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Communication

Fatty Acid Composition and Lipid Oxidation in Plant-Based Meat Analogue Chicken Schnitzels Under Different Cooking Conditions

by
Owen Miller
,
Christopher J. Scarlett
and
Taiwo O. Akanbi
*
School of Environmental and Life Sciences, College of Engineering, Science and Environment, The University of Newcastle (UON), Brush Road, Ourimbah, NSW 2258, Australia
*
Author to whom correspondence should be addressed.
Lipidology 2025, 2(4), 23; https://doi.org/10.3390/lipidology2040023
Submission received: 26 September 2025 / Revised: 31 October 2025 / Accepted: 11 November 2025 / Published: 25 November 2025
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Abstract

Background/objectives: Plant-based meat analogues (PBMAs) are designed to mimic meat products and to be cooked under similar conditions by consumers. There have been few studies into the lipid stability of PBMAs, and no published studies have investigated the effect of cooking on the lipid stability of PBMAs. Methods: This study analysed the effect of recommended cooking conditions on the lipid oxidation of three commercial chicken schnitzel PBMAs with differing fatty acid composition. Fatty acids and lipid classes were analysed using gas chromatography (GC) and capillary chromatography (Iatroscan) with flame ionisation detectors, respectively. Lipid oxidation was analysed using multiple tests, including peroxide value (POV), p-Anisidine value, acid value, and thiobarbituric acid reactive substance (TBARS) tests, which then allowed for the total oxidation (TOTOX) to be calculated. Results: Fatty acid analysis by GC showed different levels of saturated and unsaturated fatty acid contents in all PBMAs, with oleic acid (C18:1) being the most abundant (product A = 52%; product B = 62%; product C = 37%). Meanwhile, lipid class analyses by Iatroscan revealed that the oils used in the PBMAs were composed of triacylglycerol (TAG), which remained intact after cooking. Lipid oxidation tests showed no major increases between the raw and cooked PBMA. Also, the TOTOX values for each product did not increase significantly (p < 0.05) due to cooking (TOTOX values for raw/cooked product A = 9.36/9.99; product B = 5.88/6.19; product C = 11.31/11.92), suggesting a broad stability of the lipids. Conclusions: Therefore, if the on-package cooking instructions are followed for these three PBMA products, their lipid oxidation levels remain within safe limits.

