UNIVERSITI PUTRA MALAYSIA COCOA BUTTER EXTRACTION FROM COCOA NIBS USING SUPERCRITICAL CARBON DIOXIDE ASEP EDI KUSNADI FSTM
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1 UNIVERSITI PUTRA MALAYSIA COCOA BUTTER EXTRACTION FROM COCOA NIBS USING SUPERCRITICAL CARBON DIOXIDE ASEP EDI KUSNADI FSTM
2 COCOA BUTTER EXTRACTION FROM COCOA NIBS USING SUPERCRITICAL CARBON DIOXIDE ASEP EDI KUSNADI DOCTOR OF PHILOSOPHY UNIVERSITI PUTRA MALAYSIA 2009
3 COCOA BUTTER EXTRACTION FROM COCOA NIBS USING SUPERCRITICAL CARBON DIOXIDE By ASEP EDI KUSNADI Thesis Submitted to the School of Graduate Studies, Universiti Putra Malaysia, in Fulfilment of the Requirements for the Degree of Doctor of Philosophy April 2009
4 This Thesis is specially dedicated to Mamah and Apa (alayarham), Aih, Ujang, Dadan and Nanang and all their family for the unconditional patient, love and support. ii
5 Abstract of thesis presented to the Senate of Universiti Putra Malaysia in fulfillment of the requirements for the degree of Doctor of Philosophy COCOA BUTTER EXTRACTION FROM COCOA NIBS USING SUPERCRITICAL CARBON DIOXIDE By ASEP EDI KUSNADI April 2009 Chairman: Professor Jinap Selamat, PhD Faculty: Food Science and Technology Cocoa beans consist mainly of cocoa butter (50-55% w/w). High quality cocoa butter used in food, cosmetic and pharmaceutical products is obtained by mechanical press, expeller, and solvent extraction using hexane. Supercritical fluid extraction (SFE) using carbon dioxide as a solvent has provided an excellent alternative to the use of chemical solvent in the extraction of cocoa butter from different plant matrices. In comparison with established methods, SFE has some important advantages, particularly in its ability to yield products that are completely free from processing residues. SFE is also an alternative from the standpoint of time-saving and for environmental reasons such as the reduction of the use of large solvent volumes. Carbon dioxide (CO 2 ) is an ideal solvent for extraction of natural products because it is nontoxic, odorless, tasteless, non explosive, readily available, and easily removable from the products. iii
6 This work is divided into several part namely; 1) the effect of flow rate with different pressure on cocoa butter extraction using supercritical carbon dioxide (SC-CO 2 ), 2) a study on the sample matrix involving: the effect of particle size, degree of fermentation and alkalization on cocoa butter extraction using supercritical fluid; 3) a study on the effect of moisture content, roasting time and roasting temperature on cocoa butter extraction using supercritical fluid, 4) a study on the effect of cosolvents with types and concentrations of polar cosolvents and non polar cosolvents in cocoa butter extraction using SC-CO 2, 5) to evaluate the mass transfer parameters in cocoa butter extraction by SFE using the Sovova Lacks plug flow model (SLM) for the effect of flow rate with different pressure, the effects of polar and non polar cosolvents, and also using single sphere model (SSM) for the effects of particle size, degree of fermentation, ph alkalization, moisture content, roasting time and roasting temperature and 6) to determine the triacylglycerols composition and fatty acid methyl esters composition of cocoa butter extracted that resulted from extraction using SC-CO 2 and SC-CO 2 with cosolvents. The study on the effect of flow rate and pressure found that the optimum conditions for flow rate and pressure for cocoa butter extraction using SC-CO 2 were at 2 ml/min and 35 MPa, r espectively. The highest yields were obtained from cocoa nibs sample with smaller particle size (S1= 0.07 mm) with 92% yield for almost 20 h using SC- CO 2, unfermented nibs (F1) with 100% yield for almost 10 h using ethanol 25% as cosolvent in SC-CO 2 and roasting treatment of 150 o C and 35 min (R6) with 100% yield for almost 14 h using ethanol 25% (mol %) as cosolvent in SC-CO 2. Increasing roasting time and temperature have resulted in the increase of the yield. Furthermore, iv
