Effect of antioxidants on soy oil conjugated linoleic acid production and its oxidative stability

Conjugated linoleic acid (CLA)-rich soy oil can be produced by photoisomerization of soy oil linoleic acid to produce a soy oil with up to 20% CLA. Recent studies indicate that mixed soy tocopherols added to refined bleached deodorized (RBD) oil produced significant increase in soy CLA yield during soy oil linoleic acid photoisomerization. However, the effect of common synthetic free radical scavenging antioxidants and specific tocopherols on CLA yield and its oxidative stability is not known. Therefore, this investigation evaluated the effects of various antioxidant systems on soy oil CLA yield and oxidative stability. Soy oil with added antioxidants consisting of combinations of mixed tocopherols (MT), ascorbyl palmitate (AP), butylated hydroxyanisole (BHA), butylated hydroxytoluene (BHT) and tert-butylhydroquinone (TBHQ) was photoisomerized to produce CLA- rich soy oil. The CLA content was determined by GC-FID analysis and oxidative stability by peroxide value (PV). The soy oil in the presence of TBHQ, MT alone and MT with 500 ppm of AP produced significantly greater CLA yields and improved oxidative stability compared to a control without added antioxidants (p < 0.05). However, added mixed tocopherols produced the greatest CLA yield and also reduced PV relative to the control. Tocopherols in the form of α-, γ- and δ-tocopherols were then each examined as to their relative effect on CLA yields and PV. The largest increase in CLA yield was obtained with 1800 ppm of γ-tocopherols with reduced PV. Mixed tocopherols, TBHQ and γ-tocopherols can be used to increase CLA yield and reduce PV of soy oil during linoleic acid photoisomerization.     

Kinetics of photoirradiation-induced synthesis of soy oil-conjugated linoleic acid isomers

Photoirradiation of soy oil with UV/visible light has been shown to produce significant amounts of trans,trans conjugated linoleic acid (CLA) isomers through conversion of various synthesized intermediate cis,trans isomers. The objective of this study was to determine the kinetics of CLA isomers synthesis to better understand the production of various isomers. Soy oil was irradiated with UV/visible light for 144 h in the presence of an iodine catalyst and CLA isomers analyzed by gas chromatography (GC). Arrhenius plots were developed for the conversion of soy oil linoleic acid (A) to form cis-, trans/trans-, cis-CLA (B), conversion of cis-, trans/trans-, cis-CLA to form trans,trans-CLA (C) with respect to B, and formation of trans,trans-CLA isomers with respect to C. The kinetics of consumption of linoleic acid (LA) to form cis-, trans/trans-, cis-CLA was found to be of second-order with a rate constant of 9.01 x 10-7 L/mol s. The rate of formation of cis-, trans/trans-, cis-CLA isomers depends on the rate of formation from LA and its rate of consumption to form trans,trans-CLA isomers. The conversion of cis-, trans/trans-, cis-CLA isomers to trans,trans-CLA isomers was found to be of first-order with a rate constant of 2.75 x 10-6 s-1. However, the formation of thermodynamically stable trans,trans-CLA isomers (C) with respect to C was found to be a zero-order reaction with a rate constant of 10.66 x 10-7 mol/L s. The consumption of LA was found to be the rate-determining step in the CLA isomers formation reaction mechanism. The findings provide a better understanding of the mechanism of CLA isomers synthesis by photoirradiation and the factors controlling the ratio of various isomers.     

