Collaborative test of determination of iodine value in fish oils. 2. Carbon tetrachloride or cyclohexane/acetic acid as solvents

Nine laboratories participated in a collaborative test to determine the iodine value (IV) of eight samples of fish oil (four with IV<150; four with IV>150) with either carbon tetrachloride (AOCS Official Method Cd 1–25) or cyclohexane/acetic acid (AOCS Recommended Practice Cd 1d-92) as solvent and 1 h of reaction time. Laboratories received coded duplicate samples (hidden duplicates) and carried out duplicate determinations on each oil by each method (open duplicates). Replacing carbon tetrachloride with cyclohexane/acetic acid resulted in similar mean values for both low- and high-IV oils and similar estimates of repeatability and reproducibility. The repeatability standard deviation (s _r ), based on hidden duplicates, with carbon tetrachloride and cyclohexane/acetic acid were 1.71 and 1.55, respectively. The corresponding reproducibility standard deviations were 1.81 and 1.98.     

A Greener Alternative Titration Method for Measuring Acid Values of Fats,Oils, and Grease

A greener alternative method is proposed for measuring acid values (AV) of fats, oils, and grease (FOG) based on visual titration. Compared with Official Method Cd 3d-63 of the American Oil Chemists' Society (AOCS), this greener alternative method can eliminate the use of toluene, which in turn reduces toxicity and cost. A total of 44 samples of yellow and brown grease with AV ranging from 0.13 to 170.37 (mg KOH) g⁻¹ were titrated using both methods. The alternative titration method can provide accurate and reliable results to determine the AV of FOG by various statistical analyses including repeatability, linear regression, f-test, t-test, and method accuracy calibration with AOCS Cd 3d-63. This low-cost method can be recommended for routine titration in research and development, and in biodiesel plants for most FOG samples.     

A Visual Titration Method for the Determination of the Acid Number of Oils and Fats: a Green Alternative

A visual and green titrimetric method is proposed for the determination of the acid number of oils and fats. The sample is dissolved in a water–ethanol mixture (1:1 v/v) and titrated with a 0.02?mol L^?1 aqueous NaOH solution. Phenolphthalein is used as acid-basic indicator. Oils of canola, sunflower, linseed, castor, corn and soy as well as swine lard, a total of twelve real samples, were analyzed. The results obtained were compared with those from the application of the AOCS Cd 3d-63 method, through a statistical linear regression procedure at the 95?% confidence level and no evidence for systematic differences between the two sets of results was observed. The mean RSD (relative standard deviation ) for the proposed procedure is 4.8?%, whereas that of the official method was found to be 5.4?%.     

Regulatory Infrared Spectroscopic Method for the Rapid Determination of Total Isolated Trans Fat: A Collaborative Study

Using attenuated total reflection-Fourier transform infrared (ATR-FTIR) spectroscopy, collaborating scientists in ten different laboratories measured (in duplicate) the total trans fat content of ten fat or oil test samples, two of which were blind duplicates. The procedure used entailed measuring the height of the negative second derivative of the IR absorption band at 966 cm^?1. This absorption is attributed to the C?CH deformation vibration that is characteristic of isolated (non-conjugated) double bonds with the trans configuration. The precision of ATR-FTIR results in this international collaborative study was satisfactory and led to the approval of this validated procedure as official method AOCS Cd 14e-09 in late 2009. This official method is also suitable for analysis of total isolated trans fat and oil products containing, or supplemented with, trans conjugated linoleic acid (CLA) isomers. Although this method does not require derivatization of the oil or fat test materials, as required for GC, fats and oils in foods must be extracted with organic solvents before analysis. This method is also rapid (5 min) and does not require any weighing or quantitative dilution of unknown neat fat or oil test samples in any solvent. The AOCS Cd 14e-09 method is suitable for determination of test samples with zero trans fat, which is defined according to the US labeling regulations as 0.5 g trans fat per serving or 1.8% trans fat, as a percentage of total fat.     

