Journal of Molecular Structure 482483 (1999) 551556

Determination of unsaturation grade and trans isomers generated during thermal oxidation of edible oils and fats by FTIR
zquita Lo pez b, M.C.M. Moya Moreno a,*, D. Mendoza Olivares b, F.J. Ame J.V. Gimeno Adelantado a, F. Bosch Reig a
b a Department of Analytical Chemistry, Faculty of Chemistry, University of Valencia, C/Dr. Moliner 50, 46100 Burjassot, Valencia, Spain Department of Chemistry, Instrumental Laboratory, Faculty of Chemistry, University of Guanajuato, Noria Alta, 36050 Guanajuato, Mexico

Received 24 August 1998; received in revised form 30 November 1998; accepted 30 November 1998

Abstract The oxidative deterioration of culinary oils and fats during episodes of heating associated with normal usage (80C300C, 2040 min) was monitored by FTIR spectroscopy. The thermal oxidation of polyunsaturated fatty acids during heating was studied by the determination of unsaturation percentage and trans isomers at various temperatures and heating times. Oils frequently used in food frying such as olive oil, sunower oil, corn oil and seeds oil (sunower, safower and canola seed), and lard were studied. The Absorbance Correction Method is proposed to correct the spectral interference and allows the analytic use of signal which would not be initially valid for quantitative analysis. The results show that there is a decrease in unsaturation and an increase in trans isomers starting at 150C and becomes more pronounced at temperatures around 250C. This variation in unsaturation grade and conformation provides evidence of the transformation of essential polyunsaturated fatty acids and subsequent decrease in the oils nutritional value. 1999 Elsevier Science B.V. All rights reserved.
Keywords: Edible oils; Polyunsaturated fatty acids; Trans isomers; FTIR

1. Introduction Oils and fats are an important part of the human diet and more than 90% of global production is used as food or as ingredients in food products. Oils and fats in the diet are a rich source of energy, they contain certain fatty acids, which are nutritionally indispensable and their functional and textural characteristics add to the avour and acceptability of many natural and processed foods. The polyunsaturated fatty acids (PUFA) which cannot be synthesised by the human
* Corresponding author. Tel.: 34 6 386 4533; fax: 34 6 386 4436. E-mail address: montse.moya@uv.es (M.C.M. Moya Moreno)

body must be present in the diet to avoid symptoms caused by shortage. They can be considered as vitamin factors and linoleic, and linolenic acids must be considered specically as they are polyunsaturated fatty acids which are necessary for the normal development and functioning of human tissues and are known as essential fatty acids [1]. Oils and fats begin to decompose from the moment they are isolated from their natural environment when changes occur causing a disagreeable taste and smell. Atmospheric oxidation is the most important cause of deterioration in fats. This oxidative rancidity is accelerated by exposure to heat, light, humidity and the presence of trace transition metals [2]. The intense frying of oil causes an oxidizing thermal degradation

0022-2860/99/$ - see front matter 1999 Elsevier Science B.V. All rights reserved. PII: S0022-286 0(98)00937-5

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with the formation of decomposition products [36], and change in physical properties [7]. Spectroscopic methods can be employed to evaluate deterioration in oils subjected to intense heat [8]. Fourier Transform Infrared (FTIR) Spectroscopy provides a quick and accurate way of evaluating thermal degradation in these lipids. This article reports a methodology to evaluate the variation in composition in edible oils (sunower oil, corn oil, olive oil and seeds oil) and lard when it is subjected to intense heat (80C300C, 2040 min). The unsaturation percentage and trans isomers percentage was determined by FTIR at various temperatures and heating times, using n-valeronitrile as an internal standard, which quickened sample preparation and did not necessitate the determination of optical path length. Quantitative analysis by IR can be performed with techniques based on the addition of a standard to the sample in order to measure absorbance quotients so that a variable and unknown spacer may be used as measurements are independent of the optical path length. An analyte band and a standard band which are free from interference are required. When the unknown band overlaps with one of the standard bands this spectral interference may be corrected by the method developed by the authors known as the Absorbance Correction Method (ACM). 1.1. Absorbance correction method: theoretical background The ACM permits the analytic use of signals which interfere with the signals from the standard. The basis for this method is as follows: Let AA;n n1 , be the absorbance of the analyte at frequency n 1 in a prepared sample n (sample standard) and AP;n n1 be the absorbance of the standard at the same frequency, thus giving rise to a spectral interference in the measurement of the analytical signal. The total measured absorbance at n 1 is denoted here as An n1 , so that An n1 AA;n n1 AP;n n1 : 1