Photocatalytic production and processing of conjugated linoleic acid-rich soy oil

Daily intake of conjugated linoleic acid (CLA), an anticarcinogenic, antiatherosclerotic, antimutagenic agent, and antioxidant, from dairy and meat products is substantially less than estimated required values. The objective of this study was to obtain CLA-rich soybean oil by a customized photochemical reaction system with an iodine catalyst and evaluate the effect of processing on iodine and iodo compounds after adsorption. After 144 h of irradiation, a total CLA yield of 24% (w/w) total oil was obtained with 0.15% (w/w) iodine. Trans,trans isomers (17.5%) formed the majority of the total yield and are also associated with health benefits. The isomers cis-9,trans-11 and trans-10,cis-12 CLA, associated with maximum health benefits, formed approximately 3.5% of the total oil. This amount is quite significant considering that total CLA obtained from dairy sources is only 0.6%. ATR-FTIR, 1H NMR, and GC-MS analyses indicated the absence of peroxide and aldehyde protons, providing evidence that secondary lipid oxidation products were not formed during the photochemical reaction. Adsorption processing vastly reduced the iodine and iodocompounds without CLA loss. Photocatalysis significantly increased the levels of CLA in soybean oil.     

Measurement of conjugated linoleic acid (CLA) in CLA-rich potato chips by ATR-FTIR spectroscopy

A conjugated linoleic acid (CLA)-rich soy oil has been produced by photoisomerization of soy oil linoleic acid. Nutritional studies have shown that CLA possesses health benefits in terms of reducing certain heart disease and diabetes risk factors. Potato chips are snacks that are readily produced in the CLA-rich soy oil containing CLA levels similar to those of the oil used for frying. The objective of this study was to develop an FTIR method to rapidly determine the CLA content of oil in potato chips. Photoirradiated soy oil samples with ～25% total CLA were mixed with control soy oil, and 100 soy oil samples with total CLA levels ranging from 0.89 to 24.4% were made. Potato chips were fried using each of these 300 g CLA rich soy oil mixtures at 175 °C for approximately 3 min. Duplicate GC-FID fatty acid analyses were conducted on oil extracted from each batch of potato chips. The chip samples were ground and then scanned using ATR-FTIR spectroscopy with the aid of a high-pressure clamp, and duplicate spectra of each sample were averaged to obtain an average spectrum. Calibration models were developed using PLS regression analysis. These correlated the CLA isomer concentrations of potato chips obtained by GC-FID fatty acid analysis with their corresponding FTIR spectral features. The calibration models were fully cross validated and tested using samples that were not used in the calibration sample set. Calibrations for total CLA, trans,trans CLA, trans-10,cis-12 CLA, trans-9,cis-11 CLA, cis-10,trans-12 CLA, and cis-9,trans-11 CLA had coefficients of determinations (R2v) between 0.91 and 0.96 and corresponding root-mean-square error of prediction (RMSEP) ranging from 0.005 to 1.44. The ATR-FTIR technique showed potential as a method for the determination of the CLA levels in unknown potato chip samples.     

Effects of temperature and agitation rate on the formation of conjugated linoleic acids in soybean oil during hydrogenation process.

The effects of hydrogen temperature and agitation rate on the formation of total conjugated linoleic acids (CLA) and CLA isomers were studied during hydrogenation with a selective Ni catalyst. The CLA isomers were identified by using a 100-m cyano-capillary column gas chromatograph and a silver ion-impregnated HPLC. Reaction temperature and agitation rate greatly affected the quantities of total CLA and individual CLA isomers, and the time to reach the maximum quantity of CLA in the partially hydrogenated soybean oil. As the hydrogenation temperature increased, the maximum quantity of CLA in soybean oil increased, but the time to reach the maximum CLA content decreased. By increasing the hydrogenation temperature from 170 to 210 degrees C, the quantity of CLA obtained was about 2.6 times higher. As the agitation rate decreased, the CLA formation in soybean oil increased, and the time to reach the maximum CLA content also increased. The maximum CLA contents in soybean oil obtained during hydrogenation at 210 degrees C with agitation rates of 300, 500, and 700 rpm were 162.82, 108.62, and 66.15 mg total CLA/g oil, respectively. The present data showed that it is possible to produce high-CLA-content soybean oil without major modification of fatty acid composition by short-time (10 min) selective hydrogenation under high temperature and low agitation rate conditions.     

