Food Control 130 (2021) 108295

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Food Control
journal homepage: www.elsevier.com/locate/foodcont

Analysis of peroxide value in olive oils with an easy and green method
Francesco Longobardi a, *, Francesco Contillo a, Lucia Catucci a, b, Luca Tommasi c,
Francesco Caponio d, Vito Michele Paradiso d, e, **
a
Department of Chemistry, University of Bari, Via Orabona 4, I-70126, Bari, Italy
b
CNR-IPCF Institute for Physical and Chemical Processes, Bari Unit, Via Orabona 4, I-70126, Bari, Italy
c
BonassisaLab, S.S.16 Km.684,300, I-71122, Foggia, Italy
d
Department of Soil, Plant and Food Sciences, University of Bari, Via Amendola 165/a, I-70126, Bari, Italy
e
Department of Biological and and Environmental Sciences and Technologies, Food Science Laboratory, University of Salento, I-73100, Lecce, Italy

A R T I C L E I N F O A B S T R A C T

Keywords: A green, easy-to-use, and sensitive spectrophotometric iodide-dependant method for the quantitative determi­
Hydroperoxides nation of the lipidic hydroperoxides in virgin olive oils was developed. Virgin olive oils were added with 0.5%
Spectrophotometric method HCl-ethanol, with a saturated KI solution and, after incubation, the resulting solution was filtered. Hydroper­
Virgin olive oil
oxides were determined indirectly by reading the absorbance of the generated triiodide at 350 nm; the total time
Green analytical chemistry
of analysis was about 7 min. A good linearity (with correlation coefficient (r) of 0.9997) of the calibration curve
was obtained with purified olive oil spiked with tert-butylhydroperoxide at levels ranging from 1.0 to 10.0
meqO2/kg, with variation coefficients less than 5% (n = 3) and limit of detection and quantification of 0.3 and
0.9 meqO2/kg, respectively. Results obtained with the spectrophotometric method showed good correlation with
those obtained with the official method with a r of 0.9819 confirming the reliability of the developed method.

1. Introduction milliequivalents of oxygen per kilogram of sample (meqO2/kg). Never­

theless, this method presents several limitations: time-consuming and
Oxidation is of paramount importance for the quality and shelf life of labour-intensive procedure, low sensitivity, need of large amounts of
vegetable oils, a fortiori for high quality products such as virgin olive sample and to weight the sample according to the presumed number of
oils (Paradiso, Pasqualone, Summo, & ). National and international peroxides, difficulty to determine the endpoint, large amounts of wastes,
regulations (CodexAlimentarius, 2017; EEC Commission, 1991) set legal use of harmful/polluting solvents (Dobarganes & Velasco, 2002;
limits for the analytical parameters related to oil oxidation. Peroxide Schaich, 2013; Shahidi & Zhong, 2005). Besides these drawbacks, the
value, still the most representative parameter considered to measure main limitation is the co-presence of the interfering reaction between
oxidation in virgin olive oils, measures the level of primary oxidation oxygen present in solution and potassium iodide which again produce
products – hydroperoxides of fatty acids (ROOH) – deriving from H iodine causing an overestimation of the PV. In addition, absorption of
abstraction from the carbons (mainly bis-allylic and allylic) of the fatty iodine by unsaturated fatty acids (leading to PV underestimation), the
acid chain (Schaich, 2005). The official method to determine peroxide oxidation of acetic acid and lipid oxidation (source of peroxides), and
value (PV) in olive oils is the iodometric titration assay, as reported in the sodium thiosulfate decomposition are possible sources of error.
the Annex III of the Reg. (EEC) 2568/91 (EEC Commission, 1991), based To overcome these drawbacks, other analytical methods have been
on the oxidation of the iodide ion (I− ) by ROOH in acidic environment. proposed for PV measurements but neither of them is free from com­
Briefly, oil is dissolved in a mixture of chloroform/acetic acid and added plications. Some proposed methods, either being green and almost sol­
with a saturated solution of potassium iodine. The reaction of I− with ventless such as FTIR (Yu, van de Voort, & Sedman, 2007), or using
ROOH leads to the formation of molecular iodine (I2) which is titrated organic solvents, such as RP-HPLC (Yang, 1992), require expensive
with a solution of sodium thiosulfate and starch as an endpoint indicator equipment. Other methods are based on the oxidation of iron with for­
(Kiritsakis, Kanavouras, & Kiritsakis, 2002). The PV is calculated as mation of ferric ions complexes (with thiocyanate or orange xylenol)

