THE EFFECT OF CRUDE BROMELAIN FROM PINEAPPLE (Ananas comosus) EXTRACT TOWARDS TOTAL YIELD AND PHYSICOCHEMICAL QUALITY OF VIRGIN COCONUT OIL

  THE EFFECT OF CRUDE BROMELAIN FROM PINEAPPLE (Ananas comosus) EXTRACT TOWARDS TOTAL YIELD AND PHYSICOCHEMICAL QUALITY OF VIRGIN COCONUT OIL

EFEK ENZIM BROMELIN KASAR DARI EKSTRAK NANAS

  (Ananas comosus) TERHADAP TOTAL YIELD DAN KUALITAS FISIKOKIMIAWI MINYAK KELAPA MURNI THESIS

  Submitted in partial fulfillment for the requirements to obtain a Bachelor Degree in Food Technology from the Faculty of Agricultural Technology

  By : CHINDYA PARAMITHA DEVI 08.70.0122 DEPARTMENT OF FOOD TECHNOLOGY FACULTY OF AGRICULTURAL TECHNOLOGY SOEGIJAPRANATA CATHOLIC UNIVERSITY SEMARANG 2012 Perpustakaan Unika

  THE EFFECT OF CRUDE BROMELAIN FROM PINEAPPLE (Ananas comosus) EXTRACT TOWARDS TOTAL YIELD AND PHYSICOCHEMICAL QUALITY OF VIRGIN COCONUT OIL

  (Ananas comosus) TERHADAP TOTAL YIELD DAN KUALITAS

  By : CHINDYA PARAMITHA DEVI NIM : 08.70.0122 Food Technology Department

  This thesis has been approved and defended in front of the examination board of members on July 2012.

  Semarang, July 2012 Faculty of Agricultural Techology Soegijapranata Catholic University

  Advisor I, Dean, Dra. Laksmi Hartayanie, MP. Ita Sulistyawati, STP., MSc. Advisor II, Ir. Lindayani, MP., PhD.

  Perpustakaan Unika

EFEK ENZIM BROMELIN KASAR DARI EKSTRAK NANAS

FISIKOKIMIAWI MINYAK KELAPA MURNI

  Perpustakaan Unika SUMMARY

  Coconut kernel is a tropical fruit that has been intensively cultivated in Indonesia. The most essential of coconut product is coconut oil. Virgin coconut oil (VCO) is pharmaceutically innovated due to the medium chain-saturated fatty acids content (lauric acids) which has proven to have a good impact to health. VCO extraction generally uses physical and chemical methods which are relatively easy and inexpensive. However, those methods affect to nutrition loss and rancidity during the storage. This research attempted to present a method in which the addition of protease was adopted so as to improve the total yield of VCO. Protease source was obtained from pineapple fruit because of its availability and reachable cost. The aim of this research is to find the effect of crude bromelain from pineapple extract towards total yield and physicochemical characteristics of VCO, such as water content, free fatty acid, peroxide value, viscosity, and clarity. Preliminary research was done to find the concentration of crude bromelain which was later used in the primary research. Extraction of VCO was done by adding flesh and core unripe pineapple extract with concentrations of 10% and 20%. Some of 10% of core bromelain resulted the best quality of water content and free fatty acid of VCO. Meanwhile, 20% of core bromelain resulted the best viscosity of VCO. The pineapple extract resulted no peroxide in VCO due to the presumptive presence of peroxidase. There was an effect towards total yield, water content, free fatty acid, peroxide value, and viscosity of VCO by crude bromelain from pineapple extract. But, there was no effect towards refractive index. The result from 20% of flesh extract gave the highest yield of VCO, meanwhile 10% of core extract gave the best physicochemical characteristics.