Effect of extrusion parameters on conjugated linoleic acids of corn extrudates

The effects of extrusion temperature, 150-190 degrees C, and torque, 50-70%, on the content and configuration of conjugated linoleic acids (CLA) in corn extrudates were analyzed by GC and HPLC. At a temperature of 150 degrees C, CLA content increased from 1.2 mg/g of oil in feed to 7.8 mg/g of oil in corn extrudates. A decrease in total CLA (P < 0.05) was obtained when the product temperature was further increased to 190 degrees C. Alteration of CLA geometrical configuration was observed at higher extrusion temperatures. trans,trans-CLA significantly increased (P < 0.05) from 10.2% in feed to 11.9% of CLA at the extrusion condition of 190 degrees C and 70% torque. The highest expansion of extrudates was found at the product temperature of 150 degrees C and 70% torque. This extrusion condition also gave the maximum total CLA content and minimum trans,trans-CLA formation.     

Stripped corn oil controls scald and maintains volatile production potential in golden supreme and delicious apples

Effects of stripped (alpha-tocopherol < 5 mg L(-)(1)) corn oil on flesh firmness, skin color, acidity, soluble solids content (SSC), scald, and fruit volatiles during 6 months at 0 degrees C were studied using Golden Supreme and Delicious apples. Treatment with 10% oil emulsion reduced production of ethylene, alpha-farnesene, and major volatile esters in the first 3 months of storage, but this trend reversed after 5 months. After 6 months at 0 degrees C plus 7 days at 20 degrees C, oil-treated fruit were firmer and greener and had higher levels of titratable acidity than the controls. In addition, control fruit developed 27% and 42% scald in Golden Supreme and Delicious apples, respectively, whereas oil-treated fruit were free from scald. Soluble solids content and ethanol production were unaffected by oil treatment.     

Effects of rhodium heterogeneous catalyst and isomerization conditions on linoleic acid conjugation of soybean oil

Rhodium heterogeneous catalyst was used to catalyze isomerization of linoleic acid in soybean oil to conjugated linoleic acid (CLA). A central composite rotatable design with five levels of three variables, namely, reaction temperature, stirring speed, and reaction time, was used to determine the maximum CLA yield. The formation of CLA during isomerization was greatly dependent on the reaction temperature and time. The CLA content of soybean oil increased from 0.63 to 202.42 mg/g oil when isomerization was done at 200 degrees C, with a stirring speed of 200 rpm for 49 min. Analysis of triacylglycerol positions showed that linoleic acid at any position in a triacylglyceride could possibly be isomerized to CLA.     

Conjugated linoleic acid content and organoleptic attributes of fermented milk products produced with probiotic bacteria

The effect of probiotic bacteria on the formation of conjugated linoleic acid (CLA), microbial growth, and organoleptic attributes (acidity, texture, and flavor) of fermented milk products was determined. Four probiotic bacteria, Lactobacillus rhamnosus, Propionibacterium freudenreichii subsp. shermanii 56, P. freudenreichii subsp. shermanii 51, and P. freudenreichii subsp. freudenreichii 23, were evaluated individually or in coculture with traditional yogurt cultures (Lactobacillus delbrueckii subsp. bulgaricus and Streptococcus salivarius subsp. thermophilus). The lipid source was hydrolyzed soy oil. L. rhamnosus, in coculture with yogurt culture, resulted in the highest content of CLA. Growth and CLA formation of propionibacteria were enhanced in the presence of yogurt cultures. Texture and flavor attributes of fermented milks produced with propionibacteria were significantly different than the fermented milks processed with yogurt cultures. The fermented milks processed with probiotic bacteria in coculture with yogurt cultures demonstrated similar acidity, texture, and flavor as the fermented milk produced with yogurt cultures.     