7 the highest yield was produced from high ph ( ) of dark alkalized cocoa liquor (A1) with 73.70% yield for nearly 18 h extraction time using ethanol 25% as cosolvent in SC-CO 2 and high moisture content (M5 =17.64%) with 60.73% after nearly 20 h extraction using SC-CO 2. Statistically, the yield (p<0.05) significantly was affected by all the treatment, however no significant different was observed for both light alkalized cocoa liquor (ph = ) and alkalized natural cocoa liquor (ph = ), and both moisture content of 9.79 and 17.64%, respectively. Ethanol showed the best cosolvent on the yield in SC-CO 2, followed by isopropanol, acetone, hexane and cyclohexane. The yield increased with an increase in concentration of cosolvents (25%>15%>5%), except for acetone in which 15% concentration was higher than 25% and 5%. Extraction of cocoa butter using SC-CO 2 with polar cosolvents obtained a yield higher than non polar cosolvents. All treatment of cosolvents studied statistically significantly different at p<0.05. The two mathematical models are based on mass transfer were used. First model, the Sovova s lack s plug flow model (SLM) was used to describe the extraction process of effect of flow rate with different pressure and effect of polar and non polar cosolvents. The hardly accessible solute xk and the volume mass transfer coefficients in the fluid phase (F) and solid phase S were used as fitting parameters. The maximum average deviation between measured and calculated oil yield was 6.861%. Mass transfer coefficients in the fluid phase and solid phase varied between E- v
8 06 to 1.498E-04 s -1 and between E-06 to 9.144E-04 s -1, respectively. Second model, the single sphere model (SSM) was used to describe the extraction process of effect of particle size, degree of fermentation, ph of alkalization, moisture content, roasting time and roasting temperature. Adjusting of effective intraparticle diffusion coefficient (De) and estimation of parameter of coefficient of mass transfer in the fluid phase (kf) and overall mass transfer coefficient kp and parameters were evaluated. The result showed that De, kf and kp varied between 8.900E-16 to 1.850E- 09 m 2 /s, between 1.139E-10 to 2.115E-04 m/s and between 9.581E-12 to 8.992E-05 m/s, respectively. All experimental data results of extraction curves were fitted fairly well by using both of the SLM model and SSM model with average absolute relative deviation (AARD) between measured and calculated oil yield were maximum of The results showed that cocoa butter extracted from various treatments had three main triacylglycerol (TG) namely Glycerol-1,3-Dipalmitate-2-Oleate (POP), Glycerol-1-Palmitate-2-Oleate-3-Stearate (POS), and Glycerol-1,3-Distearate-2- Oleate (SOS). In general, POS was the major component with to 45.78% followed by SOS and POP with to 31.67% and to 22.79%, respectively. Furthermore, analysis of fatty acid (FA) showed that palmitic acid (C16:0), stearic acid (C18:0) and oleic acid (C18:1) were the three main fatty acids in the cocoa butter extracted with stearic acid being the major component with to 39.06%, followed by oleic acid and palmitic acid with to 31.72% and to 31.33%, respectively. Statistically, there are differences the effects of all treatment on TG and FA compositions. vi
