Kinetics in Hydrolysis of Oils/Fats and Subsequent Methyl Esterification in Two-step Supercritical Methanol Method for Biodiesel Production

Similar documents
A Novel Non-catalytic Biodiesel Production Process by Supercritical Methanol as NEDO High Efficiency Bioenergy Conversion Project

NEDO Biodiesel Production Process by Supercritical Methanol Technologies

NEDO Biodiesel Production Process by Supercritical Methanol Technologies. Shiro Saka

Conversion of Glycerol as By-Product from Biodiesel Production to Value-Added Glycerol Carbonate

COMPARISON OF TOTAL ENERGY CONSUMPTION NECESSARY FOR SUBCRITICAL AND SUBCRITICAL SYNTHESIS OF BIODIESEL. S. Glisic 1, 2*, D.

ScienceDirect. Biodiesel production in supercritical methanol using a novel spiral reactor

Effects Of Free Fatty Acids, Water Content And Co- Solvent On Biodiesel Production By Supercritical Methanol Reaction

4. Synthesis of Biodiesel from Palm Fatty Acid Distillate. Research Article

Citation Fuel Processing Technology (2014),

Production of Biodiesel from Waste Oil via Catalytic Distillation

Non-catalytic alcoholysis process for production of biodiesel fuel by using bubble column reactor

Treatment of BDF Wastewater with Hydrothermal Electrolysis

KINETIC MODEL OF ALGAL BIODIESEL PRODUCTION UNDER SUPERCRITICAL METHANOLYSIS

Some Basic Questions about Biodiesel Production

Current Situations and Prospects of Oil/Fat Resources for Biodiesel Production

Towards a Biodiesel-based Biorefinery: Chemical and Physical Properties of Reactively Extracted Rapeseed (Canola)

Biodiesel Production from Jatropha Curcas, Waste Cooking Oil and Animal Fats under Supercritical Methanol Conditions

Effect of Co-solvents on Transesterification of Refined Palm Oil in Supercritical Methanol

Biodiesel from soybean oil in supercritical methanol with co-solvent

BIODIESEL PRODUCTION IN A BATCH REACTOR 1. THEORY

Methanol recovery during transesterification of palm oil in a TiO2/Al2O3 membrane reactor: Experimental study and neural network modeling

Optimized Method for Analysis of Commercial and Prepared Biodiesel using UltraPerformance Convergence Chromatography (UPC 2 )

Phase Distribution of Ethanol, and Water in Ethyl Esters at K and K

EXCESS METHANOL RECOVERY IN BIODIESEL PRODUCTION PROCESS USING A DISTILLATION COLUMN: A SIMULATION STUDY

This presentation focuses on Biodiesel, scientifically called FAME (Fatty Acid Methyl Ester); a fuel different in either perspective.

Asian Journal on Energy and Environment ISSN Available online at

Transesterification of Vegetables oil using Sub-and Supercritical Methanol

Biodiesel production from waste vegetable oils over MgO/Al 2 O 3 catalyst

Application Note. Author. Introduction. Energy and Fuels

V.Venkatakranthi Teja. N S Raju Institute of Technology (NSRIT), Sontyam, Visakhapatnam, Andhra Pradesh , India.

Kinetics determination of soybean oil transesterification in the design of a continuous biodiesel production process

Conventional Homogeneous Catalytic Process with Continuous-typed Microwave and Mechanical Stirrer for Biodiesel Production from Palm Stearin

Quantitative Analysis of Chemical Compositions from Various Sources of Crude Glycerine

RESEARCH PROJECT REPORT. Trash to Treasure. Clean Diesel Technologies for Air Pollution Reduction. Submitted to. The RET Site. For

The Purification Feasibilityof GlycerinProduced During

Palm Fatty Acid Biodiesel: Process Optimization and Study of Reaction Kinetics

Hydrocracking of atmospheric distillable residue of Mongolian oil

TULSION BIODIESEL PRODUCTION: WET VS. DRY WHICH METHOD SHOULD YOU USE?

