Palm oil, unlike many other dietary oils, does not increase the yield of chemically-induced mammary tumors in rats when fed at high levels in the diet. This difference appears to be due to the vitamin E fraction of palm oil, which is rich in tocotrienols, since palm oil stripped of this fraction does increase tumor yields. Experiments in our laboratory have shown that tocotrienols inhibit proliferation and growth of both MDA-MB-435 and MCF-7 cells in culture much more effectively than a-tocopherol. In addition, it was found that combinations of tocotrienols with Tamoxifen, a drug widely used for treatment of breast cancer, inhibit these cells more effectively than either tocotrienols or Tamoxifen alone. The present studies have now shown synergistic effects between tocotrienols and a number of other flavonoids from various plant sources, including citrus fruits, in the inhibition of both MDA-MB-435 and MCF-7 cells (IC50s 0.05-25 and 0.02-5 μg/mL respectively). In the MCF-7 cells, 1:1:1 combinations of tocotrienols, flavonoids and Tamoxifen were even more effective, with the best combination being d-tocotrienol, hesperetin and Tamoxifen (IC50 0.0005 μg/mL). These results suggest that diets containing palm oil may reduce the risk of breast cancer, particularly when eaten with other plant foods containing flavonoids, and may also enhance the effectiveness of Tamoxifen for treatment of breast cancer.
Syrian Golden hamsters have been widely used as a experimental model for the investigation of the aetiology and development of atherosclerosis and cardiovascular disease. The responses of the hamster to dietary fat manipulations are in many ways similar to that observed in humans. The lipidaemic effect of a tocotrienol rich fraction (TRF) from palm oil on human trials has not been consistent. In this study, the cholesterolaemic effect of tocotrienols and tocopherols were differentiated by using pure tocotrienols (that were isolated from palm oil fatty acid distillate) and pure commercial tocopherols and squalene. A palm oil triacylglycerol fraction (POTG), free of all unsaponifiable matter, was used as the dietary fat in different feeding experiments. Tocotrienols added at 162 ppm to POTG (POTG-T3L) significantly (p<0.05) lowered serum total cholesterol (TC) level as compared to that of the POTG group; but the serum LDL-C , HDL-C and TG levels of the POTG-T3L group were not significantly lower than that of the POTG group (P>0.05). Increasing the level of tocotrienol supplementation to the diet (POTG-T3H) appeared to raise rather then reduce the serum TC, LDL-C and HDL-C levels as compared to that of POTG-T3L group. This observation that lower level of tocotrienol supplementation appeared to exhibit stronger hypocholesterolaemic effect than a higher level of tocotrienol supplementation is interesting; but its explanation is not yet forthcoming. When tocopherols were supplemented at 72 ppm to the POTG diet it was observed that the serum TC, LDL-C and HDL-C levels were all somewhat increased when compared to that of the POTG group. These results suggest that tocotrienols and tocopherols may have opposite cholesterolaemic effects in the hamster, and further experiments need to clarify the mode of action of these vitamin E isomers. In our second series of experiments the cholesterolaemic effects of tocotrienols and tocopherols were studied in the presence of squalene, a key intermediate in the cholesterol synthesis pathway and a controversial cholesterol lowering agent. Squalene added to the diet at 0.1% level significantly lowered (p<0.05) serum TC level when compared to that of the POTG group. The LDL-C, HDL-C and TG levels appeared to be lowered by the squalene supplementation also but the differences between the POTG-SQ and POTG groups were not statistically significant (P>0.05). When tocotrienols or tocopherols were added to the squalene-containing POTG diets, the serum TC and LDL-C levels were further reduced (p<0.01) when compared to that of the POTG and POTG-SQ groups. The HDL-C and TG levels were not affected by tocotrienol or tocopherol supplementation in the presence of squalene. These results indicate that in the presence of tocotrienols and squalene POTG exhibit hypocholesterolaemic action whereas tocopherols may have a hypercholesterolaemic effect in the hamster.