Evaluation of SafTestTM methods for monitoring frying oil quality

The feasibility of applying methods developed by Safety Associates, Inc., to monitor oil degradation products, including malondialdehydes (AldeSafe^TM), FFA (FASafe^TM), and peroxides (PeroxySafe^TM), in fresh and heat-abused deep-fat frying oil was evaluated. Based on performance qualification studies, the AldeSafe method was the most suitable SafTest^TM assay for monitoring the quality of frying oil because of its high accuracy, precision, linearity, and reproducibility, and low detection/quantitation limits. A strong correlation (r=0.924) between the AldeSafe method and its counterpart, AOCS Official Method Cd 19-90, also supported the suitability of the SafTest method for monitoring oil quality. Moreover, the FASafe method had a moderately strong relationship with AOCS Official Method Ca 5a-40 (r=0.761). Our studies suggest that this test can be applied for monitoring frying oil; however, certain method performance limitations must be considered for routine analysis purposes. In contrast, the PeroxySafe method probably should not be used to monitor heat-abused oil without further development because of high variability, low accuracy, and low correlation (r=0.062) with the AOCS Official Method Cd 8-53 assay.     

Production of free fatty acids in safflower seeds by fungi

In 1975, free fatty acids (FFA) reached 0.50% in hand-sampled seeds (achenes) from the Sacramento Valley but never exceeded 0.10% in samples from the San Joaquin Valley. High levels of FFA in safflower seeds from the Sacramento Valley were attributed to seed-borne fungi, especiallyAlternaria sp., having a lipolytic action on safflower oil. Greenhouse inoculations on representative safflower cultivars reduced yields and oil content while increasing FFA. In vitro experiments using safflower oil-potato dextrose agar medium showed that theAlternaria species and four other fungi isolated from safflower had lipolytic activity. TheAlternaria sp. produced 3.90% FFA after 20 days of growth on this medium. Separation of the FFA by thin layer chromatography and characterization by gas liquid chromatography indicated that fatty acids are released by all five fungi in proportions similar to those in safflower oil. Presented in part at the AOCS Meeting, New York, May 1977.     

FTIR estimation of free fatty acid content in crude oils extracted from damaged soybeans

A user-interactive computer-assisted Fourier transform infrared (FTIR) method has been developed for estimation of free fatty acids (FFA) in vegetable oil samples by deconvolution of the infrared (IR) absorbances corresponding to the triglyceride ester and FFA carbonyl bonds. Peak areas were used to determine FFA as a percentage of the total carbonyl areas in weighed standards of refined, bleached, deodorized soybean oil containing from 0 to 5% added oleic acid. These data for percent FFA by FTIR were compared to corresponding FFA data obtained by two titration methods-the AOCS Official Method Ca 5a-40 and the Official Method with a slight modification. Correlation coefficients were 0.999 for the Ca 5a-40, 0.999 for the modified and 0.989 for the FTIR methods. FFA in samples of crude soybean oils extracted from damaged beans (0.5 to 2.1% FFA) were measured by FTIR and compared to data obtained by titration of the same samples (correlation coefficient, 0.869). To whom correspondence should be addressed at National Center for Agricultural Utilization Research, Agriculture Research Service, U.S. Department of Agriculture, 1815 North University Street, Peoria, IL 61604. ¹The mention of firm names or trade products does not imply that they are endorsed or recommended by the U.S. Department of Agriculture over other firms or similar products not mentioned.     