Quantification and partial characterization of the residual protein in fully and partially refined commercial soybean oils

A method has been developed to determine residual protein in refined oils, a potential trigger of allergic reactions. High-pH bicarbonate or borate buffers were found to be the most effective extractants, residual oil protein comprising a mixture of proteins of M(r) 6000-100000. Extracted protein could be quantified with superior precision using 3-(4-carboxybenzoyl)quinolone-2-carboxaldehyde (CBQCA). Residual protein content determined in a set of oils using the borate extraction-CBQCA assay was positively correlated with contents determined using a bicarbonate-total amino acid analysis method. Oil refining substantially reduced the oil protein content determined by the borate-CBQCA assay with neutralized/refined, bleached, and deodorized (fully refined) oils containing 62-265 ng/g oil, whereas crude un-degummed oils contained 86000-87900 ng/g of protein. These analyses and published data on cumulative threshold doses for soybean suggest that even the most sensitive individuals would need to consume at least 50 g of highly refined oil to experience subjective symptoms.     

Rapid determination of free fatty acids in poultry feed lipid extracts by SB-ATR FTIR spectroscopy

A simple, rapid, and reproducible method has been developed for the quantitative determination of free fatty acid (FFA) content in lipids extracted from poultry feeds by Fourier transform infrared (FTIR) spectroscopy with the use of a single-bounce attenuated total reflectance (SB-ATR) accessory. An FTIR calibration curve was prepared by gravimetrically adding oleic acid (15-37%) to pure refined, bleached, and deodorized (RBD) canola oil and measuring the area of the COOH absorption band at 1710 cm-1. The oil from each of 12 poultry feed formulations was extracted using conventional Soxhlet extraction, and after evaporation of the solvent, the FFA content was determined by the conventional AOCS titrimetric procedure and by the SB-ATR/FTIR method. The SB-ATR/FTIR FFA predictions were related to those determined by the AOCS titrimetric method by linear regression, producing an R value of 0.999 and a SD of +/-0.28% FFA. Time-course spectra collected as lipids extracted into hexane indicated that a 15 min extraction was adequate to obtain a representative sample for FFA determination, with further extraction resulting in little, if any, change in the proportion of FFA in the lipid extract. Only a small volume of the hexane extract ( approximately 20 mL) yielded sufficient material for the SB-ATR/FTIR analysis. Thus, by shortening the extraction time and taking a small sample so as to reduce solvent removal time, the SB-ATR/FTIR procedure provides a very simple and rapid means of determining the FFA content of poultry feed lipids.     

Formation of conjugated linoleic acids in soybean oil during hydrogenation with a nickel catalyst as affected by sulfur addition

The effects of sulfur addition on the formation of conjugated linoleic acid (CLA) isomers were studied during the hydrogenation of soybean oil with a nonselective type nickel catalyst. Sulfur addition greatly promoted CLA formation in soybean oil during hydrogenation. As the amount of sulfur increased to a certain level, the maximal quantity of CLA in soybean oil during hydrogenation increased greatly. However, further increase in sulfur addition above the certain level decreased CLA formation. The optimal sulfur level for the promotion of CLA formation differed greatly with the amount of nickel used. It was of great interest to find that the optimal ratio of sulfur to nickel for the promotion of CLA formation was always 0.06:1, regardless of the nickel amount used. At the same ratio of sulfur to nickel, higher nickel content induced significantly higher production of CLA (p < 0.05). At the optimal sulfur to nickel ratio, an increase in the nickel amount from 0.05 to 0.15% produced approximately 1.5 times higher levels of CLA during hydrogenation under the tested conditions. The CLA isomer compositions were greatly affected by both sulfur addition and amounts of nickel used for treatment. This is the first report of the possibility that the total quantity of CLA and their isomer composition could be manipulated during hydrogenation by controlling the amounts of sulfur and nickel.     

