nutrients

Systematic Review
Influence of Heating during Cooking on Trans Fatty Acid
Content of Edible Oils: A Systematic Review and
Meta-Analysis
Saiuj Bhat 1 , Damian Maganja 2 , Liping Huang 2 , Jason H. Y. Wu 2 and Matti Marklund 2,3,4, *

1 Department of Vascular Surgery, Royal Perth Hospital, Victoria Square, Perth, WA 6000, Australia;
[email protected]
2 The George Institute for Global Health, University of New South Wales, Sydney, NSW 2050, Australia;
[email protected] (D.M.); [email protected] (L.H.);
[email protected] (J.H.Y.W.)
3 Department of Epidemiology, Johns Hopkins Bloomberg School of Public Health, Baltimore, MD 21205, USA
4 Department of Public Health and Caring Sciences, Uppsala University, SE75105 Uppsala, Sweden
* Correspondence: [email protected]

Abstract: Consumption of trans fatty acids (TFA) is associated with adverse health outcomes and is a
considerable burden on morbidity and mortality globally. TFA may be generated by common cooking
practices and hence contribute to daily dietary intake. We performed a systematic review and meta-
analysis to investigate the relationship between heating edible oils and change in their TFA content.
A systematic search of experimental studies investigating the effect of various methods of heating
on TFA content of edible oils was conducted in Medline and Embase since their inception up to



1 October 2020 without language restrictions. Comparable data were analysed using mixed multilevel

 linear models taking into account individual study variation. Thirty-three studies encompassing
Citation: Bhat, S.; Maganja, D.; twenty-one different oils were included in this review. Overall, heating to temperatures <200 ◦ C
Huang, L.; Wu, J.H.Y.; Marklund, M. had no appreciable impact on different TFA levels. Between 200 and 240 ◦ C, levels of C18:2 t (0.05%
Influence of Heating during Cooking
increase per 10 ◦ C rise in temperature, 95% CI: 0.02 to 0.05%), C18:3t (0.18%, 95% CI: 0.14 to 0.21%),
on Trans Fatty Acid Content of Edible
and total TFA (0.38%, 95% CI: 0.20 to 0.55%) increased with temperature. A further increase in
Oils: A Systematic Review and
total TFA was observed with prolonged heating between 200 and 240 ◦ C. Our findings suggest that
Meta-Analysis. Nutrients 2022, 14,
heating edible oils to common cooking temperatures (≤200 ◦ C) has minimal effect on TFA generation
1489. https://doi.org/10.3390/
nu14071489
whereas heating to higher temperatures can increase TFA level. This provides further evidence in
favour of public health advice that heating oils to very high temperatures and prolonged heating of
Academic Editor: Sheila Erin
oils should be avoided.
Fleischhacker

Received: 12 March 2022 Keywords: cooking; frying; baking; food policy

Accepted: 28 March 2022
Published: 2 April 2022

Publisher’s Note: MDPI stays neutral

1. Introduction
with regard to jurisdictional claims in
published maps and institutional affil- Trans-fatty acids (TFA) are fatty acids with at least one unsaturated, non-conjugated
iations. double bond in the trans configuration. Consumption of TFA is associated with cardiovascu-
lar disease risk factors including increased low-density lipoprotein cholesterol, triglyceride,
and lipoprotein (a) levels, decreased high-density lipoprotein cholesterol, and increased
insulin resistance, adiposity, and endothelial dysfunction [1]. Increased TFA consumption
Copyright: © 2022 by the authors. has also been associated with greater incidence of diabetes and coronary heart disease [2–4].
Licensee MDPI, Basel, Switzerland. Approximately 645,000 annual deaths globally are attributed to a diet high in TFA [5]. In
This article is an open access article
view of its adverse health effects, the World Health Organisation recommends that TFA
distributed under the terms and
intake be minimised and contribute no more than 1% of total daily energy intake [6], in line
conditions of the Creative Commons
with other food and health advisory organisations [7]. While global TFA intake appears to
Attribution (CC BY) license (https://
have gradually declined over the past three decades [8], consumption of TFA remains high
creativecommons.org/licenses/by/
in many parts of the world, accounting for up to 6.5% of total daily energy [9].
4.0/).