Optimization of the Temperature and Reaction Duration of One Step Transesterification

BIODIESEL PRODUCTION BY A CONTINUOUS PROCESS USING A HETEROGENEOUS CATALYST

Effect of Pressure, Temperature and Steam to Carbon Ratio on Steam Reforming of Vegetable Oils: Simulation Study

Reaction Parameters and Energy Optimisation for Biodiesel Production Using a Supercritical Process

OPTIMIZATION OF BIODIESEL PRODCUTION FROM TRANSESTERIFICATION OF WASTE COOKING OILS USING ALKALINE CATALYSTS

Biodiesel. As fossil fuels become increasingly expensive to extract and produce, bio-diesel is

Project Reference No.: 40S_B_MTECH_007

The preparation of biodiesel from rape seed oil or other suitable vegetable oils

Biodiesel Solutions André Y. Tremblay, P.Eng., Ph.D. Department of Chemical and Biological Engineering University of Ottawa

Simultaneous Determination of Fatty Acid Methyl Esters Contents in the Biodiesel by HPLC-DAD Method

International Journal of ChemTech Research CODEN (USA): IJCRGG ISSN: Vol.8, No.4, pp , 2015

Biodiesel Production using Reactive Distillation: A Comparative Simulation Study

Use of Ultrasound for Monitoring Reaction Kinetics of Biodiesel Synthesis: Experimental and Theoretical Studies.

Direct Production of Biodiesel from Lipid-Bearing Materials, Including Canola

Effect of Catalysts and their Concentrations on Biodiesel Production from Waste Cooking Oil via Ultrasonic-Assisted Transesterification

International Journal of ChemTech Research CODEN (USA): IJCRGG ISSN: Vol.7, No.4, pp ,

Biodiesell productionn withh Lewatit GF202 Lewatit GF202

Biodiesel production by esterification of palm fatty acid distillate

Using Response Surface Methodology in Optimisation of Biodiesel Production via Alkali Catalysed Transesterification of Waste Cooking Oil

CHAPTER 4 PRODUCTION OF BIODIESEL

Free and Total Glycerol in B100 Biodiesel by Gas Chromatography According to Methods EN and ASTM D6584

Johnson Matthey is the leading technology provider for butanediol (BDO) plants worldwide.

Experimental Investigation and Modeling of Liquid-Liquid Equilibria in Biodiesel + Glycerol + Methanol

Process units needed to make biodiesel continuously. Michael Allen Department of Mechanical Engineering Prince of Songkla University Thailand

Non-Catalytic Production of Ethyl Esters Using Supercritical Ethanol in Continuous Mode

Analysis of Glycerin and Glycerides in Biodiesel (B100) Using ASTM D6584 and EN Application. Author. Abstract. Introduction

Investigation of Single Cylinder Diesel Engine Using Bio Diesel from Marine Algae

Optimization of Biodiesel production parameters (Pongamia pinnata oil) by. transesterification process,

Application of the factorial design of experiments and response surface methodology to optimize biodiesel production

Production and Evaluation of Biodiesel from Sheep Fats Waste

Two-step biodiesel production using supercritical methanol and ethanol

A NOVEL CONTINUOUS-FLOW REACTOR USING A REACTIVE DISTILLATION TECHNIQUE FOR BIODIESEL PRODUCTION

Keywords: Simarouba Glauca, Heterogeneous base catalyst, Ultrasonic Processor, Phytochemicals.

EFFECT OF BIODIESEL IMPURITIES ON FILTERABILITY AND PHASE SEPARATION FROM BIODIESEL AND BIODIESEL BLENDS. National Biodiesel Conference 2008

Production of Biodiesel Fuel from Waste Soya bean Cooking Oil by Alkali Trans-esterification Process

Determination of phase diagram of reaction system of biodiesel

Kinetic Processes Simulation for Production of the Biodiesel with Using as Enzyme

Kinetic Study on the Esterification of Palm Fatty Acid Distillate (PFAD) Using Heterogeneous Catalyst

Research Article. Synthesis of biodiesel from waste cooking oil by two steps process transesterification and ozonation

Author: Vincenzo Piemonte, Associate Professor, University UCBM Rome (Italy)

CONVERSION OF GLYCEROL TO GREEN METHANOL IN SUPERCRITICAL WATER

Properties and Use of Jatropha Curcas Ethyl Ester and Diesel Fuel Blends in Variable Compression Ignition Engine

THERMAL PROCESSING OF LOW-GRADE GLYCEROL TO ALCOHOLS FOR BIODIESEL PRODUCTION

CHAPTER - 3 PREPARATION AND CHARACTERIZATION OF

Synthesis, Characterization and Evaluation of Sulphated Zirconias for Biodiesel Production by Triglyceride Cracking

Cataldo De Blasio, Dr. Sc. (Tech.)