加工工艺对油茶籽油氧化稳定性及酚类物质含量的影响

为了探究加工工艺对油茶籽油营养品质的影响,了解油酚类物质在加工工艺中的变化规律,该文从油茶籽油加工企业的生产线中取样,对压榨毛油和浸出毛油精炼工艺以及压榨毛油适度精炼工艺等不同工艺中油茶籽油中总酚、多酚组成及含量、抗氧化活性系数和油的氧化诱导时间等指标进行了测定。压榨毛油精炼工艺包括水洗、脱色、脱臭、冬化;浸出毛油精炼工艺包括碱炼、水洗、脱色、脱臭、冬化;压榨毛油适度精炼工艺包括脱胶、碱炼、水洗、冬化等工艺步骤。结果显示:压榨油茶籽毛油中总酚平均质量分数为103.06μg/g,显著高于浸出油茶籽油48.52μg/g(P0.05);油茶籽毛油经过精炼工艺后总酚和总酚的抗氧化活性均呈现下降趋势,三种精炼工艺后油茶籽油中总酚分别下降了88.9%,86.7%和63.81%,总酚的抗氧化活性分别下降了88.3%,93.51%和83.25%。适度精炼相比普通精炼对于保留油茶籽油中的多酚有明显优势,前者精炼后油茶籽油中总酚及其抗氧化活性系数分别为37.82μg/g和9.33%,后者仅为11.41μg/g和4.71%。通过高效液相色谱测定发现,浸出油茶籽毛油精炼后仅含有少量肉桂酸,压榨毛油传统精炼后压榨油茶籽油中测到苯甲酸、芦丁和肉桂酸等3种多酚的质量分数分别为4.7、1.58和0.22μg/g,压榨毛油适度精炼后的油茶籽油中测到了单宁酸、绿原酸、表儿茶素等9种多酚,其中单宁酸和绿原酸的质量分数最高,分别为4.57和3.26μg/g;适度精炼后油茶籽油氧化诱导时间从初始的8.56 h增加到11.66 h,增加了26.63%;压榨毛油传统精炼后油茶籽油氧化诱导时间从8.14 h增加到10.42 h,增加了21.83%。研究结果表明,压榨毛油适度精炼相比传统精炼工艺对于保留油茶籽油中多酚等成分有明显优势,所生产的茶油具有更强的氧化稳定性。研究结果为油茶籽油中营养成分的保留提供了途径,为油茶籽油加工工艺选择提供参考。     

Analysis of monoglycerides, diglycerides, sterols, and free fatty acids in coconut (Cocos nucifera L.) oil by 31P NMR spectroscopy

Phosphorus-31 nuclear magnetic resonance spectroscopy ( (31)P NMR) was used to differentiate virgin coconut oil (VCO) from refined, bleached, deodorized coconut oil (RCO). Monoglycerides (MGs), diglycerides (DGs), sterols, and free fatty acids (FFAs) in VCO and RCO were converted into dioxaphospholane derivatives and analyzed by (31)P NMR. On the average, 1-MG was found to be higher in VCO (0.027%) than RCO (0.019%). 2-MG was not detected in any of the samples down to a detection limit of 0.014%. On the average, total DGs were lower in VCO (1.55%) than RCO (4.10%). When plotted in terms of the ratio [1,2-DG/total DGs] versus total DGs, VCO and RCO samples grouped separately. Total sterols were higher in VCO (0.096%) compared with RCO (0.032%), and the FFA content was 8 times higher in VCO than RCO (0.127% vs 0.015%). FFA determination by (31)P NMR and titration gave comparable results. Principal components analysis shows that the 1,2-DG, 1,3-DG, and FFAs are the most important parameters for differentiating VCO from RCO.     