9 Abstrak tesis yang dikemukakan kepada Senat Universiti Putra Malaysia sebagai memenuhi keperluan untuk ijazah Doktor Falsafah PENGEKSTRAKAN LEMAK KOKO DARI PADA NIB KOKO MENGGUNAKAN BENDALIR SUPERKRITIK KARBON DIOKSIDA Oleh ASEP EDI KUSNADI April 2009 Pengerusi: Profesor Jinap Selamat, PhD Fakulti: Sains dan Teknologi Makanan Sebahagian besar daripada komponen biji koko terdiri daripada lemak koko (50-55%), dimana lemak koko yang berkualiti tinggi yang digunakan di dalam produk makanan, kosmetik dan farmaseutikal diperolehi melalui proses pengekstrakan menggunakan tekanan secara mekanik, penggunaan tekanan dengan kuasa putaran dan pelarut kimia seperti heksana. Proses pengekstrakan dengan cecair superkritik (SFE) yang menggunakan gas karbon dioksida sebagai pelarut telah menjadi satu alternatif yang baik bagi pengekstrakan lemak koko daripada matriks tumbuhan yang berbeza berbanding pengekstrakan menggunakan pelarut kimia. Dibandingkan dengan cara yang sedia pakai, SFE mempunyai beberapa kelebihan terutamanya dalam menghasilkan produk yang bebas dari sisa-sisa pemprosesan. SFE juga lebih baik dari segi penjimatan masa dan lebih mesra alam sekitar kerana mengurangkan penggunaan pelarut dalam isi padu yang tinggi. Karbon dioksida pula merupakan pelarut yang vii
10 ideal bagi pengekstrakan produk asli kerana ciri-cirinya yang tidak toksik, tidak berbau, tidak mempunyai rasa, tidak mudah meletup/terbakar, sedia didapati dan mudah dipisahkan dari produk. Kajian ini dimulakakan dengan mengkaji, 1) kesan kadar alir dengan tekanan yang berbeza ke atas pengekstrakan lemak koko menggunakan supercritical karbon dioksida (SC-CO 2 ), 2) kajian matriks sampel: kesan saiz partikel, darjah penapaian dan pengalkalian, 3) kajian untuk kesan kandungan kelembapan, masa pemanggangan dan suhu pemanggangan dan 4) kesan pelarut penyokong/pengubahsuai (CS) mengikut jenis dan kepekatan dari pada CS yang bersifat polar dan tidak polar semasa pengekstrakan lemak koko menggunaan SC-CO 2, 5) kajian pemindahan jisim menggunakan model Sovova yang berasaskan model lanjutan aliran palam Lack (SLM) untuk pengaruh kadar alir dengan tekanan berbeza dan kesan CS yang polar dan tak polar, dan juga model partikel bulat tunggal (SSM) untuk pengaruh saiz partikel, tingkatan penapaian, pengalkalian, massa pemanggangan dan suhu pemanggangan dan 6) kajian komposisi daripada trigliserida dan asid lemak yang terekstrak menggunakan SC-CO 2 dan CS. Kajian ini mendapati keadaan optimum bagi kadar alir dan tekanan yang digunakan untuk pengekstrakan lemak koko menggunakan SC-CO 2 adalah pada 2 ml/min dan 35 MPa. Hasil tertinggi diperolehi daripada nib koko yang mempunyai saiz partikel terkecil (S1= 0.07 mm) dengan penghasilan 92% setelah hampir 20 jam menggunakan SC-CO 2, nib koko tertapai (F1) dengan penghasilan 100% setelah hampir 10 jam menggunakan etanol sebagai CS di dalam SC-CO 2 dan juga bagi pemanggangan 150 viii