* Corresponding author. Department of Chemistry, University of Bari, Via Orabona 4, I-70126, Bari, Italy.
** Corresponding author. Department of Soil, Plant and Food Sciences, University of Bari, Via Amendola 165/a, I-70126, Bari Italy.
E-mail addresses: francesco.longobardi@uniba.it (F. Longobardi), vito.paradiso@unisalento.it (V.M. Paradiso).

https://doi.org/10.1016/j.foodcont.2021.108295
Received 30 September 2020; Received in revised form 24 May 2021; Accepted 30 May 2021
Available online 7 June 2021
0956-7135/© 2021 Elsevier Ltd. All rights reserved.
F. Longobardi et al. Food Control 130 (2021) 108295

that can be detected spectrophotometrically (Schaich, 2013). The ferric 5.2, 6.7, and 8.2 meqO2/kg; (b) by spiking the POO with increasing
thiocyanate method is sensitive and reproducible, nevertheless it re­ quantity of TBHP obtaining calibrants with PV of 1.0, 2.0, 4.0, 6.0, 8.0,
quires harmful solvents. The orange xylenol method, instead, is affected and 10 meqO2/kg. The limit of detection (LOD) was calculated as: 3 ×
by many factors, such as the amount of sample, solvent used, and source sy/x/b where sy/x is the residual standard deviation of the regression line
of xylenol orange (Shahidi & Zhong, 2005). Moreover, these methods and b is the slope of the calibration curve. The limit of quantification
were conceived for peroxides in solvents or living tissues and may show (LOQ) was calculated as: 3 × LOD.
lack of linearity in the concentration ranges expected in food samples, so For method validation a total of 23 virgin olive oils (VOOs) were
that sample size and dilution should be adjusted, particularly in shelf-life used. Ten VOOs were supplied by producers (year of production 2019),
studies (Schaich, 2013). Analogous considerations could be made for the directly sampled from storage tanks, and filled in dark glass bottles. At
methods developed using triphenylphosphine (Talpur, Sherazi, Mahe­ the time of sampling (July 2020), they had an average age of eight
sar, & Bhutto, 2010; Yu, Li, Sun, Dong, & Wang, 2015). months. Thirteen VOOs were purchased from supermarkets (dark green
Almost three decades ago, Løvaas proposed a spectrophotometric coloured bottles, year of production 2018, with best-before dates
iodide-dependant method to assess lipid hydroperoxides based on the ranging from September 2020 to February 2021). All 23 samples were
absorbance of the triiodide (I3) formed following the redox reaction produced in Italy as reported by producer or in product label. VOOs
between iodide and hydroperoxides (Løvaas, 1992). In fact, the liberated were analyzed within one week from sampling and were stored until
iodine (generated by the redox) and iodide (in excess) react to form I−3 , analysis at room temperature under dark conditions. The validation
which can be detected spectrophotometrically providing an indirect procedure was carried out by comparing the PV results obtained by
quantification of hydroperoxides. The method proposed by Løvaas was simultaneously analyzing each VOO sample, both with the spectropho­
applied to capelin oil, a fish oil, which is quite a different matrix, tometric method and with the official method.
compared to virgin olive oil, for fatty acid composition, as well as for the
presence of potentially interfering substances. Moreover, though 3. Methods
showing the feasibility of this analytical approach, the paper did not
report method validation. Therefore, in view of the application of this Titrations were performed as described by the Reg. (EEC) 2568/91,
method to virgin olive oils, possibly for in-situ analysis (e.g. in olive Annex III (EEC Commission, 1991).
mills and bottling/storage plants) and oxidative status tracking, the For spectrophotometric analysis an aliquot of 0.10 g of sample was
following issues should be considered: added with 1 mL of 0.5% HCl-ethanol, 0.10 mL of saturated KI solution
and incubated for 5 min. The resulting solution was added with 4 mL of
- Adaptation to the complexity of the virgin olive oil matrix water and transferred to a 1-cm UV cuvette with a plastic syringe
- Use of low volumes of reagents and solvents equipped with a 0.45 μm pore size regenerated cellulose (RC) filter
- Reduction of the environmental and health impact of reagents and (Levanchimica s.r.l., Bari, Italy); the absorbance was measured at 350
solvents nm against the blank (obtained by using the same procedure but without
- Method validation. sample). The total time of analysis was about 7 min. Solutions showing
absorbance values higher than 1.5 a.u. were suitably diluted and re-
In this framework, this research work was focused on the develop­ analyzed; the diluted factor was considered for the calculation of
ment of spectrophotometric iodide-dependant method for monitoring concentrations.
the oxidation of virgin olive oils during early stages of the oxidation All experiments were done in triplicate.
process aiming to create an analytical protocol easy-to-use, as sustain­
able as possible (green), with a reduced consumption of solvents and 3.1. Statistical analysis
samples, with a high sensitivity and specificity towards primary oxida­
tion products. For comparison between two or more group means, the t-test or the
one-way analysis of variance (ANOVA) were used, respectively.
2. Materials and methods Regression lines were compared by using the analysis of co-variance
(ANCOVA) (Miller, Miller, & Freund, 2014). Statistical analyses were
2.1. Reagents and apparatus performed using Statistica version 8.0 (StatSoft Italia srl, Padova, Italy).