  Perpustakaan Unika RINGKASAN

  Buah kelapa adalah buah tropis yang telah lama dibudidayakan di Indonesia. Dari berbagai manfaat, produk kelapa yang paling berharga adalah minyak kelapa. Minyak kelapa murni (MKM) mulai banyak dikembangkan sebagai suplemen karena kandungan asam lemak rantai sedang, yaitu asam laurat yang memiliki efek positif bagi kesehatan. Ekstraksi MKM yang umum dilakukan adalah dengan metode fisik dan kimia, terkait prosedurnya yang relatif mudah dan murah. Namun, metode tersebut mengakibatkan penurunan nilai nutrisi pada produk dan terbentuknya reaksi ketengikan selama proses penyimpanan. Dalam penelitian ini, ekstraksi MKM dilakukan dengan enzim protease dari ekstrak nanas untuk meningkatkan total yield MKM. Sumber protease yang digunakan adalah buah nanas yang ketersediaannya cukup melimpah dan harganya relatif murah. Penelitian ini bertujuan untuk mengetahui pengaruh ekstraksi dengan penambahan enzim bromelin kasar terhadap total yield dan karakteristik fisikokimia MKM, antara lain kadar air, asam lemak bebas, bilangan peroksida, viskositas, dan kejernihan. Uji pendahuluan dilakukan untuk menentukan konsentrasi bromelin kasar yang digunakan dalam penelitian utama. Ekstraksi MKM dilakukan dengan penambahan ekstrak daging dan inti buah nanas muda dengan konsentrasi 10% dan 20%. Hasil penelitian ini menunjukkan bahwa 10% ekstrak inti nanas menghasilkan MKM dengan karakteristik kadar air dan asam lemak bebas terbaik. Sedangkan, 20% ekstrak inti nanas menghasilkan MKM dengan karakteristik viskositas terbaik. Selain itu, ekstrak nanas mengakibatkan tidak terbentuknya senyawa peroksida lemak, yang diduga disebabkan adanya enzim peroksidase. Hasil menunjukkan bahwa ekstrak kasar bromelin pada ekstraksi MKM mempengaruhi total yield, kadar air, asam lemak bebas, bilangan peroksida, dan viskositas, tetapi tidak mempengaruhi kejernihan produk MKM. Ekstraksi dengan 20% ekstrak daging nanas menghasilkan total yield MKM terbanyak, sedangkan 10% ekstrak inti nanas menghasilkan kualitas fisikokimia MKM terbaik.

  Perpustakaan Unika ACKNOWLEDGEMENT

  Glory be to the Almighty Lord for the wonderful love and blessings, so that I can finally finish this thesis entitled “THE EFFECT OF CRUDE BROMELAIN FROM PINEAPPLE (Ananas comosus) TOWARDS TOTAL YIELD AND PHYSICOCHEMICAL QUALITY OF VIRGIN COCONUT OIL”. This thesis is submitted in partial fulfillment for the requirements to obtain a Bachelor Degree in Food Technology from the Faculty of Agricultural Technology. There is so much new knowledge and experiences that had been acknowledged during the research, particularly on the science and scientific writing that the learning experience has been useful as a new step towards my future life.

  This thesis has been done by invaluable assistance from those who give their best supports that I am very grateful for. Without them, I would not have done this thesis successfully. I would like to gratefully acknowledge to: 1.

  Jesus Christ and Holy Mary, for Thy wonderful grace and strength given to a sinner like me.

  2. Ita Sulistyawati, STP., MSc., as the Dean of Agricultural Technology Faculty of Soegijapranata Catholic University Semarang.

  3. Dra. Laksmi Hartayanie, MP., and Ir. Lindayani, MP., PhD., as the advisors who are being patient in sharing knowledge and experience and also being great teachers for me.

  4. My beloved family; Papa, Mama, Dhinar, and all of relatives, for supporting in every single second and reminding tirelessly to finish this project.

  5. Mas Dhalang Kanda Kerwin, for being my great motivator and his love in teaching much lesson of life.

  6. F. Sholeh Kuntoro, Endah P., and Supriyana, as the laborants for their huge endurance during the lab days.

  7. Bu Ekawati MD, Mas Anggara Graphic, and Wisnu Ardhi, for being my kind mentors in the thesis compilation. v

  Perpustakaan Unika 8.

