Category Archives: 2016 Vol 28 March

EVALUATION OF OIL PALM LEAF-SPECIFIC PROMOTER (LSP1) ACTIVITY FOR EXPRESSING PHB GENES IN Arabidopsis thaliana

Production of polyhydroxybutyrate (PHB), a biodegradable plastic, in plant, has been studied all over the world. We have constructed a transformation vector pLSP15 carrying the PHB biosynthesis genes. Each of PHB genes was driven by an oil palm leaf-specific promoter (LSP1). This transformation vector was later transformed into Arabidopsis thaliana and successfully produced T3 putative transgenic plants. The mature T3 generation was analysed by Reverse Transcription-qPCR (RT-qPCR) to test the capability of a monocotyledon oil palm LSP1 promoter in driving the expression of the PHB genes in a dicotyledon A. thaliana plant. Expression of all PHB genes, i.e. phbA (2.32), phbB (1.53) and phbC (1.47) relative to the wild-type, were detected in the plant. Nile blue A staining demonstrated that the PHB polymer was successfully produced throughout the developmental stage of the transgenic A. thaliana plant without any deleterious effects. The results demonstrated that the oil palm LSP1 promoter could drive the expression of the PHB biosynthesis genes in plants.

SAFETY ASSESSMENT OF bacillus thuringiensis-BASED PRODUCTS USING EYE/OCULAR IRRITATION ON RABBITS

To register with the National Pesticide Board, Malaysia, a pesticide must undergo laboratory testing for short-term and long-term health effects. Laboratory animals such as rabbits are tested with high doses of Bacillus thuringiensis (Bt) products to assess the potential to produce lesions or reversal effects on eyes. The objective of this study was to investigate the effect of Bafog-1 (S) and Ecobac-1 (EC) against young New Zealand albino rabbit’s eye or the potential of Bt products to cause eye or ocular irritation. The test was conducted according to the Guidelines of International Organisation for Standardisation 10993-10, Test for Irritation and Sensitisation. The results showed that Bafog-1 (S) is not an irritant to rabbits, whereas, Ecobac-1 (EC) produced irritation that was reversal after 120 hr of instillation. The Bt as an environmental-friendly microbial insecticide was generally non-toxic to human, domestic animals and vertebrates. Therefore, Bt is recommended to be used as a safe microbial insecticide for controlling bagworm in oil palm plantation.

EFFECTS OF RODENTICIDE ON GROWTH OF NESTLING BARN OWL, Tyto alba javanica IN OIL PALM PLANTATIONS

This study investigated the effects of the first generation anti-coagulant rodenticide, chlorophacinone and the second generation rodenticide, bromadiolone on nestling growth of barn owl, Tyto alba javanica in oil palm plantations. Forty-two nestlings were selected from rodenticide-free, chlorophacinone and bromadiolone treated plots. Growth rates of nestlings were measured by taking body weight, culmen length, tarsus length and wing length on a periodic basis. Dead nestlings were collected during daily observations. Autopsies were carried out to determine cause of mortality and internal organ (livers) were harvested for analysis of residue by using high performance liquid chromatography (HPLC). Rodenticide-free plot scored the highest fledging success, i.e. 71.4% (n = 14) at 52 days after hatching. In chlorophacinone treated plot, six out of 14 nestlings or 42.85% successfully grew to fledging age of 52 days. In bromadiolone treated plot, only five or 35.75% (n = 14) of nestlings survived to fledging age. Three out of nine nestling carcasses found in bromadiolone treated area were collected for the analysis. Of these, two were detected with bromadiolone residue measuring 0.33 μg g-1 and 0.41 μg g-1 wet weight, respectively. In chlorophacinone treated plot, three out of eight carcasses were collected for analysis. The residue of chlorophacinone detected ranged from 0.18 μg g-1 to 0.21 μg g-1. Nestlings that survived in the rodenticide-free area showed higher measurements for body weight, tarsus, culmen and wing length compared to bromadiolone and chlorophacinone treated plots.

DISSIPATION OF FLUROXYPYR IN A MALAYSIAN AGRICULTURAL SOIL WITH SIMULATION USING THE PERSIST AND VARLEACH MODEL

The study was carried out to compare the dissipation of fluroxypyr in a Malaysian agricultural soil based on field experimentation and simulation using the PERSIST and VARLEACH model. A plot situated in an oil palm plantation near Sepang, Selangor, Malaysia was selected for the field experiment. The plot was treated with fluroxypyr at the recommended dosage (72.5 g ha-1). The soil samples from the field trial were collected and analyses conducted in the laboratory according to the sampling schedule. Simulation of fluroxypyr persistence and leaching was also conducted using computer-run software, PERSIST and VARLEACH. The predicted data obtained using PERSIST for fluroxypyr was found to be higher than the observed data from field measurements. However, the predicted data for fluroxypyr using VARLEACH matched well with the observed data from the field trial.