Potassium iodide (KI, purity ≥99.0%), tert-Butyl hydroperoxide so­ 4. Results and discussion
lution 70 wt % in H2O (TBHP, Luperox® TBH70X), hydrochloric acid
37% v/v (HCl), glacial acetic acid (CH3COOH, purity ≥99.0%), chlo­ 4.1. Method development
roform (CHCl3 purity ≥99.5%), sodium thiosulfate (Na2S2O3, purity =
99%), and starch were purchased from Sigma Aldrich (Milan, Italy). 4.1.1. Preliminary evaluation of the spectrophotometric detection feasibility
Absorbance measurements were carried out with a Shimadzu (Shimadzu Preliminarily, to verify the possibility of using the spectrophoto­
Corporation, Kyoto, Japan) UV–Vis spectrophotometer model UV-1700 metric detection rather than the manual titration for the determination
PharmaSpec. of hydroperoxide contents, the absorbances of the aqueous solutions
obtained by analyzing oxidized olive oil samples with the official
2.2. Samples, calibration curves, and validation method were measured between 250 and 500 nm. As showed in Fig. 1,
two absorption peaks due to the formed chromophore at about 290 and
Oxidized olive oil (OOO) samples used for the development of the 350 nm, respectively, were observed having intensities proportional to
method were prepared by incubating a refined olive oil (ROO) at 60 ◦ C the PV of the analyzed OOOs (Løvaas, 1992). Nevertheless, also for the
for different times ranging between 3 and 14 days. The concentrations of experiment carried out in the absence of sample, the I−3 signals were still
hydroperoxide generated were determined by using the iodometric detectable, even if of low intensity. This confirms the presence of the
titration. interfering reaction between the dissolved oxygen and iodide to form
A purified olive oil (POO) having a negligible PV value was obtained again triiodide, causing PV overestimation risks. Subsequently, aiming
as reported by Paradiso et al. (Paradiso, Gomes, Nasti, Caponio, & ). to improve the analytical performance, some factors such as nature of
Two matrix-assisted calibration curves were built with calibrants organic solvent and acid, acidic concentration, and intensity of the
prepared as following: (a) by blending two OOOs having PV of 8.2 and interfering reaction absorbance were investigated studying their effect
2.2 meqO2/kg, respectively, obtaining calibrants with PV of 2.2, 3.7, on spectra and/or on the peak intensity at 350 nm.

