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ProductionofVirginCoconutOil(VCO)byCentrifugationMethod

CONFERENCEPAPER·NOVEMBER2011

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AbdurahmanNour

UniversitiMalaysiaPahang

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ChoonGekKhoo

UniversitiSainsMalaysia

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AzhariHamidNour

UniversitiMalaysiaPahang

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ICCEIB -SOMChE 2011 UNIVERSITI MALAYSIA PAHANG, KUANTAN

28th November to 1st December 2011

Production of Virgin Coconut Oil (VCO) by Centrifugation Method

N.H. Abdurahman1; C.G. Khoo1; N.H. Azhari2

1Faculty of Chemical and Natural Resources Engineering, University of Malaysia Pahang-UMP, Malaysia 2Faculty of Industrial Sciences and Technology, University of Malaysia Pahang-UMP, Malaysia

Email: chunyukhoo@gmail.com

ABSTRACT: The quality of virgin coconut oil (VCNO) produced from the water soluble or oil soluble materials and centrifugation method of coconut milk from freshly harvested coconuts was investigated and the Separation and filtration of the oil from the cream after centrifugation. The characteristic that sets this VCO apart from oil is its high Lauric acid (C12) content. The conventional methods of breaking and production of virgin coconut oil (VCO) are disadvantageous from both economic and environmental perspectives. In this study, the potentials of centrifugation in breaking VCO were investigated. The percentage of water soluble material used for this experiment was 0.5 v/v % and it has high influence in demulsification of emulsions. The centrifuge speed was varied from 2000 rpm to 8000 rpm and the separation time was varied from 30 minutes to 90 minutes. The present research found that, centrifugation method with the help of water soluble materials can enhance the demulsification of oil-in-water (O/W) coconut milk emulsion in a very short time. Key words: Demulsification; O/W emulsion; Centrifuge; Virgin coconut oil (VCO)

1. INTRODUCTION

The production of Virgin Coconut Oil (VCO) or it is also known as the demulsification (emulsion breaking) of coconut milk through the extraction of oil from coconut milk emulsion that consists of two immiscible liquids, which is coconut oil and water, with the coconut oil layers dispersed as small spherical droplets in the water layers in a continuous phase that cannot be separated easily [1]. The main components of coconut milk are water and fats, with carbohydrates, proteins and ash as minor components [2].

The unfavorable contact between coconut oil droplets and water molecules cause coconut milk emulsion exists thermodynamically unstable and readily separates into two distinct phases – a heavy aqueous phase and a lighter cream phase [3]. This is due to the protein contents and quality is not sufficient to stabilize the fat globules [4]. Since the coconut milk emulsion is known to be naturally stabilized by coconut proteins – globulins, albumins and phospholipids [5], so this concept can be apply in production of VCO, which is the coconut oil that being extracted from well matured and fresh coconut through specialized process without changing its natural nutrition as they were present in their original state in coconut milk emulsions.

In past, the conventional method used to produce VCO is via fermentation method. Fermentation is a natural separation of coconut oil from water by gravity separation, which is a time consuming and complicated process that needed at least one week. As a result, the centrifugation method that assists by

the addition of low concentration of chemicals had been introduced. The method employed is based on the demulsification of coconut milk by damaging the bond of lipoprotein in emulsion [6]. The advantage of demulsification process is it can increase the yield of VCO and at the same time to reduce the production costs and yield a shorter time frame in the production process. 2. MATERIALS AND METHODS

Raw material used is the fresh coconut milk that brought from local market. The unused coconut milk is stored in refrigerator at 4oC. Coconut milk has to be unfreezing before conducting experiment.

The chemicals used are water soluble demulsifier – TERGITOL TM NP-8 Surfactant, and oil soluble demulsifier – TERGITOL TM NP-4 Surfactant, which brought from Optimal Chemicals (M) Sdn. Bhd. These chemicals are belonging to nonionic surfactant, which will destabilized the dispersed phase of emulsion.

When conducting the research, the coconut milk emulsion stability, demulsification of coconut milk emulsion, and quality characterization of the coconut oil yield had been study.

2.1 Coconut Milk Emulsion Stability

The stability of emulsion study included the physical properties such as viscosity, interfacial and surface tension, viscosity and oil droplets size. By using Brookfield device, viscosity can be determined at four different temperatures that is 30, 50, 70 and

90oC at agitation speed of 100 RPM. Besides that, Tensiometer is used to measure the interfacial tension between oil and water phase; and surface tension for both oil and water with air. Lastly, the droplets size of emulsion is observed through optical microscope. 2.2 Demulsification of Coconut Milk Emulsion

Demulsification of coconut milk emulsion was done via centrifugation method with chemicals assisted. The concentration of chemicals used is 0.5 v/v% for three different centrifugal processing conditions.