New FTIR method for the determination of FFA in oils

A rapid, practical, and accurate FTIR method for the determination of FFA in edible oils was developed. Analogous to the AOCS titration procedure, the FTIR FFA determination is effected by an acid/base reaction but directly measures the product formed rather than utilizing an end point based on an electrode potential or color change. A suspension of a weak base, potassium phthalimide (K-phthal) in 1-propanol (1-PrOH), is used to convert the FFA present in oils to their carboxylate salt without causing oil saponification, and differential spectroscopy is used to circumvent matrix effects. Samples are first diluted with 1-PrOH, then split, with one-half treated with the K-phthal reagent and the other half with 1-PrOH (blank reagent), their spectra collected, and differential spectra obtained to ratio out the invariant spectral contributions from the oil sample. Quantification of the percentage of FFA in the oil, expressed as %oleic acid, based on measurement of the peak height of the ν (COO⁻) absorption of the FFA salt formed, yielded a calibration with an SE of <0.020% FFA over the range of 0–4%. The method was validated by standard addition and the analysis of Smalley check samples, the results indicating that the analytical performance of the FTIR procedure is as good as or better than that of the standard titrimetric procedure. As structured, the FTIR procedure is a primary method, as calibration is not dependent on reference values provided by another method, and has performance criteria that could lead to its consideration as an instrumental AOCS procedure for FFA determination. The FTIR portion of the analysis is automatable, and a system capable of analyzing ∼60 samples/h was developed that could be of benefit to laboratories that carry out a large number of FFA analyses per day.     

Determination of iodine value with a fourier transform-near infrared based global calibration using disposable vials: An international collaborative study

A method for the determination of iodine value (IV) by Fourier transform-near infrared (FT-NIR) spectroscopy was developed and evaluated in an international collaborative study. The FT-NIR analyzer employed in this work uses disposable vials for sample handling and incorporates validation protocols designed to ensure that the calibration will give accurate results from analyzer to analyzer and stability over time without any further calibration development work. The global IV calibration was developed from over 1,200 animal, marine, and vegetable oils and fats, which were obtained on a number of different instruments worldwide. The Standard Error of Cross Validation measured from a range of 0–190 IV varied from ±0.2–1.4 IV (1 sigma). The repeatability for all models was on the order of 0.1 IV, which states that most of the error was inherited from the primary methods. Finally, an international interlaboratory study was carried out with 16 samples obtained from the AOCS Smalley Laboratory Proficiency Program and an oil processor. The average reproducibility error in any one lab was better than 0.15, and the average reproducibility between labs was better than 0.33. An uncertainty of 0.45 was calculated from the average FT-NIR values obtained from the collaborative study vs. the AOCS Certified Wijs method (Cd 1d-92).     

Determination of optimal conditions for selected adsorbent combinations to recover used frying oils

The treatment of frying oils with adsorbents could practically extend the frying life of oils. Combined synthetic adsorbent treatment of used frying oils was studied the first time. The combinations of four commonly used filter aids: Britesorb (Br), Hubersorb 600 (HB), Frypowder (Fr), and Magnesol (Ma) were evaluated for frying oil recovery. AOCS official methods were used to evaluate their adsorptiveness, including free fatty acids (FFA), conjugated diene value, total polar components, oxidative stability index (OSI), and absorbance at 420 nm. The selected combinations HB+Ma+Fr and HB+Ma+Br exhibited consistent high recovery abilities on various used oil samples. A 3, 3, and 2% HB, Ma, and Fr, respectively, for the first combination (F), and 2, 3, and 2% HB, Ma, and Br, respectively, for the second combination (B) were the most effective. The optimal treatment duration was 6–9 min and 3–6 min for combinations F and B, respectively, which reduced FFA by 82.6–87.6%, absorbance by 26.8–32.6%, and Foodoil Sensor readings by 5.6–8.6%. Addition of antioxidant, such as 50 ppm butylated hydroxytoluene and 50 ppm propyl gallate, increased the OSI value by 48.9–80.8%. Such adsorbent combinations may be used in practical operation to extend frying life of frying oils and improve the healthy aspects of used frying oils.     

Estimating the content of free fatty acids in high‐oleic sunflower seeds by near‐infrared spectroscopy

The content of free fatty acids (FFA) in vegetable oils represents an important quality factor in oil crops. The objective of the investigation was to develop a near‐infrared (NIR) calibration for estimating the FFA content in high‐oleic sunflower seeds. A sample set of different varieties from the harvest of 2004 as well as of 2005 from two locations in Germany was used; additionally seeds from 2003 were stored under unsuitable conditions to obtain samples utilised for calibration with an extended FFA range. A direct titration method for FFA determination was developed and adjusted to the official AOCS method. The modified method is sufficiently reliable, much faster than the AOCS method and therefore suitable for use in the calibration of NIR spectrometers. The developed NIR spectroscopy (NIRS) calibration was calculated with a modified partial least square algorithm, standard normal variate and detrend scatter correction and the 2^nd derivative of the spectra of ground sunflower seeds. The standard error of prediction of the validated calibration was 0.20, and the multiple coefficient of determination (RSQ_val) reached 0.94. The obtained results demonstrated clearly the efficiency and how cost effective the NIRS method is for the estimation of FFA content in sunflower seeds.     