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F. Longobardi et al. Food Control 130 (2021) 108295

Fig. 1. Triiodide absorption spectra of aqueous solutions obtained as follows: Fig. 2. Triiodide median absorption spectra (n = 3) of aqueous solutions ob­
0.25 g of oxidized olive oil was added with 6 mL of chloroform/acetic acid tained as follows: 3 mL of 3% HCl-methanol solution was added with 0.25 mL of
(3:2), 0.25 mL of saturated KI solution and incubated for 5 min. After 75 mL of saturated KI solution and incubated for 5 min. After 40 mL of water addition the
water addition the resulting solution was transferred to a 1-cm UV cuvette and resulting solution was transferred with a syringe equipped with a RC (‒‒ ‒), CA
the absorbance was measured between 250 and 500 nm. Sample peroxide value (•), or no (solid line) filter to a 1-cm UV cuvette and the absorbance was
(meqO2/kg): 43.1(•), 26.9(‒ ‒ ‒), 8.2(• —); solid line was obtained measured between 250 and 500 nm. The inset displays the analogous spectra
without sample. obtained with 0.25 g of an oxidized olive oil with a peroxide value of 14.3
meqO2/kg.
4.1.2. Filtration and extraction optimization
To reduce the environmental impact of the method, the acetic acid/ 4.1.3. Effect of the concentration of hydrochloric acid
chloroform mixture used in the titrimetric method was replaced with an As known, the strongly acid conditions adopted in the iodometric
acidified alcohol. Firstly, 3% HCl-methanol solution was evaluated. methods represent a weakness of these approaches because they also
However, the use of alcoholic solutions did not lead to a clear separation accelerate the iodide background oxidation. Taking into account that
between the organic and the aqueous phase in the subsequent steps of acid cannot be omitted (being necessary for the reaction between ROOH
the analysis, and required a filtration step before carrying out the and I− ), different ethanol solutions containing decreasing amounts of
spectrophotometric measurements. Therefore, to identify a filter with HCl (3%, 2%, 1%, 0.5%), both in the presence and absence of samples
minimal retention of the chromophore of interest, experiments were (PV = 5.0 meqO2/kg), were used in order to evaluate these background
initially carried out by using 3% HCl-methanol in the absence of olive oil reactions. As expected, as the amount of acid decreases, a significant
and the absorption spectra due to the interfering reaction were recorded decrease in the intensity of absorbance was observed both in the pres­
both without the filtering operation, and using two different filters, i.e. ence and absence of sample (see Fig. 3). Specifically, it is important to
cellulose acetate (CA) and regenerated cellulose (RC) filter. As shown in note that the spectrum obtained by using the 0.5% HCl-ethanol solution
Fig. 2, similar absorbances were obtained from the unfiltered solutions in the presence of the oxidized olive oil was still well observable
and the solutions filtered with the RC with no statistically significant allowing the ROOH detection even at this low PV value. On the contrary,