1. Introduction

Plant-based meat analogues (PBMAs) are novel products that utilise plant-based ingredients in their formulations to replicate the sensory experience of consuming a traditional meat product [1]. PBMAs are designed to be treated similarly to meat products by consumers during the processes of meal preparation, cooking, and consumption [2]. A large variety of PBMAs are available commercially, with more than 6485 new products launched globally from 2015 to 2021 [3]. It has proven difficult to make an accurate general or summarising statement regarding the healthiness of these commercially available PBMAs [4]. As the vast majority of these PBMAs are produced using the high-temperature and high-pressure process of extrusion [4], it is important to study any effect these processing conditions may have on lipid oxidation.
Stability studies over the past few decades have focused on lipid oxidation, as it is an important indicator of food product quality degradation [5,6,7,8]. The oxidation of lipids can lead to a decrease in sensory value, nutritional quality, and the functional properties of food products [9,10,11]. Therefore, it is crucial to consider the stability of PBMAs due to their significant processing, which is followed by a cooking process outlined on their packaging.
A variety of methods have been used to measure the level of oxidation in foods by detecting both primary and secondary lipid oxidation products [8]. Commonly used methods include thiobarbituric acid reactive substances (TBARSs), peroxide value (POV), p-Anisidine value (p-AV), acid value (AV), and the calculation of total oxidation (TOTOX) [8,12,13,14,15,16,17].
The stability of lipids within PBMAs is a new area of research. The amount of lipid oxidation in five PBMA products, which had occurred by the last day of their shelf life, has recently been investigated [18]. Through a TBARS test, they found relatively high values for all products, ranging from 3.20 mg MDA kg−1 up to 9.71 mg MDA kg−1. Another recent study has shown that when stored under its recommended storage conditions, a commercial PBMA patty did not undergo significant lipid oxidation up to its best-before date [19]. Other studies have shown that oils with a high degree of unsaturation are more susceptible to lipid oxidation caused by the process of extrusion [20] and that the degree of oxidation increases as a function of extrusion temperature [21]. However, there have been no previous studies into the lipid oxidation of PBMAs due to cooking. A recent extensive review article has comprehensively summarised the impacts of cooking on the appearance, texture, key aroma compounds, flavour, nutritional properties, digestibility, and functional properties of proteins [22].
Current commercial PBMAs often use canola oil in their formulations, which has a high proportion of unsaturated fatty acids, making it highly susceptible to oxidation [23,24]. Canola oil can undergo oxidation through exposure to light (photo-oxidation), hydrolysis (influenced by moisture, temperature, and enzymes), and auto-oxidation (influenced by exposure to heat, light, and oxygen) [25]. To improve the lipid stability of an oil, it should have a low content of saturated fatty acids (<30%) and low content of polyunsaturated acids (linoleic and linolenic; <2%) [26]. As the temperature of canola oil increases, there is an increase in lipid oxidation as measured by peroxide value [23]. The stability of canola oil during storage and cooking has been investigated, and an increase in oxidation, as measured by POV, has been found when heated in a microwave oven for only 36 min [24]. Coconut oil is another plant-based lipid that has been used in PBMA formulations. Coconut oil is regarded as a saturated oil and is considered more stable compared to other edible oils due to its low susceptibility to autoxidation [27,28]. The stability of coconut oil during storage has been previously studied [28], and blending coconut oil with vegetable oils has been shown to increase their oxidative stability [29]. However, the effect of cooking conditions on the lipid stability of processed foods, such as PBMAs, which contain coconut oil, is sparse.
The research objective of this study was to measure the lipid oxidation for three different brands of commercially available chicken schnitzel PBMAs due to oven baking using the recommended cooking conditions as stated on their packaging. The products were chosen with varied recommended cooking times and temperatures to explore any possible effect or interaction of these variables on lipid oxidation. The products also varied in their total fat content and the type of plant-based oil used in their formulation. This variation in cooking conditions, fat content, and oil type could result in variations in lipid oxidation responses. This choice of products allowed for an overview of any possible differences that may arise due to the identified variables, rather than a comprehensive comparison of any observable lipid oxidation of these three products.

2. Materials and Methods

2.1. Chemicals and Reagents

Chemicals such as 1-butanol, isooctane, diethyl ether, p-Anisidine, potassium hydroxide, glacial acetic acid, potassium iodide, sodium lauryl sulphate, heptane, butylated hydroxytoluene, 2-Thiobarbituric acid, and isopropanol were purchased from Sigma Aldrich (Castle Hill, NSW, Australia). The capillary (Iatroscan) chromatography and gas standards were purchased from Nu-Chek Prep (Elysian, MN, USA). All other chemicals used were of analytical grade.

2.2. Plant-Based Meat Analogues and Sampling

Three freshly stocked chicken schnitzel PBMA products were bought from a local supermarket on the Central Coast of New South Wales (NSW), Australia. None of the products contained an added antioxidant. The products were purchased under refrigerated conditions, and their labels identified whether they needed to be cooked prior to consumption. Once removed from their packaging, the products were in a “raw” state. The products were oven-baked according to the recommended cooking instructions on their packaging, which specified temperature and time (Figure 1). After being removed from the oven, the products were in a “cooked” state. A single sample consisted of an entire chicken schnitzel, and three samples were completed for each condition (raw or cooked) for each product.

2.3. Lipid Extraction

To extract the lipids from each raw and cooked schnitzel, it was first broken down by hand into small chunks approximately 1 cm3 in size. Then, 10 g of sample schnitzel was placed into a 50 mL tube, and heptane was then added to bring the volume up to 50 mL. Heptane was used as a replacement for hexane, as it has been shown to be as effective at extracting lipids and is less toxic [30]. For 2 h, the tube was then placed into an auto-mixer (Intelli-mixer RM-2M, ELMI Ltd., Riga, Latvia) to allow the heptane to extract the lipid from the sample. To evaporate the heptane, a rotary vacuum (BUCHI Rotavapor R-114, BUCHI Labortechnik, Flawil, Switzerland) was set to a pressure of 120 mbar. The extracted lipid was then stored under refrigerated conditions (4 °C) until used for analysis. Lipid extraction was completed in triplicate on the three commercial PBMA products before cooking (labelled as the raw condition) and after cooking was completed under its recommended conditions (labelled as the cooked condition). The extracted lipids were kept in the refrigerator at 4 °C for no longer than 24 h before testing.