Effect of soy milk characteristics and cooking conditions on coagulant requirements for making filled tofu

The amount of coagulant added to soy milk is a critical factor for tofu-making; particularly it affects the textural properties of tofu. Earlier research indicated that the critical point of coagulant concentration (CPCC) is a characteristic parameter of soy milk and could be used as an effective indicator of optimal coagulant concentration (OCC) for making filled tofu. The objective of this study was to investigate the possible correlations between CPCC and the characteristics of soy milk made from various soybean samples and the effect of soy milk cooking and dilution conditions on CPCC. CPCC was determined by a titration method. Calcium chloride and magnesium chloride were used as coagulants. Soy milk characteristics including solid, protein, phytate, pH, titratable acidity, mineral content, and 11S/7S protein and these characteristics as affected by heating rate, heating time, and sequence of dilution and heating were studied. The results showed that the CPCC was significantly (p < 0.05) positively correlated with phytate content (grams per gram of protein), pH, and 7S protein content but negatively correlated with protein content, 11S protein content, 11S/7S ratio, titratable acidity, and original calcium content. Within the same soybean material, more proteins required more coagulant, but higher protein concentration during cooking resulted in less coagulant required by each gram of protein during coagulation. The CPCC decreased with increasing soy milk heating time or decreasing heating rate. The sequence of heating and diluting for preparing soy milk also had an effect on CPCC.     

Characterization of Moringa oleifera variety Mbololo seed oil of Kenya.

The oil from Moringa oleifera variety Mbololo seeds from Kenya was extracted using three different procedures including cold press (CP), extraction with n-hexane (H), and extraction with a mixture of chloroform/methanol (50:50) (CM). The oil concentration ranged from 25.8% (CP) to 31.2% (CM). The density, refractive index, color, smoke point, viscosity, acidity, saponification value, iodine value, fatty acid methyl esters, sterols, tocopherols (by HPLC), peroxide value, and at 232 and 270 nm and the susceptibility to oxidation measured with the Rancimat method were determined. The oil was found to contain high levels of unsaturated fatty acids, especially oleic (up to 75.39%). The dominant saturated acids were behenic (up to 6. 73%) and palmitic (up to 6.04%). The oil was also found to contain high levels of beta-sitosterol (up to 50.07%), stigmasterol (up to 17.27%), and campesterol (up to 15.13%). alpha-, gamma-, and delta-tocopherols were detected up to levels of 105.0, 39.54, and 77. 60 mg/kg of oil, respectively. The induction period (at 120 degrees C) of M. oleifera seed oil was reduced from 44.6 to 64.3% after degumming. The M. oleifera seed oil showed high stability to oxidative rancidity. The results of all the above determinations were compared with those of a commercial virgin olive oil.     

Comparison of analytical methodologies based on 1h and 31p NMR spectroscopy with conventional methods of analysis for the determination of some olive oil constituents

The present study was designed to assess the agreement between analytical methodologies based on 1H and 31P NMR spectroscopy and conventional analytical methods (titration, gas chromatography, and high performance liquid chromatography) for measuring certain minor and major constituents (free acidity, fatty acids, iodine value, and phenolic compounds) of olive oil. The standard deviations of the NMR method were comparable to those of the conventional methods, except perhaps those of the total hydroxytyrosol and total tyrosol. Linear regression analyses showed strong correlations between NMR and conventional methods for free acidity, total hydroxytyrosol, total tyrosol, total diacylglycerols, (+)-pinoresinol, (+)-1-acetoxypinoresinol, and apigenin; good correlations for linoleic acid, free hydroxytyrosol, and free tyrosol; and weak correlations for oleic acid, linolenic acid, saturated fatty acids, and luteolin. Furthermore, a method comparison study was conducted and the agreement between NMR and conventional methods was evaluated by using the Bland and Altman statistical analysis. The distribution of the data points in the bias plot showed that 96.4% and 100% of the measurements of free acidity and iodine value, respectively, were within the limits of agreement of the two methods. For the remaining constituents of olive oil, the percentage of measurements, located within the limits of agreement, ranged from 94% to 98.5%.     