SAFETY ASSESSMENT OF TOCOTRIENOL SUPPLEMENTAT ION IN SUBJECTS WITH METABOL IC SYNDROME: A RANDOMISED CONTROL TRIAL

Previous studies have reported that tocotrienols (T3) possess many distinct properties such as antioxidant, cardioprotective, neuroprotective, anti-cancer, anti-inflammatory and anti-angiogenic, which are beneficial for the improvement of human health. However, there is limited data available on the safety assessment of T3 compared to tocopherols (T). A randomised, double-blinded, cross-over and placebo-controlled human clinical trial was conducted to determine the safety and tolerance of T3 supplementation in 31 subjects with metabolic syndrome. The subjects were supplemented with tocotrienol-rich fraction (TRF) 200 mg or placebo capsules twice daily for two weeks followed by a post-intervention visit. Results showed that T3 supplementation had no significant adverse effect on the red blood cell (RBC), white blood cell (WBC) and platelet counts between TRF (5.10 ± 0.78 x 1012 litre-1, 7.35 ± 1.59 x 109 litre-1, 279.45 ± 73.86 x 109 litre-1, respectively) and placebo interventions (5.13 ± 0.76 x 1012 litre-1, 7.25 ± 1.95 x 109 litre-1, 267.45 ± 68.72 x 109 litre-1, respectively). Measures of serum aspartate aminotransferase (AST), serum alanine aminotransferase (ALT)) and albumin did not differ between TRF (25.68 ± 10.72 IU litre-1, 38.26 ± 24.74 IU litre-1, 43.61 ± 2.26 g litre-1, respectively) and placebo interventions (27.39 ± 16.44 IU litre-1, 42.23 ± 33.58 IU litre-1, 43.68 ± 2.15 g litre-1, respectively).This study indicated that supplementation with T3 at the dosage of 400 mg per day for 14 days did not induce haematoxicity and hepatotoxicity in subjects with metabolic syndrome.

In situ CHARACTERISATION OF PALM OLEIN CRYSTALLISATION BEHAVIOUR BY FOCUSED BEAM REFLECTANCE MEASUREMENT (FBRM)

This article examines the in situ characteristics and behaviour of particles during palm olein crystallisation using Focused Beam Reflectance Measurement (FBRM). Melted palm olein was isothermally crystallised at 9ºC, 13ºC and 15ºC for 6 hr in a stirred, jacketed vessel and changes in crystal size, population and behaviour were tracked using FBRM. Results indicate that the induction time for primary nucleation for palm olein increased while crystal population was significantly lower at higher temperatures, as expected. Crystallisation occurred in a two-step manner at 9ºC and 13°C while a single step crystallisation was observed at 15°C. These occurrences were plausibly due to the lower super cooling and fewer species of triacylglycerols capable of crystallising at higher temperatures. FBRM detected major agglomeration of crystals at 9°C due to increased crystal population and contact between crystals. Chord length distributions (CLD) showed that the final mode crystal size was 32 μm at 9°C while significantly larger crystals of 108 μm were observed at 15°C. Microscopic observations revealed that crystals within the slurry appear gel-like below 13°C. This study extends current knowledge on FBRM application for in situ particles characterisation in real time within a palm-based crystallisation system.

THE PRODUCTION OF ACETONE-BUTANOL-ETHANOL FROM OIL PALM WASTES BY CLOSTRIDIUM ACETOBUTYRICUM AND UTILISATION OF THE WASTEWATER FOR POLYHYDROXYBUTYRATE PRODUCTION

Oil palm wastes including palm oil mill effluent (POME), empty fruit bunch (EFB), palm pressed fibre (PPF) and palm kernel shell (PKS) were collected and utilised as substrate for acetone-butanolethanol (ABE) production. Each waste was pre-treated before use. POME was centrifuged to obtain the supernatant. EFB, PPF and PKS were pre-treated by steam explosion and hydrolysed by cellulase from Aspergillus niger. The cultivation of Clostridium acetobutyricum under 180 ml of diluted POME supplemented with 10 g litre-1 of pre-treated EFB gave the maximum ABE and total acids at 3.8 g litre-1 and 2.0 g litre-1, respectively. The ABE production was then recovered by distillation. The wastewater after distillation was collect and characterised. Wastewater contained organic acid profiles such as acetic acid, particularly propionic and butyric acid in the amount of 0.1-1.0 g litre-1. Acetic and butyric present in wastewater can possibly be used for polyhydroxybutyrate (PHB) production. Therefore, the wastewater was utilised as substrate for PHB production by Cupriavidus necator. The maximum PHB (48.4% of dry cell weight, DCW) was obtained after 60 hr of cultivation. The polymer was identified by gas chromatography to be PHB when compared to commercially available product.

