Comparison of Nutritional Composition between Palm Kernel Fiber and the Effect of the Supercritical Fluid Extraction on Its Quality

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Comparison of Nutritional Composition between Palm Kernel Fiber and the Effect of the Supercritical Fluid Extraction on Its Quality Nik Nurolaiani. Ab. Rahman 1, Moftah. M. Ben Nama 1, Sawsan.S. Al-Rawi 2, Ahmad. H. Ibrahim 3, Mohd. O. Ab Kadir 2 1 Department of biology, School of Distance Education, Universiti Sains Malaysia, Penang, Malaysia (norulain@usm.my)(mohacettepe1@yahoo.com) 2 Department of environmental technology, School of Industrial Technology, Universiti Sains Malaysia, Penang, Malaysia. (akmomar@usm.my), (swsanrawi@yahoo.com) 3 Department of Pharmacology, School of Pharmaceutical Sciences, Universiti Sains Malaysia, Penang, Malaysia (abraihimi@yahoo.com) ABSTRACT High fibre foods start to gain much attention in the industrial world, due to its importance in human health for treating and prevention wide range of diseases. Finding new and cheap source of dietary fibre is highly needed. Palm kernel cake which is wastes produced after oil extraction from palm kernels is increasing in volume as the oil palm industry expands in many parts of Asia and Africa. The aim of this study is to compare the nutritional composition of palm kernel fibre derived from PKC samples: Supercritical CO 2 (SC-CO 2 ) palm kernel cake with testa (SC-PKt), SC-CO 2 -Palm kernel cake without testa (SC-PK), and palm kernel cake from palm oil mill. Supercritical carbon dioxide (SC-CO2) was used to extract the residual oil from Palm kernel with and without testa at temperature of 80 C and pressure 41.36 MPa. Total dietary fibre, crude fiber, crude protein, ash and moisture were determined in these samples and compared with the Palm kernel cake from palm oil mill. The results showed that the palm kernel with testa contains the highest amount of the total dietary fibre with 63.06g/ 100g sample, crude fibre 8.49g/100g, crude protein content 14.40 g/100g, ash 4.43g/100g sample with p<0.05. The mean moisture content of the Palm Kernel with testa was 3.26 which is least than the other samples. The results of this study suggest that, extracting oil using supercritical can minimize the waste by producing a new and cheap source of fibre with good nutrition. Moreover, the defatted palm kernel with testa considered superior than the defatted palm kernel without testa and palm kernel cake, and might be used as a good and cheap source of dietary fiber for human consumption. However, still others tests are in progress in order to confirm it validity for human consumption. Keywords: Supercritical; Palm; kernel; Nutritional; composition. INTRODUCTION High fibre foods has gained much attention in the food industries, due to it health benefits in decreasing the risks of many diseases such as cancer, blood sugar, lowering cholesterol levels, weight loss management, and including the prevention of constipation and many more. The recommended total daily fibre intake for adults is 30 to 40 grams. But most people get only about 10 grams of fibre a day. Finding a new cheap source of fibre that can be utilized as a source for high fibre food such as bakery product, pasta, cake and many other products has become a target for food companies to enhance their products and attract customers. Palm kernel cake (PKC), is a waste product after oil extraction from palm kernels using screw pressed technique. The global production of this by-product (PKC) is increasing in volumes as the oil palm industry expands in many parts of Asia and Africa in recent years. More than four million metric ton of PKC produced globally in the year 2002 [1]. Malaysia, as one of the world s greatest palm oil producers generates about 2 million metric ton of PKC in 2004 as wastes. The only usage for this cake is as a feed ingredient for ruminant animals in the livestock feed industry [2]. Ramachandran [3] reported that PKC has good nutrient content, and a useful source of protein with 18.6 % of crude protein and with high quality of amino acids with a total crude dietary fiber of 37% and an average residual oil content of 10%. However, this amount of residual oil in the cake is giving the cake a bad small by time and hindering the use of this cake. It is well known that the chemical composition of PKC depends essentially on the oil extraction method. Supercritical fluid extraction (SFE) has proven that it is powerful technique as an extraction method to remove the oil from the fibre [4].The supercritical carbon dioxide (SC-CO2) has been employed as a solvent and has replaced other traditional solvents in food industries due to its characteristics being nontoxic, non-flammable and available at reasonably low

