Food Chemistry 278 (2019) 144–162

Contents lists available at ScienceDirect

Food Chemistry
journal homepage: www.elsevier.com/locate/foodchem

Review

Food fingerprints – A valuable tool to monitor food authenticity and safety T

Sonia Medina , Jorge A. Pereira, Pedro Silva, Rosa Perestrelo, José S. Câmara
⁎ ⁎

CQM – Centro de Química da Madeira, Universidade da Madeira, Campus da Penteada, 9020-105 Funchal, Portugal

ARTICLE INFO ABSTRACT

Keywords: In recent years, food frauds and adulterations have increased significantly. This practice is motivated by fast
Food fingerprints economical gains and has an enormous impact on public health, representing an important issue in food science.
Authenticity In this context, this review has been designed to be a useful guide of potential biomarkers of food authenticity
Food safety and safety. In terms of food authenticity, we focused our attention on biomarkers reported to specify different
Adulteration
botanical or geographical origins, genetic diversity or production systems, while at the food safety level, mo-
Biomarkers
lecular evidences of food adulteration or spoilage will be highlighted. This report is the first to combine results
from recent studies in a format that allows a ready overview of metabolites (< 1200 Da) and potentially mo-
lecular routes to monitor food authentication and safety. This review has therefore the potential to unveil im-
portant aspects in food adulteration and safety, contributing to improve the current regulatory frameworks.

1. Introduction
Food fingerprints can be defined as molecular markers that re-
present a characteristic food state or condition, allowing more effective
product discrimination. Essentially, it is a marker or set of markers that
allow us to answer many questions about food authenticity, as “Are
those organic carrots truly organic? Does this saffron really originate
from Spain? Can we discriminate between orange juice and pulp wash?
(Cubero-Leon, De Rudder, & Maquet, 2018; Le Gall, Puaud, &
Colquhoun, 2001; Rubert, Lacina, Zachariasova, & Hajslova, 2016).
Unfortunately, the search for these fingerprints is not only related with
the quality of the products, but also with their safety to human health.
Although the deliberate adulteration of food and beverages to deceive
consumers is illegal in any part of the world, the promise of fast eco-
nomical incomes is becoming very prevalent and affects a broad range
of products, from dietary food products to beverages. Food products
adulteration is carried out to increase volume and weight, mask inferior
quality and replace authentic substances with cheaper ones, with ben-
efits for sellers and possible harms to consumers. It is therefore a major
concern for consumers, food producers, companies, and regulatory
agencies. The food frauds most often detected are adulteration (e.g.,
addition of water), sophistication processes (e.g., use of sugars, ad-
ditives, or flavours), and counterfeiting (e.g., use of prohibited additives
and colorants, incorrect botanical and geographical declarations, reuse
of expired products, or incorrectly naming something an organic pro-
duct) (Avula, Smillie, Wang, Zweigenbaum, & Khan, 2015; Granato,
Koot, Schnitzler, & van Ruth, 2015; Tette, Guidi, Bastos, Fernandes, &
Gloria, 2017; van Ruth, et al., 2011; Xue et al., 2013). Therefore, when
there is a food suspected of adulteration, rapid and robust methods, as
well as specific and reliable markers, must be available to support the
withdraw of such products from the food chain and act against the
facilitators of that frauds (Llano, Muñoz-Jiménez, Jiménez-Cartagena,

Abbreviations: ABA, abscisic acid; CINs, cytoplasmic invertases; CLA, conjugated linoleic acids; CQA, caffeoylquinic acids; DART, direct analysis in real time; DART-
q-TOF-MS, direct analysis in real time-quadrupole-time of flight-mass spectrometry; DUFA, diunsaturated fatty acids; DXP, 1-deoxy-D-xylulose-5- phosphate; EFSA,
European Food Safety Authority; FAME, fatty acyl methyl ester; FDA, Food and Drug Administration; FSMA, Food Safety Modernization Act; FT-ICR-MS, Fourier
transform-ion cyclotron resonance-mass spectrometry; GC-O, gas chromatography-olfactometry; GLVs, green leaf volatiles; HPLC-ESI-MS/MS, high-performance
liquid chromatography-electrospray ionization-tandem mass spectrometry; HPLC-q-TOF-MS, high-performance liquid chromatography-quadrupole-time of flight-
mass spectrometry; HS-SPME-GC–MS, headspace solid phase microextraction gas chromatography-mass spectrometry; LC, liquid chromatography; LOOCV, leave-
one-out cross validation; MDGC, multidimensional gas chromatography; MS, mass spectrometry; MUFA, monounsaturated fatty acids; MVA, mevalonic acid; NIRS,
near-infrared spectroscopy; NMR, nuclear magnetic resonance; O/C, organic and conventional; OPLS, orthogonal partial least square; PCA, principal component
analysis; PDO, protected designation of origin; PLOT, porous layer open tubular; PTR-MS, proton transfer reaction mass spectrometry; PUFA, polyunsaturated fatty
acids; SBSE, stir bar sorptive extraction; SIMCA, soft independent modelling of class analogy; SIFT-MS, selected ion flow tube mass spectrometry; SLDA, stepwise
linear discriminant analysis; TMA-N, trimethylamine nitrogen; TVB-N, total volatile bases nitrogen; UPLC-HR-MS, ultra-performance liquid chromatography-high
resolution-mass spectrometry; VIN, vacuolar invertases; VOCs, volatile organic compounds
⁎
Corresponding authors.
E-mail addresses: sonia.escudero@staff.uma.pt (S. Medina), jsc@staff.uma.pt (J.S. Câmara).