Biodiesel Production from Used Cooking Oil using Calcined Sodium Silicate Catalyst

Transesterification of Waste Cooking Oil into Biodiesel Using Aspen HYSYS

Introduction During a time of foreign fuel dependency and high green house gas emissions, it is

Synthesis of biodiesel from palm oil with dimethyl carbonate and methanol as reagent variation using KOH and enzyme catalyst

Aspen HYSYS Simulation for Biodiesel Production from Waste Cooking Oil using Membrane Reactor

Energy requirement estimates for two step ethanolysis of waste vegetable oils for biodiesel production

Study of Transesterification Reaction Using Batch Reactor

PROJECT REFERENCE NO.: 39S_R_MTECH_1508

Proposal to Determine Various Properties of Biodiesel Fuels Based on Methyl Ester. Composition. Jason Freischlag. Dr. Porter Chem /25/2013

BIODIESEL PRODUCTION USING SUPERCRITICAL ALCOHOLS AND DIFFERENT VEGETABLE OILS IN BATCH AND CONTINUOUS REACTORS

Production of Biodiesel from Used Groundnut Oil from Bosso Market, Minna, Niger State, Nigeria

Simulation Approach to Biodiesel Production from Palm Oil by Conventional and Reactive Distillation Processes

FATTY ACID METHYL ESTERS SYNTHESIS FROM TRIGLYCERIDES OVER HETEROGENEOUS CATALYSTS IN PRESENCE OF MICROWAVES. C. Mazzocchia, G. Modica R.

Published in Offshore World, April-May 2006 Archived in

Determination of Free and Total Glycerin in Pure Biodiesel (B100) by GC in Compliance with EN 14105

Goran Jovanovic Oregon State University School of Chemical, Biological, and Environmental Engineering

Transcription:

Kinetics in Hydrolysis of ils/fats and Subsequent Methyl Esterification in Two-step Supercritical Methanol Method for Biodiesel Production Eiji Minami and Shiro Saka * Graduate School of Energy Science, Kyoto University, Kyoto, Japan Abstract: For high-quality biodiesel fuel production, our research group has developed the catalyst-free two-step supercritical methanol method (Saka-Dadan process). This process consists of oils/fats hydrolysis to fatty acids in subcritical water and subsequent methyl esterification to fatty acid methyl esters in supercritical methanol. In this study, kinetics in these reactions was studied to optimize reaction conditions for this process. Simple mathematical models were applied to hydrolysis and methyl esterification reactions, assuming that fatty acid acts as acid catalyst. Based on the models, regression curves fitted well with experimental results in both cases. Fatty acid was, thus, found to play an important role in the two-step method. From this result, efficient reaction conditions for hydrolysis and methyl esterification were discussed. Keywords: Biodiesel, Esterification, Supercritical Methanol, Hydrolysis, Subcritical Water 1. INTRDUCTIN Biodiesel (fatty acid methyl esters; FAME) can be produced from oils/fats through transesterification of triglycerides (TG) with methanol. At present, most of the methods in transesterification use alkaline catalysts, even though this process needs sophisticated purification steps for removal of the catalyst and saponified products from free fatty acids. ur research group has, therefore, developed catalyst-free processes employing supercritical methanol; one-step method (Saka process) and two-step method (Saka- Dadan process) [1-6]. In the former method, transesterification reaction of TG proceeds without any catalyst due to high ionic product of methanol in supercritical state. Furthermore, free fatty acids in oil/fat feedstocks can be also converted into FAME due to simultaneous methyl esterification, thus achieving a higher yield of FAME than that by the conventional alkali-catalyzed method. However, severe reaction conditions (35 o C/~5MPa) are required in the one-step method. The latter method involves, on the other hand, hydrolysis of oils/fats to fatty acids (FA) in subcritical water and subsequent methyl esterification of FA to FAME in supercritical methanol. This process can allow more moderate reaction conditions (27 o C/7~MPa) than those of the one-step method [4]. In this paper, a kinetic study was made on each reaction to optimize reaction conditions and propose more efficient process design for the two-step method. 2. EXPERIMANTAL METHDS Refined rapeseed oil and oleic acid were used as TG and FA samples, respectively. High-performance liquid chromatography (HPLC)-grade water and methanol were used as solvents. They were purchased from Nacalai Tesque, Inc., Kyoto. The treatments of rapeseed oil and oleic acid in subcritical water and supercritical methanol were, respectively, conducted using a flow-type reactor [5] under conditions between 25 o C and 3 o C with a constant pressure of 1 or MPa. The reaction time t (sec) was calculated by dividing the reaction tube volume (V=285mL) by volumetric flow rate of reactants at designated reaction conditions as in the following equation: t = V ρ ' + S F S F ρ S ρ ' ρ (1) where F s, ρ s and ρ s are setting flow rate (ml/sec), specific density (g/ml) and one at reaction temperature and pressure (g/ml) for solvent, respectively, while subscript o means those for oil. In this study, however, it was assumed that the value of ρ o was almost equal to that of ρ o. After the treatment, in case of oil/fat hydrolysis in subcritical water, reaction mixture was evaporated at 7 o C for min to remove water at reduced pressure. Remaining portion was then separated into oil phase and glycerol (G) by decantation. The obtained oil phase, which contained FA as well as unreacted TG and intermediate compounds, diglycerides (DG) and monoglycerides (MG), was analyzed by HPLC carried out with a Shimadzu LC-1A system under the following conditions: i) column, Asahipak GF31-HQ; flow rate, 1.mL/min; eluent, acetone; detector, refractive index detector (RID); temperature, o C and ii) column, STR DS-II; flow rate, 1.mL/min; eluent, methanol; detector, RID; temperature, o C. Although rapeseed oil consists of various fatty acids, the major one is oleic acid constituting about % of all. Therefore, calibration curves for the quantitative determination of TG, DG, MG and FA were made using triolein, 1,3-diolein, 1-monoolein, which were purchased from Aldrich-Sigma, saka, and oleic acid, respectively. In case of methyl esterification of FA, a similar experimental procedure was employed as described above. Corresponding author: saka@energy.kyoto-u.ac.jp This is an excerpt from the original paper published in Fuel 85 (6) 2479-2483. 1