11 o C dan 35 min (R6) dengan 100% hasil setelah hampir 14 jam menggunakan 25% etanol (%mol) sebagai CS di dalam SC-CO 2. Peningkatan suhu dan masa pemanggangan telah menningkatkan hasil yang diperolehi. Hasil yang tinggi juga diperolehi daripada likur koko hitam yang beralkali pada ph tinggi (A1) dengan 73.70% hasil setelah pengekstrakan selama 18 jam menggunakan 25% etanol sebagai CS di dalam SC-CO 2 dan juga pada kandungan kelembapan yang tinggi (M5 = 17.64%) dengan 60.73% hasil setelah pengekstrakan menggunakan SC-CO 2 selama 20 jam. Secara statistiknya, hasil yang diperolehi dipengaruhi oleh semua perlakuan penyelidikan pada p<0.05, tetapi tiada perbezaan yang signifikan diperolehi bagi kedua-dua koko likur beralkali rendah (ph = ) dan koko likur beralkali asli (ph = ) dan juga pada kedua-dua kandungan kelambapan masing-masing 9.79 dan 17.64%. Etanol telah menunjukkan ciri terbaik sebagai CS bersama SC-CO 2, diikuti oleh isopropanol, aseton, siklohesana dan heksana. Peningkatan hasil diperolehi bersama peningkatan kepekatan (mol%) daripada CS (25%>15%>5%), kecuali bagi aseton yang mana pada kepekatan 15% mempunyai hasil yang lebih tinggi dari 25 dan 5%. Pengekstrakan lemak koko dengan SC-CO 2 bersama CS yang polar mempunyai hasil yang tinggi berbanding CS yang tidak polar. Semua perlakuan dalam penyelidikan tentang pengaruh CS secara statistik menunjukkan perbezaan yang signifikan pada p<0.05. ix
12 Kedua dua model berasaskan pemindahan jisim dapat dilakukan. SLM dapat digunakan untuk menjelaskan terjadinya pengaruh kadar alir dengan tekanan berbeza dan efek dari pada jenis dan konsentrasi CS yang polar dan non polar. Parameter dari xk, F dan S dapat digunakan untuk penilaian parameter pemindahan jisim dalam fasa bendalir dan padatan dari pada data eksperimen. AARD maksimum adalah 6.861%. Koefisien pemindahan jisim fasa bendalir dengan padatan adalah beragam masingmasing diantara E E-04 s -1 dan E E-04 s -1. Untuk model kedua (SSM), hasil kajian menunjukkan bahawa nilai-nilai De, kf dan kp beragam, dimana masing-masing berada diantara 8.900E E-09 m 2 /s, 1.139E E-04 m/s dan 9.581E E-05 m/s. Semua rajah dari data eksperimen memberikan hasil yang bersesuaian dengan model, dengan AARD maksimum pada Keputusan menunjukkan lemak koko yang diekstrak dari proses yang berbeza mengandungi tiga jenis trigliserida (TG) utama yaitu 1,3-Dipalmitoyl-2-Oleoyl- Gglycerol (POP), 1-Palmitoyl-2-Oleoyl-3-Stearoyl-Glycerol (POS) dan 1,3- Distearoyl-2-Oleoyl-Glycerol (SOS). Secara amnya, POS didapati dalam kandungan yang tertinggi sebanyak % diikuti oleh SOS dan POP masing-masing sebanyak % dan %. Analisa asid lemak (FA) menunjukkan bahawa asid palmitik (C16:0), asid stearik (C18:0) dan asid oleik (C18:1) merupakan tiga asid lemak utama didalam lemak koko yang diekstrak dengan kandungan tertinggi asid strearik sebanyak %, diikuti oleh asid oleik dan asid palmitik masingmasing sebanyak dan %. Secara statistik terdapt pengaruh yang berbeda dari semua perlakukan terhadap kandungan TG dan FA lemak. x
13 ACKNOWLEDGEMENTS Alhamdulillahirabbil'alamin, first of all I would like to express my utmost thanks and gratitude to the Almighty Allah S.W.T, the Sustainer, the most Gracious and most Merciful; without whose will no one can achieve anything. My salawat and salam is addressed to His righteous messenger, prophet Muhammad S.A.W. I would like to take this opportunity to express my appreciation and gratitude to the chairman of my supervisory committee, Prof. Dr. Jinap Selamat for her invaluable guidance, suggestions, discussions and patience throughout the research and preparation of this thesis. I am also grateful to the members of my supervisory committee, Prof. Dr. Russly Abdul Rahman, Assoc. Prof. Dr. Harcharan Singh and Dr. Nazimah Sheikh Abdul Hamid for their constructive comments and suggestions. My sincere gratitude is also extended to the financial support provided by the Intensification of Research in Priority Area (IRPA) fund for this research which was awarded to Prof. Dr. Jinap Selamat. I am also indebted to all the staffs of the Department of Food Science for their generous cooperation. An acknowledgement is also due to Prof. Dr. H. Adang Kadarusman (Alayarham) and Prof Dr. H. Sutarman, Dean Faculty of Industrial Engineering, University of Pasundan, Bandung. Special thanks also to all her graduate friends, especially cocoa group members and food Safety; Tan Teng Ju, Yusep Ikrawan, Willy Pranata Widjaja, Liza bt Mad xi