Nutrients 2022, 14, 1489. https://doi.org/10.3390/nu14071489 https://www.mdpi.com/journal/nutrients

Nutrients 2022, 14, 1489 2 of 11

Trans fatty acids in diet originate from naturally occurring TFA from ruminants or
industrially produced TFA formed by partial hydrogenation of vegetable oils. In addition,
thermal treatment of cooking oils can alter their physicochemical properties and potentially
generate TFA from cis-unsaturated fatty acids during cooking procedures (e.g., deep-, pan-,
or stir-frying) [10–12]. While some previous studies have reported increases in TFA content
of edible oils during cooking, others demonstrate little or no change [13–15]. Furthermore,
amongst studies that report increases in TFA content of oil during cooking, the magnitude
of the reported increase is variable. No systematic review has assessed the impact of heating
on TFA content in cooking oils. Therefore, the aim of our study was to perform a systematic
review of experimental studies to determine how thermal treatment influences the TFA
content of edible oils. We hypothesised that an increase in heating temperature and time
would be associated with an increase in the TFA content of edible oils, irrespective of the
method of heating.

2. Materials and Methods

This systematic review followed the Preferred Reporting Items for Systematic Reviews and
Meta-Analyses (PRISMA) guidelines and was registered on PROSPERO [CRD42020223609].
Two reviewers independently screened studies for eligibility (LH, SB, or DM) and ex-
tracted data (SB, DM). Discrepancies were resolved by consensus or involvement of a
third reviewer (MM).

2.1. Search Strategy

A systematic literature search, up to 1 October 2020, was conducted in Medline
and Embase without date or language restrictions. Full search terms are available in
Appendix A. Reference lists of eligible studies were manually scanned to identify additional
relevant publications.

2.2. Eligibility Criteria

Experimental studies quantifying the TFA content of edible, commercially available
cooking oils before and after various common cooking practices, including heating, deep-
frying, stir-frying, pan-frying, and baking, were included. Studies that did not measure
changes in TFA composition of oils were excluded.

2.3. Data Extraction

The following data were extracted from included studies: publication information,
type of oil, type of foods (if any) used in the experiment, type of TFA studied, experi-
mental conditions including cooking method, temperature, heating time, heating cycle,
and heating vessel, method of quantifying fatty acid composition, TFA level and units of
measure, estimates of uncertainty, and number of replicates. Cooking oils that contained
mixtures of different oils (e.g., sunflower, safflower, and canola oils) were consolidated into
a category labelled “blend”; vanaspati, bakery shortening, and other hydrogenated oils
were consolidated into the “hydrogenated vegetable fat” category.

2.4. Risk of Bias Assessment

Included studies were evaluated against the following criteria to determine study
quality: reporting of analytical variability (i.e., reporting of coefficient of variation) (yes
or no), more than one replicate (yes or no), and reporting of measures of uncertainty (SD,
SE, or 95% CI) (yes or no). Studies scored a point for each of the criteria fulfilled. Studies
that did not report measures of uncertainty for some outcomes received a score of 0 for
that criterion.

2.5. Data Analysis

The primary outcome was change in level of TFA (% of total fatty acids), calculated
as the mean difference between TFA levels before and after the experiment. Comparable
Nutrients 2022, 14, 1489 3 of 11

data from included studies were analysed using mixed multilevel linear models, with
intercepts for individual studies specified as random effects to account for variation in
experimental conditions and oils between studies. Differences in mean TFA levels between
predefined heating temperature intervals were assessed in models with temperature coded
as an ordinal variable (room temperature [control], <200 ◦ C, 200–240 ◦ C, and >240 ◦ C). The
temperature intervals were defined based on usual cooking temperatures (i.e., common
cooking methods rarely utilise temperatures above 200 ◦ C) and smoke points of commonly
used cooking oils (i.e., the smoke points of most commonly used cooking oils do not exceed
240 ◦ C). To assess linear associations of heating temperature and TFA content within tem-
perature intervals, we utilised linear splines with knots at 200 ◦ C and 240 ◦ C. Two models
were evaluated to assess the impact of cooking temperature on TFA content: (1) crude,
without adjustment; and (2) adjusted for heating time and type of oil used. Subgroup analy-
ses for each type of oil were not conducted given the small number of data points available
for individual cooking oils. We investigated the interaction between heating temperature
and heating time for those temperature intervals where a significant change in TFA levels
was observed. All statistical tests were performed using STATA 16 (Stata Corp, College
Station, TX, USA), with two-tailed alpha of 0.05. Data are presented as mean ± standard
deviation (SD) or median [interquartile range, IQR] unless stated otherwise.