https://doi.org/10.1016/j.foodchem.2018.11.046
Received 13 July 2018; Received in revised form 2 November 2018; Accepted 8 November 2018
Available online 09 November 2018
0308-8146/ © 2018 Elsevier Ltd. All rights reserved.
S. Medina et al.
Londoño-Londoño, & Medina, 2018). This is particularly relevant when,
besides the economic loss, the adulteration involves health risks for the
consumers. Hence, the ability to trace and authenticate food products is
a great concern for the food industry, but also for the public authorities.
For this reason, there is extensive legislation regarding food safety and
authenticity all over the world, being USA and EU responsible for the
most comprehensive and strict guidelines. In USA, the Food Safety
Modernization Act (FSMA) (https://www.fda.gov/Food/
GuidanceRegulation/FSMA/, accessed October 19, 2018) defined by
the Food and Drug Administration (FDA) establishes several rules to
prevent foodstuffs adulteration. In turn, the European Food Safety
Authority (EFSA) (https://www.efsa.europa.eu/en/aboutefsa, accessed
October 19, 2018) includes a set of laws and guidelines for the food
quality and safety assurance, designed as General Food Laws.
In food authentication field, there are targeted and untargeted
methods of analysis. The first approach focuses on the analysis of a
specific metabolite or group of metabolites, while in the untargeted
approach, the major goal is discrimination of patterns of metabolites
that may change in response to environmental, genetic or human al-
terations (adulteration) (Llano et al., 2018). In this context, application
of metabolomics to food science for assessing safety and authenticity
has gained much interest (Arbona, Iglesias, & Gómez-Cadenas, 2015;
Bonte, et al., 2014; Eisenmann, et al., 2016; Souard et al., 2018). Me-
tabolomics studies are mainly discriminative and predictive, aiming to
find differences between samples (e.g., for the oranges origin dis-
crimination, for the variety and vintage differentiation of wines or for
the civet coffees authentication) and to create statistical models to
predict class memberships (e.g., for the classification of green coffee
beans or wines according to botanical and geographical origins). Food
fingerprints may be obtained for food quality, safety and authenticity
purposes. To achieve this, an experimental design should be carefully
established, being the sample preparation and the analytical platforms
selected critical parameters to evaluate. A wide range of instrumental
techniques, as chromatography (gas and liquid), mass spectrometry or
spectroscopy is used to monitor food authenticity and safety. To support
this review, it was queried PubMed and Scopus databases with the
keywords “biomarkers” and “food authentication” to obtain the re-
search published in the last five years in food authenticity. This has
retrieved us hundreds of reports, essentially involving mass spectro-
metry (MS)-based studies (mainly gas chromatography coupled with
mass spectrometry (GC–MS) and liquid chromatography mass spectro-
metry with quadrupole time of flight technology (LC-q-TOF-MS))
(Arbona, et al., 2015; Bonte, et al., 2014; Klockmann, Reiner,
Bachmann, Hackl, & Fischer, 2016), spectroscopy (nuclear magnetic
resonance (NMR), Raman, UV–Visible, etc.) and spectrofluorimetry
methods (Farag, Labib, Noleto, Porzel, & Wessjohann, 2018; Li, Zhang,
Jin, Liu, & Wang, 2016; Longobardi, et al., 2017; Nekvapil, et al.,
2018). Nevertheless, more than 75% of the publications cited in this
review were performed by MS technology (Tables 1–5). The popularity
of MS approaches is due to ability to distinguish small differences be-
tween samples in complex matrices. In contrast, the development of
spectroscopy and spectrofluorimetry methods may be related with their
non-destructive nature, which is very relevant for expensive samples.
Other analytical strategies, as stable isotope and DNA analyses are
being also applied routinely for food authenticity purposes. However, in
this review, we will focus mainly on studies involving small molecules
(< 1200 Da) identified by chromatographic methodologies and labelled
as biomarkers given their ability to discriminate different samples.
Moreover, we will only consider metabolites certified through robust
statistical models. Hence, the main objective of this work is to assign
high significant biomarkers to food authenticity and safety monitoring
according to their geographical and botanical origin, genetic diversity,
production systems, adulteration and spoilage or freshness indication.
This practical guide will allow an easy search of metabolites, con-
tributing to eventually identify new molecular routes for food authen-
tication and safety assessment, fostering the improvement of the
145
Food Chemistry 278 (2019) 144–162
current regulatory frameworks.
2. Potential biomarkers to monitor food authenticity and safety
Food chemistry research is a very dynamic field and multiple studies
aiming food products authenticity have performed, producing many
high quality and robust classification models. However, in many of
these reports the discriminant metabolites were not unequivocally
identified, because this is a very time-consuming step, still constituting
nowadays the major bottleneck in the metabolomics workflow. For this
reason, high-throughput approaches have been developed to detect
specific metabolites contributing for samples discrimination. These
metabolites may be used as promising biomarkers of authentication,
but also monitor biochemical changes in many foodstuffs. The main
biomarkers and biological pathways involved in food authenticity and
safety according to different varieties/cultivars, production systems,
geographical/botanical origin, adulteration practices, as well as spoi-
lage/freshness indicators are detailed below. As already referred, due to
the extensive scientific literature publish on this subject, we only focus
on studies that were able to identify discriminant metabolites (bio-
markers) using consistent statistical models (that are also referred in
Fig. 1 and in the respective Tables 1–5).
2.1. Species, varieties or cultivars
There are serious economic and quality reasons to certify the au-
thenticity of the varieties used in different foodstuffs. However, as food
processing progresses, it becomes extremely difficult to distinguish
among species, varieties or cultivars and this fact is often used to carry
out food adulterations. This problem may be solved by the detection
and eventually quantification of specific metabolites able to dis-
criminate the adulteration. In Table 1 are referred relevant examples of
potential biomarkers of food adulteration in different foodstuffs, being
the respective matrices, methodology and statistical analyses identified
for the reader guidance. Grapes (Vitis vinifera) have a critical im-
portance in wine properties and quality and different studies have in-
vestigated a putative discrimination between cultivars. Different me-
tabolites, as organic acids, amino acids, carbohydrates,
phenylpropanoids, and flavonoids, have been previously pointed as
potential biomarkers for such discrimination. In this context, Ali et al.
(2009) reported the use of 1H NMR technique and multivariate analyses
for the differential characterization of plant samples based on their
metabolic composition. This study also demonstrates that the sample
storage conditions and the solvent extraction may produce artifacts,
leading to the detection of outliers in the statistical analysis. Never-
theless, small variations as the age of the plant samples (young versus
old leaves of the same plant) may result in a different metabolic profile
and so the interpretation of the results should consider these inter-
ferences (Ali, et al., 2009). Related with this work, another study pre-
sented C6-alcohols (1-hexenol, (E)-3-hexenol and (Z)-3-hexenol) and
the ratios among them as varietal markers in grapes. It allowed the
discrimination in six different white grapes varieties (Alvarinho, Arinto,
Avesso, Azal, Loureiro, Trajadura) and three red grapes varieties
(Amaral, Borraçal and Vinhão) from Portugal (Oliveira, Faria, Sá,
Barros, & Araújo, 2006). In the same context, five monovarietal red
wines made from Negramoll, Listán negro, Tintilla, Vijariego negro and
Baboso negro were analysed with gas chromatography-olfactometry
(GC-O) and multidimensional gas chromatography (MDGC). In this
report, several compounds were identified as potential markers and
some of them were detected as important odorants in only one variety.
This was the case of 2,3-pentanedione and 3-ethylphenol (present only
in Negramoll), ethyl valerate and 3-isopropyl-2-methoxypyrazine (de-
tected in Tintilla) and ethyl 4-methylpentanoate (Vijariego negro). This
fact indicates that there is strong variability among wine varietals and
GC-O and MDGC is a powerful technology to detect these differences
(Culleré, Escudero, Pérez-Trujillo, Cacho, & Ferreira, 2008). A very
 Table 1
Potential biomarkers for the differentiation among species, varieties and cultivars of different foodstuffs.
Biomarkers Matrix (number of Methodology Statistical analysis References
S. Medina et al.