2.4. Analysis of Lipid Classes by Capillary Chromatography (Iatroscan-FID)

Lipid classes were analysed by capillary chromatography with flame ionisation detector (Iatroscan, Iatron Laboratories Inc., Tokyo, Japan) following a previously reported method [31]. The Iatroscan settings which were used in this study were as follows: air flow rate of 2.0 L/min, hydrogen flow rate of 160 mL/min, and scan speed of 30 s/scan. Also, to clean the chromarods, they were scanned twice under those conditions. In brief, a sample of ten milligrams (10 mg) of oil was dissolved in 5 mL of heptane, then an autopipette was used to spot 1 μL of the sample onto the chromarods at the origin along the rod holder. This was then developed for 22 min in a heptane/diethyl ether/acetic acid mix (60:17:0.2 v/v/v). Lipid classes were identified by TLC standards obtained from Nu-Chek Prep. The percentage composition of each lipid class was determined using the in-built SIC-480 II software, version MK-7s for multiple chromatogram processing.

2.5. Analysis of Fatty Acid Composition by Gas Chromatography (GC)

A previous methodology was followed for the analysis using gas chromatography with flame ionisation detection (GC-FID) to analyse the composition of fatty acids in each of the three products [31]. Fatty acid methyl esters (FAMEs) of the samples were analysed through the use of a Shimadzu GC-2030 gas chromatograph with flame ionisation detector (Shimadzu Corporation, Tokyo, Japan), which was equipped with a BPX70 SGE column (30 m, 0.25 mm I.d., 0.25 µm film thickness). The oven was programmed from 140 °C (5 min hold) to 200 °C (5 min hold) at a rate of 4 °C/min with a total run time of 20 min. A volume of 2 µL of solution was injected using an auto-injector (injector temperature of 250 °C). The carrier gas was nitrogen (1.3 mL/min, constant flow), with the detector gases being 30 mL/min hydrogen, 200 mL/min air, and 30 mL/min nitrogen. LabSolution software version 5.93 was used to integrate the peak areas, and the known reference standards purchased from Nu-Chek Prep (Elysian, MN, USA) were used to identify fatty acids by comparing their retention times. Fatty acids were quantified as a percentage as previously reported [19].

2.6. Oxidation Stability Tests

A range of testing methodologies was undertaken as previously recommended to give an accurate measure of the overall oxidative state of lipid products [32]. Each lipid oxidation measurement was taken in triplicate for each of the three products. All lipid oxidation tests were performed following previously reported methodology [19]. The lipid oxidation tests performed included peroxide value (POV), thiobarbituric acid reactive substance (TBARS), p-Anisidine value (p-AV), and acid value (AV), where the wavelength for absorption for TBARS was measured at 532 nm and p-AV at 350 nm. TOTOX was calculated using a previously reported methodology [19].

2.7. Statistical Analysis

Statistical Product and Service Solutions (SPSS version 30.0.0.0) was used for all statistical analyses performed, and graphs were produced using Microsoft Excel 365. An independent samples t-test (2-sided) was performed, using a significance level of p < 0.05, to determine any significant differences between the raw and cooked conditions for each product. Standard deviation has been represented as error bars on all graphs.