14 Saleh, Muh. Zuhkrufuz Zaman, Bakti Kumara, Misnawi, Rashidah Sukor, Noor Sofalina, Elham Moazami, Ismail Fitri, and Mohd. Safzan for sharing the literature and invaluable assistance. The time spent and memorable memories will always be cherished. Last but not least, I also wish to express my deepest appreciation to my beloved parents, sister and brother for all over their moral support, understanding, endless love, patience, and never ending encouragement and support in anyway during the many years of my seemingly ending pursue for knowledge. xii
15 I certify that a Thesis Examination Committee has met on 28 April 2009 to conduct the final examination of Asep Edi Kusnadi on his thesis entitled Cocoa Butter Extraction from Cocoa Nibs Using Supercritical Carbon Dioxide in accordance with the Universities and University Colleges Act 1971 and the Constitution of the Universiti Pertanian Malaysia [P.U.(A) 106] 15 March The Committee recommends that the student be awarded the Doctor of Philosophy. Members of the Thesis Examination Committee were as follows: Roselina Karim, PhD Lecturer Faculty of Food Science and Technology Universiti Putra Malaysia (Chairman) Robiah Yunus, PhD Associate Professor Faculty of Engineering Universiti Putra Malaysia (Internal Examiner) Lai Oi Ling, PhD Associate Professor Faculty of Food Science and Technology Universiti Putra Malaysia (Internal Examiner) Mohd Omar Abdul Kadir, PhD Professor Faculty of Industrial Technology University of Sains Malaysia Country of Malaysia (External Examiner) BUJANG KIM HUAT, PhD Professor and Deputy Dean School of Graduate Studies Universiti Putra Malaysia Date: 17 September 2009 xiii
16 This thesis was submitted to the Senate of Universiti Putra Malaysia and has been accepted as fulfilment of the requirement for degree of Doctor of Philosophy. The members of the Supervisory Committee were as follows: Jinap Selamat, PhD Professor Faculty of Food Science and Technology Universiti Putra Malaysia (Chairman) Russly Abdul Rahman, PhD Professor Faculty of Food Science and Technology Universiti Putra Malaysia (Member) Harcharan Singh, PhD Associate Professor Faculty of Engineering Universiti of Malaya (Member) HASANAH MOHD. GHAZALI, PhD Professor and Dean School of Graduate Studies Universiti Putra Malaysia Date: 16 October 2009 xiv
17 DECLARATION I declare that the thesis is my original work except for quotations and citations which have been duly acknowledged. I also declare that it has not been previously, and is not concurrently, submitted for any other degree at Universiti Putra Malaysia or at any other institutions. ASEP EDI KUSNADI Date: 15 September 2009 xv