samples)

1
Adenine, alanine, caffeic acid, fructose, inositol, linolenic acid, proline, quercetin-3-O-glucoside, succinic Grape (6) H NMR HCA; PCA; PLS-DA Ali et al. (2009)
acid
2-Methoxyphenol, 2, 3-pentadione, 3-isopropyl-2-methoxypyrazine, β-damascenone, ethyl cinnamate, ethyl Wine (5) GC-O, MDGC ANOVA Culleré et al. (2008)
4-methylpentanoate, 2-furfural, octanal
1-Hexenol, (E)-3-hexenol, (Z)-3-hexenol Wine (43) GC–MS ANOVA; Mann–Whitney test Oliveira et al. (2006)
1,1,6-Trimethyl-1,2-dihydronaphtalene, α-terpineol, β-damascenone, (E,E)-farnesal, geraniol, linalool, Wine (36) GC–MS PCA; LDA; LOOCV Câmara et al. (2007)
nerolidol, trans-furan linalool oxide, vitispirane
1
Alanine, citramalic acid, citrate, epicatechin, galactose, isoleucine, myo-inositol, quinic acid, xylose Apple (140) H NMR PCA Eisenmann et al. (2016)
1-Hexanol, 2-methylbutyl acetate, 6-methyl-5-hepten-2-ol, butyl 2-methylbutanoate, ethyl 2- Apple juice (50) GC–MS LDA; PCA; SLDA; LOOCV Guo et al. (2012)
methylbutanoate, ethyl hexanoate, hexyl acetate, hexyl butanoate, hexyl propanoate, pentyl 2-
methylbutanoate
1-Butanol, 1-hexanol, 2-methylbutyl acetate; 2-methylpropyl acetate, acetaldehyde, butyl acetate, (E)-3- Apple juice (60) APCI-MS, GC–MS LDA; PCA; SIMCA; LOOCV Gan et al. (2014)
hexenol, ethanol, hexanal, hexyl acetate, (Z)-2-hexenal
Abscisic acid, ferulic acid hexoside, linonoate A-ring lactone, naringin, neohesperidin, nomilin glucoside, Citrus juice (36) LC-ESI-q-TOF-MS HCA; PLS-DA; ANOVA Arbona et al. (2015)
obacunone, tryptophan
3-O-Trans-feruloyl tormentic acid Shrub chaste, Agnus castus LC-MS, NMR OPLS-DA Yahagi et al. (2016)
fruits (8)
C16:0, C18:0, C18:1, C20:0, C20:2, α-tocopherol, β-sitosterol, ɣ-aminobutyric acid, glucose, isoleucine, Rice (23) GC-FID, GC–MS HCA; PCA Frank et al. (2012)
methionine, myo-inositol, phosphoric acid, raffinose, sucrose
4-Guanidinobutanoate, agmatine, alanine, arginine, asparragine, ɣ-aminobutyric acid, glutamate, glycine, Rice (100) UHPLC-MS/MS, GC–MS ANOVA; PCA; ICV Hu et al. (2014)
pipecolate, serine, trans-4-hydroxyproline, tyrosine
Alanine, ɣ-aminobutyric acid, leucine, oxalic acid, valine Rice (48) GC–MS PCA Danial et al. (2013)
2-Methoxybenzoic acid, 2-methylbenzofuran, 2,6,6-trimethyl-2-cyclohexene-1,4-dione, 2′- Honey (9) UHPLC-PDA-MS/MS, PCA Beitlich et al. (2014)
hydroxyacetophenone, 2′- methoxyacetophenone, 3-hydroxy-1-(2-methoxyphenyl)-penta-1,4-dion, GC–MS