3. RESULTS AND DISCUSSIN 3.1 Hydrolysis of oils/fats Hydrolysis reaction of TG consists of three stepwise forward and backward reactions as shown in Fig.1. At each forward reaction step, one molecule of water is consumed producing one molecule of FA. For the backward reaction, on the other hand, G reacts with FA to return to MG. As in a similar manner, DG and MG also reverse to TG and DG, respectively, consuming one molecule of FA. Fig.2 shows the yield of FA obtained from rapeseed oil in subcritical water at various conditions. A higher reaction temperature resulted in a higher rate of FA formation, though the hydrolysis reaction always came to equilibrium at around 9wt% in yield of FA when the volmetric ratio of water to rapeseed oil was 1/1. This incomplete result might be originated from the backward reaction mentioned above. Under the conditions of 27 o C/MPa, the yield of FA reached 9wt% after the subcritical water treatment for min. With regard to time course of the yield, FA tended to increase very slowly in early stage of the reaction, especially at low temperature region between 25 o C and 27 o C. The rate of FA formation was, then, gradually increased as the treatment was prolonged, and finally reached plateau. This phenomenon implies that FA produced by hydrolysis reaction acts like an acid catalyst. The mechanism for acid-catalyzed hydrolysis of TG can be briefly described as shown in Fig.3. In this process, a proton elimination of FA causes the protonating carbonyl oxygen of TG, thus allowing carbonyl carbon to receive nucleophilic attack by water, which promotes hydrolysis reaction of TG. A similar route can be also given for DG and MG. Assuming that the hydrolysis reaction of oils/fats proceeds by such autocatalytic mechanism of FA, the rate of FA formation could be represented as below: dc dt FA ( kccw k C CFA ) CFA = ' ' (2) where C FA, C W, C and C refer to the concentrations of FA, water, TG+DG+MG and DG+MG+G (mol/m 3 ), respectively. By the equation (2), time courses of FA formation in Fig.2 could be successfully explained. For further validation of the effect of FA on hydrolysis reaction, FA-added rapeseed oil was also treated in subcritical water. When 1wt% of FA was added in rapeseed oil before the treatment, as shown in Fig.4, hydrolysis reaction reached equilibrium only in min, while in case of neat rapeseed oil, it took about min. In the former case, furthermore, eventual yield of FA was slightly higher than that in the latter case. Regression curves, which are represented by dotted lines in Fig.4, fitted well with experimental results in both cases. Based on these lines of evidence, it was concluded that FA has an important role as acid catalyst on hydrolysis reaction of oils/fats in subcritical water. TG + H 2 DG + FA DG + H 2 MG + FA MG + H 2 G + FA Fig. 1 Three stepwise reactions for hydrolysis of triglycerides (TG) to fatty acids (FA) Fatty acids (wt%) 3 o C 3 o C 29 o C 27 o C 25 o C Fig.2 Hydrolysis of rapeseed oil (TG) to fatty acids (FA) in subcritical water at MPa (Water/TG = 1/1 (v/v)) 2