Influence of postharvest hot water treatment on nutritional and functional properties of kumquat (Fortunella japonica Lour. Swingle Cv. Ovale) fruit

The present study investigated the influence of a hot water dip (HWD) for 2 min at 50 degrees C, a standard and effective treatment for postharvest decay control of citrus fruit, on the nutritional and health-related properties of kumquats. The results show that most of the parameters examined, including titratable acidity, soluble solids content, maturity index, glucose, fructose, sucrose, ascorbic acid, dehydroascorbic acid, alpha- and gamma-tocopherols, beta-carotene, zeaxantin, rhoifolin, and antioxidant activity, were not significantly affected by treatment. The levels of beta-cryptoxanthin, narirutin, and total flavonoids increased after HWD, whereas lutein and total phenols decreased. The concentration of the essential oil and the relative percentage of the individual components of the essential oil were not affected by HWD except for the minor compound p-menta-1,5-dien-1-ol, which increased after HWD. After storage, lower levels of glucose, total sugars, beta-carotene, beta-cryptoxanthin and lutein were recorded in HWD fruit. A decrease in antioxidant activity and increases in alpha-tocopherol and total vitamin E were found both in control and HWD fruit. The influence of HWD at 50 degrees C for 2 min on individual nutraceuticals and health-related properties was thus generally low and may depend on storage conditions.     

Antioxidant evaluation and oxidative stability of structured lipids from extravirgin olive oil and conjugated linoleic acid

Structured lipid (SL) was synthesized from extravirgin olive oil (EVOO) and conjugated linoleic acid (CLA) via a lipase-catalyzed reaction. CLA provides a variety of health benefits, but it is not consumed in free fatty acid form. The synthesized SL olive oil contained 42.5 mol % CLA isomers, and the major isomers were cis-9,trans-11-CLA (16.9 mol %) and trans-10,cis-12-CLA (24.2 mol %). The antioxidant activity determined by the radical scavenging capacity with the 2,2-diphenyl-1-picrylhydrazyl radical was lower in SL olive oil than in EVOO. The oxidative stability was also lower in SL olive oil since it had a higher peroxide value, rho-anisidine value, and 2-thiobarbituric acid reactive substances values during 20 days of storage at 60 degrees C. This observation could be due to the reduction in the natural phenolic compounds (97%) and tocopherols (56%), and the incorporated CLA with two conjugated double bonds in the SL olive oil. The oxidative stability of SL olive oil was increased by added rosemary extracts at concentrations of 100, 200, and 300 ppm. The present study suggests that the SL olive oil may be a suitable way to incorporate or deliver CLA into human diets. However, the addition of a proper antioxidant would be required for improving its oxidative stability.     

Methylation methods for the quantitative analysis of conjugated linoleic acid (CLA) isomers in various lipid samples

Precise methylation methods for various chemical forms of conjugated linoleic acid (CLA), which minimize the formation of t,t isomers and allylmethoxy derivatives (AMD) with the completion of methylation, were developed using a 50 mg lipid sample, 3 mL of 1.0 N H(2)SO(4)/methanol, and/or 3 mL of 20% tetramethylguanidine (TMG)/methanol solution(s). Free CLA (FCLA) was methylated with 1.0 N H(2)SO(4)/methanol (55 degrees C, 5 min). CLA esterified in safflower oil (CLA-SO) was methylated with 20% TMG/methanol (100 degrees C, 5 min), whereas CLA esterified in phospholipid (CLA-PL) was methylated with 20% TMG/methanol (100 degrees C, 10 min), followed by an additional reaction with 1.0 N H(2)SO(4)/methanol (55 degrees C, 5 min). Similarly, CLA esterified in egg yolk lipid (CLA-EYL) was methylated by base hydrolysis, followed by reaction with 1.0 N H(2)SO(4)/methanol (55 degrees C, 5 min). These results suggest that for the quantitative analysis of CLA in lipid samples by GC, proper methylation methods should be chosen on the basis of the chemical forms of CLA in samples.