COMPATIBILITY OF BIODIESEL FUEL WITH METALS AND ELASTOMERS IN FUEL DELIVERY SYSTEM OF A DIESEL ENGINE

This article intends to evaluate available literature findings and determine if they are representative of the actual compatibility between fuel delivery materials (FDM) and biodiesel fuel in the physical system. The study will also evaluate current test standards on their effectiveness of representing the physical fuel delivery system. Although the compatibility of a number of materials with biodiesel fuel has been reported, there is a need to establish the exact materials present in the fuel delivery system. This is especially true for elastomers since their resistance is mainly dependent on their elemental compositions. While typical standards such as the ASTM G31 and ASTM D471 for metals and elastomers, respectively are deemed suitable for evaluating the effects of water content, total acid number and oxidised products in biodiesel on FDM degradation based on laboratory immersion studies, none of these standards resemble the actual engine operating conditions such as varying fuel pressure/temperature as well as presence of a wide range of materials in a typical diesel engine’s fuel delivery system. As such, findings from existing studies so far are inadequate to conclusively determine the compatibility between FDM and biodiesel fuel in the actual fuel delivery system of a diesel engine.

ACUTE ECOTOXICITY (48-HR EC50) ASSESSMENT OF PALM-BASED METHYL ESTER SULPHONATES (MES) TOWARDS Daphnia magna

Palm-based methyl ester sulphonates (MES) is an anionic surfactant derived from palm oil through sulphonation of fatty acid methyl esters. It has good surface-active properties and biodegradability, excellent detergency and is less sensitive to water hardness. It has been used in powder and liquid detergent formulations and other cleaning applications. The evaluation of MES in aquatic ecosystems is vital as they are discharged in large volumes into the environment. An attempt was made to evaluate the acute effect of palm-based MES on Daphnia magna through acute immobilisation studies (48 hr EC50). MES of various chain lengths were selected for the investigation. Based on the results, the EC50 of MES (based on % immobilisation) decreased (became more toxic) with increasing carbon chain length of the surfactant. This trend was also observed in many other anionic surfactants. MES is well suited for environmental-friendly detergent due to its good biodegradability and its toxicity which will not pose any environmental effect on aquatic organisms. The present study provides relevant data concerning the effects of MES surfactant on freshwater invertebrate, which are useful to establish water quality criteria in a regulatory framework.

FUEL AND ASH PROPERTIES OF BIOCHAR PRODUCED FROM MICROWAVE-ASSISTED CARBONISATION OF OIL PALM TRUNK CORE

Oil palm trunk core (OPTC) is the biomass left over from plywood industry that utilises oil palm trunk as feedstock. This research aims to investigate the fuel and ash properties of biochar converted from OPTC biomass through microwave-assisted carbonisation process. The raw OPTC contains high moisture (~64.3%) with low fixed carbon (~6.4%) and calorific value (4.6 MJ kg-1). Microwave-assisted carbonisation of OPTC was investigated under several sizes, temperature and microwave power variables. The results showed that biochar yield decreased with the increasing carbonisation temperature. The optimum conditions yielding 8.4% biochar was obtained at 350oC, microwave power 800 W and 6 cm cube. Compared to raw OPTC, biochar has higher fixed carbon (~20-64%) and about four times higher calorific value than the raw OPTC. In general, all of the biochars produced in this study showed higher alkali index and total alkali values than those obtained from raw OPTC. Feedstock size seemed to affect biochar ash properties in this study. Biochar obtained from 6 cm cube tend to have lower slagging and fouling tendency than those produced with 4 cm cube.

ALTERNATIVE LUBRICANTS: STUDY ON PALM OIL-BASED LUBRICANTS IN METAL FORMING PROCESS

Mineral oil-based lubricants are always the preferred choice for industrial application due to their excellent properties and reasonable price. However, due to environmental and depletion issues, a number of studies have been carried out to explore alternative lubricants such as plant oils. In metal forming processes, reducing frictions, improving tool life and product quality are some of the important issues to be considered. In our study, palm oil-based lubricants were tested for cold extrusion processes and mineral oils were used for comparison purposes. It was found that refined bleached deodorised (RBD) palm kernel oil and palm olein performed an enhanced extrusion load than mineral oil-based lubricant, and they showed no severe wear on product surface. Based on the results, renewable oil-based lubricants can be considered as a substitute to common mineral oil based lubricants used in the industry.