standard by relating it with the absorbance at another auxiliary frequency n 2 at which only the standard absorbs and not the analyte, i.e., knowing the absorbance arising exclusively from the standard at another frequency n 2, it is possible to predict the absorbance at the frequency of interest n 1, as the relation between the both is constant. Therefore, the Standard Correction Factor F c n1 ; n2 is dened at the frequency n 1 using n 2 as auxiliary frequency from the infrared spectrum of the pure standard compound, F c n1 ; n2 AP;0 n1 =AP;0 n2 ; 2

where, AP;0 n1 and AP;0 n2 are the measured absorbance values at n1 and n2, respectively, in the spectrum of the pure standard compound. The Standard Correction Factor multiplied by the absorbance at the auxiliary frequency n 2 in the prepared sample n; AP;n n2 , gives the absorbance at the frequency of interest n 1 in the same sample which is only because of the standard, AP;n n1 AP;n n1 AP;n n2 F c n1 ; n2 : 3

Arranging the terms in Eq. (1) and substituting Eq. (3) will provide the corrected absorbance at n 1 which is only because of the analyte AA;n n1 An n1 AP;n n1 AA;n n1 An n1 AP;n n2 F c n1 ; n2 : 4

Once the corrected absorbance value is obtained, the chosen quantitative method can then be applied.

2. Experimental 2.1. Instrumentation Perkin-Elmer 1600, Model 1605 FTIR Spectrometer with deuterated triglycine sulphate (DTGS) detector. Number of the co-added scans:16; Resolution: 4 cm 1. Data acquisition and processing software Spectrum for Windows, Perkin-Elmer. 2.2. Reagents n-Valeronitrile Aldrich analysis grade; nujol Aldrich IR grade; methyl linoleate Aldrich analysis

Note that in the infrared spectrum the absorbance bands owing to the same compound show a constant relation even when the concentration of this compound or sample thickness is varied, it is possible to predict the absorbance at n 1 resulting from the

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Fig. 1. Infrared spectra of sunower oil unheated () and heated at 300C during 40 min ().

grade; methyl elaidate Aldrich analysis grade; and chloroform Baker analysis grade. 2.3. Samples Oils from the Mexican and Spanish market were analysed: Virgin Olive oil (Domeq), Corn oil (Mazola), Sunower (Cristal), Seeds oil (Sunower, Safower and Canola, As), and lard. 2.4. Procedure Samples were heated in a porcelain capsule and were subjected to increasing heat on an electric device, up to a temperature of 300C, during this process heating was maintained for 20 and 40 min at the following temperatures: 80C, 150C, 200C, 250C and 300C ^ 5C. The samples and internal standard n-valeronitrile were mixed in a test tube, weighed on an analytical scale and the sample was shaken manually to ensure total homogenisation. The proportion in weight of n-valeronitrile in the prepared samples was between 35% and 65%. To obtain the infrared spectra one or two drops of the prepared samples were placed, using a Pasteur pipette, between two circular pieces of well polished potassium bromide crystals without using a spacer so sample thickness was not controlled but varied in

each prepared sample. In the IR spectra of the prepared samples absorbance was measured as the peak height at 3008 cm 1 with respect to the base line tangent at 31162718 cm 1, at 970 cm 1 with respect to the baseline 990944 cm 1 and the peak 2246 cm 1, corresponding to n-valeronitrile, with respect to the base line tangent at 24082144 cm 1. Eight mixtures of known composition of n-valeronitrile, methyl linoleate and nujol were prepared to obtain the calibration straight line for the determination of unsaturation grade. In these mixtures, the absorbance quotient 3008 cm 1/2246 cm 1 was measured with respect to the earlier mentioned baselines. In the same way eight mixtures of known composition of n-valeronitrile, methyl elaidate and nujol were prepared to obtain the calibration straight line for the determination of trans isomers and the absorbance quotient (970 cm 1/2246 cm 1) was measured.

3. Results and discussion The infrared spectra show how the intense frying produces a variation in composition of the oils nature, causing a decrease in unsaturated components and an increase in trans isomers (Fig. 1). The

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Fig. 2. Calibration straight line to obtain the trans isomers percentage as methyl elaidate, using ACM.