3. Results
3.1. Study Characteristics
Of the 234 studies identified by our search, 33 were included in this review (Figure 1
and Supplementary Table S1). Collectively, the studies analysed 21 different cooking
oils, with corn [11,13,16–23], soybean [15,20,21,23–29], sunflower [11,16,20,21,30–34], and
hydrogenated vegetable fat [15,24,27,29,31,35–37] being the most commonly assessed; fewer
studies investigated other oils including Aleppo pine seed [38], blend [16,17,23,33,39,40],
canola [17,41], coconut [42], cottonseed [35], groundnut [24], linseed [20], olive [11,12,16,20,24,32,34],
palm [33,36,39,42,43], peanut [20,28,36], peony seed [20], rapeseed [20,24,44], rice bran [17,20,40],
safflower [17,23], and sesame [17,20] oil; or solid fats such as ghee [24] and lard [16].
The relatively high baseline (preheating) levels of TFA in hydrogenated vegetable fats
compared to the two most commonly studied cooking oils (corn and soybean) are shown
in Supplementary Table S2. While all studies evaluated the change in TFA content of
cooking oils, only seven investigated the change in TFA content of foods—chicken, fish,
or potato—cooked in various oils [18,21,27,32,35,43,45]. Quality assessment of included
studies is shown in Supplementary Table S3. Only 3 studies (9%) were deemed to have the
highest quality score (3/3) in our assessment, 15 (45%) scored 2 out of 3, 9 studies (27%)
scored 1 out of 3, and 6 studies (18%) scored 0. The major quality limitation was lack of
reporting of analytic variability across the included studies.
Trans fatty acids investigated included trans-palmitoleic acid (C16:1t), elaidic
acid (C18:1t), trans-linoleic acid (C18:2t), trans-linolenic acid (C18:3t), trans-eicosenoic
acid (C20:1t), monounsaturated TFA, polyunsaturated TFA, and total TFA. The me-
dian TFA level at baseline was 0.23% of total fatty acids [0.04% to 0.70%]. Five
different cooking methods were employed: baking [13], heating [19,24,30,33,36,38],
deep-frying [11,13,17,18,20,21,23,24,27–29,32–35,37,39–43], pan-frying [11,13,20], and stir-
frying [13,20]. Heating temperatures, heating times, and heating cycles corresponding to
the various cooking methods and for the various TFA studied are summarised in Supple-
mentary Tables S4 and S5. Across studies and experiments, the median heating temperature
was 180 ◦ C [IQR: 175–200 ◦ C], the median heating time was 45 min [IQR: 6–480 min], and
the median cooking cycle was 1 [IQR: 1–2 cycles].
Nutrients 2022, 14, 1489 4 of 11

Figure 1. Flow diagram of study selection.

3.2. Heating Temperature and Change in Level of TFA

The median TFA levels (shown separately for subtypes of TFA and total TFA) at differ-
ent temperature intervals are depicted in Table 1. Compared to room temperature, heating
oils to temperatures <200 ◦ C (i.e., common cooking temperatures) had no significant impact
on TFA levels (Figure 2 and Supplementary Table S6). However, within this temperature
range, C18:3t levels increased significantly, albeit modestly, with higher temperature (0.02%
increase per 10 ◦ C rise in temperature, 95% CI: 0.01 to 0.02%). While mean TFA levels in oils
heated to between 200 and 240 ◦ C did not differ significantly from TFA levels in unheated
or less heated oils, the levels of C18:2t (0.05% increase per 10 ◦ C rise in temperature, 95%
CI: 0.02 to 0.05%), C18:3t (0.18%, 95% CI: 0.14 to 0.21%), and total TFA (0.38%, 0.20 to 0.55%)
increased with temperature within this temperature range.
Nutrients 2022, 14, 1489 5 of 11

Table 1. Levels of different trans fatty acid (TFA) (% of total fatty acids) as a function of heating temperature. Changes in TFA levels resulting from heating oils
above room temperature as well as the difference in TFA levels in oils heated to high (>200 ◦ C) versus usual cooking temperatures (≤200 ◦ C) are depicted for each
TFA studied.