costs [5]. Additionally, it is environmentally friendly due to its ability to produce products free of solvent residues [6]. Thus in this study, we are proposing the use supercritical extraction to extract the oil and to produce defatted palm kernel fiber that can be use as a source of fiber for high fiber food product. The aims of this study is to investigate and compare the nutritional compositions of palm kernel fiber derived from three types of PKC samples: Supercritical CO 2 palm kernel cake with testa (SC-PKt), Supercritical CO 2 Palm kernel cake without testa (SC-PK), and palm kernel cake (PKC) from palm oil mill. MATERIALS & METHODS Commercial liquid carbon dioxide gas with a purity of 99 g kg -1 was purchased locally from Malaysian Oxygen, Penang, Malaysia in a gas cylinder at temperature below -5 C. Palm Kernel Cake samples were purchased from Fleet Palms Sdn Bhd factory in Seberang Prai in Penang, Malaysia. SC-CO2 Extraction Before the supercritical extraction, the testa were removed from the palm kernels as follows: the palm kernels were soaked in a flask with 6MHCL in a water bath for 5 minutes then washed and cooled to room temperature and the testa removed using forceps. The samples were labeled as pkt (palm kernels with testa) and the pk for (palm kernels without testa ). The pkt and pk samples were ground using analytical mill model IKA(R) A11 from Retsch, and then passed through an aperture sieve to get particle sizes 500µm. The samples were packed in polyethylene zipped bags and kept at -20 C until ready for extraction. Supercritical extraction parameters were selected to extract the highest oil yield based on previous experiments by our group. In these experiments, the selected operating parameter were temperature 40, 60 and 80 C and pressure 27.57, 34.47 and 41.36MPa using particle size 500µm and CO 2 flow rate of 1 ml/ min. The supercritical extractions were performed using extraction system ISCO, Inc., Lincoln, NE. USA model SFX 220. The extraction was run with 2 g of sample placed in the vessel for extraction at the selected operating temperature and pressure. At the end of the extraction, the supercritical fluid was depressurized to atmospheric pressure, and the oil was collected in a pre weighed glass vial. The oil yield was calculated at the end of the extraction and kept at -20 C until ready for analysis and the palm kernels with and without testa is now oil free or are defatted cakes. Determination of oil removed: gram oil obtained ---------------------------- X 100 = g oil/g sample Gram sample Analytical Methods. The three defatted samples, SC-pkt, SC-pk and PKC were tested for protein content, crude fiber and dietary fiber content, ash and moisture content according to the following method; protein content was analyzed using the traditional method (AOAC, 1984) for the three samples using a Tecator Kjeltec autosampler system 1035 analyzer. To estimate the nitrogen content, 10.0g Na 2 SO 4 and 1.0g CuSO 4 were added to the digestion mixture and the distillate was collected in 4% boric acid, It was then titrated with 0.1 N HC1 using a few drops of methyl red and methylene blue indicators. The percentage of nitrogen was calculated on a dry matter basis and was then multiplied by 6.25 for percentage of protein. Fiber content was estimated according to an ISO method (5983). A finely ground sample (2.5 g) of meal was weighed and freed from residual fat using 15-mL n-hexane extraction. The test portion was boiled with sulfuric acid (0.255 mol L 1), followed by separation and washing of the insoluble residue. The residue was then boiled with sodium hydroxide (0.313 mol L 1 ), followed by separation, washing, and drying. The dried residue was weighed and ashed in a muffle furnace (TMF-2100; Eyela) at 600 C, and the loss of mass was determined. Ash content was determined according to an ISO method (749). Two grams of the test portion was taken and carbonized by heating on a gas flame. The carbonized material was then ashed in an electric muffle furnace at 550 C until a constant mass was achieved and this will give the ash content. Moisture content was measured using the Moisture Balance from SHIMADZU type MOC-120H. The same analyses were conducted for supercritical palm kernel cake with and without testa and palm kernel cake from oil mill that have not undergone supercritical extraction and still contain oil.

