Abstract: Cholesterol Oxidation Products (COPS) are well known for their negative biological effects. Till now,there is no study reported on solvent extraction used for COPs. In this study, two solvent extractiontechniques, namely chloroform/methanol (2:1, v/v) or Folch method and n-hexane/2-propanol (3:2, v/v) orHara-Radin method of cholesterol and COPs in beef tallow and lard were compared. Cholesterol and COPs(5-cholestane, 7-ketocholesterol and 25-hydroxycholesterol) contents in both fats also analyzed. The analysispassed through four major steps; extraction of lipids, saponification, enrichment of COPs and quantificationby gas chromatography mass spectrometry triple quadrupole. All standards showed good linearity withcorrelation coefficient (r ) of 0.9998 (5-cholestane), 0.9999 (cholesterol), 0.9873 (25-hydroxycholesterol) and20.9693 (7-ketocholesterol). Data obtained by this method was analyzed based on precision and recovery criteria.Precision measured as standard deviation (SD) was between 0.0040 and 2.2460; and no significant different(P>0.05) for the recovery testing using 5 -cholestane in both method. Recovery by Hara Radin method in lard(27.23%) and beef tallow (8.73%) was higher than the Folch method, 14.25% and 2.97% respectively. This studyimplies that Hara-Radin method can be an alternative method to avoid the use of harmful solvent such aschloroform in Folch method. Key words:Cholesterol Oxidation Products Gas Chromatography Mass Spectrometer Triple Quadrupole Fat Solvent Extraction

Abstract: Blood plasma contain transglutaminase (TGase) enzyme - catalyst the reaction of cross-linkingbetween proteins which has a significant impact on properties of protein gel capacity, thermal stability, waterholding capacity thereby protein characteristics elasticity, mouth feel, flavour, texture, binding force. Theobjectives of this study are to design and analyze the specificity of oligonucleotide primers of blood plasmatransglutaminase from chicken, bovine and procine blood and to detect the presence of blood plasmatransglutaminase in eight samples of fish surimi based products using PCR method. In this study, PolymeraseChain Reaction (PCR) method has been used in detecting the existence of blood transglutaminase enzyme DNAin surimi based products. Specific primers for chicken (Gallus gallus), cow (Bos taurus) and pig (Sus scrofa)blood transglutaminase enzyme were designed for positive detection. Two of the six primers designed for thechicken blood tranglutaminase, G3 and G5 have shown 99 % significant identity to the sequence of G. gallussimilar to XP-C repair complementing (transglutaminase) and the latter to the hypothetical LOC428804(transglutaminase) sequence. However, there was no positive result using the six primers designed forBos taurus transglutaminase and the six primers designed for S. scrofa transglutaminase. PCR amplification withG3 and G5 primers in surimi based products also showed negative results. Based on this study, G3 primersequence for chicken showed 99 % of G. gallus similar XP-C repair complementing (transglutaminase) followedby G5 primer which also obtained 99 % G. gallus hypothetical LOC428804 (transglutaminase) of significantidentity. However, for primer of cow and pig, there were no positive results. On the contrary, PCR amplificationon surimi based products had not showed positive bands of chicken blood transglutaminase, G3 and G5 in thesamples. Further research should be done to verify the consistency of this result and to redesign the primersfor cow and pig’s blood transglutaminase enzymes in order to increase its specificity which is vital in assuringthe reliability of the detection method in food products in accordance to the Halal food guidelines andregulations. Key words: Transglutaminase Fish Surimi Primer Design Polymerase Chain Reaction

Enzyme catalysis is most attractive for the synthesis of fine organic compounds, which are difficult to prepare and to handle by conventional means. In this work, green synthesis of lauryl palmitate, a wax ester was successfully carried out by lipase-catalyzed esterification of palmitic acid and lauryl alcohol. In this study, commercial immobilized lipase from Candida antartica (Novozym 435) was used as biocatalyst. The effect of various reaction parameters were optimized to obtain a high yield of wax esters. The optimum condition to produce lauryl palmitate was at reaction time (RT); 10 min, temperature (T); 40°C, amount of enzyme (E); 0.4 g, molar ratio of substrate (N); 2:1 and organic solvents of log P>3.5. The product was then subjected to characterize using Fourier-transform infrared spectroscopy (FT-IR) and Gas chromatography spectroscopy (GC) to ensure the purity of product obtained. Analysis of yield showed that at optimum condition, lauryl palmitate was produced in short time with high purity, >90%.