FA FA - + H + TG + H + TG + TG + + H 2 DG + FA + FA + FA + H + Fig.3 Autocatalytic mechanism by FA for hydrolysis reaction of TG in subcritical water. (H+, proton; TG+, protonated TG; FA+, protonated FA; a similar route can be given for hydrolysis of DG and MG) Fatty acids (wt%) FA 1wt% FA wt% Theoretical lines Fig.4 Hydrolysis of FA-added TG in subcritical water at 27 o C/1MPa (Water/oil = 1/1(v/v)) 3.2 Methyl esterification of fatty acid Methyl esterification of FA (Fig.5) is a major reaction to produce FAME in the two-step supercritical methanol method, whereas transesterification of TG is a major one in the conventional alkali- and acid-catalyzed methods. This esterification reaction is, therefore, an important step for high quality biodiesel fuel production. Fig.6 shows typical changes in the yield of FAME as oleic acid treated in supercritical methanol at various conditions. In a similar manner to hydrolysis reaction, a higher temperature resulted in a faster rate of FAME formation. However, the reaction did not reach equilibrium since the yield was still slightly increasing even after the treatment over min. In addition, the yield of FAME tended to increase quickly in early stage of the reaction. Then the rate of FAME formation became slow when the reaction proceeded. It seems apparent that the rate of FAME formation is nearly connected with concentration of FA in the reaction system. n the other hand, Fig.7 shows the effect of volumetric ratio of methanol on methyl esterification reaction in supercritical methanol. Contrary to our expectation, it was obviously found that a higher yield of FAME was achieved when a lesser amount of methanol was added. For example, about 94wt% of FAME can be obtained with a volumetric ratio of.9/1, while only wt% in case of 5.4/1 (methanol/fa) as treated at 27 o C/MPa for 3min. According to the common sense for the methyl esterification reaction, an excessive amount of methanol should be added to prevent the backward reaction of FAME to FA. However, assuming that FA acts as acid catalyst, a small amount of methanol could enhance the methyl esterification reaction since it makes the concentration of FA to be high in the reaction system. As well as hydrolysis reaction, therefore, the autocatalytic mechanism by FA was also applied for methyl esterification reaction as in the following equation: dc dt FAME ( kcfacm k CFAMECW ) CFA = ' (3) where C FAME, C M, C FA and C W refer to the concentrations of FAME, methanol, FA and water (mol/m 3 ), respectively. Based on the equation (3), regression curves fitted well with experimental results as represented by dotted lines in Fig.7. Dependence of the volumetric ratio of methanol on FAME yield could be, thus, explained by the autocatalytic mechanism of FA. Similarly, time courses of the yield in Fig.6 were also explained: i.e., a faster conversion of FA to FAME should be originated from a higher concentration of FA in early stage of the reaction. 3

FA + MeH FAME + H 2 Fig.5 Methyl esterification reaction of fatty acids (FA) to fatty acid methyl esters (FAME) 3 o C 29 o C 27 o C 25 o C Methyl ester (wt%) Fig.6 Methyl esterification of FA to its methyl ester (FAME) in supercritical methanol at MPa (MeH/FA = 1.8/1(v/v)).9/1 Methyl ester (wt%) Theoretical lines 1.8/1 3.6/1 5.4/1 MeH / FA (v/v) Fig.7 Methyl esterification of FA to FAME in supercritical methanol at 27 o C/MPa with various volumetric ratios of MeH/FA 3.3 Back-feeding of FA for further efficient hydrolysis According to the autocatalytic mechanism by FA mentioned above, further efficient reaction conditions were discussed for both of hydrolysis and methyl esterification reactions. In case of the hydrolysis reaction, it can be suggested that the addition of FA to oil/fat feedstocks before subcritical water treatment is expected to realize more efficient hydrolysis as previously noted in Fig.4 due to the acidic character of FA. As a similar purpose, we can feed a part of produced FA back to the inlet of the tubular reactor. For the feasibility of this process, as shown in Fig.8, a comparison was theoretically made between hydrolysis reactions in subcritical water a) without and b) with back-feeding of FA, assuming that the inner volume of tubular reactor is L for both cases. In the former case (a), about 91wt% purity of FA is produced with a throughput of 219kg/h as treated at 29 o C/15MPa for min. In the latter case (b), on the other hand, about 94wt% purity of FA can be expected with a slightly higher throughput of 254kg/h even under more moderate reaction conditions, 27 o C/15MPa for 25min. In this way, the back-feeding process for hydrolysis reaction is found to be an efficient tool to realize a good conversion of oils/fats into FA with milder reaction conditions. For the methyl esterification reaction, on the other hand, we can also suggest an efficient reaction conditions. In case of the conventional esterification employing mineral acid catalyst, a large amount of methanol should be added to FA to obtain a high yield of FAME as previously noted. In case of the supercritical methanol treatment, however, a lesser amount of methanol is more desirable to get a faster rate of FAME formation, thus achieving higher energy efficiency for biodiesel fuel production. These findings will contribute to development of more efficient process for the two-step supercritical methanol method. 4