RESULTS & DISCUSSION Supercritical extraction was used to remove the oil from the palm kernels with testa, palm kernel without testa so that to produce defatted samples free of oil. Carbon dioxide under supercritical conditions will be in fluid form and behaves as a solvent to extract the oil. Supercritical CO 2 extraction parameters were selected based on previous experiments by our group. The extraction tested parameter were temperature 40, 60 and 80 C, pressure 27.57, 34.47 and 41.36 MPa and particle size was fixed to 500µm and CO 2 flow rate of 1ml/min. In these experiments, it was found that the best condition that gave the highest extraction yield was at operating temperature of 80 C with operating pressure of 41.36 MPa, particle size of 500µm and CO 2 flow rate of 1ml/min. However, at this condition the highest amount of extracted yield was with 54.9% for the sample pkt as shown in figure 1.a, and 33.9 % extraction yield for the pk as shown in figure 1.b. The explanation behind this result due to the extraction condition as at higher pressures, the solubility increased with temperature increment as it was reported previously [7]. Figure 1a. Effect of supercritical operating parameter on the extraction yield from palm kernel with testa at operating temperature 40, 60 and 80 C, operating pressure 27.57, 34.47 and 41.36MPa, CO 2 flow rate of 1ml/min and particle size of 500µm Figure 1b. Effect of supercritical operating parameter on the extraction yield from palm kernel without testa at operating temperature 40, 60 and 80 C, operating pressure 27.57, 34.47 and 41.36MPa, CO 2 flow rate of 1ml/min and particle size of 500µm. The nutritional composition of all samples SC-PKt, SC-PK and PKC from the oil mill were compared in order to determine the most suitable defatted cake samples for human dietary food fiber source. It was found from the results of our experiment that it is a disadvantage to remove the testa, as it did not significantly affect the nutritional composition of the palm kernel fiber but alternatively some components were negatively affected such as the total dietary fiber. It was found that the total dietary fiber for SC-pkt, was 61.58 % which is higher than SC-pk at 57.78 % and SC-PKC 60.71%. However, supercritical fluid extraction (SFE) enhanced the total dietary fiber content for all the three samples as shown in table 1. The total dietary fiber for PKt was increased from 61.58% to 63.03% for SC-PKt, while for the palm kernel without testa was increased from 57.78 % to 58.69 % as shown in figure 3. These results give evidence that supercritical extraction is a viable technique to extract the oil and enhance the fiber content in the raw material. Additionally to the dietary fiber content the crude fiber in these sets of samples were tested. The results of this experiment shows that the supercritical extraction did not have great affect on the crude fiber content as it affects the total dietary fiber. It was found that the palm kernel cake has the highest crude fiber of 15.7 %, while the crude fiber content for PKt before and after the supercritical extraction was 8.99 %, 8.49 % respectively, and for the PK was 7.29 and 7.23% before and after SFE respectivly as shown in Table 1.

Figure 3. The total dietary fibre content in three sets of samples before and after the supercritical extraction; Palm kernel with testa (PKt), palm kernel without testa (PK) and palm kernel cake (PKC). Figure 4. The moisture content before and after the supercritical extraction for the Palm kernel with testa (PKt), palm kernel without testa (PK), and compared to palm kernel cake (PKC). However, the sample moisture content was affected greatly by the supercritical extraction, and its values were varied among the three sets of samples. It was found that the moisture content for pkt was 10.51% before the extraction and reducd to 3.26% after supercritical extraction, the moisture content for the pk were 11.86% and was reduced to 3.44% after supercritical extraction, whereas it was found that the moisture content for the palm kernel cake from the oil mill was 6.84% which is higher from the result for the other two samples that were extracted using supercritical extraction. These result revealed that the moisture content for the samples that were extracted using supercritical extraction were affected greatly by the supercritical extraction and was reduced to more than one third and was less than the moisture content for the palm kernel cake that was produced from the palm oil mill as shown in figure 4. The protein content before and after the supercritical extraction for the three samples were measured using Kjeltec autosampler system, the experiment results show that the PKt have the highest protein content 15.61 % compared with the other samples PK 15.01% and PKC 13.56% as shown in Table 1. Nonetheless, it was found that the protein content before the supercritical extraction was 15. 