ENVIRONMENTAL PERFORMANCE OF PALM-BASED METHYL ESTER SULPHONATES PRODUCTION USING LIFE CYCLE APPROACH

Palm-based methyl ester sulphonates (MES) derived from palm oil through sulphonation process offer an interesting alternative to petroleum-based surfactants. This study was carried out to identify any potential environmental impacts that could be associated with the production of palm-based MES. Data on pilot plant production were obtained from the Malaysian Palm Oil Board (MPOB)’s MES pilot plant with 20 kg hr-1 capacity. Life cycle assessment (LCA) performance was based on representative inventory data of MES processes for three years of production. The results were analysed using Eco-indicator 99 methodology operated under SimaPro version 8.0.2 software. The single score results showed that the main impact contributors for MES production at pilot plant scale were from production of methyl ester, steam and methanol. Based on weighted score, the most significant impact categories from this production were fossil fuels, respiratory inorganics and climate change. However, there was no significant impact value on land use category since this study was conducted with best management practices in oil palm plantation in Malaysia. From this study, it can be concluded that the production of MES at pilot plant using the best approach used in the oil palm industry are more sustainable and environmental-friendly.

REPRODUCIBILITY OF PALM-BASED POLYOLS PRODUCTION

The purpose of this article is to verify the reproducibility of palm-based polyols from the optimised reaction conditions. The reproducibility studies on the production of the palm-based polyols were conducted via alcoholysis of epoxidised palm olein with isobutanol, catalysed by K10 montmorillonite under the optimised reaction conditions (reaction temperature at 60°C for 2 hr). It was proven that palm-based polyols with similar properties can be repeatedly produced when a set of standard synthesis steps was followed. This indicates that palm-based polyol can be produced with specific range of properties and this enables a feasible commercial production of palm-based polyols. The study on the recyclability of used K10 montmorillonite in the production of palm-based polyols will be carried out in the future.

PRODUCTION OF TOCOLS NANOEMULSION BY ULTRASONICATION

Nanoemulsion has proven to be one of the efficient approaches to improve tocols (tocopherols and tocotrienols) solubility in aqueous solution. High energy emulsification method such as ultrasonication and microfluidisation has been widely applied. The objective of this study was to prepare palm-based tocols nanoemulsion using ultrasonication to produce small average droplet size (<100 nm) and narrow droplet size distribution with low polydispersity index (PDI) value. The optimal ultrasonication duration and amplitude of 80 min and 100% produced nanoemulsion of an average droplet size of 104.1 ± 2.9 nm with PDI value of 0.215 ± 0.008. The mixing of Tween 80-Brij 35 (50:50 w/w) with concentration of 1.5% w/v produced an average droplet size of 54.8 ± 1.2 nm and PDI of 0.266 ± 0.006. Although the increase of emulsifier concentrations to 3% w/v from 0.75% w/v produced an average droplet size of 45.4 ± 1.0 nm, PDI obtained was high 0.404 ± 0.002 indicating that the droplet size distribution was becoming less monodisperse. The produced nanoemulsion will be extruded into gelling solution to form calcium alginate beads. Total tocols encapsulate within the bead and release of tocols from the bead will be evaluated and reported in future study.

SYNTHESIS OF GLYCERYL CARBONATE VIA MICROWAVE IRRADIATION

Supported by increasing awareness for environment and sustainability, glyceryl carbonate (Gly-C) has gained much interest over the last 20 years because of its versatile reactivity and as a way to valorise waste glycerol. In this study, synthesis of Gly-C via microwave irradiation was conducted as an alternative method to synthesise the glycerol derivative. Reaction parameters which include the type of carbonating agent, glycerol to carbonating agent mole ratio, reaction temperature as well as the reaction time have been examined. By setting microwave power at the lowest power (100 W), results from this study show that the synthesis of Gly-C under microwave irradiation is best carried out using ethylene carbonate as the carbonating agent with glycerol to ethylene carbonate molar ratio of 3:1 for 10 min at 80°C under atmospheric pressure. Gly-C was successfully obtained with 62% yield and purity of 96%. This study implies the significant use of microwave technique in the synthesis of Gly-C.