a) Without back-feeding Water 247kg/h ils/fats 225kg/h Separator Reactor Volume: L 29 o C/15MPa/min Fatty acids 219kg/h Purity: 91wt% Waste water 253kg/h b) With back-feeding Water 416kg/h 113kg/h Separator Fatty acids 254kg/h Purity: 94wt% ils/fats 262kg/h Reactor Volume: L 27 o C/15MPa/25min Waste water 424kg/h Fig.8 A theoretical comparison between hydrolysis reactions of oils/fats in subcritical water a) without back-feeding and b) with bac k-feeding of FA 4. CNCLUDING REMARKS In this study, a kinetic study was made on hydrolysis of oils/fats in subcritical water and the following methyl esterification of FA in supercritical methanol, respectively, to optimize reaction conditions and propose more efficient process design for the two-step supercritical methanol method. For the hydrolysis reaction, the rate of FA formation was increased with the amount of FA produced, though it was very slow in early stage of the reaction. This phenomenon can be explained by assuming that FA acts as acid catalyst. For the esterification reaction, similarly, the reaction was found to proceed by the autocatalytic mechanism due to dissociation of FA itself in supercritical methanol. In this way, it was found that the produced FA has an important role in the two-step method. In case of the transesterification of TG in supercritical methanol (one-step method), on the other hand, the reaction proceeded according to a typical pseudo-first-order reaction, since there is no FA in the reaction system. That is a main reason why the two-step method can realize more moderate temperature and pressure than those of the one-step method. To achieve a further efficient process of the two-step method, back-feeding of the produced FA can be proposed for the hydrolysis step, while a smaller amount of methanol is desirable for the methyl esterification step. 5. ACKNWLEDGMENTS This work has been done in the Kyoto University 21 CE program Establishment of CE on Sustainable-Energy System, Grant-in-Aid for Scientific Research (B) (2) (No.135558, 1.4~3.3) from the Ministry of Education, Science, Sports and Culture, Japan, and in part in NED High Efficiency Bioenergy Conversion Projects, for all of which the authors are highly acknowledged. 6. REFERENCES [1] Saka, S. and Kusdiana, D. (1) Biodiesel fuel from rapeseed oil as prepared in supercritical methanol, Fuel,, pp. 225-231. [2] Kusdiana,.D. and Saka, S. (1) Kinetics of transesterification in rapeseed oil to biodiesel fuel as treated in supercritical methanol, Fuel,, pp. 693-698. [3] Kusdiana, D. and Saka, S. (1) Methyl esterification of free fatty acids of rapeseed oil as treated in supercritical methanol, J. Chem. Eng. Jpn., 34, pp. 383-387. [4] Kusdiana, D. and Saka, S. (4) Two-step preparation for catalyst-free biodiesel fuel production: Hydrolysis and methyl esterification, Appl. Biochem. Biotechnol, 115, pp. 781-791. [5] Saka, S., Minami, E., Yamashita, K., Toide, Y., Miyauchi, H. and Hattori, M. (5) NED High Efficiency Bioenergy Conversion Project - R & D for biodiesel fuel production by two-step supercritical methanol method -, Proceedings of the 14th European Biomass Conference & Exhibition, pp. 156-159. [6] Saka, S. and Minami, E. (5) A novel non-catalytic biodiesel production process by supercritical methanol as NED High Efficiency Bioenergy Conversion Project, Proceedings of the 14th European Biomass Conference & Exhibition, pp. 1419-1422. 5