3. Results and Discussion

3.1. Lipid Content and Fatty Acid Composition of the PBMA Chicken Schnitzels

The relevant on-package nutritional information for the three commercial chicken schnitzel PBMA products is outlined in Table 1. All products have high protein contents, with product A having the highest. All the products contained canola oil, with coconut fat/oil present in the coating of product A and as the first listed lipid ingredient for product C. The high amount of saturated fat in product C is likely due to coconut oil being the major lipid included in this formulation, as coconut oil is known to have a high saturated fat content [28].
GC analysis for product A identified the major fatty acids as oleic—C18:1 (58.4%), linoleic—C18:2 (19.7%), and linolenic—C18:3 (9.0%) (Figure 2). This proportion of major fatty acids is similar to that of canola oil [33]. The presence of the shorter chain fatty acids myristic—C14:0 (2.3%) and lauric—C12:0 (1.1%) is likely due to the inclusion of coconut fat in the coating for this product [34].
The profile of major fatty acids using GC for product B shows a high proportion of oleic—C18:1 (62.2%), linoleic—C18:2 (19.8%), and linolenic—C18:3 (6.8%) (Figure 2), which is consistent with the fatty acid profile of canola oil [33].
The profile of major fatty acids using GC for product C identified the presence of oleic—C18:1 (37.4%), lauric—C12:0 (16.8%), and linoleic—C18:2 (12.3%) (Figure 2). Both sunflower and canola oil have high proportions of oleic and linoleic acid, whereas coconut oil has a high proportion of lauric acid [35]. This fatty acid profile matches that expected based on the oils listed in its ingredients (Table 1).

3.2. Effect of Cooking on Lipid Classes of the PBMA Chicken Schnitzels

The Iatroscan-FID results presented in Figure 3 show that the lipid extracted from the chicken schnitzel PBMAs was composed of triacylglycerol (TAG), which suggests that the oils used in these products have no other lipid classes. There was only a small change in signal (mV) observed due to cooking for each of the PBMA products (Figure 3).

3.3. Effect of Cooking on Lipid Classes and Oxidation for the PBMA Chicken Schnitzels

For products A and B, the only statistically significant difference (p < 0.05) due to cooking was an increase in TBARS value (Table 2). However, the magnitude of the increase was low for both products; for product A, it increased from 1.0 to 1.3 mg MDA/kg lipid, and for product B, it increased from 1.0 to 1.2 mg MDA/kg lipid. These low levels of lipid oxidation after cooking for both of these PBMA products, as measured using TBARS, fall into the “not rancid” category (<1.5 mg/MDA/kg) and would therefore have no impact on the safety of consuming these products, regarding lipid oxidation [36].
For product C, the effect of cooking caused a statistically significant increase in p-AV, from 2.8 to 11.1 (Table 2). It has been previously shown that an oil with a high proportion of oleic acid (C18:1) can oxidise rapidly, which could cause an increase in p-AV due to an increase in secondary oxidation products, as seen in product C [37]. Although p-AV generally correlates well with POVs and TBARSs, it has been suggested that the TBARS methods can provide a more accurate measure of oxidation deterioration than p-AV [32]. This is due to the high likelihood that p-AV can give an overestimation with high oleic oils, being at least 10 to 60 times more sensitive to alkenals compared to alkanals [38]. However, there was no significant increase in p-AV for products A and B, and they had higher proportions of oleic acid than product C. Also, p-AV unsaturated aldehydes have a higher colour response than saturated aldehydes, and p-AV can interact with coloured oils [39]. Therefore, as there was no significant increase in TBARS, which is an alternative measurement for secondary oxidation, the significant increase in p-AV may not be an accurate indicator by itself of any possible lipid oxidation that occurred in product C.
The increases observed for products A and B were of much lower magnitude compared to studies of lipid oxidation in meat [16,40] and stability studies in plant oils [23,24,28]. This is most likely due to the significantly shorter cooking time and lower temperature used for the PBMAs in this study compared to previous studies. One possibility for the increase in lipid oxidation as measured by TBARSs could be due to the higher proportion of unsaturated fatty acids in products A and B, as compared to product C, which did not have a significant increase in TBARSs after cooking.
Coconut oil has a higher degree of saturation than canola oil, and it has been suggested that oils with a higher degree of saturation can help limit degradation under high temperature conditions [20]. Coconut oil has also been shown to provide oxidative stability to vegetable oils, including sunflower and rice bran oil [29]. This is supported by this study, where product C, which contains the highest proportion of coconut oil among plant oils in its formulation, showed no significant increase in TBARS, in contrast to products A and B.
For both raw and cooked products, product C had the highest TOTOX value, followed by product A and then product B, in the same order as seen for POV (Table 2). It is difficult to identify the cause(s) for these differences in POV for these products. One possibility is that these differences may be due to variations in their processing conditions during manufacturing. However, since these were commercial PBMA products, their exact processing conditions are unknown. These products were manufactured using the extrusion process, and extrusion conditions, such as the temperature of the extruder barrel, have been shown to affect lipid oxidation in PBMA products [20,21]. A second possibility could be due to differences in the packaging of these PBMA products. A decrease in CO2 concentration within the packaging of a mince PBMA product has been observed due to its high moisture content [18]. This reaction between CO2 and the moisture within the PBMA was shown to lead to the formation of carbonic acid, which could have an impact on lipid oxidation [41]. However, the composition of the gases within the packaging of PBMA products was beyond the scope of this study.
The relationship between the fatty acid profiles of the three PBMA products in this study and their lipid oxidation, as determined using their TOTOX value, could not be further explored in the present study, as there was no significant increase in the TOTOX value for any of the three products due to the cooking conditions. The TOTOX value showed no significant increase for any of the products, which is a strong indicator that, for these PBMA products, there was no significant increase in lipid oxidation due to cooking (Table 2).