Firstly, 200 mL of coconut milk is prepared. Then, 0.5 v/v% water soluble demulsifier is added. The mixture was stirred until it is homogeneously mixed. After that, the well-mix mixture was poured into centrifugal tube for centrifugation process. A total of 9 samples was centrifuge at centrifugal speed 2000, 5000 and 8000 RPM for 30, 60 and 90 minutes. The above process was repeated for oil soluble demulsifier. Lastly, the sample was poured into separated funnel for gravity settling process for 24 hours. Observation for the emulsion separation process was done for each 30 minutes for the first 3 hours. Then, the amount of oil recovery was recorded and extracted after 24 hours. 2.3 Quality Characterization of Coconut Oil Yield

The coconut oil yield from the demulsification part has been analyzed through Gas Chromatography – Mass Spectrometer (GC-MS). The analysis of fatty acid components results were compared with Asian Pacific Coconut Community (APCC) Standard.

3. RESULTS AND DISCUSSION 3.1 Coconut Milk Emulsion Stability

3.1.1 Microstructure of Coconut Milk Emulsions

Fig. 1 Optical micrograph for Batch 1 emulsion

Fig. 2 Optical micrograph for Batch 2 emulsion

From Fig. 1 and 2, the optical micrographs using

reflected fluorescent light showing oil-in-water (O/W) emulsion, which is the continuous water phase (darker phase) shows dispersed oil droplets (bright droplets). The emulsions are stabilized by coconut milk proteins, such as casein micelles, which form a membrane around the oil droplets [7]. This cause the oil droplets are approximately in spherical shape. On the other hand, the fluid flowing has impact on the shape of dispersed phase. The shape of oil droplets may deform in shear flow since they are not rigid spheres [8]. 3.1.2 Droplets Size Distribution

Since the emulsions are polydisperse, so they will undergo some aggregation leading to a distribution of aggregate sizes. The characterization size - dispersed species is in terms of an average size.

Fig. 3 Droplets size distribution of Batch 1 coconut milk emulsion

Fig. 4 Droplets size distribution of Batch 2 coconut milk emulsion

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From both histograms above, the distribution is skew to left. The statistical values were tabulated in below: Table 1 Statistical values for size distribution of coconut milk emulsions

Statistical Value Batch 1 Batch 2

Minimun (µm) 0.04 0.04 Maximum (µm) 0.95 0.85

Mode (µm) 0.11 0.13

Mean (µm) 0.21 0.17 Median (µm) 0.19 0.15

From the results obtained, it was found that the

average fat droplets diameter for these two batches of coconut milk emulsion was 0.21 and 0.17 µm, while, the most frequent droplet size was 0.11 and 0.13 µm, respectively. The distribution illustrated an opposite result for average size and most frequent size due to the higher oil content in Batch 1 emulsion. Since the droplets size larger than 0.1 µm and smaller than 180 µm, these emulsions were considered as Stoke’s model [8].

Other than this, size distribution has an important influence on the emulsion viscosity. For smaller size distribution, the dispersion viscosity will be higher. This is due to the increased in interfacial area and thinner films increased the resistance to the flow of fluid [9]. Thus, it was predicted that the second batch of coconut milk emulsion will have higher viscosity. 3.1.3 Density of Coconut Milk Emulsions

Since the size distribution of these coconut milk emulsions were different, so they result in the different for their density. The density is 0.952 and 0.926 g/cm3, respectively.

Coconut milk is an O/W emulsion make up by water, fats, carbohydrates, protein and ash. Hence, the different in the compositions percentage had contributed to the different of density for each emulsion. Besides that, from visual observation, it was found that the second batch emulsion is creamier, cause lighter density. While from the point of view of size distribution, it was found that the mode of droplets size for Batch 1 emulsion was smaller, which contributed to a larger interfacial contact area in dispersed phase, causing the increase in density of emulsion. 3.1.4 Effect of Temperature to Viscosity of Coconut Milk Emulsions

Viscosity is a temperature dependent property and it is inversely proportional to density. At ambient condition, the viscosity for emulsion is 54.30 and 65.30 cP, for Batches 1 and 2, which match with the result from density part. Despite of this, the general trend for the viscosity and temperature also should be inversely proportional to each other.

