) terhadap kandungan kurkumin dan aktivitas antioksidan pada temulawak segar dan temulawak kering yang dikeringkan dengan menggunakan Solar Tunnel Dryer (STD). Beberapa perlakuan awal yang dilakukan, antara lain pengukusan selama 3, 5, 10 menit, perendaman 0.5% dan 1% larutan asam sitrat selama 10, 20, 30 menit. Temulawak dikeringkan menggunakan Solar Tunnel Dryer hingga kadar airnya mencapai di bawah 7%. Setelah itu, temulawak diekstrak dengan methanol selama 30 menit. Senyawa kurkumin dianalisa menggunakan High Performance Liquid Chromatography (HPLC) dengan panjang gelombang 425 nm dan spektrofotometer pada 421 nm. Aktivitas antioksidan dianalisa dengan menggunakan metode DPPH dengan spektofotometer pada 517 nm. Hasil penelitian menunjukkan perendaman temulawak dengan larutan asam sitrat dan steam blanching dapat mempercepat proses pengeringan dan meningkatkan aktivitas antioksidan (%inhibition). Perendaman temulawak dengan larutan 1% asam sitrat selama 30 menit merupakan pretreatment terbaik untuk mempercepat proses pengeringan dan menghasilkan kandungan aktivitas antioksidan (92,25±2,41%) dan senyawa kurkumin sebanyak 15054,21±1939,29 ppm untuk metode HPLC dan 17759,35±214,30 ppm untuk metode spektrofotometer. ii

6 PREFACE Praise and gratitute to God Almighty for His blessing and mercy, the author would have the opportunity to undergo this experiment and finish the thesis with the title is THE EFFECT OF PRETREATMENTS ON THE CURCUMIN CONTENTS AND ANTIOXIDANT ACTIVITIES OF FRESH AND SOLAR TUNNEL DRIED CURCUMA (Curcuma xanthorrhiza Roxb.). This report is submitted to partial fulfillment of the requirements for obtaining the Bachelor Degree in Soegijapranata Catholic University Semarang. This research was supported by Dana Hibah Bersaing DIKTI SK No 011/K6/SP2H/RISETTERAPAN/2016 entitled Optimasi Kondisi Proses pada Solar Tunnel Dryer untuk Simplisia Rimpang dan Standarisasi Mutu Produknya. The author would not be able to finish these thesis alone, and only by huge support and guidance given by the great and very helpful people around the author, these thesis could be finished. Special thanks for : 1. God Almighty that always blessed, saved, and guided author in every step to finish this experiment and thesis. 2. Dr. Victoria Kristina Ananingsih, ST., MSc. as the Dean and also author s supervisor of Faculty of Agricultural Technologi Soegijapranata Catholic University, Semarang for always patient, understanding, giving me best advice and support. 3. Dr. R. Probo Y. Nugrahedi, STP, MSc as the author s co-supervisor who is always giving his time to help the author with her advise and motivation, who has influenced author during the learning process in the Food Technology Department. 4. My family (Setiyawan Leonardi, Arlita Setiyawan, and Elma Setiyawan) who always support me and cheers me everyday. 5. My dearest friends Pamela, Devi, Novi, Donna, Hans, Arief, Alan, Rainier, Adri Novita Wulandari, Michael, who always supported and cheered me for the thing I had gone through. 6. Mr. Suparman as chief of Klaster Biofarmaka Karanganyar that gave information and supplay the sample (curcuma) for this research. iii

7 7. All the beloved lectures of FTP for being great family and team to pass on the knowledge, experience, and education during all year. So, I could finish this thesis. 8. The laboratory assistants (Mas Soleh, Mas Supriyana, Mas Lylyx, and Mbak Agatha) for always helping me and give me their motivation throughout the laboratory life in Food Technology Department. 9. The administrative staffs and employees (Mbak Susi, Pak Andre, Pak Joko, Bu Tutik, and Pak Surono) for always providing a great service during the study. The author realized that this report is still far from perfect and there are still mani shortcomings due to the limitation of the author. However, the author hoped that this report can still be an inspiration and provide useful information for all the readers. Semarang, January 2017 Riana Natalia Setiyawan Author iv