  My lecturers in FTP, especially Pak Sumardi, Bu Kartika Puspa, and Pak Binardo, for their supports and provisions during my study period. They mean too much for me! 9. My friends in FTP: Merry, Intan, Nita, Melita, Jece, Vaut, Nug, Tommy, Onon,

  Devina, Jeffry, Mona, Dewi, Cucok, and other FTP’ers, for the sweet memories during this 4 years. Never will I forget our togetherness.

  10.

  “GLORY” – The SUPERB-mate: Silvani Andalita, Dede Anjang, Bagus Mukti, Dita Anindia, Dhimas Rangga, Doni Wahyu, Arprisca Andhina, Yudha Krisdhanarto, Nugroho Fajar, Shinta Elok, Eris Putra, Radith Casandra, and the others, for inspiring and creating precious moments together. Be sure, we will always be GLORY in the future! 11. Anne Margaretha, Paskalia Marlina, Maya Tundjoeng, Georgius Tjitra, Yuliana, for the inspiration and endurable spirit given to me.

  12. My kind fathers: Pak Ferdinandus Hindiarto and Pak Benny Danang Setianto, for the abundant motivation in Awakening the Giant within myself.

  13. Staffs of Vice Rector III: Pak Tikno, Pak Eko, Pak Totok, for making their room like my house.

  14. FTP administration staffs and employees, for providing valuable help during study period.

  15. Last but not least, I wish to express my deepest gratitude for all the people who made this thesis finally complete with their kindness and support, which I can not possibly mention one by one. I realize that this thesis report is still far from perfect. I still require advice and criticism to improve this report, so I welcome any encouraging comments. I hope that this research could give a valuable contribution to the food science knowledge and can be implemented usefully in food industries.

  Semarang, July 2012 Chindya

  Perpustakaan Unika CONTENTS

  page SUMMARY ................................................................................................................ ii

  RINGKASAN ................................................................................................................. iii

  ACKNOWLEDGEMENT ............................................................................................ iv CONTENTS ................................................................................................................ vi LIST OF TABLES ....................................................................................................... viii LIST OF FIGURES ...................................................................................................... ix LIST OF APPENDICES ............................................................................................. x

  1. INTRODUCTION .................................................................................................... 1

  1.1. Background ....................................................................................................... 1

  1.2. Literature Review ............................................................................................. 3 1.2.1. Proteolytic Enzyme .................................................................................

  3

  1.2.2. Virgin Coconut Oil (VCO) ...................................................................... 4

  1.3. Aim of this Research ......................................................................................... 6

  2. MATERIALS AND METHODS ............................................................................. 7

  2.1. Time and Place of Experiment .......................................................................... 7

  2.2. Materials and Instruments ................................................................................. 7 2.2.1. Materials ..................................................................................................

  7 2.2.2. Instruments ..............................................................................................

  7

  2.3. Preliminary Research ........................................................................................ 8 2.3.1.Crude Bromelain Extraction ....................................................................

  8

  2.3.2. Proteolytic Activity Assay ...................................................................... 9 2.3.3.

  VCO Extraction .......................................................................................

  10

  2.4. Primary Research .............................................................................................. 12

  2.4.1. Crude Bromelain Extraction ................................................................... 13

  2.4.2. Proteolytic Activity Assay ...................................................................... 13 2.4.3.

  VCO Extraction ......................................................................................

  13

  2.4.4. Quality Analyses of VCO Products ........................................................ 13

  2.4.4.1. Water Content ............................................................................ 14

  2.4.4.2. Free Fatty Acid ........................................................................... 14

  2.4.4.3. Peroxide Value ........................................................................... 14 2.4.4.4. Viscosity .....................................................................................

  15 2.4.4.5. Clarity .........................................................................................

  15 2.4.5. Data Analysis ..........................................................................................

  16

  3. RESULT .................................................................................................................. 17

  3.1. Preliminary Research ........................................................................................ 17

  3.1.1. Crude Bromelain Extract ........................................................................ 17 3.1.2.

  VCO Products .........................................................................................