Although dietary trans fatty acids can affect plasma lipoproteins negatively in humans, no direct comparison with specific saturated fatty acids has been reported, even though trans fatty acids were designed to replace saturates in foods and food processing. In this study, dietary trans 18:1 [elaidic acid at 5.5% energy (en)] was specifically exchanged for cis 18:1, 16:0 or 12:0 + 14:0 in 27 male and female subjects consuming moderate fat (31% en), low cholesterol (<225 mg/d) whole food diets during 4-wk diet periods in a crossover design. The trans-rich fat significantly elevated total cholesterol and LDL cholesterol relative to the 16:0-rich and 18:1-rich fats and uniquely depressed HDL cholesterol relative to all of the fats tested. Trans fatty acids also elevated lipoprotein (a) [Lp(a)] values relative to all dietary treatments. Furthermore, identical effects on lipoproteins were elicited by 16:0 and cis 18:1 in these subjects. The current results suggest that elaidic acid, one of the principal trans isomers produced during industrial hydrogenation of edible oils, adversely affects plasma lipoproteins. Thus, the negative effect of elaidic acid on the lipoprotein profile of humans appears to be unmatched by any other natural fatty acid(s).
This article examines the fat and fibre intakes of Malaysian adults and highlights discrepancies and practical limitations if these intakes are to match the levels for these nutrients advocated in the World Health Organisation (WHO) and American Heart Association (AHA) ‘diet models’. Local data on food consumption showed that the total fat intakes amongst Malaysian adults, contrary to common perception, were not high and the mean values obtained fell within the range of 40-66g or 22-26% kcal. As such, the dietary target of 30% kcal total fat or its intermediate target of 30-35% kcal, advocated by WHO and AHA mainly to address the problem of a high consumption of dietary fats in western populations, should not be adopted indiscriminately by Malaysians. Dietary fatty acid (FA) analysis by high performance liquid chromatography (HPLC) coupled with the use of food composition tables, showed that the typical Malaysian diet prepared with palm olein or palm olein-groundnut oil blends as cooking oil contained 3.2-4.0% kcal polyunsaturated fatty acids (PUFA), mainly as the w-6 linoleic acid, which is also the predominant essential fatty acid (EFA) in humans. This level of linoleic acid, with an ω-6/ ω--3 FA ratio approximating 10, is adequate for basal PUPA and EFA needs but fell short of the 4-10% kcal linoleic acid recommended by WHO (1993) to counter the effects of the cholesterol-raising saturated fatty acids (SFA). This raised upper limit of 10% kcal linoleic acid (previously 7% kcal), which equals the level of PUFA implied in the AHA diet model, appears unnecessarily high considering that the cholesterol-lowering potential of linoleic acid is maximum at about 6% kcal, while the health hazards associated with long-term high intakes of PUPA have never been completely dismissed. The new WHO lower limit for dietary linoleic acid (4% kcal) would have a controversial impact of raising the previous minimal 3% kcal EFA to above 4% kcal (linoleic + alpha-linolenic acids). Similarly, the WHO recommendation for total dietary fibre of 27-40g (equivalent to a daily combined intake of 400g of vegetables and fruits, 30g of which should come from pulses) appears at present, too high a dietary target for the average Malaysian adult whose habitual daily diet was estimated to contain about 180g of vegetables plus fruits, providing only about 13-16g total dietary fibre. Appropriately, an expert panel on Malaysian Dietary Guidelines has recommended instead, 20-30% kcal total fat containing 3-7% kcal PUFA, and 20-30g total dietary fibre for the local population.
AIM: To evaluate the anthropometric measurements and dietary intake of the free-living elderly in a Chinese community.
METHOD: A survey was carried out on 48 male and female subjects aged 60 to 96 years using anthropometric, dietary intake and questionnaire techniques.
RESULTS: The study indicated female subjects to have more health problems, like pain at the joints (33%), hypertension (17%) and diabetes (27%) as compared to the male subjects. Dietary intake analysis showed the Chinese male subjects to have a higher energy intake (1,623 kcal) compared to the females (1,197 kcal) even though they did not fulfill the recommended dietary intake. The intake for energy, fats and carbohydrates, was found to be significantly different (p < 0.05) between both sexes. Anthropometric measurements indicated male elderly subjects to be significantly heavier (p < 0.05, 55.4 kg) and taller (161.8 cm) than female elderly subjects (49.5 kg; 146.2 cm respectively). About half of the elderly were normal in their BMI (male 55.6%; female 50%) and only 6.6% of the female subjects were obese. More male subjects were found to be underweight (33%) compared to female subjects (17%). Waist hip ratio was 0.92 for male and 0.87 for female.