Activated Kaolin Minerals as Bleaching Clays for Prolonging the Useful Life of Palm Oil in Industrial Frying Operations

During frying operations, vegetable oils break down and compounds with undesirable flavors are produced. Various procedures have been developed to extend the useful life of frying oils, including treatment with bleaching clays. In this article, we describe the activation of kaolin minerals by a combination of grinding and chemical treatments, and report their performance in removing breakdown products generated in palm oil that had been used for 20 hours continuous frying. There was little influence of the original kaolin mineral form on the ability to reduce the free fatty acid (FFA) contents, and grinding only changed FFA reduction from ∼32% to ∼36%. However, much greater FFA reductions were obtained after chemical treatment of the ground clays, and the best performing kaolin product gave similar FFA reduction to a commercial bleaching clay (∼76% and ∼77%, respectively). This activated kaolin also produced a reduction in viscosity at 40 °C from ∼73 to 48.4 cSt (compared to 45.5 cSt in the unused oil), and in the peroxide value from 30.0 to 22.0 meq/kg (compared to 10.0 meq/kg in the initial oil). Thus, activated kaolin samples represent a cheap and convenient alternative to conventional bleaching clay for improving common quality parameters in used palm oil, although we also found that the optimum kaolin preparation conditions were different from those that have been reported for raw rice bran oil refining.     

Collaborative test of determination of iodine value in fish oils. 1. Reaction time and carbon tetrachloride or cyclohexane as solvents

Twenty-two laboratories participated in a collaborative test to determine the iodine value (IV) of eight samples of fish oil (four with IV<150, four with IV>150) with either carbon tetrachloride (AOCS Official Method Cd 1–25) or cyclohexane (AOCS Recommended Practice Cd 1b-87) as solvent and either 1 or 2 h of reaction time. Laboratories received coded duplicate samples (hidden duplicates) and carried out duplicate determinations on each oil by each solvent-time combination (open duplicates). Replacing carbon tetrachloride with cyclohexane resulted in a lower IV (P<0.001). The decrease averaged 1.6 IV units for low-IV oils and 3.8 IV units for high-IV oils; this difference in response of 2.2 IV units between low- and high-IV oils was significant (P<0.001). Increasing the reaction time had a relatively small effect (0.34±0.18). There was no interaction of reaction time with solvent or oil type. Cyclohexane caused emulsions, which made it difficult to titrate residual iodine and thus increased the variability of the determination. The repeatability standard deviations (s _r ), based on hidden duplicates, for 1-h reaction time with carbon tetrachloride and cyclohexane were 2.17 and 3.35, respectively. The corresponding reproducibility standard deviations were 2.73 and 4.53.     

Comparative study of methods of determining oil content of sunflower seed

An extraction-gravimetric method (AOCS Official Method Ai 3-75) was compared with 2 instrumental techniques, near-infrared reflec-tance (NIR) spectroscopy and wide-line nuclear magnetic resonance (NMR), for the determination of the oil content of oilseed-type hybrid sunflower seed. Eight sunflower seed samples of varying oil contents, replicated 5 times, were analyzed by the 3 procedures. The overall mean oil contents and standard deviations for the 8 samples were: AOCS method, 44.5% ± 0.33%; NMR, 44.8% ± 0.27%; and NIR, 44.2% ± 0.81%. Analysis of variance of the means of the 3 methods of analysis indicated no difference (p>0.05) in oil content due to the method. However, there was a difference (p>0.001) in total oil content due to replicated analyses of the same sample with the NIR method. With the AOCS and NMR methods, no effect (p>0.05) of replicated analyses of the same sample was found. The NMR method was more precise and repro-ducible than the other 2 methods. Although the NIR mean oil contents were not significantly different from the means of the other 2 methods, the coefficient of variations for all samples were consistently higher for the NIR analyses than for the AOCS and NMR analyses.     