Fig. 5 Effect of viscosity to temperature at 100RPM

From Figure 5 presented above, the results obtained followed two different trends, in which it followed the normal trend at low temperature, but viscosity kept on increasing at higher temperature. The minimum viscosity achieved by Batch 1 coconut milk emulsion at temperature 64oC was around 25.0 cP, while Batch 2 at 50oC was 48.8 cP. At this temperature, the emulsion changed from stabilizing Oil-in-Water (O/W) to Water-in-Oil (W/O) emulsion. This transition temperature is termed as Phase Inversion Temperature (PIT), where phase inversion is occurs. Since coconut milk is a food emulsion, where the protein existed is function as surfactant that will stabilised the O/W emulsion at low temperature. Hence, as temperature increased, W/O emulsion was preferred, where now the coalescence rate will be slow, a more stable emulsion is formed [10]. 3.1.5 Surface Tension and Interfacial Tension of Coconut Milk Emulsions

The surface tension for both oil and water was 30.431 and 70.023 mN/m, respectively. For the interfacial tension of oil and water, it was less than the individual surface tension of each liquid, recorded a value 7.987 mN/m. This is due to the mutual attraction had been moderated by all the molecules involved [11]. 3.2 Demulsification of Coconut Milk Emulsions

Demulsification of coconut milk emulsion was carried by centrifugation and assisted with nonionic surfactant, which will self-associate and solubilize themselves to destabilize emulsion. For water soluble material or hydrophilic chemical will dissolve in water, while the oil soluble material or lipophilic chemical will dissolve in oil phase. Emulsions will undergo some aggregation by their own, leading to a distribution of aggregate (structured protein globes) sizes, and assisted by the nonionic surfactants. Hence, a low concentration, which is 0.5 v/v % is needed to promote aggregation. A large numbers of molecules from these organize aggregates will lead to the formation of casein micelles [12]. By homogeneous

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mixing, it can promote the destabilization process by breaking the stabilizing film surrounding both fat droplets and casein micelles [13].

By centrifugation, it can accelerate the creaming process in minutes. High rotation frequencies allowed an effective acceleration, which induced a faster separation compared to simple gravity separation. The centrifuge works according to the sedimentation principle, where the centripetal acceleration is used to separate substance of greater and lesser density. By using centrifuge, it is possible to break down emulsions and to separate dispersions droplets; which will coalesce following separation of coconut oil from the water phase [14]. Sometimes, when the densities different for the emulsion is too low that cause the association forces holding the components together. It leads gravity separation to be time consuming [3].

The percentage of phase separation can be calculated by using the formula below:

Layer   % =

The  height  of  separated  layerTotal  height  of  emulsion x  100%          (1)

According to Stoke’s law, settling velocity of oil

droplets through water is given by:

𝑣! =ρ! − ρ! x  rω

!x  D!

18  µ!                                                                                                          (2)

Where vo is settling velocity; ρw is water density; ρo is oil density; r is radius of rotation; ω is angular velocity of centrifugation; d is diameter of droplets; and µw is viscosity of water phase. 3.2.1 Demulsification of Coconut Milk Emulsions via Water Soluble Materials and Centrifugation Method

Fig. 6 Phase separation of coconut milk emulsion under centrifugal force for 30 minutes

Fig. 7 Phase separation of coconut milk emulsion under centrifugal force for 60 minutes

Fig. 8 Phase separation of coconut milk emulsion under centrifugal force for 90 minutes

Fig. 6, 7 and 8, were showed that cream layer was kept increasing when the centrifugal processing speed increased from 2000 to 8000 RPM. Cream is the fat-rich portion of coconut milk that separated by skimming, and it still remains in the form of an O/W emulsion [15]. Hence, as centrifugal processing time increase, the time for coalescence to happen will be longer. This increased the opportunity for the cream formation. Since Batch 2 emulsion has lower oil content was used for this part of experiment, so there was no significant oil recovery.

Fig. 9 VCO yield with the effect of centrifugal processing time

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Fig. 10 VCO yield with the effect of centrifugal processing speed

For 5000 and 8000 RPM, there was no oil

recovery for all centrifugal processing time used. However, there was a yield of 20.7% oil at 2000 RPM after centrifuge for 90 minutes, but still remain no separation for 30 and 60 minutes centrifuge. This was known as its optimum condition for the separation to occur by using Batch 2 emulsion, which has a larger most frequent droplets size of 0.13 µm that has larger inter distance between droplets due to weak attraction force. As centrifugal processing time increase, the time for coalescence to happen will be longer, and causes destabilization of the fat globes, consequent aggregation and yield an oil separation. Due to creamier composition, which is a high viscosity emulsion, the oil content was reduced. Therefore, accelerate emulsion at higher speed caused the droplets size to decrease further, make them difficult to coalesce and settle down. 3.2.2 Demulsification of Coconut Milk Emulsions via Oil Soluble Materials and Centrifugation Method