CONCLUSION: Our study showed that female elderly subjects had more health problems compared to male elderly subjects. On the whole, the elderly did not fulfill the recommended amount for energy intake while the percentage for carbohydrates, fats and protein from the total calorie intake were not in accordance with the healthy diet guidelines. Anthropometrically, male subjects were heavier and taller than female subjects. Looking at body mass index, most of the male elderly subjects were in the normal to underweight range. With these results, more research is warranted to give a clearer picture of the Chinese elderly in the country.
Study site: Kampung Baru Tabuh Naning, Alor Gajah, Malacca
A rapid and direct liquid chromatographic (LC) technique is described for the determination of a eutectic mixture of diphenyl oxide and biphenyl such as Dowtherm A thermal heating fluid (THF) in oleochemicals and palm olein. Analysis is performed with an RP-18 column with fluorescence detection (excitation at 247 nm and emission at 310 nm). The isocratic mobile phase (1.0 mL/min) consists of methanol and water (90 + 10, v/v). A linear calibration model (correlation coefficient = 0.9999) was developed directly from used Dowtherm A THF with the biphenyl peak (4.70 min) as a marker. Average recoveries from spiked glycerin, fatty alcohol mixture, methyl ester mixture, fatty acids, and palm olein were 90.9-108.7%, with a detection limit of 0.1 microgram/mL. The technique requires no prior sample cleanup nor extraction steps and is good for quality assurance purposes.
The application of membrane separation in palm oil refining process has potential for energy and cost savings. The conventional refining of crude palm oil results in loss of oil and a contaminated effluent. Degumming of crude palm oil by membrane technology is conducted in this study. The objective of this research is to study the feasibility of membrane filtration for the removal of phospholipids in the degumming of crude palm oil, including analyses of phosphorus content, carotene content free fatty acids (as palmitic acid), colour and volatile matter. A PCI membrane module was used which was equipped with polyethersulfone membranes having a molecular weight cut off of 9,000 (type ES209). In this study, phosphorus content was the most important parameter monitored. The membrane effectively removed phospholipids resulting in a permeate with a phosphorus content of less than 0.3 ppm The percentage removal of phosphorus was 96.4% and was considered as a good removal. Lovibond colour was reduced from 27R 50Y to 20R 30Y. The percentage removal of carotene was 15.8%. The removal of colour was considered good but the removal of carotene was considered insignificant by the membrane. Free fatty acids and volatile matter were not removed. Typical of membrane operations, the permeate flux decreased with time and must be improved in order to be adopted on an industrial scale. Membrane technology was found to have good potential in crude palm oil degumming. However, an appropriate method has to be developed to clean the membranes for reuse.
Hydrolysis of palm oil has become an important process in Oleochemical industries. Therefore, an investigation was carried out for hydrolysis of palm oil to fatty acid and glycerol using immobilized lipase in packed bed reactor. The conversion vs. residence time data were used in Michaelis-Menten rate equation to evaluate the kinetic parameters. A mathematical model for the rate of palm oil hydrolysis was proposed incorporating role of external mass transfer and pore diffusion. The model was simulated for steady-state isothermal operation of immobilized lipase packed bed reactor. The experimental data were compared with the simulated results. External mass transfer was found to affect the rate of palm oil hydrolysis at higher residence time.
There is little evidence to show that strict dietary modification alone confers any significant impact on cardiac events in primary and secondary prevention of coronary heart disease. Given the efficacy of the statins, the need for strict dietary modification in patients on statin therapy has been questioned. This study was performed to assess 1) the added benefit on serum lipid levels of a strict low-fat dietary regimen in patients with hypercholesterolaemia already treated with simvastatin; 2) the efficacy of simvastatin on the lipid profile of our sample Asian population; and 3) the tolerability and side-effect profile of simvastatin. This study was a prospective evaluation of 60 patients with hypercholesterolaemia treated with simvastatin who were subjected to either a normal diet or a dietitian guided low-fat diet. Assessment of the effects on serum lipid levels, side-effects, biochemical and haematological markers were performed. After 24 weeks of treatment, a strict dietitian guided low-fat diet conferred no additional benefit over and above what was achieved by simvastatin alone. Furthermore, a higher dose of simvastatin was needed in the dietitian guided diet group to achieve the target LDL-cholesterol level. Simvastatin resulted in a significant positive alteration of lipid profiles in all patients. The drug was well tolerated, with no significant change in either haematological or biochemical indices. Simvastatin is a highly effective cholesterol-lowering drug with a beneficial effect on the entire lipid spectrum in a cross-section of Asian patients, and is well tolerated. A dietitian guided dietary approach confers no additional advantage once statin therapy has been initiated.
