Comparative study of mycotoxin occurrence in Andean and cereal grains cultivated in South America and North Europe

Abstract

The consumption of high-quality Andean grains (a.k.a. pseudocereals) is increasing worldwide, and yet very little is known about the susceptibility of these crops to mycotoxin contamination. In this survey study, a multi-analyte liquid chromatography–tandem mass spectrometry (LC–MS/MS) method was utilised to determine mycotoxin and fungal metabolite levels in Andean grains (quinoa and kañiwa) in comparison to cereal grains (barley, oats and wheat), cultivated in both South American (Bolivia and Peru) and North European (Denmark, Finland and Latvia) countries. A total of 101 analytes were detected at varying levels, primarily produced by Penicillium spp., Fusarium spp. and Aspergillus spp., depending on the type of crop, geographical location and agricultural practices used. Generally, Andean grains from South America showed lower mycotoxin contamination (concentration and assortment) than those from North Europe, while the opposite occurred with cereal grains. Mycotoxin contamination profiles exhibited marked differences between Andean and cereal grains, even when harvested from the same regions, highlighting the need for crop-specific approaches for mycotoxin risk mitigation. Lastly, the efficacy of grain cleaning in respect to total mycotoxin content was assessed, which resulted in significantly lower levels (overall reduction approx. 50%) in cleaned samples for the majority of contaminants.

Full text

Food Control 130 (2021) 108260
Available online 29 May 2021
0956-7135/© 2021 The Author(s). Published by Elsevier Ltd. This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/).
Comparative study of mycotoxin occurrence in Andean and cereal grains
cultivated in South America and North Europe
J.M. Ramos-Diaz
a
,
*
, M. Sulyok
b
, S.E. Jacobsen
c
, K. Jouppila
a
, A.V. Nathanail
a
a
Department of Food and Nutrition, University of Helsinki, P.O. Box 66 (Agnes Sj¨
obergin Str. 2), FI-00014, Helsinki, Finland
b
Department of Agrobiotechnology, IFA-Tulln, Institute of Bioanalytics and Agro-Metabolomics, University of Natural Resources and Life Sciences, Vienna (BOKU),
Konrad-Lorenz Str. 20, 3430, Tulln, Austria
c
Quinoa Quality ApS, Teglvaerksvej 10, 4420, Regstrup, Denmark
ABSTRACT
The consumption of high-quality Andean grains (a.k.a. pseudocereals) is increasing worldwide, and yet very little is known about the susceptibility of these crops to
mycotoxin contamination. In this survey study, a multi-analyte liquid chromatography–tandem mass spectrometry (LC–MS/MS) method was utilised to determine
mycotoxin and fungal metabolite levels in Andean grains (quinoa and ka˜
niwa) in comparison to cereal grains (barley, oats and wheat), cultivated in both South
American (Bolivia and Peru) and North European (Denmark, Finland and Latvia) countries. A total of 101 analytes were detected at varying levels, primarily
produced by Penicillium spp., Fusarium spp. and Aspergillus spp., depending on the type of crop, geographical location and agricultural practices used. Generally,
Andean grains from South America showed lower mycotoxin contamination (concentration and assortment) than those from North Europe, while the opposite
occurred with cereal grains. Mycotoxin contamination proles exhibited marked differences between Andean and cereal grains, even when harvested from the same
regions, highlighting the need for crop-specic approaches for mycotoxin risk mitigation. Lastly, the efcacy of grain cleaning in respect to total mycotoxin content
was assessed, which resulted in signicantly lower levels (overall reduction approx. 50%) in cleaned samples for the majority of contaminants.
1. Introduction
Quinoa (Chenopodium quinoa) and ka˜
niwa (Chenopodium pallidicaule)
are grains that were widely cultivated in the Andean mountains by Pre-
Hispanic civilizations, there comes its name Andean grains (a.k.a. pseu-
docereal; Andean grains belong to fam. Amaranthaceae while conven-
tional cereals to fam. Poaceae). Despite changes in dietary traits during
colonial and republican times, the rural consumption of quinoa and
ka˜
niwa was relatively common until the early 20th century, mostly, on
the Andean plateau. However, the massive importation of wheat
severely affected local farmers, leading to reduced cultivation and
consumption (Tapia, 1979). Additionally, ethnic discrimination,
involving indigenous communities and their traditional food, may have
restrained climate-resilient quinoa and ka˜
niwa to areas where no other
crop could grow (Hellin & Higman, 2005; Martinez-Zu˜
niga, 2007), thus
becoming staple crops for subsistence farming (Vassas and Viera Pak,
2010). After the revaluation of ancient knowledge, quinoa and ka˜
niwa
were found to be formidable food alternatives that could contribute to
the world food security (Bazile et al., 2016; FAO, 2011; Jacobsen, 2017;
Rodriguez et al., 2020). These gluten-free grains contain not only a
complete pool of amino acids, but also a high amount of essential
micronutrients and minerals. Those are key features for their nutritional
revalorization and growing popularity in the Western World (Repo--
Carrasco et al., 2003).
Even though Andean grains are as susceptible to fungal growth and
mycotoxin contamination as cereal grains (e.g. maize, wheat), there is
scarce information on the contaminating fungi and mycotoxin occur-
rence in quinoa and ka˜
niwa. The few available studies focused primarily
on the investigation of the mycoora present in Andean grains and not
on mycotoxin contamination. Case in point, the presence of Ascohyta,
Altenaria, Phoma, Fusarium, Bipolaris, Cladosporium and Pyronochaeta
genera in seeds of Chenopodium quinoa from Bolivia, Brazil, Czech Re-
public and Peru have been reported, but no accompanying mycotoxin
data were provided (Boerema et al., 1977; Spehar et al., 1997;
Dˇ
rímalkov´
a, 2003). Amaranth grains from Argentina were analysed to
examine mycoora, which was found to be dominated by the
mycotoxin-producing fungal species A. avus, A. parasiticus,
P. chrysogenum and F. equiseti (Bresler et al., 1995). Additionally, Pap-
pier et al. (2008) reported that Penicillium and Aspergillus were the most
frequently encountered genera in quinoa harvested from three locations
in Argentina. In the same study, processing of the grains for removal of
saponins (wet method) caused a decrease in Aspergillus incidence, whilst
increased the proportion of Penicillium, Eurotium, Mucor and Rhizopus
that was characterised as internal mycobiota.
* Corresponding author.