8 LIST OF CONTENT SUMMARY... i RINGKASAN... ii PREFACE... iii LIST OF CONTENT... v LIST OF TABLES... vii LIST OF FIGURES... viii LIST OF APPENDIX...ix 1. INTRODUCTION Background Literature Review Curcumin in Curcuma xanthorixa Roxb Antioxidant Activity in Curcuma zanthorrhiza Roxb Citric Acid and Steam Blanching Drying Process Using Solar Tunnel Dryer (STD) High Performance Liquid Chromatography (HPLC) to Identify Curcumin Compound Purpose of Research MATERIALS AND METHODS Place and Time Research Materials Equipments Materials Research Methods Preparation of Sample Water Content Analysis Preparation of Standard Extraction of Curcumin Linearity and Range Study Determination of Curcumin Using HPLC Determination of Curcumin Using Spectrophotometric Antioxidant Activity Data Analysis Research Design RESULTS Temperature Profile Water Content Antioxidant Activities (% Inhibition) Curcumin Contents Using HPLC and Spectrophotometer Water Activity DISCUSSION Pretreatments for The Curcuma The Drying Process Using Solar Tunnel Dryer Antioxidant Activities of Curcuma (% Inhibition) Curcumin Contents Using HPLC and Spectrophotometric Water Activity (a w ) CONCLUSIONS AND SUGGESTION Conclusions v

9 5.2. Suggestion REFERENCES APPENDIX Antioxidant Activities (% Inhibition) in Dry Basis Chromatograms of Curcumin Contents Using HPLC Chromatograms of Curcumin Before Drying Process Chromatograms of Curcuma After Drying Process Output of Test Of Normality Output of Post Hoc One Way Anova Output of T Test Linearity Curve of Curcumin Standard Using HPLC Linearity Curve of Curcumin Standard Using Spectrophotometer vi

10 LIST OF TABLES Table 1. The Water Content Every 15 Minutes Table 2. The Antioxidant Activities (% Inhibition) of Curcuma Before and After Drying Process Table 3. Curcumin Contents Before and After Drying Process Using HPLC and Spectrophotometer vii

11 LIST OF FIGURES Figure 1. Curcuma xanthorhiza Roxb Figure 2. The chemical structure of (a) curcumin, (b) demethoxycurcumin, (c) bisdemethoxycurcumin... 3 Figure 3. Flow chart of sample preparation... 8 Figure 4. Research design Figure 5. Temperature profile during drying process Figure 6. Water content of curcuma control and soaked in 0.5% citric acid Figure 7. Water content of curcuma soaked in 1% citric acid Figure 8. Water content of curcuma steam blanched Figure 9. Chromatograms of curcumin standard Figure 10. Chromatograms of curcumin contents in curcuma before drying process Figure 11. Chromatograms of curcumin contents in curcuma after drying process viii

12 LIST OF APPENDIX Appendix 1. Antioxidant Activities (% Inhibition) in Dry Basis Appendix 2. Chromatograms of Curcumin Standards Appendix 3. Non Pretreatment Curcuma (C0) Appendix 4. Curcuma soaked in 0.5% citric acid solution for 10 minutes Appendix 5. Curcuma soaked in 0.5% citric acid solution for 20 minutes Appendix 6. Curcuma soaked in 0.5% citric acid solution for 30 minutes Appendix 7. Curcuma soaked in 1% citric acid solution for 10 minutes Appendix 8. Curcuma soaked in 1% citric acid solution for 20 minutes Appendix 9. Curcuma soaked in 1% citric acid solution for 30 minutes Appendix 10. Curcuma steam blanched for 3 minutes Appendix 11. Curcuma steam blanched for 5 minutes Appendix 12. Curcuma steam blanched for 10 minutes Appendix 13. Non Pretreatment Dried Curcuma (C0) Appendix 14. Dried curcuma soaked in 0.5% citric acid solution for 10 minutes (C1A) Appendix 15. Dried curcuma soaked in 0.5% citric acid solution for 20 minutes (C1B) Appendix 16. Dried curcuma soaked in 0.5% citric acid solution for 30 minutes (C1C) Appendix 17. Dried curcuma soaked in 1% citric acid for 10 minutes (C2A) Appendix 18. Dried curcuma soaked in 1% citric acid solution for 20 minutes (C2B). 45 Appendix 19. Dried curcuma soaked in 1% citric acid solution for 30 minutes (C2C). 45 Appendix 20. Dried curcuma steam blanched for 3 minutes (C3A) Appendix 21. Dried curcuma steam blanched for 5 minutes (C3B) Appendix 22. Dried curcuma steam blanched for 10 minutes (C3C) Appendix 23. Antioxidant Activities Appendix 24. Curcumin Contents using HPLC Appendix 25. Curcumin Contents using Spectrophotometric Appendix 26. Water Activity Appendix 27. Antioxidant Activity Appendix 28. Curcumin Contents using HPLC Appendix 29. Curcumin Contents using Spectrophotometric Appendix 30. T Test of Antioxidant Activities Before and After Drying Appendix 31. T Test of Curcumin Contents Before and After Drying Appendix 32. T Test of Water Activity Before and After Drying Appendix 33. Linearity Curve of Curcumin Standard using HPLC Appendix 34. Linearity Curve of Curcumin Standard Using Spectrophotometer ix

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