Analysis of Sesamin, Asarinin, and Sesamolin by HPLC with Photodiode and Fluorescent Detection and by GC/MS: Application to Sesame Oil and Serum Samples

New sensitive and specific analytical methods are needed for the analysis of sesamin, asarinin, and sesamolin in sesame seed oils, sesame dietary supplements, as well as in serum samples from clinical studies involving sesamin, asarinin, and sesamolin. The objective of this study was to develop a high performance liquid chromatographic (HPLC) method with photodiode array and fluorescent detectors and a gas chromatography mass-spectrometry (GC/MS) method for the analysis of sesamin, asarinin (episesamin), and sesamolin in sesame oil and in serum samples. Sesame oil samples were extracted with methanol whereas the serum samples were extracted with ethyl acetate or n-hexane. The individual lignans were analyzed by HPLC using reversed phase C18 columns. Analytical recoveries of sesamin, asarinin, and sesamolin from sesame oil were 92–94?% with two extractions. Recoveries from serum ranged from 87 to 97?%. The limit of quantitation with the fluorometric detector was 0.1?ng compared to 0.1?μg with the PDA detector. The concentrations of sesamin, asarinin, and sesamolin in Orchids and Sigma sesame oil were 0.4, 0, and 0.15?% and 0.19, 0.09, and 0?%, respectively. The identities of the individual lignans obtained by HPLC were confirmed by GC/MS and the concentrations of sesamin, asarinin, and sesamolin obtained with the fluorometric detector correlated with those obtained by GC/MS (r ²?=?0.94, P?<?0.001). The HPLC and GC/MS methods permit simple and efficient procedures for the analysis of sesamin, asarinin, and sesamolin in sesame oil samples as well as in serum samples.     

A modified method for determining free fatty acids from small soybean oil sample sizes

A modification of the AOCS Official Method Ca 5a-40 for determination of free fatty acids (FFA) in 0.3 to 6.0-g samples of refined and crude soybean oil is described. The modified method uses only about 10% of the weight of oil sample, alcohol volume, and alkali strength recommended in the Official Method. Standard solutions of refined and crude soybean oil with FFA concentrations between 0.01 and 75% were prepared by adding known weights of oleic acid. The FFA concentrations, determined from small sample sizes with the modified method, were compared with FFA percentages determined from larger sample sizes with the Official Method. Relationships among determinations obtained by the modified and official methods, for both refined and crude oils, were described by linear functions. The relationship for refined soybean oil had an R ² value of 0.997 and a slope of 0.99±0.031. The values for crude soybean oil are defined by a line with R ²=0.9996 and a slope of 1.01±0.013.     

Optimization of Enzymatic Hydrolysis of Sacha Inchi Oil using Conventional and Supercritical Carbon Dioxide Processes

Sacha inchi (Plukenetia volubilis) oil has high polyunsaturated fatty acids content. The hydrolysis of this oil is an efficient way to obtain desirable free fatty acids (FFA). The optimization of parameters was carried out according to the maximum production of FFA using two enzymatic hydrolysis processes. The effect of enzyme concentration (5–40 % based on weight of oil), temperature (40–60 °C), and oil:water molar ratio (1:5–1:70) were studied for the conventional enzymatic hydrolysis process, while pressure (10–30 MPa) and oil:water molar ratio (1:5–1:30) were studied for the enzymatic hydrolysis in supercritical carbon dioxide (SC-CO₂) media. The hydrolysis in SC-CO₂ media resulted in higher production of FFA (77.98 % w/w) at 30 MPa and an oil:water molar ratio equal to 1:5 compared to the conventional process (68.40 ± 0.98 % w/w) at 60 °C, oil:water molar ratio equal to 1:70, and 26.17 % w/w, enzyme/oil. The only significant parameter on the production of FFA for conventional enzymatic hydrolysis was enzyme concentration, while for the hydrolysis in SC-CO₂ media both pressure and the molar ratio of oil:water were significant. Lipid class analyses showed that with both methods, FFA, monoglycerides, and diglycerides content in the final product increased compared to pure oil, while triglycerides content decreased. Fatty acid composition analysis showed that the content of fatty acids in the FFA form were similar to their triglyceride form.     