Experiments were performed to determine the thermal resistance of hepatitis A virus (HAV) in three types of dairy products containing increased amounts of fat content (skim milk, homogenized milk; 3.5% MFG, and table cream; 18% MFG). HAV-inoculated dairy products were introduced into custom-made U-shaped microcapillary tubes that in turn were simultaneously immersed in a waterbath, using custom-made floating boats and a carrying platform. Following exposure to the desired time and temperature combinations, the contents of each of the tubes was retrieved and was tested by plaque assay to determine the reduction in virus titer. Our data indicated that < 0.5 min at 85 degrees C was sufficient to cause a 5-log reduction in HAV titer in all three dairy products, whereas at 80 degrees C, < or = 0.68 min (for skim and homogenized milk), and 1.24 min (for cream) were needed to cause a similar log reduction. Using a nonlinear two-phase negative exponential model (two-compartment model) to analyze the data, it was found that at temperatures of 65, 67, 69, 71, and 75 degrees C, significantly (P < 0.05) higher exposure times were needed to achieve a 1-log reduction in virus titer in cream, as compared to skim and homogenized milk. For example, at 71 degrees C, a significantly (P < 0.05) higher exposure time of 0.52 min (for cream) was needed as compared to < or = 0.18 min (for skim and homogenized milk) to achieve a 1-log reduction in virus titer. A similar trend of inactivation was observed at 73 and 75 degrees C where significantly (P < 0.05) higher exposure times of 0.29 to 0.36 min for cream were needed to cause a 1-log reduction in HAV in cream, as compared to < or = 0.17 min for skim and homogenized milk. This study has provided information on the heat resistance of HAV in skim milk, homogenized milk, and table cream and demonstrated that an increase in fat content appears to play a protective role and contributes to the heat stability of HAV.
The aims of this study were to investigate (1) platelet phospholipid (PL) polyunsaturated fatty acid (PUFA) composition in subjects who were the Melbourne Chinese migrants, compared with those who were the Melbourne Caucasians and (2) the relationship between platelet PL PUFA and intake of fish, meat and PUFA.
In Singapore. there exists differences in risk factors for coronary heart disease among the three main ethnic groups: Chinese, Malays and Indians. This study aimed to investigate if differences in dietary intakes of fat, types of fat, cholesterol, fruits, vegetables and grain foods could explain the differences in serum cholesterol levels between the ethnic groups. A total of 2408 adult subjects (61.0% Chinese, 21.4% Malays and 17.6% Indians) were selected systematically from the subjects who took part in the National Health Survey in 1998. The design of the study was based on a cross-sectional study. A food frequency questionnaire was used to assess intakes of energy, total fat, saturated fat, polyunsaturated fat, monounsaturated fat, cholesterol, fruits, vegetables and cereal-based foods. The Hegsted score was calculated. Serum total cholesterol, low density lipoprotein cholesterol, high density lipoprotein cholesterol were analysed and the ratio of total cholesterol to high density lipoprotein cholesterol was computed. The results showed that on a group level (six sex-ethnic groups), Hegsted score, dietary intakes of fat, satutrated fat, cholesterol, vegetables and grain foods were found to be correlated to serum cholesterol levels. However, selected dietary factors did not explain the differences in serum cholesterol levels between ethnic groups when multivariate regression analysis was performed, with adjustment for age, body mass index, waist-hip ratio, cigarette smoking, occupation, education level and physical activity level. This cross-sectional study shows that while selected dietary factors are correlated to serum cholesterol at a group level, they do not explain the differences in serum cholesterol levels between ethnic groups independently of age, obesity, occupation, educational level and other lifestyle risk factors.
Anhydrous milkfat (AMF) was fractionated to obtain a series of high-melting milkfat fractions (HMF). Solid fat content (SFC) of HMF as determined by nuclear magnetic resonance (NMR) was in the range 37.6-43.6% and 21.2-27.5% measured at 20 and 30 degrees C, respectively. The HMF have a higher melting characteristic compared to AMF as analyzed by differential scanning calorimetry (DSC) with melting enthalpies of 92.2-105.0 J/g and melting peak temperatures of 39.3-41.5 degrees C. The AMF was also blended with soft palm stearin (SPOs and/or hard palm stearin (HPOs)) according to a three conventional component mixture design which providing suitable formulations for HMF. This represented three selected blends of AMF:SPOs:HPOs at three different proportions (70:15:15, 60:30:10 and 50:45:5), having SFC and DSC melting characteristics of HMF. The study revealed that higher-melting characteristics of AMF could be achieved equally well by using both fractionation and blending techniques.