percentage of unsaturation expressed as a percentage in weight of methyl linoleate (l) and trans isomers expressed as a percentage in weight of methyl elaidate (e) was obtained for each type of oil at different heating. The internal standard method was used with n-valeronitrile (v) as internal standard. The infrared spectrum of n-valeronitrile shows an absorbance band at 2246 cm 1, owing to the stretching vibration of the triple CxN link which can be used as reference band as it does not interfere with any of the sample bands. The analytical signal used to determine the unsaturation was at the peak 3008 cm 1, caused by the stretching vibration of the CH link adjoining the double CyC link. From eight mixtures of

n-valeronitrile/methyl linoleate/nujol, the calibration straight line was obtained representing the quotient [A(3008)/A(2246)] against the Cl =Cv concentration quotient, where Cl is methyl linoleate concentration and Cv the n-valeronitrile concentration both expressed as %(w/w). Linear regression provided the following behaviour with a correlation index 0.998: A3008=A2246 0:3888 2:0326 Cl =Cv . For determination of the trans isomers, n-valeronitrile gives rise to a spectral interference in the measurement of the peak at 970 cm 1, which can be corrected by the proposed ACM. The Standard Correction Factor F c(970, 2246), was obtained as 0.0175, providing the corrected absorbance value

Fig. 3. Infrared spectrum of a prepared sample of the unheated corn oil.

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Table 1 Unsaturation percentage (%Un), expressed as methyl linoleate (%w/w) and trans isomers percentage (%t-I), expressed as methyl elaidate (%w/w) in oils and lard, obtained by internal standard method in FTIR spectroscopy Sample Heating (C/min) Olive oil %Un 18.91 18.83 18.24 18.30 18.53 16.71 12.41 11.85 11.54 9.81 5.65 %t-I 0.56 0.29 0.41 0.42 0.53 0.62 0.64 1.08 1.73 4.23 4.37 Sunower oil %Un 29.59 29.34 29.71 29.21 29.22 28.34 26.86 26.34 24.89 19.03 11.89 %t-I 0.23 0.21 0.25 0.23 0.29 1.05 1.10 2.01 3.20 7.19 11.45 Corn oil %Un 27.43 27.84 27.01 27.10 26.39 24.60 24.12 24.00 23.20 18.67 18.29 %t-I 0.35 0.70 0.85 0.92 0.62 0.87 0.96 0.87 1.24 3.19 5.32 Seeds oil %Un 23.23 23.32 22.51 22.04 21.17 20.71 19.65 18.83 18.09 16.41 10.94 %t-I 0.47 0.43 0.45 0.46 0.84 0.86 0.83 2.48 2.13 5.57 5.41 Lard %Un 11.21 10.99 10.86 11.53 11.17 11.07 10.90 8.73 8.85 8.78 8.59 %t-I 0.79 0.62 0.92 1.14 1.29 1.59 1.55 1.26 2.23 2.79 3.62

1 2 3 4 5 6 7 8 9 10 11

Tamb 80-20 8040 15020 15040 20020 20040 25020 25040 30020 30040

A C(970 cm 1). In the same way, from eight mixtures of n-valeronitrile/methyl elaidate/nujol, the calibration equation obtained was: A C(970)/A(2246) 0.0161 1.3423 Ce/Cv, r 0.998, where Ce is the methyl elaidate concentration (Fig. 2). The unsaturation percentage and trans isomers were calculated from the spectra of the samples prepared with n-valeronitrile (Fig. 3) using the corresponding calibration equations, the values obtained are shown in Table 1. The results obtained show how the intense heating of oils causes a decrease in unsaturation and increase in trans isomers. It is

possible to establish that in all the oils studied this degradation starts becoming considerable after 150C and at higher temperatures, a more signicant variation is apparent. The peroxide index obtained for each sample by the iodimetric method [9] reveals that these changes coincide with the decomposition of hydroperoxides (Fig. 4).

4. Conclusions FTIR Spectroscopy provides a very useful

Fig. 4. Variation in composition (unsaturation grade, trans isomers and peroxide value) of sunower oil at different thermal treatment.

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information on the composition and the extent of thermal self-oxidation in oils and lard subjected to intense heat, equivalent to that used in the ordinary preparation of food. The methodology developed in this study permitted determination of the percentage of unsaturated components and trans isomers in the samples at different heating temperatures, with no prior treatment. The use of absorbance quotient measurements permitted quantitative analysis without requiring a constant and known cell thickness thus facilitating sample handling and reducing analysis time. Further it permits quantication in cases where it is impossible to know the optic path length accurately. ACM, permits the analytical use of signals which would initially not be valid because of the spectral interference and in this particular case permits the measurements of signals which are signicant for the information provided on the state of decomposition of triglycerides. It can be said that when an edible oil is subjected to a heating process or frying, the initial nature of the trigliceride changes signicantly and the decrease in unsaturation and increase in trans isomers is evidence of the transformation occurring in the essential polyunsaturated fatty acids which the oil initially contains, thus indicating a decrease in the oils nutritional value.

Acknowledgements Authors are very grateful to the Department of Foreign Affairs in Mexico for a research grant for one of the authors (Moya Moreno, M.C.M.).

References
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