146
3,4,5-trimethylphenol, 4-methoxyphenyllactic acid, 5-methyl-3-furancarboxylic acid, acetyl-2-hydroxy-
4-(2-methoxyphenyl)-4 oxobutanal, cis-linalool, kojic acid, leptosin, lumichrome, methyl syringate, p-
anisaldehyde, p-anisic acid, phenethyl alcohol
2,3-Butanediol, cinnamaldehyde, hexanal, hexanol Date (13) GC–MS HCA; PCA; OPLS-DA Khalil et al. (2017)
Chrysoeriol rhamnosyl hexoside, fructose, glucose, isoquercetin, luteolin rhamnosyl dihexoside, rutin, Date (21) UPLC-PDA-ESI-q-TOF-MS HCA; PCA Farag et al. (2014)
sucrose
Caffeoyl shikimic acid, chrysoeriol glucoside, fructose, glucose, luteolin glycoside, quercetin conjugates Date (18) UPLC-q-TOF-MS PCA; OPLS-DA Farag et al. (2016)
13
Caffeine, caffeoylquinic acids, choline, citrate, malate, sucrose, trigonelline Coffee (12) C NMR PCA; OPLS-DA; ICV Wei et al. (2012)
Caffeoylquinic acid, dicaffeoylquinic acid, feruloylquinic acid, linoleic acid, palmitic acid, sucrose Coffee (23) ESI-FT-ICR-MS PCA; PLS-DA; LOOCV Garrett et al. (2013)
1
16-O-mehylcafestol, kahweol Coffee (101) H NMR ANOVA; PCA; ICV Monakhova et al. (2015)
Caffeine, ent-kaurane-diterpenoid derivatives (C20) Coffee (150) LC-q-TOF-MS PCA; PLS-DA Souard et al. (2018)
Caffeine, inositol, pyroglutamic acid Kopi Luwak (21) GC–MS PCA; OPLS-DA; ICV Jumhawan et al. (2013)
1-Octen-3-ol, 1-octen-3-one, 3-octanol, 3-octanone, octanal Truffle (6) GC–MS, GC-O – Culleré et al. (2013)
1,2-Dimethoxybenzene, 2-phenyl-2-buten-1-al, 5-methyl-2-phenyl-2-hexenal Truffle (25) GC–MS CA; Mann–Whitney test Splivallo et al. (2007)
5-(12-Nonadecenyl)-resorcinol, 5-heptadecylresorcinol, 5-(heneicosenyl)-resorcinol, 5-heneicosylresorcinol, Wheat (52) UHPLC-q-TOF-MS PCA; OPLS-DA; ROC curve Righetti et al. (2018)
5-nonadecanylresorcinol, 5-pentacosylresorcinol, 5-tricosylresorcinol, digalactosyl diglyceride,
monogalactosyl diglyceride, triacylglycerols
2-Methoxyhydroxytyrosol glucoside, dimethyl oleuropein, hydroxytyrosol glucoside, methoxytyrosol Olive tree (70) HPLC-ESI-MS/MS LDA; SIMCA; KNN; ICV Di Donna et al. (2010)
glucoside
2-Methyl-6-methyleneoctan-2-ol, 3- octanone, 4-octadecylmorpholine, 2,2,6-Trimetylcyclohexanone, (Z)- Tomato (5) GC–MS ANOVA; LSD; PCA; LOOCV Figueira et al. (2014)
methyl-3-hexenoate
2-Methyl-2-methylpropyl butyrate, 2-pentyl acetate, 3-methylhexyl butyrate, heptan-2-ol, isopentyl Banana (5) GC–MS PCA; SLDA; ICV Pontes et al. (2012)
hexanoate, n-butyl butyrate, (Z)-2-hexenyl butyrate
1-Octanol, 1-pentanol, 3-octanol, 3-octanone, (E)-2-octen-1-ol, (E)-2-octenal, hexanal, hexanol, linalool, ρ- Mushroom (18) GC–MS PCA Malheiro et al. (2013)
anisaldehyde
2-Heptanone, 2-methyl-4-propyl-1,3-oxathiane, α-terpinolene, benzyl pentanoate, butyl hexanoate, ethyl Passion fruits (9) GC–MS PCA; PLS-DA; LOOCV Porto-Figueira, Freitas, Cruz, Figueira,
crotonate, ethyl hexanoate, ethyl hexyl carbonate, furaneol, hexyl acetate, isopentyl acetate, methyl and Câmara, (2015)
butanoate, methyl hexanoate, methyl-(E)-2-hexenoate, methyl pentanoate, propyl hexanoate
(continued on next page)
Food Chemistry 278 (2019) 144–162
S. Medina et al. Food Chemistry 278 (2019) 144–162