E-mail address: jose.ramosdiaz@helsinki. (J.M. Ramos-Diaz).
Contents lists available at ScienceDirect
Food Control
journal homepage: www.elsevier.com/locate/foodcont
https://doi.org/10.1016/j.foodcont.2021.108260
Received 22 March 2021; Received in revised form 29 April 2021; Accepted 17 May 2021

Food Control 130 (2021) 108260
2
However, analysis of mycotoxins is essential for all types of grains, as
these low-molecular-weight toxins can contaminate crops in all climatic
regions. Importantly, mycotoxins have been associated with a broad
range of toxic effects to both humans and animals, including acute
toxicity, immunotoxicity, hepatotoxicity, nephrotoxicity, carcinogenic-
ity and reproductive toxicity (Bhat et al., 2010). Safety of food- and
feedstuffs is paramount to consumers and thus, complex regulatory
frameworks and monitoring systems have been developed globally that
rely on the latest scientic knowledge and analytical tools. In the Eu-
ropean Union (EU), maximum levels have been established for a number
of mycotoxins in cereals and cereal-derived products (EU Commission
Regulation (EC) No 1881/2006). However, no such levels specic to
Andean grains exist.
Liquid chromatography–tandem mass spectrometry (LC–MS/MS) is
the most widely used method for accurate and reliable determination of
multiple mycotoxins at even minute concentrations in complex matrices
such as cereal grains and cereal-based foods (Malachov´
a et al., 2018).
Recent applications of LC–MS/MS methods for the simultaneous deter-
mination of multiple mycotoxins and modied forms include analysis of
wheat, barley, maize and cereal-derived products, among others, uti-
lising a variety of matrix-dependent sample preparation techniques
(Spaggiari et al., 2019; Ekwomandu et al., 2020; Ostry et al., 2020;
Drakopoulos et al., 2021; Rausch et al., 2021).
From the very limited number of studies that have measured my-
cotoxins in non-cereal grains, zearalenone (ZEN) was determined at
levels up to 1980
μ
g/kg in two samples of Amaranthus cruentus grains,
which had been stored moist (Bresler et al., 1991). No aatoxins,
ochratoxin A (OT-A) or sterigmatocystin were found, however, the
number of samples analysed was very limited. No mycotoxin contami-
nation was reported in the previously mentioned study of Pappier et al.
(2008), although the method was only capable of analysing aatoxins
and citrinin. In 2014, Arroyo-Manzanares et al. developed and validated
an ultra-high performance liquid chromatography (UHPLC)–MS/MS
method for the determination of 15 mycotoxins. It was used to analyse
quinoa and amaranth samples purchased from local markets in Spain,
but again none were found positive to any of the mycotoxins included in
the method. Lastly, commercially available quinoa our from Italy was
recently analysed with enzyme-linked immunosorbent assay (ELISA)
and found to contain 4.4 ng/g total aatoxins (B
1
, B
2
, G
1
and G
2
) and ca.
370 ng/g total fumonisins (B
1
and B
2
) (Sacco et al., 2020). In the same
study, total aatoxins at the level of 1.6 ng/g, and fumonisins at 111
ng/g were reported for amaranth (grain).
The aim of the present study was to determine mycotoxins present in
commercial varieties of quinoa and ka˜
niwa cultivated in South America
and North Europe. This work constitutes the most comprehensive survey
of mycotoxin content in Andean grains to date. Data generated herein
facilitate comparisons of mycotoxin content between regions and grain
types that can be valuable in the identication of mycotoxin-producing
fungi and mycotoxins of concern, thus contributing to the safe con-
sumption of Andean grains worldwide.
2. Materials and methods
2.1. Chemicals and reagents
HPLC gradient grade acetonitrile (HiPerSolv Chromanorm) was ob-
tained from VWR Chemicals (Vienna, Austria) and LC–MS Chromasolv
grade methanol from Honeywell (Seelze, Germany). LC–MS grade
ammonium acetate and glacial acetic acid (p.a.) were purchased from
Sigma-Aldrich (Vienna, Austria). Purication of reverse osmosis water
was performed using a Purelab Ultra system (ELGA LabWater, Celle,
Germany). Analytical standards of mycotoxins and fungal metabolites
were isolated in-house at the Department of Agrobiotechnology, IFA-
Tulln (Tulln, Austria), received as gifts by external collaborators or
purchased from commercial suppliers. The complete list of the analytical
standards’ details is provided in Sulyok et al. (2020).
2.2. Grains and sample preparation
Grain samples were obtained from Finland, Denmark, Latvia, Peru
and Bolivia (latitude descending order; Fig. 1). Andean and cereal grains
were collected from plots in close proximity, within the same cultivation
area (i.e. Denmark, Peru). Where applicable, a simple random sampling
was conducted with resulting specimens mixed into a pool. Grain sam-
ples were pre-treated as follows: Uncleaned, grains went through me-
chanical pre-cleaning (removal of large debris, leaves, twigs, etc.) but
rinsing was not conducted. Traditionally washed grains went through
mechanical pre-cleaning (removal of large debris, leaves, twigs, etc.)
and rinsing (water at room temperature) until foam was no longer
formed (indicative of saponin removal). Pearled grains were exposed to
an abrasive surface to remove saponin-containing outer layers. Me-
chanically cleaned grains were winnowed and screened but rinsing was
not conducted. Detailed information regarding individual pre-
treatments and cultivation areas is shown in Table 1 and Fig. 1. All
grains were eventually milled using an ultra-centrifugal mill (Retsch ZM
200, Haan, Germany) at 10,000 rpm, weighed (5 g), sorted (3–6 repli-
cates) and stored in falcon tubes at −20 ◦C. Prior to analysis, Andean and
cereal grain samples were extracted using 20 mL of the extraction so-
lution acetonitrile:water:acetic acid (79:20:1, v/v/v) and shaken for 90
min with a rotary shaker (GFL 3017, GFL; Burgwedel, Germany). The
supernatants (300
μ
L) were transferred into HPLC vials and diluted with
300
μ
L acetonitrile:water:acetic acid (20:79:1, v/v/v).
2.3. LC–MS/MS analysis
The method used for analysis of the Andean and cereal grains was
recently published by Sulyok et al. (2020). Briey, samples were ana-
lysed with a 1290 series Agilent Technologies UHPLC system (Wald-
bronn, Germany) coupled to a QTrap 5500 MS/MS that was equipped
with a TurboV electrospray ionisation (ESI) source (Sciex, California,
USA). Chromatographic separation was performed on a Gemini C18
column (150 mm ×4.6 mm, 5
μ
m particle size; Phenomenex, California,
USA) with a C18 security guard cartridge (4 mm ×3 mm; Phenomenex).