18 TABLE OF CONTENTS ABSTRACT ABSTRAK ACKNOWLEDGEMENTS APPROVAL DECLARATION LIST OF TABLES LIST OF FIGURES LIST OF ABBREVIATIONS Page iii vii xi xiii xv xxi xxv xxix CHAPTER 1 GENERAL INTRODUCTION 1 2 LITERATURE REVIEW Origin and Botany of Cocoa Processing of Cocoa Bean Fermentation Roasting Alkalization Grinding Cocoa Butter Extraction Methods The Conventional Liquid Solvent Extraction Advantages of Conventional Solvent 27 Extraction Disadvantages of Conventional Solvent 28 Extraction 2.5 The Supercritical Fluid Extraction The Supercritical Fluid Properties of Supercritical Fluid Historical Background of Supercritical Fluid 38 Extraction Advantages of Supercritical Fluid Extraction The Experimental Parameters in Supercritical Fluid 42 Extraction Extraction Time Supercritical Flow Rate Extraction Cell Supercritical Fluid Flow Direction Pressure Temperature Sample Matrix and Analyte Properties Nature and Composition of Supercritical fluids 57 xvi
19 2.7 Cosolvent or Modifier in SFE Modeling on Supercritical Fluid Extraction Sovova s Lack s Plug Flow Model (SLM) Single Sphere Model (SSM) 69 3 MATERIALS AND METHODS Materials Methods Supercritical Fluid Extraction Method with 84 Pure Carbon Dioxide (SC-CO 2 ) Supercritical Fluid Carbon Dioxide (SC-CO 2 ) 86 with Cosolvents Method Determination of Yeld Measurement of Real and Bulk Density of 88 Cocoa Nibs Particles Modeling of Supercritical Fluid Extraction 88 with Sovova s Lacking Plugs Flow Model (SLM) Modeling of Supercritical Fluid Extraction 90 with Single Sphere Model (SSM) Triacylglycerol (TG) Profile Analysis by using 91 High Performance Liquid Chromatography (HPLC) Fatty Acid (FA) Profile Analysis by using Gas 91 Chromatography (GC) Statistical Analysis 92 4 EFFECT OF FLOW RATE WITH DIFFERENT 93 PRESSURE ON COCOA BUTTER EXTRACTION USING SC-CO Introduction Materials and Methods Materials Methods Statistical Analysis Results and Discussion Effect of Flow Rate with Different Pressure on 97 the Experimental Yield and Comparison with Calculated Mass Transfer Values Effect of Flow Rate with Different Pressure on 106 Triacylglycerol Effect of Flow Rate with Different Pressure on 110 Fatty Acid Methyl Ester 4.4 Summary 113 xvii
20 5 EFFECT OF PARTICLE SIZE, DEGREE OF 115 FERMENTATION AND ALKALIZATION ON COCOA BUTTER EXTRACTION USING SUPERCRITICAL FLUID 5.1 Introduction Materials and Methods Materials and Treatments Methods Statistical Analysis Results and Discussion Effect of Particle Size on the Experimental 121 Yield and Comparison with Calculated Mass Transfer Values Effect of Fermentation Degree on the 127 Experimental Yield and Calculated Mass Transfer Values Effect of Alkalization on the Experimental 133 Yield and Calculated Mass Transfer Values Effect of Particle Size, Degree of Fermentation 138 and Alkalization on Triacylglycerol Effect of Particle Size, Degree of Fermentation 143 and Alkalization on Triacylglycerol Effect of Particle Size, Degree of Fermentation and Alkalization on Fatty Acid Methyl Ester 5.4 Summary EFFECT OF MOISTURE CONTENT, ROASTING 151 TIME AND ROASTING TEMPERATURE ON COCOA BUTTER EXTRACTION USING SUPERCRITICAL FLUID 6.1 Introduction Materials and Methods Materials and Treatments Methods Statistical Analysis Results and Discussion Effect of Moisture Content on the 158 Experimental Yield and Comparison with Calculated Mass Transfer Values Effect of Roasting Time and Roasting 163 Temperature on the Experimental Yield and Calculated Mass Transfer Values Effects of Moisture Content, Roasting Time 169 and Roasting Temperature on Triacylglycerol Effects of Moisture Content, Roasting Time 172 and Roasting Temperature on Fatty Acid Methyl Ester 6.4 Summary 176 xviii