spectrometry; ESI-FT-ICR-MS: electrospray ionization Fourier transform ion cyclotron resonance mass spectrometry; GC-FID: gas chromatography-flame ionization detector; GC-O: gas chromatography-olfactometry; HCA;
hierarchical cluster analysis; ICV: internal cross validation; GC–MS: gas chromatography mass spectrometry; GC-O: gas chromatography-olfactometry; KNN: K-nearest neighbours; MDGC: multidimensional gas-chro-
matography; MS/MS: tandem mass spectrometry; NMR: nuclear magnetic resonance; LC-ESI-q-TOF-MS: liquid chromatography-electrospray ionization-quadrupole-time of flight-mass spectrometry; LC-MS: liquid

analysis; PCA: principal component analysis; PDA: photodiode array; PLS-DA: partial least squares-discriminant analysis; ROC: receiver operating characteristics; SIMCA: soft independent modeling of class analogy; SLDA:
ANOVA: analysis of variance; APCI-MS: atmospheric-pressure chemical ionization-mass spectrometry; CA: cluster analysis; DA: discriminant analysis; DART-TOF-MS: direct analysis in real time- time of flight mass

chromatography mass spectrometry; LDA: linear discriminant analysis; LOOCV: leave-one-out cross validation; LSD: least square difference; OPLS-DA: orthogonal projections to latent structures modeling discriminant
recent study carried out in two white wines (Feteasca Regala and Sau-
vignon Blanc), indicates tannins as discriminant markers between these
wines. This was a little surprising because tannins are usually asso-
ciated to red wines. However, Feteasca Regala is a wine characterized by
a higher content of tannins when compared with other white wines
References

(Magdas, Guyon, Feher, & Pinzaru, 2018). Moving to fruit juices, Guo
Ferreira et al. (2009)
Avula et al. (2015)

Farag et al. (2018)

and colleagues performed a method for an efficient discrimination of

apple juice according to apple varieties used in their production in
China (Starkrimson, Qinguan, Gala, Jonagold, Golden Delicious, and Fuji).
The authors used headspace solid phase microextraction gas chroma-
tography-mass spectrometry (HS-SPME-GC–MS) and processed the
chromatographic data obtained using different chemometrics tools. In
ANOVA; PCA; OPLS; LOOCV

this work, stepwise linear discriminant analysis (SLDA) revealed sa-

tisfactory discrimination of apple juices and provided 100% success
Statistical analysis

rate in terms of prediction ability. Hence, this discriminative model is a

valuable tool to control the quality of the apple juices produced in
ANOVA; PCA

China and detect eventual adulterations (Guo, Yue, & Yuan, 2012).
Several citrus juices obtained from 12 commercial varieties (lemon
(Eureka and Fino), grapefruit (Marsh and Star Ruby), mandarin (Fortune,
PCA

Nadorcott, Pixie, and Hernandina), blonde orange (Sucrenya, Midknight)

and navel orange (Washington and Lane Late)) were also evaluated by
non-targeted metabolite profiling (Arbona et al., 2015). The results
obtained showed that several metabolites (abscisic acid (ABA), limo-
Methodology

DART-q-TOF-MS

noids, flavonoids and amino acids) were able to discriminate the citrus
varieties used in the study. Accordingly, Sucrenya variety showed a
specific accumulation of obacunone, nomilin glucoside and limonoate
GC–MS
NMR

A-ring lactone, as well as the highest ABA levels among all varieties,
suggesting a different ABA metabolite fingerprinting for each variety. In
turn, naringin and neohesperidin were exclusively present in grape-
fruits, while tryptophan was found in two grapefruits cultivars with
Matrix (number of

values four-fold higher than the average levels. In contrast, ferulic acid
samples)

hexoside was scarce in the juice of Sucrenya but it was found in sig-
Cherimoya (12)
Cinnamon (35)

Cinnamon (10)

nificantly higher levels in Fortune and Midknight varieties. These com-

pounds have gained therefore scientific and commercial interest as
important phylogenetic markets, but also due to their beneficial effects
on human health (Arbona et al., 2015).
Rice is a widely grown crop, endowed with a rich genetic diversity,
furan-2-one, α-terpeneol, benzoic acid, butyl propanoate, furaneol, linalool, methyl 2-furoate, methyl 3-

but the harvested seeds are often adulterated with others of lower
3-Methoxybutan-1-ol, 3-methylbutanoate, 3-methylbutan-1-ol, 3-methylbutyl butanoate, 5-methyl-(3H)-

stepwise linear discriminant analysis; UHPLC: ultra-high performance liquid chromatography.

Cinnamaldehyde, coumarin, dehydro-sesquiterpene, dehydro-sesquiterpene-oxide, methyl cinnamate,