Quantication was based on external calibration (linear, 1/x weighed)
using a serial dilution of a multi-analyte working solution. Results were
corrected using apparent recoveries obtained through spiking experi-
ments (Sulyok et al., 2020). The accuracy of the method is veried on a
continuous basis by participation in a prociency testing scheme orga-
nized by BIPEA (Gennevilliers, France) with a current rate of z-scores
between −2 and 2 of >94% (>1500 results submitted).
2.4. Data processing
Data corresponding to the mycotoxin contaminants from all samples
was primarily sorted by cleaning method (cleaned and uncleaned) and
collection year (2015 and 2017). Standard normal variate (SNV) was
used as pre-treatment method due to its effectiveness in scattering
correction. Subsequently, principal component analysis (PCA) was used
to observe potential correlations with-in/among samples (a.k.a. load-
ings) and mycotoxins (a.k.a. scores). Data pre-processing and plotting
was done using SIMCA 15.0 software package (v. 13, Umetrics, Sweden).
The degree of variation was assessed via Hotelling’s T-squared distri-
bution (T
2
) at three condence intervals: 50% (HT
2
_50%), 75%
(HT
2
_75%) and 99% (HT
2
_99%). The construction of calibration curves
and peak integration were performed using MultiQuant™ v. 2.0.2
software by Sciex.
It is worth noting that siccanol (SIC, 57), dihydrotrichotetronine
(DHTTT, 75) and trichotetronine (TTT, 77) were expressed as peak area
values, as no analytical standards were available at the time of analysis.
J.M. Ramos-Diaz et al.

Food Control 130 (2021) 108260
3
3. Results and discussion
3.1. Method performance
The method was transferred to the new matrices of this study, ac-
cording to our suggestion in Sulyok et al. (2020), by spiking different
individual samples on one concentration level. As considers compliance
to ofcial performance criteria, similar results were obtained (Supple-
mentary Table S1). The 70–120% criterion for “recovery” was met for
52–63% and for 83–88% of all investigated analytes for apparent re-
coveries and recoveries of the extraction step, respectively, whereas the
RSD <20% criterion for reproducibility was met for 92–98% of analytes
despite using different individual samples for spiking.
Fig. 1. Cultivation areas of the samples listed in Table 1.
Section 1F: geographic coordinates corresponding to Uyuni Salt Flats (Potosí, Bolivia); exact cultivation area is unknown. Source of the images was Google Inc.
(California, USA).
J.M. Ramos-Diaz et al.

Food Control 130 (2021) 108260
4
Table 1
List of grain samples and their corresponding varieties harvested in 2015 and 2017. Cleaning methods (W, traditional washing; P, pearling; M, mechanical cleaning) and cultivation areas (e.g. 1A =Fig. 1A) are specied.
2015 2017
Sample Cleaning methods
a
Cultivation area Varietal
code
Sample Cleaning methods
a
Cultivation
area
Varietal code
U
b
W P M U
b
W P M
Quinoa (Chenopodium quinoa) Quinoa (Chenopodium quinoa)
minttumatilda
c
● ● 1A QMM minttumatilda
c
● ● 1A QMM
kancolla ● 1E QKA Kancolla ● ● 1E QKA
kuchivila ● 1E QKU Kuchivila ● ● 1E QKU
mistura ● 1E QM Mistura ● ● 1E QMM
negra Collana ● 1E QNC negra collana ● ● 1E QNC
pasankalla ● 1E QP Pasankalla ● ● 1E QP
real
d
● 1F QR real
d
● ● 1F QR
rosada taraco ● 1E QRT rosada taraco ● ● 1E QRT
salcedo INIA ● 1D QSI salcedo INIA ● ● 1D QSI
titicaca Denmark ● 1B QTID puno ● ● 1B QPU
titicaca Denmark ● ● 1B QTID
Ka˜
niwa (Chenopodium pallidicaule) vikinga ● ● 1B QVI
titicaca Latvia ● ● 1C QTIL
cupi INIA ● 1D KCI Ka˜
niwa (Chenopodium pallidicaule)
illpa INIA ● 1D KII
cupi INIA ● ● ● 1D KCI
Barley (Hordeum vulgare) illpa INIA ● ● ● 1D KII
ramis ● ● 1D KRA
commercial
e
● ● 1E BC Barley (Hordeum vulgare)
Oat (Avena sativa)
commercial
e
● ● 1E BC
Landsort ● ● 1B OL Oat (Avena sativa)
commercial
e
● ● 1E OC
Riegel ● ● 1B OR commercial
e
● ● 1E OC
Wheat (Triticum L.) Wheat (Triticum L.)
commercial
e
● ● 1E WC commercial
e
● ● 1E WC
a
Some grain samples went through more than one cleaning procedure.
b
Uncleaned samples.
c
Population variety.
d
Variety cultivated on the Bolivian side of the Andean Plateau. Exact location is unknown.
e
Cereal grains whose variety could not be specied.
J.M. Ramos-Diaz et al.

Food Control 130 (2021) 108260
5
3.2. Mycotoxin proles
A total of 101 metabolites were detected in all grains (Table 2). The
largest array of mycotoxins were produced by Penicillium spp. (32 me-
tabolites), followed by Fusarium spp. (26 metabolites), Aspergillus spp.
(10 metabolites), Alternaria spp. (6 metabolites), Trichoderma spp. (3
metabolites), Claviceps spp. (3 metabolites), Ascochyta spp. (2 metabo-
lites), Cladosporium spp. (2 metabolites), Metharhizium spp. (2 metabo-
lites), Beauvaria spp. (1 metabolite) and Ramularia spp. (1 metabolite).
For 13 analytes, no producing species could be attributed and were thus
labelled as unspecic metabolites. From all Andean and cereal grains
analysed, only two exceeded the maximum levels for mycotoxins in
established unprocessed cereals, based on the EU Commission Regula-
tion (EC) No 1881/2006. More specically, OC from the 2017 harvest
contained 64
μ
g/kg OT-A (22) and 377
μ
g/kg ZEN (58), both exceeding
the limits of 5
μ
g/kg OT-A (22) and 100
μ
g/kg ZEN (58), and QTIL
contained 5.6
μ
g/kg OT-A (22). In this section, mean mycotoxin con-
centration values from both 2015 and 2017 harvest are reported.