4. Conclusions

In conclusion, the lipid oxidation tests performed in this study indicated no significant increases in lipid oxidation of concern when the PBMAs were prepared according to their on-package instructions. However, it is noteworthy that product C displayed a potential indicator of lipid oxidation related to cooking, as evidenced by the p-AV measurement.
A follow-up study is being designed to investigate the cooking conditions that would be required to cause lipid oxidation to occur at an unsafe level for human consumption. This would provide knowledge of the lipid oxidation pathways that may occur during the complete degradation of PBMA products. Although the products studied in this work were only oven-baked, alternative cooking methods, including shallow frying, air frying, and microwaving, are also being investigated. Comparing these different recommended cooking methods could determine which one causes the greatest lipid degradation, or whether any lipid oxidation occurs. Additionally, the three products studied here were all chicken schnitzel analogues. We are expanding our investigations into the stability of other PBMA categories under recommended cooking conditions, such as burger patties, sausages, or minces.

Author Contributions

Conceptualization, O.M. and T.O.A.; methodology, O.M. and T.O.A.; software, O.M. and T.O.A.; validation, O.M., T.O.A., and C.J.S.; formal analysis, O.M.; investigation, O.M.; resources, O.M. and T.O.A.; writing—original draft preparation, O.M.; writing—review and editing, O.M. and T.O.A.; supervision, T.O.A. and C.J.S. All authors have read and agreed to the published version of the manuscript.

Funding

This research received no external funding.

Data Availability Statement

Data are available upon request.

Acknowledgments

Owen Miller acknowledges the PhD scholarship award by the University of Newcastle, Australia, under the Strategic Engagement Scheme.

Conflicts of Interest

The authors declare no conflicts of interest.

Abbreviations

The following abbreviations are used in this manuscript:
AVAcid value
DAGDiacylglycerol
FFAsFree fatty acids
FIDFlame ionisation detector
GCGas chromatography
MAGMonoacylglycerol
MDAMalondialdehyde
p-AVp-Anisidine value
PBMAPlant-based meat analogue
POVPeroxide value
TAGTriacylglycerol
TBARSThiobarbituric Acid Reactive Substance
TOTOXTotal oxidation
UPFUltra-processed food