Fig. 11 Phase separation of coconut milk emulsion under centrifugal force for 30 minutes

Fig. 12 Phase separation of coconut milk emulsion under centrifugal force for 60 minutes

Fig. 13 Phase separation of coconut milk emulsion under centrifugal force for 90 minutes

Batch 1 coconut milk emulsion was used in this part of experiment. The water phase for all the nine samples has almost the same amount. Besides that, it was observed that the creaming part was kept reducing as centrifugal speed increases at constant centrifugal processing time. Cream is the fat-rich portion of coconut milk that separated by skimming, and it still remains in the form of an O/W emulsion [15]. Thus, it can yield coconut oil via gravity separation. As a result, the coconut oil yield increasing as centrifugal speed increases at constant centrifugal processing time.

Fig. 14 VCO yield with the effect of centrifugal processing time

From Fig. 14, it illustrated at 5000 and 8000 RPM,

the highest coconut oil yield is occurs at 60 minutes, with 40.6 and 45.2 v/v % while for 2000 RPM is at 90 minutes with 25.1 v/v %. This is due to the

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different in droplets size produces by centrifugal acceleration. As the centrifugal speed increase, the droplets size become smaller since more energy was created to the emulsion in order to break the droplets film. Therefore, at lower centrifugal speed, longer processing time is need for large droplets to coalesce. As for high centrifugal speed, the optimum centrifugal processing time was 60 minutes since the increasing of processing time has no much effect to the creaming process.

Fig. 15 VCO yield with the effect of centrifugal processing speed

By centrifugation process, heat is generated through centrifugal rotation. Since density and viscosity are temperature dependence properties, so when temperature increased, viscosity will decrease faster that density different and results in an increase in droplets diameter. As a result, centrifugation will increase the settling velocity of oil droplets, which will accelerate the separation of emulsion in return [3]. Therefore, the oil recovery was highest at 8000 RPM for all the processing time involved. As for the highest oil yield (optimum condition), it occurs at 8000 RPM after centrifuge for 60 minutes with a 45.2 v/v% from Fig. 15. 3.3 Quality Characterization of Coconut Oil Yield

VCO produced by different centrifugal processing speed and time will yield some differences in quality properties. The free fatty acid components were determined by GC-MS and tabulated below: Table 2 Quality characteristics of VCO samples

Fatty acid

Content (%) water

soluble & centrifugatio

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Centrifuge condition

8000 RPM, 90 min

8000 RPM, 60 min ---

Caproic acid, C6:0 0.25 0.23 0.4-0.6

Caprylic acid, C8:0 4.05 3.83 5.0-10.0

Capric acid, C10:0 28.37 28.95 4.5-8.0

Lauric acid, C12:0 0.00 0.00 43.0-

53.0

Myristic acid, C14:0 0.00 0.00 16.0-

21.0 Palmitic acid,

C16:0 4.91 5.23 7.5-10.0

Palmitoleic acid, C16:1 0.01 0.01 2.0-4.0

Stearic acid, C18:0 1.40 1.56 5.0-10.0

Oleic acid, C18:1 2.88 2.89 0.0 Linoleic acid,

C18:2 0.61 0.67 1.0-2.5

C 18:3 – C 24:1 0.04 0.05 < 0.5

Others dominant components (%) methyl

tetradecanoate 10.81 10.39 0.00

6,6-dimethyltetrahydro-2H-pyran-2-one

27.24 25.80 0.00

From Table 2, the distribution of samples’ straight

chain fatty acid components does not reach the target range value of APCC standard VCO except for Capric acid, C10:0 with 28.37 and 28.95%, respectively. Besides that, Lauric acid, C12:0 and Myristic acid, C14:0 cannot be detected in the samples analysed. Other than this, the existence of two others dominant components – methyl tetradecanoate and 6,6-dimethyltetrahydro-2H-pyran-2-one that do not exist in standard VCO.