Detailed mycotoxin levels are provided in Supplementary Table S2 and
S3.
3.2.1. High contamination levels (>1000
μ
g/kg)
Flavogaucin (FG, 17) was the Penicillium-produced metabolite
exhibiting the highest concentration in uncleaned grains; it was mostly
abundant in QTIL (18 mg/kg) (Fig. 2A, centre right). Regarding Fusa-
rium metabolites, antibiotic Y (AB-Y, 33) and aurofusarin (AUR, 35)
were detected in large concentrations in uncleaned OC (3638
μ
g/kg) and
QPU (1041
μ
g/kg), respectively. Concerning metabolites from Alternaria
spp., infectopyron (INFE, 72) was present in oat var. landsort (OL; 1962
μ
g/kg), whereas tenuazonic acid (TeA, 74) was mostly detected in QPU
(2218
μ
g/kg) and QTIL (1213
μ
g/kg) (Fig. 2A, upper left section). The
unspecic metabolites neoechinulin A (NC-A, 98; Fig. 2A, right section),
asperphenamate (AsP, 90; Fig. 2A, centre) and N-benzoyl-phenylalanine
(NBP, 97; Fig. 2A, centre right) were detected in QTIL in the following
concentrations: 9784
μ
g NC-A/kg, 3258
μ
g AsP/kg and 1062
μ
g NBP/kg.
Asperglaucide (AsG, 89; Fig. 2, centre) was primarily present in quinoa
var. real (QR; 1008
μ
g/kg).
3.2.2. Medium contamination levels (100–1000
μ
g/kg)
A large number of mycotoxins produced by Fusarium spp. were
detected in concentrations between 100 and 1000
μ
g/kg. Fusarium
mycotoxins such as butenolid (BU, 38), chlamydosporol (ChlaD:iol, 40),
culmorin (CULM, 42), enniatin A (ENN-A, 45), enniatin A1 (ENN-A1,
46), enniatin B (ENN-B, 47), enniatin B1 (ENN-B1, 48), equisetin (EQ,
51), moniliformin (MON, 54) and nivalenol (NIV, 55) were measured at
levels within the 100–1000
μ
g/kg range. Despite their high concentra-
tions in OC, ENN-A1 (46), ENN-B (47) and ENN-B1 (48) were three out
of the only ve mycotoxins detected in ka˜
niwa var. cupi INIA (KCI), illpa
INIA (KII) or ramis (KRA) (Fig. 2A, lower left section). ENN-A (45) was
found in OC and minimally detected in KCI (Fig. 2A, lower right section).
QPU and QTIL were the only grains where BU (38) was found, whilst
ChlaD:ol (40) was only present in OC. CULM (42) was detected in QPU,
QVI, QTIL, quinoa pop. var. minttumatilda (QMM) and QTID; NIV (55)
was found in oat (OL, OC and OR) (Fig. 2A, upper left section). The
highest concentrations of EQ (51) were measured in BC, QPU, OC, and to
a lesser extent in QVI (Fig. 2A, upper right section). ZEN (58) was only
found in OC at a mean concentration of around 190
μ
g/kg (Fig. 2a), and
no fumonisins were detected in any of the samples.
Altersetin (ALT, 71), produced by Alternaria spp., was solely detected
in QPU and QTIL (Fig. 2A, far right section). Citreohybridinol (CHOL, 7)
and viridicatol (VOH, 32), produced by Penicillium spp., were only found
in OC (Fig. 2a). Furthermore, mycophenolic acid (MPA, 20) was present
in OC, QTIL and to a lesser extent in quinoa var. kuchivila (QKU). Pyr-
enocin A (Pyre-A, 28) was only detected in OL. Calphostin (CAL, 83),
attributed to Cladosporium spp., was identied at descending levels of
concentration in QPU, QTIL, OL, QVI, QTID and QMM. Lastly,
unidentied metabolites such as emodin (EMO, 94) and tryptophol (3-
IE, 101) were found in almost every sample. For instance, EMO (94) was
observed in oats (OL, OR and OC), barley (BC), quinoa (QTIL, QTID,
QPU, QMM, QR, QVI and QKU) and ka˜
niwa (KRA) (Fig. 2A, upper left
section). 3-IE (101) was found in every sample except for OL and OR
(Fig. 2A, lower left section).
3.2.3. Low contamination levels (<99
μ
g/kg)
A larger array of Penicillium mycotoxins were detected at this con-
centration range (Fig. 2a), in comparison to those from Fusarium spp.
The occurrence of prominent Penicillium mycotoxins, such as OT-A (22)
and ochratoxin B (OT-B, 23), in uncleaned grains was relatively low
(Fig. 2a, lower right section). QKU and QTIL were contaminated with a
mean level of around 5
μ
g OT-A/kg, whilst OC with 30
μ
g/kg. The
concentration of OT-B (23) in QKU and OC was considerably lower than
that of OT-A (22). Similar OT-A concentrations have been previously
reported in milled quinoa products, obtained from Canadian markets,
where 39% of the analysed samples were found to contain OT-A at a
mean level of 1.7
μ
g/kg (Kolakowski et al., 2016). Among other Pen-
icillium-produced contaminants, atlantinol A (AT-A, 5), citrinin (CIT, 8),
cyclopenol (COH, 12), viridicatin (VIN, 15), griseophenone B (GSP-B,
19), andrastin A (A-A, 22) and dihydrocitrinone (DH-CIT, 31) were –in
some cases– uniquely identied in OC. At lower levels, 7-hydroxypesta-
lotin (7HP, 1), agroclavine (AC, 2), chanoclavin (ChC, 6) and ques-
tiomycin A (Qu-A, 29) were detected in QTIL. QKU had only minor
concentrations of AT-A (5) and Qu-A (29).