Acetic acid, (E)-cinnamic acid; (Z)-cinnamic acid, eugenol, (E)-cinnamaldehyde, (E)-cinnamaldehyde

quality. So, it is very complicated to differentiate seeds of various rice

varieties based on visual observation. In this context, the metabolite
hydroxy-3-methylbutanoate, methyl-3-hydroxybutanoate, (Z)-2-penten-1-ol, (Z)-3-hexen-1-ol

profiling of rice (Oryza sativa L.) revealed an increase in the content of

fatty acid methyl ester (FAME), free fatty acids, organic acids and
amino acids and allowed the discrimination of red, black and non-co-
loured indica and japonica rice varieties (Frank, Reichardt, Shu, & Engel,
2012). These results were in agreement with another study that also
reported stigmasterol and ɣ-aminobutyrate as biomarkers to distinguish
between indica and japonica rice cultivars using a non-targeted meta-
bolomics approach (Hu et al., 2014). Moreover, in this recent work, 92
metabolites including 28 amino acids, 23 carbohydrates, 22 lipids, 12
CPGECs (cofactors, prosthetic groups and electron carriers), five nu-
Biomarkers

dimethyl acetal, glycerol, o-hydroxy-cinnamaldehyde

cleotides and two secondary metabolites exhibited statistically sig-

nificant differences between the referred cultivars. They observed that
metabolites, particularly amino acids, can be used to differentiate be-
tween different rice cultivars, with asparagine ranked the highest (Hu
et al., 2014). Using principal component analysis (PCA), it was also
possible to observe differences among rice cultivars (MR 219, MR220,
MR232 and Intani-2). So, high levels of leucine and oxalic acid appear
to be associated with a good tillering ability (Danial, et al., 2013). These
results highlighted the potential role of metabolites as early predictor
sesquiterpene oxide

biomarkers that may be very useful in crop breeding programmes and

Table 1 (continued)

can serve as a basis for the future improvement of rice quality via
metabolic engineering. Honey is a food product facing a great pressure
by the consumers. As it production shortages, honey price increases as
well as the number of adulterations of this product in the international
market. Moreover, counterfeiting honey is analytically very difficult to
discriminate. In this context, a recent work reported that 18 metabolites

147
 Table 2
Metabolites as potential biomarkers for food authenticity according to production systems.
Production systems Biomarkers Matrix (number of Methodology Statistical analysis References
S. Medina et al.

samples)

Farming practices (organic vs.
conventional)
Glutamylphenylalanine, N-malonyltryptophan Tomato (15) HPLC-q-TOF-MS ANOVA Vallverdú-Queralt et al.
(2011)
β1-Tomatine, dehydrotomatine, tomatidine Tomato (11) HPLC-LTQ- – Caprioli et al. (2014)
Orbitrap-MS
2-Furanone, α-ketobutyric acid, alanine, ascorbic acid, catechol, citramalic acid, citric Tomato, pepper (40, DART-TOF-MS PCA; LDA; LOOCV Novotná et al. (2012)
acid, dehydroascorbic acid, ferulic acid, fructosyl glutamate, furfural, glucoheptonic 24)
acid, gluconolactone, glutaric acid, glycerate, hydroxymethylfural, itaconic acid,
lactate, lactic acid, leucine, malic acid, proline, phenylalanine, piruvic acid,
pyroglutamic acid, quinic acid, succinic acid, threonine, valine;
Alanine, β-alanine, glycerate, hydroxyglutarate, myo-inositol, panthotenic acid, urea, Wheat (20) GC–MS Tukey test Zörb et al. (2006)
valine
4-Aminobutanoate, alanine, adenosine, arabinose, asparagine, aspartate, Wheat (22) GC–MS Studentś t-test; PCA Bonte et al. (2014)
ethanolamine, malate, myo-inositol, tryptophan, urea
Protocatechuic acid Wheat (36) LC-HRMS ANOVA; PCA Weesepoel, et al. (2016)
2-Piperidinecarboxylic acid, 2,3-dihydroxypropanoic acid, α-solanine, allantoin, Potato (48) GC–MS ANOVA; PCA Shepherd et al. (2014)
arginine; asparagine, aspartic acid, β-alanine, ɣ-aminobutyric acid, fumaric acid, LC-MS
galactinol, galactose, glucose, glutamine, glutathione, glutamic acid, glycerol,
glycine, histidine; inositol; isoleucine, leucine, L-alanine; lysine; methionine, oxalic
acid, oxoproline, phenylalanine; proline, quinic acid, serine; solasonine, sucrose,
thiamine, threonine, trihydroxypentanoic acid, tryptophan, tyrosine, valine
Phosphate, malic acid, myo-inositol Maize (54) GC–MS ANOVA; PCA Röhlig and Engel (2010)
Arabinofuranosyl-[a-L-arabinofuranosyl]-L-arabinose, chlorogenic acid, citric acid, L- Carrot (140) UHPLC-MS PCA; OPLS-DA; ROC curve; ICV; Cubero-Leon, et al. (2018)
arginine, sedoheptulose ECV
1-Acetyl-3,14,20-trihydroxywitha-5,24-dienolide 3-glucoside, butyl (S)-3- Goldenberry (30) UHPLC-q-TOF-MS Studentś t-test; PCA Llano et al. (2018)