TFA Concentration (% of Total Fatty Acids)

Unheated <200 ◦ C 200–240 ◦ C >240 ◦ C
n studies Median (IQR)/ n studies Median (IQR)/ n studies Median (IQR)/ n studies Median (IQR)/
Fatty acid
(n samples) Estimate (95% CI); p (n samples) Estimate (95% CI); p (n samples) Estimate (95% CI); p (n samples) Estimate (95% CI); p
16:1t Median 0 (0) - 4 (60) 0.02 (0.01; 0.03) 2 (30) 0.02 (0.01; 0.03) 1 (16) 0.02 (0.00; 0.02)
Difference from usual
- Reference 0.00 (−0.00, 0.01); p = 0.62 −0.01 (−0.01, 0.00); p = 0.052
cooking temperatures
18:1t Median 3 (14) 0.35 (0.06, 0.56) 13 (306) 0.24 (0.03, 1.49) 5 (79) 0.07 (0.01, 3.72) 3 (37) 1.08 (0.08, 3.20)
Difference from unheated Reference 0.71 (−1.11, 2.54); p = 0.45 0.61 (−1.26, 2.48); p = 0.52 2.06 (0.10, 4.02); p = 0.04
Difference from usual
Reference −0.10 (−0.97, 0.77); p = 0.82 1.33 (0.07, 2.60); p = 0.039
cooking temperatures
18:2t Median 1 (1) 0.13 (-) 10 (178) 0.31 (0.01, 0.50) 4 (69) 0.42 (0.03; 0.62) 1 (48) 0.48 (0.21; 0.98)
Difference from unheated Reference 0.33 (−0.51, 1.18); p = 0.45 0.38 (−0.47, 1.23); p = 0.38 0.76 (−0.10, 1.61); p = 0.084
Difference from usual
Reference 0.05 (−0.07, 0.18); p = 0.42 0.43 (0.28, 0.57); p < 0.001
cooking temperatures
18:3t Median 2 (7) 0.01 (0.00, 0.20) 4 (56) 0.01 (0.00, 0.27) 3 (27) 0.01 (0.00, 0.30) 1 (12) 0.70 (0.53, 1.48)
Difference from unheated Reference 0.11 (−0.23, 0.45); p = 0.52 0.20 (−1.14, 0.53); p = 0.25 0.80 (0.44, 1.15); p < 0.001
Difference from usual
Reference 0.05 (−0.11, 0.21); p = 0.52 0.62 (0.37, 0.88); p < 0.001
cooking temperatures
Total TFA Median 1 (1) 0.09 (-) 10 (117) 0.97 (0.62, 1.53) 5 (52) 1.42 (0.87; 3.70) 2 (17) 1.54 (0.97; 4.10)
Difference from unheated Reference 2.14 (−8.24, 12.52); p = 0.69 2.49 (−7.93, 12.90); p = 0.64 3.78 (−6.84, 14.40); p = 0.49
Difference from regular
Reference 0.34 (−1.42, 2.10); p = 0.70 1.64 (−1.22, 4.49); p = 0.26
cooking temperatures
Nutrients 2022, 14, 1489 6 of 11

Figure 2. Change in C18:1t (A), C18:2t (B), C18:3t (C), and total TFA (D) (% of total fatty acids)
content of cooking oil as a function of heating temperature. Data were fitted using a mixed multilevel
linear regression model adjusted for heating time and oil type with random intercepts for studies
and spline knots at 200 and 240 ◦ C. Slopes represent change in TFA per 10 ◦ C change in heating
temperature within a particular spline range (<200 ◦ C, 200–240 ◦ C, >240 ◦ C). Data point colours
represent different studies.