Regarding Fusarium-produced metabolites, the type-A trichothe-
cenes HT-2 toxin (HT-2, 53) and T-2 toxin (T-2, 57) were detected in OL
and oat var. riegel (OR) (Fig. 2A, upper left section). OL was found to
contain around 50
μ
g HT-2/kg and 70
μ
g T-2/kg. OR, on the other hand,
contained around 30
μ
g HT-2/kg and 10
μ
g T-2/kg. Apicin (APIC, 34),
beauvericin (BEA, 36) and bikaverin (BIKA, 37) were mostly found in OL
and OR (Fig. 2A, upper section). Deoxynivalenol (DON, 43) was only
detected in OL (Fig. 2A, upper left section) and fungerin (FUN, 52) only
in OC. Cladosporium-produced cladosporin (CLADO, 84) was identied
in OC, QTIL and QSI (Fig. 2A, centre). Aspergillus-produced 3-nitropro-
pionic acid (3-NA, 66) and Metarhizium-produced destruxin B (D-B,
86) were detected in OC and QTIL (Fig. 2A–a, right section). In contrast
to Sacco et al. (2020), who reported aatoxin contamination in both
amaranth and quinoa, no aatoxins were detected in any of the samples
analysed in this study, most likely due to unfavourable geographic and
climatic conditions. Trichoderma-produced trichodimerol (TCOH, 76)
and Claviceps-produced ergometrine (ERG, 78) were found in OC and
QTIL, respectively. Finally, unspecic metabolites such as cyclo
L-Pro-L-Tyr (CDP-Tyr, 92) and cyclo L-Pro-L-Val (CDP-Val, 93) were
detected in all the uncleaned grains, whereas citreorosein (91) and fal-
lacinol (96) were mostly found in QTIL.
At trace level concentrations (<10
μ
g/kg), Penicillium-produced
mycotoxins represented the largest proportion, followed by mycotoxins
produced by Aspergillus spp. (mostly in BC and OC), Alternaria spp.
(mostly in QMM, QPU, QTID and QVI), Fusarium (only found in QTIL),
Ascochyta (OC and QSI), Metarhizium (OC), Romularia (BC) and Beau-
varia (QTIL and OL). In this concentration range, only two unspecic
metabolites were identied: norlichexanthone (NX, 99) and skyrin (SKY,
100). Most of these metabolites are depicted in Fig. 2a (right section).
3.3. Post-harvest cleaning
Noticeable differences in the content and distribution of mycotoxins
were observed by comparing samples before and after cleaning (Figs. 2
and 3). For instance, uncleaned QMM, located on the extreme upper left
side of PCA plot (Fig. 2), was initially contaminated with Pyre_A (28),
CULM (42), NIV (55), EMO (94) and ENC (95), all of which became
nearly undetectable after cleaning, as evidenced by the QMM relocation
to the right side of the PCA plot (Fig. 3). Despite this, QMM still con-
tained certain Fusarium [e.g. CULM (42)] and Alternaria [e.g. ALT (71)]
J.M. Ramos-Diaz et al.

Food Control 130 (2021) 108260
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Table 2
Codied list of detected mycotoxins sorted by pre-treatment and year.
Numerical code Origin Mycotoxin Cleaned Uncleaned
2015 2017 2015 2017
1 Penicillium spp. 7-Hydroxypestalotin 7HP 7HP
2 Agroclavine AC AC
3 Anacin AN AN
4 Andrastin A A-A A-A
5 Atlantinon A AT-A AT-A
6 Chanoclavin ChC ChC
7 Citreohybridinol CHOL CHOL
8 Citrinin
a
CIT CIT
9 Communesin B COM-B COM-B
10 Curvularin CURV CURV
11 Cyclopenin CIN CIN
12 Cyclopenol COH COH
13 Cyclopeptine CP CP
14 Dechlorogriseofulvin DCGSF DCGSF
15 Dihydrocitrinone DH-CIT DH-CIT
16 Festuclavine FC
17 Flavoglaucin FG FG
18 Griseofulvin GSF GSF
19 Griseophenone B GSP-B GSP-B
20 Mycophenolic acid MPA MPA MPA
21 Mycophenolic acid IV MPA-4
22 Ochratoxin A
a
OT-A OT-A OT-A
23 Ochratoxin B
a
OT-B OT-B
24 Okaramine B Ok-B Ok-B
25 O-Methylviridicatin OMV OMV
26 Pestalotin PES PES
27 Pinselin PIN PIN
28 Pyrenocin A Pyre_A Pyre_A
29 Questiomycin A Qu-A Qu-A Qu-A
30 Quinolactacin A QuL-A QuL-A
31 Viridicatin VIN VIN
32 Viridicatol VOH VOH
33 Fusarium spp. Antibiotic Y AB-Y AB-Y
34 Apicidin APIC APIC APIC APIC
35 Aurofusarin AUR AUR AUR AUR
36 Beauvericin BEA BEA BEA BEA
37 Bikaverin BIKA BIKA
38 Butenolid BU BU
39 Chlamydospordiol ChlaD:iol ChlaD:iol
40 Chlamydosporol ChlaD:ol ChlaD:ol
41 Chrysogin Chry Chry Chry Chry
42 Culmorin CULM CULM
43 Deoxynivalenol DON DON
44 Diacetoxyscirpenol DAS DAS
45 Enniatin A ENN-A ENN-A ENN-A ENN-A
46 Enniatin A1 ENN-A1 ENN-A1 ENN-A1 ENN-A2
47 Enniatin B ENN-B ENN-B ENN-B ENN-B
48 Enniatin B1 ENN-B1 ENN-B1 ENN-B1 ENN-B1
49 Enniatin B2 ENN-B2 ENN-B2
50 Epiequisetin epi-EQ epi-EQ
51 Equisetin EQ EQ EQ EQ
52 Fungerin FUN FUN
53 HT-2 toxin HT-2 HT-2
54 Moniliformin MON MON MON MON
55 Nivalenol NIV NIV NIV NIV
56 Siccanol SIC SIC SIC
57 T-2 toxin T-2 T-2
58 Zearalenone ZEN ZEN
59 Aspergillus spp. Averantin AVN AVN
60 Averun AVR AVR
61 Methoxysterigmatocystin MST MST
62 Norsolorinic acid NA NA
63 Sterigmatocystin ST ST
64 Versicolorin A Ver-A Ver-A
65 Versicolorin C Ver-C Ver-C
66 3-Nitropropionic acid 3-NA 3-NA
67 Sydonic acid SA SA
68 Territrem B T-B T-B
69 Alternaria spp. Alternariol AOH AOH AOH
70 Alternariolmethylether AME AME AME
71 Altersetin ALT ALT
72 Infectopyron INFE INFE INFE INFE
73 Tentoxin TEN TEN TEN TEN
(continued on next page)
J.M. Ramos-Diaz et al.