difference at 350 nm (p ≤ 0.05; n = 3); on the other hand, the CA filter in the same acidic condition, the absorbance obtained without sample
negatively affected the signal intensity reducing it significantly. Subse­ was drastically reduced. Consequently, in the following experiments the
quently, the same experiments were repeated in the presence of an OOO acid concentration was fixed at 0.5%.
having PV = 14.3 meq O2/kg. Also, in this case, a signal intensity
decrease was observed with the CA compared to the RC filter. Therefore, 4.1.4. Baseline evaluation for interfering reaction removing
the latter was chosen as the optimal filter and used in subsequent ex­ To completely remove the influence on the absorbance due to the
periments. It is important to highlight that even if the filtration step interfering reaction, the mixture prepared following the analytical
slightly complicates the analysis protocol, it simultaneously reduces the protocol but without olive oil could be used as spectrophotometric blank
contact time between the iodine produced and the sample limiting the and consequently its absorption spectrum as baseline. However, to be
risk of undesirable absorption of iodine by unsaturated fatty acids. sure that this approach did not introduce a source of error, it was
Besides 3% HCl-methanol, also 3% HCl-ethanol was evaluated. Re­ considered necessary that the blank spectrum should have never
sults obtained analyzing olive oil samples (PV = 14.3 meqO2/kg) recorded an absorbance reading significantly varying during time. To
revealed that the maximum intensity at 350 nm was increased by 211 ± test the stability of the saturated solution of KI over time, so as to
10% (n = 3) by using ethanol rather than methanol. This absorbance determine the same extent of the interfering reaction and consequently a
increment could be explained considering two aspects. On the one hand, constant baseline, starting from the same saturated solution of KI,
I−3 showed a greater extinction coefficient in ethanol than in methanol, different blank solutions were prepared at different times from KI so­
on the other hand, even if the reaction of iodine and iodide is strongly in lution preparation, i.e. 10, 20, 30, 60, 120, 240, and 360 min, and their
favor of I−3 formation, the equilibrium constant (K) is inversely related to absorbance quickly read. The experiments were carried out in triplicate:
the dielectric constant of the solvent determining a K value greater in the mean values at 350 nm and their standard deviations were calcu­
ethanol (ε = 25.16) than in methanol (ε = 33.64) (Gregory & Clarke, lated and showed in Fig. 4. By applying an analysis of variance ANOVA,
2005; Løvaas, 1992). Therefore, in the following studies HCl-ethanol no significant differences (p ≤ 0.05) were found among the means,
was used as organic solvent. highlighting that the absorbance could be considered constant at least
during the investigated time range, i.e. up to 6 h from the preparation of
the KI solution. Therefore, in the following experiments the contribution