21 7 EFFECT OF POLAR COSOLVENTS ON COCOA 177 BUTTER EXTRACTION USING SC-CO Introduction Materials and Methods Materials Methods Statistical Analysis Results and Discussion Effects of Ethanol, Isopropanol and Acetone Concentration on the Experimental Yield and Comparison with Calculated Value of Mass Transfer Effects of Ethanol, Isopropanol and Acetone 195 Concentration Levels on Triacylglycerol Profile and Selectivity (α) Effects of Ethanol, Isopropanol and Acetone 205 Concentration Levels on Fatty Acid Methyl Ester Profile and Selectivity (α) 7.4 Summary EFFECT OF NON POLAR COSOLVENTS ON 215 COCOA BUTTER EXTRACTION USING SC-CO Introduction Materials and Methods Materials Methods Statistical Analysis Results and Discussion Effect of Hexane and Cyclohexane Concentration on the Experimental Yield and Comparison with Calculated Value of Mass Transfer Effect of Hexane and Cyclohexane 229 Concentration on Triacylglycerol Profile and Selectivity ( Effects of Hexane and Cyclohexane 235 Concentrations on Fatty Acid Methyl Ester Profile and Selectivity (α) 8.4 Summary CONCLUSIONS AND RECOMMENDATIONS Conclusions Recommendations 250 xix
22 BIBLIOGRAPHY 252 APPENDICES 273 BIODATA OF STUDENT 282 LIST OF PUBLICATIONS 283 xx
23 LIST OF TABLES Table 2.1 Characteristic of three different roasting methods Lipid compositions (wt %) of cocoa butter Fatty acids distribution (wt %) of cocoa butter Triacylglycerol compositions (%) of cocoa butter Comparison of the properties of supercritical CO 2, supercritical fluids, liquids and gases 2.6 Critical Conditions for Various Solvents The model based on differential mass balance and applied to the extraction of natural products with near critical CO Calculated and estimated SLM model parameters of mass transfer for cocoa butter extraction using supercritical carbon dioxide at T = 60 o C with different flow rate and pressure 4.2 Triacylglycerol composition (area %) of cocoa butter extracted by supercritical carbon dioxide (SC-CO 2 ) for effect of flow rate (f) with different pressure 4.3 Fatty acids composition (area %) changes of cocoa butter extracted by supercritical carbon dioxide (SC-CO 2 ) for effect of flow rate (f) with different pressure 5.1 Calculated and estimated model (SSM) parameters of mass transfer for cocoa butter extraction using SC-CO 2 at T = 60 o C and flow rate = 2 ml/min with different of particle size Page Calculated and estimated model (SSM) parameters of mass transfer for cocoa butter extraction using SC-CO 2 at T = 60 o C and flow rate = 2 ml/min with different of fermentation 5.3 Calculated and estimated model (SSM) parameters of mass transfer for cocoa butter extraction using SC-CO 2 at T = 60 o C and flow rate = 2 ml/min with different of alkalization 5.4 Triacylglycerol composition (area %) of cocoa butter extracted by supercritical carbon dioxide (SC-CO 2 ) at 35 MPa, 60 o C and 2 ml/min of flow rate with different particle size and extraction time xxi
24 5.5 Triacylglycerol composition (area %) of cocoa butter extracted by supercritical carbon dioxide (SC-CO 2 ) and 25% ethanol as cosolvent 35 MPa, 60 o C and 2 ml/min of flow rate with different fermentation treatment and extraction time 5.6 Triacylglycerol composition (area %) of cocoa butter extracted by supercritical carbon dioxide (SC-CO 2 ) and 25% ethanol as cosolvent 35 MPa, 60 o C and 2 ml/min of flow rate with different ph level of alkalization and extraction time 5.7 Fatty acids composition (area %) of cocoa butter extracted by supercritical carbon dioxide (SC-CO 2 ) at 35 MPa, 60 o C and 2 ml/min of flow rate with different particle size and extraction time 5.8 Fatty acids composition (area %) of cocoa butter extracted by supercritical carbon dioxide (SC-CO 2 ) and 25% ethanol as cosolvent at 35 MPa, 60 o C and 2mL/min of flow rate with different fermentation treatment and extraction time 5.9 