Food Control 130 (2021) 108260
7
mycotoxins. As a matter of fact, the content of 3-IE (101) increased
consistently in various grains after cleaning. Presence of mycotoxins
after cleaning could be attributed to internal mycobiota that often re-
mains capable of producing mycotoxins even after post-harvest clean-
ing, as reported in Pappier et al. (2008). Regarding uncleaned QKU,
contaminants were mostly unspecic metabolites such as AsG (89), AsP
(90) and NBP (97), whose concentrations also reduced dramatically
after cleaning. A newly positioned QKU, from the centre (Fig. 2) to the
extreme left side (Fig. 3), reects drastic changes in the mycotoxin
prole. In KCI, mycotoxins were practically absent in both cleaned or
Table 2 (continued )
Numerical code Origin Mycotoxin Cleaned Uncleaned
2015 2017 2015 2017
74 Tenuazonic acid TeA TeA
75 Trichoderma spp. Dihydrotrichotetronine DHTTT DHTTT
76 Trichodimerol TCOH TCOH
77 Trichotetronine TTT
78 Claviceps spp. Ergometrine ERG ERG
79 Ergometrinine ERGOE ERGOE
80 Ergine LSA LSA
81 Ascochyta spp. Ascochlorin Ach Ach
82 Ascofuranone AF AF
83 Cladosporium spp. Calphostin CAL CAL CAL CAL
84 Cladosporin CLADO CLADO
85 Metarhizium spp. Destruxin A D-A D-A
86 Destruxin B D-B D-B D-B
87 Beauvaria spp. Bassianolide BASS BASS
88 Ramularia spp. Rubellin D R-D R-D
89 Unspecic Asperglaucide AsG AsG AsG
90 Asperphenamate AsP AsP AsP AsP
91 Citreorosein CTO CTO CTO CTO
92 cyclo(L-Pro-L-Tyr) CDP-Tyr CDP-Tyr CDP-Tyr CDP-Tyr
93 cyclo(L-Pro-L-Val) CDP-Val CDP-Val CDP-Val CDP-Val
94 Emodin EMO EMO EMO EMO
95 Endocrocin ENC ENC ENC ENC
96 Fallacinol FOH FOH
97 N-Benzoyl-Phenylalanine NBP NBP NBP
98 Neoechinulin A NC-A NC-A NC-A
99 Norlichexanthone NX NX
100 Skyrin SKY SKY SKY SKY
101 Tryptophol 3-IE 3-IE 3-IE 3-IE
a
These mycotoxins have been attributed to Penicillium spp. as the most likely producing species in the samples analysed.
Fig. 2. Principal component analysis bi-plot for mycotoxins detected from uncleaned ka˜
niwa, quinoa, barley, oats and wheat grains (total variance, 82.8%).
Numerically coded mycotoxins were colour-labelled based fungal origin. The symbol diameter was set to vary depending on the total occurrence (
μ
g/kg) of a
particular mycotoxin in the sample set. The meaning of alphanumerical and numerical codes corresponding to grain varieties and mycotoxins, respectively, are
explained in Tables 1 and 2 Plot resulting from the data combination of 2015 and 2017. Siccanol (56), dihydrotrichotetronine (75) and trichotetronine (77) values
expressed as absolute peak area. (For interpretation of the references to colour in this gure legend, the reader is referred to the Web version of this article.)
J.M. Ramos-Diaz et al.

Food Control 130 (2021) 108260
8
uncleaned grains. However, SIC (56) was still present in cleaned KCI.
Cross-contamination cannot be dismissed given the presence of SIC (56)
in barley, oats and quinoa. In Fig. 2, KCI is located in the lower left side
of the PCA plot, which indicated a degree of association with Fusarium
mycotoxins (<1
μ
g/kg) and unspecic metabolites (<10
μ
g/kg). After
cleaning, KCI migrated to the opposite side of the plot (Fig. 3). Prior to
cleaning, KRA presented a very similar mycotoxin prole to KCI (both
located on the left side of the PCA plot, Fig. 2). However, upon cleaning,
the few contaminants of KRA were reduced [e.g. ENN-B2 (49)]. SIC (56)
was not detected in either cleaned or uncleaned KRA. Clearly, KRA
moved from the outskirts of the PCA plot (beyond HT
2
_99%) towards the
centre of the plot (just below HT
2
_50%).
Fusarium and Alternaria mycotoxins were detected in uncleaned
QTID. QTID was initially located on the right side of the PCA plot (just
below HT
2
_75%) and moved to the opposite side of the plot (beyond
HT
2
_50%) after cleaning. This occurred in response to a drastic reduc-
tion in the concentration of mycotoxins. For instance, the peak of SIC
(56) disappeared in cleaned QTID. QPU, QVI and QTIL moved towards
the centre of the plot (below HT
2
_50%) following cleaning, due to a
reduction (though minimal) in the content of Fusarium and Alternaria
mycotoxins. The cleaning of OR and OL was linked to a reduction in the
type and concentration of Fusarium and Alternaria mycotoxins. The
differences are noticeable if one compares the strong association of OR
and OL with various mycotoxins prior to cleaning (upper left side,
Fig. 2), against their newly formed mycotoxin associations (upper left
side, Fig. 3). Other samples stayed mostly within the HT
2
_50%, meaning
that variations in the content of mycotoxins, as a consequence of
cleaning, could not be statistically veried. Overall, post-harvest
cleaning of cereal grains has been broadly characterised in the litera-
ture as an efcient and cost-effective mitigation strategy to signicantly
reduce grain mycotoxin content (Neme & Mohammed, 2017). The
cleaning methods were found to reduce the overall concentration of
mycotoxins in tested grains from 2017 by roughly 50% (SIC was omitted
from the calculation). In the case of quinoa and ka˜
niwa, where tradi-
tional washing is mainly applied for saponin removal, mycotoxin
content was signicantly reduced, in some cases dropping below the
detection level [e.g. FC (16), MPA-4 (21), BASS (87)]; a fact that con-
rms the effectiveness of this simple mycotoxin mitigation technique
also for non-cereal grains.
3.4. South American contaminants
In general, South American samples presented low mycotoxin con-
tent as observed in the PCA plot (Fig. 4). South American samples (blue)
clearly dominated the centre of the plot, meaning that their differences,
in terms of mycotoxins, was minimal. Conversely, KRA, KCI and QKU
were located beyond HT
2
_75%, indicating differences from the rest of
South American samples (Fig. 4). For instance, Fusarium mycotoxins
[ENN-A1 (46), ENN-B (47) and ENN-B1 (48)] and unspecic metabolites
[CDP-Tyr (92), CDP-Val (93) and 3-IE (101)] were detected in uncleaned
KRA and KCI (Fig. 4A, cluster b’). After cleaning, KRA and KCI moved to
the centre of the PCA plot, as a consequence of the decrease in myco-
toxin levels. On the other hand, QKU moved from the centre to the
outskirts of the PCA plot after cleaning. This meant that, unlike the rest,
QKU remained highly associated to mycotoxins like NBP (97) or AsP
(90) (Fig. 4B, lower left section).