148
hydroxybutyrate [arabinosyl-(1-greater than6)-glucoside], physagulin D
2-Oxoglutarate semialdehyde, 4-oxoglutarate semialdehyde, β-myrcene, Orange juices (19) UPLC-HR-MS; PCA; PLS-DA; HCA; LOOCV; PT; RT Cuevas et al. (2017)
dehydroascorbic acid, ethyl butanoate, ɣ-terpinene, hexanal, limonene, (Z)-3-hexanal GC–MS
(+)-Catechin, chlorogenic acid, (−)-epicatechin, gallic acid, myricetin, p-coumaric Grape juices (96) HPLC-DAD Mann-Whitney test; ANOVA; PCA; Granato et al. (2015)
acid quercetin HPLC-FLD PLS-DA; SIMCA; KNN; LOOCV;
ECV
Delphinidin 3-arabinoside, delphinidin 3-galactoside, delphinidin 3-glucoside, Blueberry (10) HPLC-DAD ANOVA Wang et al. (2008)
malvidin 3-arabinoside, myricetin 3-arabinoside, petunidin 3-galactoside, petunidin
3-glucoside, quercetin 3-glucoside
Caffeine, chlorogenic acid, trigonelline Coffee HPLC-DAD Newman-Keuls test; ANOVA do Carmo Carvalho et al.
(2011)
β-apo-carotenal, β-carotene, β-cryptoxanthin, canthaxanthin, luteoin/zeaxanthin Egg (84) HPLC-DAD ANOVA; PCA; LSD van Ruth et al. (2011)
α-carotene, β-carotene, chlorogenic acid, cis-β-carotene, gallic acid, kaempferol, Pepper (9) HPLC-DAD ANOVA Hallmann and
quercetin, quercetin-D-glucoside Rembiałkowska (2012)
1-Pentenol, 1-penten-3-ol, 2-heptyl butanoate, 2-hexanol, 3-hydroxy-2-butanone, 3- Banana (120) GC–MS t-test Dong et al. (2014)
methoxy-1-propanol acetate, 4-methyl-2-pentyl acetate, decanal, eugenol,
hexadecane, isobutyl isovalerate, isopropyl myristate, (Z)-3-hexenyl acetate
Phytanic acid, pristanic acid Cheese, milk, cream, GC–MS ANOVA Vetter and Schröder (2010)
butter (32)

Rearing practices (wild vs.
cultured)
Cholesterol, choline, C22:6 n-3, C20:5 n-3, fumaric acid, glutamine, malic acid, Sea bass (7) NMR ANOVA; PCA Mannina et al. (2008)
phosphatidylethanolamine, trimethylamine oxide
C14:0, C15:0, C16:0, C17:0, C18:0, C16:1 n-7, C18:1 n-9, C20:1 n-9, C18:2 n-6, C20:4 Sea bass (27) HPLC; GC–MS ANOVA; LSD Fuentes et al. (2010)
n-6, C20:5 n-3, C22:6 n-3, arginine, alanine, asparagine, glutamic acid, glycine,
histidine, methionine, phenylalanine, serine, taurine, tyrosine

(continued on next page)

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S. Medina et al. Food Chemistry 278 (2019) 144–162

liquid chromatography-fluorescence detector; HPLC-q-TOF-MS: high performance liquid chromatography-quadrupole- time of flight mass spectrometry; ICV: internal cross validation; LC-HRMS: liquid chromatography
high resolution mass spectrometry; LC-MS: liquid chromatography mass spectrometry LDA: linear discriminant analysis; LOOCV: leave-one-out cross validation; LSD: least square difference; LTQ: linear trap quadrupole;

permutation test; ROC: receiver operating characteristic; RT: randomization test; SLDA: stepwise linear discriminant analysis; SWDA: stepwise discriminant analysis; UHPLC-MS: ultra-high performance liquid chro-
ANOVA: analysis of variance; CANDISC: canonical discriminant analysis; CLA: conjugated linoleic acids; DA: discriminant analysis; DART-TOF-MS: direct analysis in real time- time of flight mass spectrometry; ECV:
external cross-validationGC-MS: gas chromatography mass spectrometry; HCA; hierarchical cluster analysis; HPLC-DAD: high performance liquid chromatography -diode array detector; HPLC-FLD: high performance

NMR: nuclear magnetic resonance; OPLS-DA: orthogonal projections to latent structures modeling discriminant analysis; PCA: principal component analysis: PLS-DA: partial least squares-discriminant analysis; PT:
(8 volatiles and 10 non-volatiles compounds) allowed a clear differ-
entiation among three kinds of honey derived from monocultures
(Manuka (8), Hanuka (7), and Jelly bush (1)) harvested in 2012.

Marseglia et al. (2013)

Caligiani et al. (2016)
Povolo et al. (2009)
Segato et al. (2017)
References

Nevertheless, in order to achieve a definitive classification of these

Vasta et al. (2011)

kinds of honey, further research is still necessary, particularly with a
larger number of samples, as well as with samples from different years
(Beitlich, Koelling-Speer, Oelschlaegel, & Speer, 2014). Date fruits have
been also subjected to an interesting discriminant analysis. Initially,
Farag, Mohsen, Heinke, and Wessjohann, (2014) reported differences
among 21 date varieties from Egypt, being flavonols glycosides (rutin
and isoquercetin), flavones glycoside (luteolin rhamnosyl dihexoside
ANOVA; SWDA; CANDISC; DA

and chrysoeriol rhamnosyl hexoside) and sugars (fructose, glucose, and

Kolmogorov-Smirnov’s test
Statistical analysis

sucrose) the principal contributors for the discrimination of the vari-

eties studied. Then, the same authors analysed 18 date cultivars from
ANOVA; SWDA; ICV

PCA; CA; LOOCV

Saudi Arabia using a metabolomics approach. The results obtained

using PCA and orthogonal partial least square (OPLS) analyses revealed
that the ratios flavones versus flavonol and sugars levels, were the main
ANOVA

contributors for the cultivar differentiation (Farag, Handoussa, Fekry, &

Wessjohann, 2016). Finally, using SPME-GC–MS, Khalil, Fekry, and
Farag, (2017) analysed the volatile fraction of date fruits and identified
a total of 89 VOCs in 13 Egyptian date varieties. Among these, lipid-
GC–MS; 1H NMR
Methodology

derived volatiles and phenylpropanoid derivatives were the major

HPLC; GC–MS

components of date fruit aroma and volatile markers as 2,3-butanediol,

GC–MS
GC–MS

cinnamaldehyde, hexanal, and hexanol, are among the ones with the
higher contribution for the discrimination of the date fruits varieties
GC

analysed. Coffee is globally one of the most widely used food products
and in the past two decades, there has been continuous growth in the
Matrix (number of