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F. Longobardi et al. Food Control 130 (2021) 108295

above, as well as the aims of this research work, it was decided to adopt
the following analytical protocol. An aliquot of 0.10 g of sample was
added with 1 mL of 0.5% HCl-ethanol, 0.10 mL of saturated KI solution
and incubated for 5 min. The resulting solution was filtered on a RC filter
and analyzed spectrophotometrically. The absorbance was measured at
350 nm against the blank (obtained by using the same procedure but
without sample). This protocol proved to be a good compromise be­
tween the desired characteristics in terms of sensitivity, quantity of
sample and solvent, eco-compatibility of the solvent, and some practical
aspects such as the maximum measurable absorbance fixed at 1.5 a.u.
The chemical reactions for the I−3 formation, the factors investigated for
the method development, and the final protocol were summarized in
Fig. 5.

4.3. Method validation

4.3.1. Calibration Curves and Detection and quantification limits

Two calibration curves were built following the developed protocol.
A first matrix calibration curve (Fig. 6a), obtained by blending refined
olive oils, showed a good linear response (r = 0.9991) obtained in the
analyzed concentration range (2.2–8.2 meqO2/kg), with variation co­
efficients less than 5% (n = 3) and limit of detection (LOD) and quan­
Fig. 3. Triiodide median absorption spectra (n = 3) of aqueous solutions ob­ tification (LOQ) of 0.4 and 1.2 meqO2/kg, respectively. A second matrix
tained as follows: 0.10 g of oxidized olive oil (peroxide value = 5.0 meqO2/kg)
calibration curve (Fig. 6b), obtained by spiking a purified olive oil with
was added with 1.2 mL of 3% (• —), 2% (‒ ‒ ‒), 1% (•), 0.5% (solid line) HCl-
TBHP (concentration range 1.0–10.0 meqO2/kg), showed a good linear
ethanol solution, 0.10 mL of saturated KI solution and incubated for 5 min.
After 16 mL of water addition the resulting solution was transferred with a response (r = 0.9997), with variation coefficients less than 5% (n = 3)
syringe equipped with a RC filter to a 1-cm UV cuvette and the absorbance was and limit of detection (LOD) and quantification (LOQ) of 0.3 and 0.9
measured between 250 and 500 nm. The inset displays the analogous spectra meqO2/kg, respectively. By carrying out an Analysis of covariance
obtained without sample. (ANCOVA) between the two linear regressions, it was found that the
regressions are not statistically different (p ≤ 0.05); therefore, the choice
to use one method or the other for the preparation of the calibrants
mainly depends on experimental considerations. In particular, the
fortification approach of a purified oil has several advantages compared
to the mixture approach. Fortification, in fact, allows to freely choose
both the minimum level of fortification and, within the limits of line­
arity, the concentration range to be used for calibration. Otherwise, in
the case of blending, the lowest point that can be analyzed depends on
the availability of a poorly oxidized oil. In addition, the concentration
values achieved through the use of the standard solution can be
considered more accurate because, as mentioned above, they do not
depend on the quantifications obtained by the official method; the latter,
in fact, is affected by an higher uncertainty. For the reasons described
above, the subsequent quantification analyses were carried out using the
calibration curve obtained by fortification.

4.4. Methods comparison

For the validation, 23 olive oil samples were analyzed by the pro­
posed method and by the official method based on iodometric titration.
As showed in Fig. 7, a very good correlation (r = 0.9819) with a slope
Fig. 4. Absorbance measurements at 350 nm of blank solutions prepared at very close to 1 and an intercept close to 0 was obtained confirming the
different times, after the preparation of the saturated KI solution, obtained as reliability of the developed method.
follows: 1.2 mL of 0.5% HCl-ethanol solution was added with 0.10 mL of
The method proposed shows different advantages in comparison
saturated KI solution and incubated for 5 min. After 4 mL of water addition, the
with the official one. In particular, the substitution of 25 mL of chloro­
resulting solution was transferred to a 1-cm UV cuvette and the absorbance was
form/acetic acid (2:3) with 1 mL of ethanol considerably increases the
measured at 350 nm. Each value represents mean ± standard deviation of
triplicates. sustainability of the method for the reduction of the volumes itself but
above all considering the nature of the solvents used. In fact, as reported
in a selection guide, based on a survey of publicly available solvent se­
of the interfering reaction in the absorption of the analysis of oil samples
lection guides and aligned with the Global Harmonized System (GHS)
was removed easily by using the spectrum obtained from the analysis of
and European regulations, a greenness evaluation between chloroform/
the saturated KI solution as baseline which was subtracted from subse­
acetic acid and ethanol would lead to choosing the latter (Prat et al.,
quent sample readings.
2016). In detail, adopting a methodology based on a simple combination
of a set of safety, health and environment criteria for the ranking of
4.2. Analytical protocol solvents, acetic acid, chloroform and ethanol are indicated as “Prob­
lematic”, “Highly Hazardous”, and “Recommended” solvent, respec­
Considering the results obtained from the experiments reported tively, demonstrating as stated before. Moreover, the replacement of

4
F. Longobardi et al. Food Control 130 (2021) 108295

Fig. 5. From top to the bottom: the chemical reactions used for the indirect determination of the lipidic hydroperoxides (solid box); the experimental factors studied
for the development of the method (dashed boxes); the steps of the developed analytical protocol (circular box).