  18

  3.2. Primary Research .............................................................................................. 18

  3.2.1. Crude Bromelain Extract ........................................................................ 18 3.2.2.

  VCO Products .........................................................................................

  19

  3.2.3. Quality Analyses of VCO Products ........................................................ 22

  vii

  Perpustakaan Unika

  4. DISCUSSION .......................................................................................................... 26

  4.1. Crude Bromelain Extraction ............................................................................. 26

  4.2. VCO Products ................................................................................................... 27

  4.3. Quality Analyses of VCO Products .................................................................. 29

  5. CONCLUSION AND SUGGESTION ................................................................... 35

  5.1. Conclusion ........................................................................................................ 35

  5.2. Suggestion ......................................................................................................... 35

  6. REFERENCES ........................................................................................................ 36 APPENDICES ............................................................................................................. 40

  Perpustakaan Unika LIST OF TABLES

  page Table 1. Physicochemical Characteristics of VCO Products by APCC ................... 5 Table 2. Fatty Acids Composition of VCO by APCC .............................................. 6 Table 3. The Characteristics of Pineapple Extract from Flesh and Core ................. 17 Table 4. Total Yield of VCO Products ..................................................................... 18 Table 5. The Characteristics of Pineapple Extract from Flesh and Core ................. 19 Table 6. Total Yield of VCO Products ..................................................................... 21 Table 7. Physicochemical Characteristics of VCO Products ................................... 22

  Perpustakaan Unika LIST OF FIGURES

  page Figure 1. Morphology of a pineapple plant (adapted from Bartholomew et al., 2002) .........................................................................................................

  4 Figure 2. The diagram of extracting crude bromelain enzyme ................................. 9 Figure 3. The diagram of analyzing proteolytic activity .......................................... 10 Figure 4. The process of VCO extraction ................................................................. 11 Figure 5. The diagram of primary research .............................................................. 12 Figure 6. VCO extraction process in separatory funnels with treatments of:

  (a)control; (b)flesh 10%; (c)flesh 20%; (d)core 10%; and (e)core 20% ... 20 Figure 7. VCO products from treatments of: (a)control; (b)flesh 10%;

  (c)flesh 20%; (d)core 10%; and (e)core 20% ............................................ 20 Figure 8. Total yield of VCO products ..................................................................... 21 Figure 9. Water content of VCO products ................................................................ 23 Figure 10.Free Fatty Acid and Peroxide Value of VCO Products ............................ 23 Figure 11.Viscosity of VCO Products ....................................................................... 24 Figure 12.Refractive Index of VCO Products ........................................................... 25 Figure 13.Coconut milk emulsion system (adapted from Asriani, 2006) ................. 27 Figure 14.Peptide hydrolisis by sulfihydril (-SH) active site of protease

  (adapted from Ketaren, 1986 in Raharja & Dwiyuni, 2005) .................... 29 Figure 15.Lipid hydrolisis mechanism (adapted from Ketaren, 1986 in

  Raharja & Dwiyuni, 2005) ........................................................................ 30 Figure 16.Free radicals formation by heating of unsaturated fatty acids

  (adapted from Winarno, 2004) .................................................................. 32 Figure 17. The molecule structure of pectin (modified from Winarno, 2004) ........... 33 Figure 18. The endpoint of titration in free fatty acid assay is shown by a slight pink colouration ........................................................................................

  47 Figure 19. Blank solution and samples appearance in some steps of peroxide value assay: (a) addition of chloroform, glacial acetic acid, and saturated Potassium Iodide; (b) addition of starch soluble indicator; (c) the endpoint of titration process with transparent colouration ............ 47

  Perpustakaan Unika

LIST OF APPENDICES

  page Appendix 1. Data of Preliminary Research .................................................................. 40 Appendix 2. Data of Primary Research ........................................................................ 41 Appendix 3. Primary Research Data Analysis by SPSS .............................................. 44 Appendix 4. Result of Free Fatty Acid Assay and Peroxide Value Assay ................... 47