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Figure 1. Recommended cooking conditions (temperature and time) for the three commercial PBMA chicken schnitzel products.
Figure 1. Recommended cooking conditions (temperature and time) for the three commercial PBMA chicken schnitzel products.
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Figure 2. Identification of major fatty acids from three different commercial plant-based meat analogues using gas chromatography analysis. Extracted lipid samples are inset: (A) product A, (B) product B, and (C) product C. Major fatty acids identified are caproic (C6:0), caprylic (C8:0), lauric (C12:), myristic acid (C14:0), palmitic (C16:0), stearic (C18:0), oleic (C18:1n9), linoleic (C18:2n6), linolenic (C18:3n3), and arachidic (C20:0). Mean values shown with error bars showing standard deviation. Letter labels show significant differences between products from each major fatty acid at p < 0.05.
Figure 2. Identification of major fatty acids from three different commercial plant-based meat analogues using gas chromatography analysis. Extracted lipid samples are inset: (A) product A, (B) product B, and (C) product C. Major fatty acids identified are caproic (C6:0), caprylic (C8:0), lauric (C12:), myristic acid (C14:0), palmitic (C16:0), stearic (C18:0), oleic (C18:1n9), linoleic (C18:2n6), linolenic (C18:3n3), and arachidic (C20:0). Mean values shown with error bars showing standard deviation. Letter labels show significant differences between products from each major fatty acid at p < 0.05.
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Figure 3. Iatroscan lipid class analysis for commercial PBMA products, comparing raw and cooked samples, (a) product A; (b) product B; (c) product C.
Figure 3. Iatroscan lipid class analysis for commercial PBMA products, comparing raw and cooked samples, (a) product A; (b) product B; (c) product C.
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Table 1. Package information from the nutrition panels for the three commercial PBMA chicken schnitzel products.
Table 1. Package information from the nutrition panels for the three commercial PBMA chicken schnitzel products.
ProductProtein (g/100 g)Total Fat (g/100 g)Saturated Fat (g/100 g)Lipids
Included *
Product A15.021.22.7Internal: Canola oil (3rd)
Coating: Canola oil, coconut fat powder
Product B9.78.21.3Vegetable oil (Sunflower, canola; 3rd)
Product C9.014.35.9Internal: Coconut oil (4th), canola oil (15th)
Coating: Canola oil (7th), olive oil (9th), rice bran oil (14th)
* The number in parentheses is the order in which the lipid was listed in the ingredients panel on the product’s packaging. The ingredients are listed in the order of percentage composition, so the earlier it is on the list, the higher the proportion of the ingredient in the product’s formulation.
Table 2. Lipid oxidation results from primary and secondary oxidation tests for each raw and cooked product.
Table 2. Lipid oxidation results from primary and secondary oxidation tests for each raw and cooked product.
ProductConditionTBARSp-AVAVPOVTOTOX
ARaw1.04 ± 0.03 a9.33 ± 0.72 a2.04 ± 0.64 a4.16 ± 0.29 a9.36 ± 0.55 a
Cooked1.31 ± 0.15 b11.82 ± 2.48 a1.93 ± 0.54 a4.34 ± 0.27 a9.99 ± 0.64 a
BRaw1.03 ± 0.07 a9.46 ± 1.78 a3.33 ± 0.55 a2.43 ± 0.36 a5.88 ± 0.80 a
Cooked1.19 ± 0.05 b9.84 ± 0.88 a3.22 ± 0.67 a2.50 ± 0.23 a6.19 ± 0.45 a
CRaw1.03 ± 0.11 a2.76 ± 1.87 a1.83 ± 0.95 a5.14 ± 0.24 a11.31 ± 0.58 a
Cooked1.06 ± 0.11 a11.06 ± 1.17 b2.06 ± 0.65 a5.43 ± 0.53 a11.92 ± 1.14 a
Superscript letters represent statistically significant differences between the raw and cooked conditions at p < 0.05.
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MDPI and ACS Style

Miller, O.; Scarlett, C.J.; Akanbi, T.O. Fatty Acid Composition and Lipid Oxidation in Plant-Based Meat Analogue Chicken Schnitzels Under Different Cooking Conditions. Lipidology 2025, 2, 23. https://doi.org/10.3390/lipidology2040023

AMA Style

Miller O, Scarlett CJ, Akanbi TO. Fatty Acid Composition and Lipid Oxidation in Plant-Based Meat Analogue Chicken Schnitzels Under Different Cooking Conditions. Lipidology. 2025; 2(4):23. https://doi.org/10.3390/lipidology2040023

Chicago/Turabian Style

Miller, Owen, Christopher J. Scarlett, and Taiwo O. Akanbi. 2025. "Fatty Acid Composition and Lipid Oxidation in Plant-Based Meat Analogue Chicken Schnitzels Under Different Cooking Conditions" Lipidology 2, no. 4: 23. https://doi.org/10.3390/lipidology2040023

APA Style

Miller, O., Scarlett, C. J., & Akanbi, T. O. (2025). Fatty Acid Composition and Lipid Oxidation in Plant-Based Meat Analogue Chicken Schnitzels Under Different Cooking Conditions. Lipidology, 2(4), 23. https://doi.org/10.3390/lipidology2040023

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