acetic acid with hydrochloric acid removes the risk of having a PV min), it showed a much less performing analytical sensitivity. In fact, the
overestimation coming from the possible oxidation of acetic acid to Løvaas method was applied to analyze oils whose lowest value (87.6
peroxides. Unlike the official method, for the method developed herein meqO2/kg) was almost three orders of magnitude larger than that
the amount of sample to be weighted does not depend on the expected measured in the method proposed here (1.0 meqO2/kg).
peroxide value but is constant. This last feature makes the PV evaluation As regards the mixture of solvents used by Løvaas, it can only be
not influenced by the amount of sample, increasing both the perfor­ considered partially “Recommended” because, even if these solvents
mance and the ease to use of the method. Advantages regarding these could be bio-derived, they are currently mostly synthetized by the
last two aspects also derive from the substitution of titration with petrochemical industry. Furthermore, methanol presents non-negligible
spectrophotometric detection. Indeed, the use of the instrumental health hazards with a significantly lower daily exposure limit than
determination leads to a higher inherent precision, a reduction of the ethanol (Prat et al., 2016). Finally, as previously mentioned, the vali­
total time of analysis (7 vs. 20–30 min), no thiosulphate decomposition dation of the method proposed by Lovaas appeared to be incomplete
risk, no need of trained operators, and, thanks to the reduced volumes because it had not been applied for the analysis of naturally oxidized
required for the spectrophotometric measurement, a reduction of the samples and no comparison with the official method was carried out.
used quantity of sample of about one order of magnitude, i.e. 0.1 g
against a value ranging from 1.2 to 5.0 g according to the expected 5. Conclusions
peroxide value. Regarding the interfering reaction, the use of the white
absorbance as a baseline allows to remove, or in any case drastically In summary, a green and sensitive spectrophotometric iodide-
reduce, this interference from atmospheric oxygen without degassing, dependant method for the quantitative determination of the lipidic hy­
further facilitating the analysis protocol. droperoxides in virgin olive oils was developed in the present research.
Comparing the method proposed herein with that of Løvaas, several The method proposed shows different advantages in comparison with
differences could be highlighted, such as the type of sample, the solvents the official method in terms of analytical performances (time, accuracy,
and reagents used, the range values of peroxides analyzed, the valida­ reproducibility, LOQ, and LOQ), sustainability of the solvent (substitu­
tion procedure adopted. In particular in the Løvaas method, capelin oil tion of chloroform/acetic acid with 0.5% HCl-ethanol), ease of use (no
was analyzed, a mixture of methanol and butanol was used as solvents titration), reduction of sample amounts and solvent volumes, and
and a Fe(II) salt was introduced to catalyze the reaction between the reduction of interferences from side reactions.
peroxides and iodide, allowing to use lower amount of HCl, thus mini­ Future studies will be focused on further validation of the proposed
mizing the interfering reaction due to oxygen. While the protocol pro­ method carrying out by interlaboratory studies.
posed by Løvaas allowed to measure absorbance without prior filtration
of the solution, on the other hand it required a greater number of sol­ Declaration of competing interest
vents and reagents. Above all, despite the analysis time turned out to be
about double compared to the method proposed herein (15 against 7 The authors have no competing interests to declare.

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F. Longobardi et al. Food Control 130 (2021) 108295

CRediT authorship contribution statement

Francesco Longobardi: Conceptualization, Methodology, Valida­

tion, Formal analysis, Writing – original draft, Writing – review &
editing, Supervision, Project administration, Funding acquisition.
Francesco Contillo: Formal analysis, Investigation, Writing – review &
editing, Visualization. Lucia Catucci: Conceptualization, Writing – re­
view & editing, Funding acquisition. Luca Tommasi: Conceptualiza­
tion, Validation, Investigation, Resources, Writing – review & editing,
Funding acquisition. Francesco Caponio: Conceptualization, Writing –
review & editing, Funding acquisition. Vito Michele Paradiso:
Conceptualization, Methodology, Validation, Resources, Writing –
original draft, Writing – review & editing, Supervision, Project admin­
istration, Funding acquisition.

Acknowledgements

The work was carried out within the M3O3 project (“Multifunctional
microsystems for the monitoring of olive oils oxidation” – project code
XMPYXR1) founded by Puglia Region (POR Puglia FESR-FSE 2014–2020
- Innonetwork call).

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