Medium chain fatty acid – C8:0, C10:0 and C12:0 are the major components in VCO. However, the sudden increase of C10:0 can be contributed by the absence of C12:0, which have been reacted with C14:0 to form methyl tetradecanoate by cleavage its methyl group. Methyl tetradecanoate, also known as methyl myristate, is methyl esters of saturated branched-chain fatty acids with anteiso-series. Despite of this, 6,6-dimethyltetrahydro-2H-pyran-2-one or 5-methyl-5-hydroxyhexanoic acid lactone, which is a branched-chain lactone (cyclic ester). Unlike the straight chain fatty acids, both of them are produced biosynthetically via the conventional mechanisms for the synthesis of saturated fatty acids in bacteria [16]. The present of bacteria is due to the storage condition for unfreezing process of raw material before conducting experiment. The coconut milk emulsion is directly exposed to ambient condition for a long period. Humidity in air will increase the moisture content and provide a suitable breeding ground to bacteria at this optimum condition. Therefore, it degraded the quality of coconut oil yield. 4. CONCLUSION

By using centrifugation with the assisted of oil soluble material, a higher yield of coconut oil was achieved. The stability of coconut milk emulsion such as droplets sizes and viscosity will affect the rate of separation process in O/W emulsion and thus reflect on the results of coconut oil recovery. However, the quality of the coconut oil yields by centrifugation, whether with the addition of hydrophilic or lipophilic chemicals, their quality are

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almost the same. Improper storage conditions for raw material will affect the quality of coconut oil yield. Although oil recovery for centrifugation via oil soluble material was higher, it is not encourage using it. This was due to the lipophilic chemicals will create environmental problems as well as various side effects on consumers health since VCO that is being extracted had been polluted by chemicals.

NOMENCLATURE µ : Viscosity [cP] T : Temperature [oC] D : Droplets diameter [µm] vo : Settling velocity of oil droplets [m/s] ρw : Density of water [kg/m3] ρo : Density of oil [kg/m3] r : Radius of centrifugal rotation [m] ω : Angular velocity of centrifugation [rad/s] µw : Viscosity of water [cP] REFERENCES [1] Friberg, S.E. (1997). Emulsion Stability. In:

Food Emulsions, Friberg, S.E. and Larrson, K. (Eds.). Marcel Dekker, New York, pp: 1-55.

[2] Tansakul, A. and Chaisawang, P. (2005). Thermophysical properties of coconut milk. J. Food Eng., pp: 73, 276 – 280.

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[4] Monera, O.D. and del Rosario, E.J. (1982). Physico-chemical evaluation of the natural stability of coconut milk emulsion. Ann. Trop. Res., pp: 4, 47 – 54.

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[6] Nik Norulaini, N.A., Setianto, W.B., Zaidul, I.S.M., Nawi, A.H., Azizi, C.Y.M. and Mohd Omar, A.K. (2009). Effects of Supercritical Carbon Dioxide Extraction Parameters on Virgin Coconut Oil Yield and Medium-Chain Triglyceride Content. Food Chemistry, pp: 116, 193 – 197.

[7] Dickinson, E. (1989). Colloid Surfactant. 42, 191 – 204.

[8] Schramm, L.L. (2005). Emulsions, Foams, and Suspensions: Fundamentals and Applications. Laurier, L.L. (ed.), Wiley-VCH, Weinheim, Germany.

[9] Becher, P. (2001). Emulsion: Theory and Practice, 3rd ed., American Chemical Society. Washington.

[10] Wadle, A., Tesmann, H., Leonard, M., and Förster, T. (1997). Phase Inversion in

Emulsions: CAPICO – Concept and Application. Surfactants in Cosmetics, 2nd ed., Rieger, M.M., Rhein, L.D. (Eds.), Dekker: New York, pp. 207 – 224.

[11] Moules, C., Camtel Ltd. [Brochure]. The Role of Interfacial Tension Measurement in the Oil Industry.

[12] Hartley, G.S. (1936). Aqueous Solutions of Paraffin Chain Salts. Hermann and Cie: Paris.

[13] Dalgleish, D.G. (1990). Aspects of Stability in Milk and Milk Products in Food Colloids. Bee, R.D., Richmond, P., and Mingins, J. (Eds.), Royal Society of Chemistry, Cambridge, pp. 295 – 305.

[14] Coulson, J.M. and Richardson, J.F. (1991). Chemical Engineering, 4th Edn, Particle Technology and Separation Processes, USA.

[15] Sogo, Y., Taneya, S., and Kako, M. (1989). Whipping Cream Emulsifiers in Food Emulsifiers: Chemistry, Technology, Functional Properties and Applications. G. Charalambous and G. Doxastakis (Eds.), Elsevier, Amsterdam, pp. 449 – 472.

[16] Christie, W.W. (2011). The AOCS Lipid Library. Fatty Acids: Branched Chain – Structure, Occurrence and Biosynthesis. James Hutton Institute (and Mylnefield Lipid Analysis), Invergowrie, Dundee (DD2 5DA), Scotland.