The effects of dietary fat during pregnancy and lactation on growth performance of pups, milk composition and very low density lipoprotein composition in rats were studied. A total of 33 dams were used in this study and each litter was adjusted to 8 pups per dam. The dams were fed on high fat (150 g fat/kg diet, HF), medium fat (75 g fat/kg of diet, MF) and low fat (2.5 g fat/kg diet, LF) diets. The body weights of dams increased during pregnancy and decreased after pregnancy. The HF pups had a higher body weight and higher weight gain than those of LF pups. The amount of feed intake of HF dams was significantly higher than LF and MF dams. The HF dams had significantly higher milk fat and water concentrations than LF dams. The milk protein was not significantly different among the treatment groups. All dams showed hypertriacylglycerolaemia in their very low density lipoprotein (VLDL) in late pregnancy. The VLDL-protein concentrations increased during the first week after parturition. The HF dams showed a greater response to the dietary fat than that of LF and MF dams. The findings suggest that addition of fat in the diet during pregnancy and lactation may improve the milk quality through modifying the composition of VLDL contents, leading to better growth of pups.
Partial hydrogenation of oil results in fats containing unusual isomeric fatty acids characterized by cis and trans configurations. Hydrogenated fats containing trans fatty acids increase plasma total cholesterol (TC) and LDL-cholesterol while depressing HDL-cholesterol levels. Identifying the content of trans fatty acids by food labeling is overshadowed by a reluctance of health authorities to label saturates and trans fatty acids separately. Thus, it is pertinent to compare the effects of trans to saturated fatty acids using stable isotope methodology to establish if the mechanism of increase in TC and LDL-cholesterol is due to the increase in the rate of endogenous synthesis of cholesterol. Ten healthy normocholesterolemic female subjects consumed each of two diets containing approximately 30% of energy as fat for a fourweek period. One diet was high in palmitic acid (10.6% of energy) from palm olein and the other diet exchanged 5.6% of energy as partially hydrogenated fat for palmitic acid. This fat blend resulted in monounsaturated fatty acids decreasing by 4.9 % and polyunsaturated fats increasing by 2.7%. The hydrogenated fat diet treatment provided 3.1% of energy as elaidic acid. For each dietary treatment, the fractional synthesis rates for cholesterol were measured using deuterium-labeling procedures and blood samples were obtained for blood lipid and lipoprotein measurements. Subjects exhibited a higher total cholesterol and LDL-cholesterol level when consuming the diet containing trans fatty acids while also depressing the HDL-cholesterol level. Consuming the partially hydrogenated fat diet treatment increased the fractional synthesis rate of free cholesterol. Consumption of hydrogenated fats containing trans fatty acids in comparison to a mixtur e of palmitic and oleic acids increase plasma cholesterol levels apparently by increasing endogenous synthesis of cholesterol.
An experiment was conducted to evaluate the interactive effects of dietary crude palm oil (CPO) concentration and water temperature on lipid and FA digestibility in rainbow trout. Four isolipidic diets with 0, 5, 10, or 20% (w/w) CPO, at the expense of fish oil, were formulated and fed to groups of trout maintained at water temperatures of 7, 10, or 15 degrees C. The apparent digestibility (AD) of the FA, measured using yttrium oxide as an inert marker, decreased with increasing chain length and increased with increasing unsaturation within each temperature regimen irrespective of CPO level fed to the fish. PUFA of the n-3 series were preferentially absorbed compared to n-6 PUFA in all diet and temperature treatments. Except for a few minor FA, a significant (P < 0.05) interaction between diet and temperature effects on FA digestibility was found. Increasing dietary levels of CPO lead to significant reductions in the AD of saturates and, to a lesser extent, also of the other FA. Lowering water temperature reduced total saturated FA digestibility in trout regardless of CPO level. Based on the lipid class composition of trout feces, this reduction in AD of saturates was due in part to the increasing resistance of dietary TAG to digestion. Increasing CPO level and decreasing water temperature significantly increased TAG content in trout fecal lipids, with saturates constituting more than 60% of the FA composition. Total monoene and PUFA digestibilities were not significantly affected by water temperature in fish fed up to 10% CPO in their diet. The potential impact of reduced lipid and FA digestibility in cold-water fish fed diets supplemented with high levels of CPO on fish growth performance requires further research.