Fatty acids composition (area %) of cocoa butter extracted by supercritical carbon dioxide (SC-CO 2 ) and 25% ethanol as cosolvent at 35 MPa, 60 o C and 2mL/min of flow rate different ph level of alkalization and extraction time 6.1 Calculated and estimated model (CSM) parameters of mass transfer for cocoa butter extraction using SC-CO 2 at T = 60 o C and flow rate = 2 ml/min with different of moisture content 6.2 Calculated and estimated model (CSM) parameters of mass transfer for cocoa butter extraction using SC-CO 2 at T = 60 o C and flow rate = 2 ml/min with different of roasting 6.3 Triacylglycerol composition (area %) of cocoa butter extracted by supercritical carbon dioxide (SC-CO 2 ) at 35 MPa, 60 o C and 2 ml/min of flow rate with different of moisture content and extraction time 6.4 Triacylglycerol composition (area %) of cocoa butter extracted by supercritical carbon dioxide (SC-CO 2 ) and 25% ethanol as cosolvent at 35 MPa, 60 o C and 2 ml/min of flow rate with different of roasting treatment and extraction time 6.5 Fatty acids composition (area %) changes of cocoa butter extracted by supercritical carbon dioxide (SC-CO 2 ) at 35 MPa, 60 o C and 2 ml/min of flow rate with different of moisture content and extraction time xxii
25 6.6 Fatty acids composition (area %) changes of cocoa butter extracted by supercritical carbon dioxide (SC-CO 2 ) and 25% ethanol as cosolvent at 35 MPa, 60 o C and 2 ml/min of flow rate with different of roasting treatment and extraction time 7.1 Composition (%) and flow rate of solvent mixture based on mol% of polar cosolvents in SC-CO 2 at 35 MPa pressure, 60 o C temperature and 2 ml/min flow rate 7.2 Triacylglycerols and fatty acids composition (%) of cocoa butter obtained by soxhlet method from cocoa liquor 7.3 Calculated and estimated model (SLM) parameters of mass transfer for cocoa butter extraction using SC-CO 2 with cosolvents at T = 60 o C, P = 35 MPa, f = 2 ml/min with different in the concentration of polar cosolvents 7.4 Triacylglycerols composition (area %) changes of cocoa butter extracted from cocoa liquor with ethanol as cosolvent in SC-CO 2 at 35 MPa, 60 o C and 2 ml/min with different ethanol concentration and extraction time 7.5 Triacylglycerols composition (area %) changes of cocoa butter extracted from cocoa liquor with isopropanol as cosolvent in SC-CO 2 at 35 MPa, 60 o C and 2 ml/min with different isopropanol concentration and extraction time 7.6 Triacylglycerols composition (area %) changes of cocoa butter extracted from cocoa liquor with acetone as cosolvent in SC-CO 2 at 35 MPa, 60 o C and 2 ml/min with different acetone concentration and extraction time 7.7 Fatty acids composition (area %) changes of cocoa butter extracted from cocoa liquor by ethanol as cosolvent in SC- CO 2 at 35 MPa, 60 o C and 2 ml/min of flow rate with different ethanol concentration and extraction time 7.8 Fatty acids composition (area %) changes of cocoa butter extracted from cocoa liquor by isopropanol as cosolvent in SC-CO 2 at 35 MPa, 60 o C and 2 ml/min of flow rate with different isopropanol concentration and extraction time 7.9 Fatty acids composition (area %) changes of cocoa butter extracted from cocoa liquor by acetone as cosolvent in SC- CO 2 at 35 MPa, 60 o C and 2 ml/min of flow rate with different acetone concentration and extraction time 8.1 Composition (%) and flow rate of solvent mixture based on mol% of non polar cosolvents in SC-CO 2 at 35 MPa pressure, 60 o C temperature and 2 ml/min flow rate xxiii
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