Esterification of sludge palm oil using trifluoromethanesulfonic acid for preparation of biodiesel fuel

Trifluoromethanesulfonic acid (TFMSA) was used to reduce the high free fatty acids (FFA) content in sludge palm oil (SPO). The FFA content of SPO was converted to fatty acid methyl ester (FAME) via esterification reaction. The treated sludge palm oil was used as a raw material for biodiesel production by transesterification process. Several working parameters were optimized, such as dosage of catalyst, molar ratio, reaction temperature and time. Less than 2% of the FFA content was the targeted value. The results showed that the FFA content of SPO was reduced from 16% to less than 2% using the optimum conditions. The yield of the final product after the alkaline transesterification was 84% with 0.07% FFA and the ester content was 96.7%. All other properties met the international standard specifications for biodiesel quality such as EN 14214 and ASTM D6751.     

Effect of temperature programming on the performance of urea inclusion compound-based free fatty acid fractionation

The effect of cooling rate on the degree of removal of saturated acyl groups from FFA derived from canola oil and the isolation of di- and polyunsaturated acyl groups from FFA derived from vegetable and fish oil, respectively, during urea inclusion compound (UIC)-based fractionation was investigated. Traditionally, slow cooling has been used (ca.−1°C min⁻¹). A more rapid cooling rate (−47°C min⁻¹) produced UIC crystals of similar morphology and thermodynamic properties, but of a size an order of magnitude smaller than the UIC formed during slow cooling. Fractionations used only renewable materials (urea, FFA, and 95% ethanol as solvent) and benign operating conditions (ambient pressure, 25–75°C, and neutral pH). When the recovery of FFA (in the solvent-rich phase) was relatively high (>60%), the selectivity of UIC-based fractionation toward the inclusion of saturated FFA and against polyunsaturated FFA was not affected by the cooling rate. In contrast, when the FFA recovery was low, representing cases in which an increase of the PUFA purity is a more important economic goal, a slower cooling rate resulted in a significantly greater discrimination against PUFA groups, hence to a FFA product with a measurably greater purity. This paper was presented at the 96th AOCS Anual Meeting and Expo, Salt Lake City, Utah, on May 4, 2005.     

A Highly Efficient Automated Flow Injection Method for Rapid Determination of Free Fatty Acid Content in Corn Oils

In the present study, a new flow injection (FI) methodology for determination of free fatty acid (FFA) content in corn oil samples is proposed. The proposed method is based on monitoring the absorbance changes at 580 nm, λ_max, as a result of neutralization of FFA in oil samples by KOH in the reagent. The analyses were performed with a single‐line FI manifold system created by modification of high performance liquid chromatography. The main parameters, such as sample and reagent volumes, reaction coil, reagent concentration and temperature were all optimized. With the newly developed sampling strategy, the oil sample and reagent at micro level were directly injected together without any pre‐treatment. The proposed flow injection analysis method was validated statistically and it was found to be linear (between 0.09 and 1.50 FFA %), accurate (recovery 87.19–122.22 %), and precise (relative standard deviation <1 % for both intra‐day and inter‐day precision). The limit of detection and limit of quantification were found to be 7.41 × 10^?3 and 2.24 × 10^?2 oleic acid %, respectively. The results were also compared with those obtained by the American Oil Chemists Society (Ca‐5a‐40) method using statistical t and F tests, and a significant difference was not observed between the methods at the 95 % confidence level. These results strongly suggest that this method is suitable for automated routine analysis of FFA in edible oils due to its simplicity, reliability, speed, and economy of solvents and sample.