61g/100g sample for the pkt and this value was not affected greatly after the supercritical extraction as it was reduced to 14.40g / 100g sample. While the protein content for the pk was 15.01 before supercritical extraction and was reduced to 14.06 after supercritical extraction. The lowest value among the tested group for the protein content was recorded for the palm kernel cake from the palm oil mill which was 13.56 g /100g sample. These results show that supercritical extraction does not affect the protein content in the extracted sample and can give defatted palm kernels fiber free of oil with higher nutritional values than from screw pressed palm kernel cake. Table 1. The composition of the palm kernel with testa, without testa and the palm kernel cake Samples Palm kernel with testa Palm kernel without testa Palm kernel cake Before SFE PKt After SFE SC-PKt Before SFE PK After SFE SC-PK Without SFE Composition Average Average Average Average Average Totally dietary fiber % 61.58 63.03 57.78 58.96 60.71 Crude fiber % 8.99 8.49 7.29 7.23 15.17 Moisture % 10.51 3.26 11.86 3.44 6.84 Crude protein % 15.61 14.40 15.01 14.06 13.56 Ash % 8.58 4.34 3.96 3.55 13.92

Ash content followed the moisture trends in these sets of sample as the PKC gave the highest ash content among the three sets of samples 13.92 %, followed by the PK give the least amount of ash. It was observed that the ash content for the PKt was greatly influenced by the supercritical extraction and was reduced from 8.58% before the supercritical extraction to 4.43% after the extraction. On the other hand, the supercritical extraction did not have great affect on the ash content for the PK, as the ash content result were very close before and after the extraction and was 3.96%, 3.55% respectively. This result demonstrated that the supercritical extraction effectively reduced the ashes same as the removal of testa as the result for the ash content for the PKt after the supercritical extraction were close to the result of the ash content for the PK without affecting it nutritional value of protein and other nutritional components as shown in figure 5. Figure 5. The ash content before and after the supercritical Extraction for the palm kernel with testa (PKT) Palm kernel without testa(pk) compared to palm kernel cake(pkc) CONCLUSION Supercritical extraction is a powerful environmentally technique to enhance the production of fiber and the separation of oil without affecting its nutrient composition. Furthermore, defatted palm kernel with testa extracted using SFE, were superior compared to defatted palm kernel without testa and PKC sample from the palm oil mill. The defatted cake composition after the supercritical extraction has enhanced and become higher in dietary fiber and protein while the moisture and the ash were reduced significantly. Yet, there are more tests and experiments are needed to approve its safety and acceptance for human consumption that is in progress in our laboratory and among our research group.

REFERENCES [1] (Atasie, V. N. & Akinhanmi, T. F. 2009. Extraction, Compositional Studies and Physico -Chemical Characteristics of Palm Kernel Oil. Pakistan Journal of Nutrition, 8 (6), 1680-5194. [2] (Akpan, H. D., Udofia, A. A., Akpan, E. J. & Joshua, A. A. 2005. The effect of phytase and zink supplementation on palm Kernel Cake toxicity in sheep. Pakistan Journal of nutrition, 4(3), 148-153. [3] (Ramachandran, S., Singh, S.K., Larroche, C., Soccol, C.R., and Pandey, A. 2007. Oil cakes and their biotechnological applications A review. Bioresource Technology.98, 2000 2009. [4] Lau, H.L.N., Y.M. Choo, Ma,A. N., and Chuah,C.H. 2006. Quality of Residual Oil from Palm-Pressed Mesocarp Fiber (Elaeis guineensis) Using Supercritical CO2 With and Without Ethanol. Journal of the American Oil Chemists' Society (JAOCS) Vol. 83, (no. 10). [5] (Nik Norulaini, N. A., Md Zaidul, I. S., Anuar, O. and Omar, M. A. K. 2004. Supercritical enhancement for separation of lauric acid and oleic acid in palm kernel oil (PKO). Journal of Separation and Purification Technology. 35, 55 60. [6] (Sihvonen, M., Jarvenpaa E., Hietaniemi V. and Huopalahti R., 1999. Advances in supercritical carbon dioxide technologies - Phase Equilibrium and Morphology Study. Trends in Food Science & Technology. 10, 217-222. [7] Hassan, M. N., Norulaini, N. A. N., Omar, A. K. M., & Ibrahim, M. H. (2000). Simple fractionation through the supercritical carbon dioxide extraction of palm kernel oil. Journal of Separation and Purification Technology, 19, 113 120.