From cleaned South American samples (Fig. 4A), those on the
farthest right side of the PCA plot (Fig. 4B) contained the largest
assortment of mycotoxins. Thus, an in-depth observation was conducted
on OC, BC, QSI and KCI (Fig. 5). OC and, to a lesser extent, BC presented
a wide array of mycotoxins, including Fusarium-, Metarhizium- or Asco-
chyta-produced metabolites. It was hard to understand the remarkable
presence of mycotoxins in OC, if we consider that it was cultivated in
close proximity to other South American samples (Fig. 1D and E). On the
other hand, QSI and KCI showed minimal variation in terms of myco-
toxins, mostly Fusarium-produced metabolites and unspecic metabo-
lites (Fig. 5). Despite the discrepancies, the peak of SIC (56) was still
present in OC, BC, QSI and KCI. Interestingly, Trichoderma-produced
mycotoxins were only found in OC.
Fig. 3. Principal component analysis bi-plot for mycotoxins detected from cleaned ka˜
niwa, quinoa, barley, oats and wheat grains (total variance, 80.9%).
Numerically coded mycotoxins were colour-labelled based fungal origin. The symbol diameter was set to vary depending on the total concentration (expressed as
μ
g/
kg) of a particular mycotoxin in the sample set. The meaning of alphanumerical and numerical codes corresponding to grain varieties and mycotoxins, respectively,
are explained in Tables 1 and 2 Plot resulting from the data combination of 2015 and 2017. Siccanol (56), dihydrotrichotetronine (75) and trichotetronine (77) values
expressed as absolute peak area. (For interpretation of the references to colour in this gure legend, the reader is referred to the Web version of this article.)
J.M. Ramos-Diaz et al.

Food Control 130 (2021) 108260
9
3.5. North European contaminants
Most samples obtained from North Europe were associated with a
large array of mycotoxins, predominantly from Fusarium spp., Alternaria
spp. and Penicillium spp.; unspecic metabolites were present in modest
amounts (Fig. 4). Unlike South American samples, all North European
samples were located outside the centre (beyond HT
2
_50%) of the PCA
plot (Fig. 4A), denoting that there was a large variation in the content
and type of mycotoxins. Among the uncleaned North European samples,
two groups were clearly observed: a low contamination group, located on
Fig. 4. Principal component analysis bi-plot for mycotoxins detected from uncleaned (A; total variance, 82.8%) and cleaned (B; total variance, 80.9%) ka˜
niwa,
quinoa, barley, oats and wheat seeds; theses were colour-labelled based on their continental origin (South America, SA; North Europe, NE). The meaning of
alphanumerical and numerical codes corresponding to grain varieties and mycotoxins, respectively, are explained in Tables 1 and 2 Plot resulting from the data
combination of 2015 and 2017. (For interpretation of the references to colour in this gure legend, the reader is referred to the Web version of this article.)
Fig. 5. Mycotoxin prole of cleaned grains with the largest presence and/or concentration of mycotoxins in accordance with the cluster shown in Fig. 4. Results are
divided based on the grains’ geographical origin. Numerically coded mycotoxins are colour-labelled based fungal origin. Mycotoxin concentration: 0–10
μ
g/kg (*);
10–10
2
μ
g/kg(**); 10
2
–10
3
μ
g/kg(***); 10
3
–10
4
μ
g/kg(****); 10
4
–10
5
μ
g/kg(*****); 10
5
–10
6
μ
g/kg(******). Siccanol (56), dihydrotrichotetronine (75) and tri-
chotetronine (77) values expressed as absolute peak area. (For interpretation of the references to colour in this gure legend, the reader is referred to the Web version
of this article.)
J.M. Ramos-Diaz et al.

Food Control 130 (2021) 108260
10
the left upper section of the PCA plot (Fig. 4, cluster a’) and a high
contamination group, located on the right section of the PCA plot (Fig. 4).
Uncleaned OR, OL and QMM were found in the low contamination
group, mostly characterised by the presence of Alternaria, Fusarium and
unspecic mycotoxins. Upon cleaning, a noticeable migration was
observed. For instance, QMM and OL moved from the far-left side to the
centre of the PCA plot, below HT
2
_50%. This is in line with a consid-
erable reduction in the content of mycotoxins. On the other hand, the
minor changes in OR reect unremarkable reductions in the content of
mycotoxins after cleaning (Fig. 4).
QTIL, QPU, QTID and QVI were allocated in the high contamination
group due to their strong association with a wide array of mycotoxins,
produced mostly by Fusarium spp., Alternaria spp. and Penicillium spp.
(Fig. 4A). Interestingly, QTIL was the only sample where Claviceps-
produced mycotoxins [ERG (78), ERGOE (79) and LSA (80)] were
detected. After cleaning, there was considerable reduction in the content
of mycotoxins that was reected in the movement (towards the centre of
the PCA plot, below HT
2
_50%) of QTIL, QPU, QVI and QMM
(Figs. 4B–5). Despite the reduction, QTIL remained strongly associated
to various Penicillium-produced mycotoxins such as FG (17) and MPA
(20) (Fig. 5). In line with their cultivating conditions (Denmark, Fig. 1B),
QPU and QVI showed similar mycotoxin prole (Fig. 5). Cleaned QPU
and QVI contained mostly Fusarium-produced [e.g. AUR (35), CULM
(42)] and, to a lesser extent, Alternaria mycotoxins [e.g. TeA (71) and
ALT (74)]. Despite the observable lower concentrations, QMM also
showed adherence to mycotoxins from Fusarium and Alternaria spp.
Differences in the weather, cultivating/harvesting conditions or
post-harvest treatment could help elucidate the reasons behind the
remarkable differences among quinoa samples cultivated in North
Europe. At rst glance, it seems that the farther north quinoa was
cultivated, the less contaminated it became. However, this hypothesis
could not be applied to samples cultivated in Denmark and Latvia, where
the latitudes of the cultivating elds were very similar (Fig. 1B and C),
yet they possessed different mycotoxin proles. Characteristics of the
cultivating methods and post-harvest treatments could provide more
plausible explanations on mycotoxin variations.