As expected, the change in TFA was more striking, with an increase in heating time at
temperatures between 200 and 240 ◦ C (Supplementary Table S7). For example, when heated
for 6 h, total TFA levels increased by 0.86% for every 10 ◦ C rise in heating temperature
between 200 and 240 ◦ C compared to no significant change after 15 or 45 min of heating
(Figure 3).
Although very few (n = 3) studies evaluated temperatures above 240 ◦ C, mean levels
of individual TFA (but not total TFA) in oils heated to these temperatures were overall
significantly greater compared to oils heated to <200 ◦ C. Within this temperature range,
C18:1t (0.72% increase per 10 ◦ C rise in temperature, 95% CI: 0.59 to 0.84%) and C18:2t
(0.15%, 95% CI: 0.01 to 0.23%) levels increased with heating, while levels of C18:3t de-
creased (−0.74%; 95% CI: −0.93 to −0.54%). There was no significant interaction between
heating temperature and time for C18:1t. While significant interactions were observed for
C18:2t and C18:3t at various temperature intervals, the interaction effect was quite modest
(Figure 3).
Nutrients 2022, 14, 1489 7 of 11

Figure 3. Margins plot demonstrating the interaction between heating temperature and heating time
for various TFA. In subfigures (A), (B), and (D), the data label for 480 min is 0.31, 0.18, and 0.86,
respectively, and in (C) it is 0.04. Data were fitted using mixed multilevel linear regression with
intercepts for studies specified as random effects and splines at 200 and 240 ◦ C. Slopes represent
change in C18:1t (A), C18:2t (B), C18:3t (C), and total TFA (D) per 10 ◦ C (% of total fatty acids) change
in heating temperature. Margins plots were created for those temperature splines where there was a
statistically significant change in TFA.

4. Discussion
In our systematic review of 33 studies investigating the effect of heating edible oils on
their TFA levels, we found that heating to temperatures most commonly used in cooking
(≤200 ◦ C) had minimal impact on TFA levels but heating to higher temperatures (>200 ◦ C)
could increase levels of TFA. The effect of heating on formation of individual types of
TFA appeared to be largely consistent, leading to gradually increasing TFA levels with
increasing temperature above 200 ◦ C. Furthermore, levels of some TFA subtypes increased
further with prolonged heating, especially at temperatures above 200 ◦ C.
Our novel findings suggest that even at the temperature range >200 ◦ C (where signifi-
cant increase in TFA was observed), the magnitude of the increase appears to be relatively
small. For instance, the expected increase in total TFA for an increase in temperature be-
tween 25 ◦ C (room temperature) and 220 ◦ C is 7.4% (after a median cooking time of 45 min
of the included studies), whereas the level of TFA in partially hydrogenated vegetable oil,
the major target of global TFA elimination, is typically around 25–40% [46]. Nonetheless,
these findings affirm recommendations from several European countries that to minimise
formation of harmful TFA, frying oil should not exceed common cooking temperatures
(i.e., <200 ◦ C) and support other public health recommendations that prolonged and/or
repeated use of cooking oils should be avoided, which may be particularly relevant for
some informal food sectors in low- and middle-income countries where such practices
Nutrients 2022, 14, 1489 8 of 11