We have previously reported an interaction between -514C>T polymorphism at the hepatic lipase (HL) gene and dietary fat on high-density lipoprotein-cholesterol (HDL-C) metabolism in a representative sample of white subjects participating in the Framingham Heart Study. Replication of these findings in other populations will provide proof for the relevance and consistency of this marker as a tool for risk assessment and more personalized cardiovascular disease prevention. Therefore, we examined this gene-nutrient interaction in a representative sample of Singaporeans (1324 Chinese, 471 Malays and 375 Asian Indians) whose dietary fat intake was recorded by a validated questionnaire. When no stratification by fat intake was considered, the T allele was associated with higher plasma HDL-C concentrations (P = 0.001), higher triglyceride (TG) concentrations (P = 0.001) and higher HDL-C/TG ratios (P = 0.041). We found a highly significant interaction (P = 0.001) between polymorphism and fat intake in determining TG concentration and the HDL-C/TG ratio (P = 0.001) in the overall sample even after adjustment for potential confounders. Thus, TT subjects showed higher TG concentrations only when fat intake supplied >30% of total energy. This interaction was also found when fat intake was considered as continuous (P = 0.035). Moreover, in the upper tertile of fat intake, TT subjects had 45% more TG than CC individuals (P < 0.01). For HDL-C concentration, the gene-diet interaction was significant (P = 0.015) only in subjects of Indian origin. In conclusion, our results indicate that there are differences in the association of -514C>T polymorphism with plasma lipids according to dietary intake and ethnic background. Specifically, the TT genotype is associated with a more atherogenic lipid profile when subjects consume diets with a fat content > 30%.
The palm fruit (Elaies guineensis) yields palm oil, a palmitic-oleic rich semi solid fat and the fat-soluble minor components, vitamin E (tocopherols, tocotrienols), carotenoids and phytosterols. A recent innovation has led to the recovery and concentration of water-soluble antioxidants from palm oil milling waste, characterized by its high content of phenolic acids and flavonoids. These natural ingredients pose both challenges and opportunities for the food and nutraceutical industries. Palm oil's rich content of saturated and monounsaturated fatty acids has actually been turned into an asset in view of current dietary recommendations aimed at zero trans content in solid fats such as margarine, shortenings and frying fats. Using palm oil in combination with other oils and fats facilitates the development of a new generation of fat products that can be tailored to meet most current dietary recommendations. The wide range of natural palm oil fractions, differing in their physico-chemical characteristics, the most notable of which is the carotenoid-rich red palm oil further assists this. Palm vitamin E (30% tocopherols, 70% tocotrienols) has been extensively researched for its nutritional and health properties, including antioxidant activities, cholesterol lowering, anti-cancer effects and protection against atherosclerosis. These are attributed largely to its tocotrienol content. A relatively new output from the oil palm fruit is the water-soluble phenolic-flavonoid-rich antioxidant complex. This has potent antioxidant properties coupled with beneficial effects against skin, breast and other cancers. Enabled by its water solubility, this is currently being tested for use as nutraceuticals and in cosmetics with potential benefits against skin aging. A further challenge would be to package all these palm ingredients into a single functional food for better nutrition and health.
A method for species identification from pork and lard samples using polymerase chain reaction (PCR) analysis of a conserved region in the mitochondrial (mt) cytochrome b (cyt b) gene has been developed. Genomic DNA of pork and lard were extracted using Qiagen DNeasy(®) Tissue Kits and subjected to PCR amplification targeting the mt cyt b gene. The genomic DNA from lard was found to be of good quality and produced clear PCR products on the amplification of the mt cyt b gene of approximately 360 base pairs. To distinguish between species, the amplified PCR products were cut with restriction enzyme BsaJI resulting in porcine-specific restriction fragment length polymorphisms (RFLP). The cyt b PCR-RFLP species identification assay yielded excellent results for identification of pig species. It is a potentially reliable technique for detection of pig meat and fat from other animals for Halal authentication.