3.6. Andean vs. cereal grain contamination
Cleaned cereal grains were more likely to contain fungal
contaminants than cleaned Andean grains, particularly those from South
America (Fig. 5). In 2015, conspicuous levels of mycotoxins produced by
Fusarium spp. [HT-2 (53), MON (54), NIV (55) and T-2 (57)], as well as
INFE (72) and some unspecic metabolites [CTO (91), CDP-Tyr (92),
CDP-Val (93), EMO (94) and ENC (95)] were detected in OR (Fig. 6A,
cereal cluster I) and BC (Fig. 6A, cereal cluster II). These ndings are in
line with previous surveys indicating high prevalence of Fusarium my-
cotoxins in oats and barley cultivated in Nordic countries (Brodal et al.,
2020; Nathanail et al., 2015). On the other hand, cleaned Andean grains
presented remarkably low contents of fungal metabolites except from
QMM and KCI (Fig. 6A, Andean grains cluster I). From the 2017 harvest
samples, cleaned QMM was mainly associated with various Fusarium and
a few Alternaria mycotoxins, but not SIC (56) (Fig. 6B, Andean grains
cluster II). QKU was strongly contaminated with certain unspecic
metabolites [AsG (89), AsP (90) and NBP (97)]. Cleaned BC and espe-
cially OC, both from 2015 to 2017 harvests, were found to contain
mycotoxins produced by almost all fungal genera identied in this study
(Table 2), except from Claviceps, Beauvaria and Ramularia.
North European cereal grains were found to be consistently less
contaminated than Andean grains of the same region, whilst the exact
opposite occurred with those from South America (Fig. 5). This outcome
could be attributed to the existence of extensive mycotoxin control
programmes in European countries, and the implementation of effective
mycotoxin contamination prevention strategies for cereal grains (e.g.
crop rotation, fertilization, pesticide application) (Agriopoulou et al.,
2020). Furthermore, the potentially more favourable climatic/envir-
onmental conditions for fungal growth and mycotoxin production of
Andean grains cultivated in Europe, in addition to less developed risk
mitigation approaches specic to Andean grains, may have be the
reasoning behind higher contamination levels. Conversely, cereal grains
cultivated in South America were evidently more prone to mycotoxin
contamination than South American Andean grains. Inadequate pre-/-
post-harvesting methods of fungal control or insufcient adaptability of
the grains to the environment might explain increased cereal contami-
nation in those regions. Apparently, the resilience of South American
Andean grains to the growth of mycotoxin-producing fungi could be due
to their formidable biological adaptation to Peruvian mountainous re-
gions (>3000 m.a.s.l.). Something that may drastically change if culti-
vated away from their natural environment. It could also be argued that
saponin-containing Andean grains may prevent the growth of fungi
Fig. 6. Principal component analysis bi-plot for mycotoxins detected from cleaned ka˜
niwa quinoa, barley, oats and wheat seeds in 2015 (A; total variance, 67.3%)
and 2017 (B; total variance, 90.3%). Mycotoxin occurrence in Andean grains (blue) or cereal grains (green) were highlighted via clusters. Numerically coded
mycotoxins were colour-labelled based fungal origin. (For interpretation of the references to colour in this gure legend, the reader is referred to the Web version of
this article.)
J.M. Ramos-Diaz et al.

Food Control 130 (2021) 108260
11
(Woldemichael Wink 2001). Herein, the well-documented existence of
saponins in various European and South American Andean grains did
not seem to at least drastically inhibit fungal growth, particularly in
Europe, although the investigation of saponin effects on fungal growth
and mycotoxin contamination was out of the scope of this study.
4. Conclusions
A comparative study concerning the natural occurrence of myco-
toxins and fungal metabolites was conducted in Andean grains (ka˜
niwa
and quinoa) and cereal grains (barley, oats and wheat) cultivated in
South America and North Europe. A state-of-the-art LC–MS/MS method
was utilised in this study that is capable for the simultaneous determi-
nation of several hundreds of analytes within a single run. Signicant
discrepancies were observed in the contamination proles between
Andean/cereal grains, South America/North Europe and 2015/2017
harvests, attributable to differences in crop physiology, climatic condi-
tions, geographic characteristics, as well as mycotoxin contamination
prevention strategies. Moreover, cleaning of grains resulted in signi-
cant reductions in the concentration of the majority of mycotoxins, even
though certain metabolites, likely produced by internal mycobiota,
remained detectable. The present study comprises the most extensive
mycotoxin survey of Andean grains to date, providing crucial informa-
tion on contamination patterns, prevalence of fungal populations and
the effect of cleaning in mycotoxin levels. In conclusion, as the value of
Andean grains in global food trade increases, more targeted research on
this agricultural commodity is needed for the identication of risks,
enabling development of effective prevention and mitigation strategies
to enhance food safety and promote food security.
CRediT authorship contribution statement
J.M. Ramos-Diaz: Conceptualization, Investigation, Formal anal-
ysis, Writing – original draft. M. Sulyok: Methodology, Validation,
Resources, Writing – review & editing. S.E. Jacobsen: Resources,
Writing – review & editing. K. Jouppila: Resources, Writing – review &
editing. A.V. Nathanail: Conceptualization, Investigation, Writing –
review & editing.
Declaration of competing interest
Please check the following as appropriate:
All authors have participated in (a) conception and design, or anal-
ysis and interpretation of the data; (b) drafting the article or revising it
critically for important intellectual content; and (c) approval of the nal
version.
This manuscript has not been submitted to, nor is under review at,
another journal or other publishing venue.
The authors have no afliation with any organization with a direct or
indirect nancial interest in the subject matter discussed in the
manuscript.
The following authors have afliations with organizations with
direct or indirect nancial interest in the subject matter discussed in the
manuscript:
Acknowledgements
The authors warmly thank Mrs Orfelina Chuquipiondo Alvis, Listail
Diaz Chuquipiondo and Bejamina Gonzalo Nina for their outstanding
support during the acquisition of quinoa and ka˜
niwa from Puno, Peru.
Special thanks to our esteemed colleagues from the Faculty of Food
Technology (Latvia University of Life Sciences and Technologies) led by
Dr Martins Sabovics for providing quinoa var. titicaca.
Appendix A. Supplementary data
Supplementary data to this article can be found online at https://doi.
org/10.1016/j.foodcont.2021.108260.
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