may be common [47–50]. However, there appears to be a lack of formal guidance on the
repeated use of cooking oils at both an international and national level, with the Food
Safety and Standards Authority of India being a notable exception [47,51]. There may be a
need to support vendors in both the formal and informal food sectors to avoid the practice
of reusing cooking oils, such as through targeted education programs or subsidised access
to fresh oils and used oil waste removal.
Though we have found some evidence to suggest that under various heating condi-
tions each of the subtypes of TFA assessed here and total TFA increase, C18:3t was found
to be the TFA that most readily increased (i.e., even below 200 ◦ C, it showed significant
increase). This may indicate that the precursor fatty acid (C18:3, alpha-linolenic acid) is
more susceptible to the effects of heating than other mono- and polyunsaturated fatty
acids (i.e., oleic acid and linoleic acid for C18:1 and C18:2, respectively). This is a novel
finding which suggests that as such, the avoidance of cooking oils that contain high levels
of C18:3, such as various seed oils, in cooking methods reaching high temperatures may
be a useful additional way to avoid the generation and consumption of TFA. It should be
noted that 240 ◦ C is above the smoking point of most cooking oils, i.e., the temperature at
which oil starts to vaporise, which is typically undesirable for appearance, taste, and utility.
Heating oils above their smoking point would not only impact the fat quality and lead to
the generation of TFA but could also increase the levels of carcinogenic compounds [52].
Thus, cooking at temperatures much lower than 240 ◦ C is important for avoidance of
generation of such compounds in addition to TFA [43].
To our knowledge, this is the first systematic review that synthesised data on the
relationship between TFA levels and cooking temperature and time. Strengths of this
study include its relatively large sample size, standardisation of outcome measures that
enabled pooling of data across studies, adjustment for individual study variation by use
of mixed multilevel models with random intercepts for individual studies, adjustment for
oil types, and analysis of changes in individual TFA isomers. Nevertheless, the results
of this study should be interpreted in the context of some limitations. First, the varied
results for different TFA isomers at temperatures above 240 ◦ C reflect an overall paucity
of data at this temperature range and should be interpreted with caution, although such
cooking temperatures may have little real-world relevance as most common household
cooking practices occur at temperatures between 160 and 200 ◦ C. Second, pooled analyses
could not be performed for individual oil types given relatively few data points. Rather, we
adjusted for oil type in our overall mixed models, thereby retaining statistical power of our
dataset. Third, given the relatively few data points investigating the change in TFA content
of food items cooked in edible oils, we were unable to meaningfully pool data investigating
change in TFA content of foods. This is an important consideration as the TFA level in
frying materials and the frying oils have been shown to affect each other by the exchange
of fats during cooking [43]. As such, more studies should evaluate how heating edible
oils changes the TFA content of various foods cooked in those oils. Fourth, owing to the
considerable heterogeneity in reported units between studies (e.g., % of total fatty acids,
% of 16:0, % C18:1-9c, % of C18:2, % of C18:3, or mg/g of chicken leg meat), and despite
consolidation of some categories, we were unable to pool data from studies that reported
different units. Future studies should report consistent units, for example “% of total fatty
acids”, to facilitate pooling across studies. Finally, for some TFA subtypes, the number of
studies and experiments including controls at room temperature was few, reducing the
statistical power of comparisons of mean TFA at different temperature intervals.

5. Conclusions
Our systematic review of the literature suggests that while heating edible oils to
commonly used cooking temperatures has little effect on TFA generation, heating to higher
temperatures and for a longer period of time can increase TFA levels. These findings
provide further evidence that prolonged heating of edible oils to very high temperatures
may be harmful and should be avoided to reduce dietary intake of TFA.
Nutrients 2022, 14, 1489 9 of 11

Supplementary Materials: The following supporting information can be downloaded at: https:
//www.mdpi.com/article/10.3390/nu14071489/s1, Table S1: Characteristics of included studies;
Table S2: Trans fatty acid concentration of two most commonly studied cooking oils (corn and
soybean) and hydrogenated vegetable fat at predetermined temperature intervals; Table S3: Quality
assessment of included studies; Table S4: Summary measures of cooking temperature, cooking
time, and cooking cycle according to method of cooking; Table S5: Summary measures of cooking
temperature, cooking time, and cooking cycle according to type of trans fatty acid; Table S6: Change
in TFA content of cooking oils (% of total fatty acids) as a function of heating temperature; Table S7:
Change in TFA as a function of the interaction between heating temperature and heating time
(per 1 ◦ C increase).
Author Contributions: Conceptualisation, L.H. and J.H.Y.W.; methodology, J.H.Y.W. and M.M.;
software, S.B.; validation, S.B. and M.M.; formal analysis, S.B.; investigation, D.M. and L.H.; data
curation, L.H. and S.B.; writing—original draft preparation, S.B.; writing—review and editing, D.M.,
L.H., J.H.Y.W. and M.M.; visualisation, S.B.; supervision, J.H.Y.W. and M.M. All authors have read
and agreed to the published version of the manuscript.
Funding: This research received no external funding.
Institutional Review Board Statement: Not applicable.
Informed Consent Statement: Not applicable.
Data Availability Statement: The data presented in this study are available on request from the
corresponding author.
Conflicts of Interest: The authors declare no conflict of interest.

Appendix A
Search strategy used for systematic review of the literature.
(cook* or fry* or baking or heat* or hydrogen* or thermal or processing)
AND
(cooking oil or vegetable oil or edible oil or canola oil or peanut oil or coconut oil or
palm oil or olive oil or avocado oil or seed oil or grapeseed oil or walnut oil or sesame oil
or flaxseed oil or sunflower oil or rice bran oil or rapeseed oil or sunflower oil or safflower
oil or ghee or clarified butter or vanaspati)
AND
(trans fat or trans fatty acids or trans isomer).

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