Asiago PDO Cheese

Milk, Cheese (220)

global consumption. Consequently, a huge number of articles have been

Milk, Cheese (43)
samples)

published concerning the coffee adulteration and some of them have

Cheese (304)

focused on identifying biomarkers for different species or varieties. In

Meat (100)

this context, Monakhova et al. (2015) reported that kahweol and 16-O-
(81)

methylcafestol may be used to authenticate arabica and robusta coffees,

respectively. Furthermore, arabica coffee was also reported to contain
significantly higher levels of sucrose, citrate, trigonelline, and malate,
whereas robusta coffee was characterized by higher levels of caffeine,
choline and caffeoylquinic acids (CQA) (Table 1) (Wei, et al., 2012). In
2-methyl-1-Butanol, 3-undecanone, cuminic alcohol, germacrene D, skatole
Anteiso-C15:0, C9:0, C18:3 n-6, C20:3 n-6, C20:4 n-6, cis-9, trans-11 CLA

another report, phenolic compounds, fatty acids, and sucrose were

cyclopropane fatty acids, Σ trans/trans CLA, trans-11, cis-13 CLA, retinol

described as cultivars biomarkers for eight arabica coffee cultivars,

what constitutes a valuable tool for the certification processes and
traceability of coffee beans (Garrett, et al., 2013). Very recently, coffees
leaves from nine species grown in the same environmental conditions
were subjected to discriminant analysis and caffeine and ent-kaurane
diterpenoid (C20) derivatives were found to be key metabolites in the
Biomarkers

discrimination of the selected coffee species. Caffeine concentration

Cyclohexyl tridecanoic acid, lactobacillic acid

was found approximately 800 times higher in C. arabica leaves com-

pared to C. canephora, what makes it a good marker to authenticate
Dihydrosterculic acid, lactobacillic acid
1-Phytene, 2-phytene, neophytadiene

coffee species and ent-kaurane diterpenoid (C20) derivative was found

in three different species (C. arabica, C. canephora and C anthonyi),
exhibiting similar metabolic profiles in the analytical conditions de-
scribed (Souard et al., 2018). Finally, it is worthwhile to refer the pe-
culiar arabica kopi luwak, an exotic Indonesian coffee and eventually the
world́s most expensive. The coffee berries from kopi luwak are eaten by
Asian palm civet, being the coffee pericarp completely digested and the
beans excreted. Then, they are collected, cleaned, wet-fermented, sun-
dried and roasted. A recent study allowed the discrimination between
robusta and arabica kopi luwak, being caffeine, inositol and pyr-
oglutamic acid the discriminant markers. Moreover, this was the first
matography mass spectrometry.
Feed practices (pasture vs. silage)

work contributing for the authentication of this expensive coffee and in

fact citric acid, malic acid, and the ratio inositol/pyroglutamic acids
Production systems

were able to differentiate among authentic and fake kopi luwak samples
Table 2 (continued)

(Jumhawan et al., 2013). Another very expensive matrix highly sus-

ceptible to frauds are the truffles, which can be morphologically very
similar, although having very different aromas. The characterization of
the volatile composition of different truffle species constitutes there-
fore, a very relevant opportunity to define biomarkers to detect frauds.
This has been reported by Splivallo, Bossi, Maffei, and Bonfante,

149
 Table 3
Potential biomarkers for the food authentication according to geographical and botanical origin.
Biomarkers Matrix (number of samples) Methodology Statistical analysis References
S. Medina et al.

Lepteridine Manuka honey (27) (New Zealand) HPLC Studentś t test Lin et al. (2017)
Synephrine Honeys (34) Monofloral citrus, ‘assa-peixe’, eucalyptus LC-MS/MS ANOVA Tette et al. (2017)
and ‘aroeira’ honey. Wildflower (polyfloral) honeys (São
Paulo, Minas Gerais)
Heptanal, hexanal, octanal, p-cymene Honeys (374) (Corsican, non-Corsican-France, Italy, GC–MS DPLS; LDA; SIMCA; SVM; RMSCV; Stanimirova et al. (2010)
Austria, Ireland, and Germany) LOOCV
1,5,8-p-Menthatriene, 1,8-nonadiyne, 2-furanmethanol, 2-hydroxy-3,5,5-trimethyl- Honeys (55) (Coffee, longan, lychee, sunflower and wild GC–MS ANOVA Pattamayutanon et al. (2017)
2-cyclohexen-1,4-dione, 2,2-dimethyl butanal, 2,6,6-trimethyl-2-cyclohexen-1- honeys of five provinces of Thailand)
ol, 3,4,5-trimethyl-phenol, 5-hydroxymethylfurfural, 10-oxodecanoate,
anisaldehyde, anise alcohol, benzaldehyde, butyryl lactone, diethyl
decanedioate, diethyl hexanedioate, ethyl heptanoate; hexadecane, lilac
alcohol B, lilac alcohol C, lilac alcohol D, methyl caproate, methyl oleate,
pentadecane, phenylmethanol, terpineol, (Z)-5,6-dimethyl-1,3-cyclohexadiene
4-Terpineol, α-terpineol, hotrienol, lilac aldehydes, linalool Honeys (25) (Acacia, sunflower, linden, rapeseed, GC–MS – Spanik et al. (2014)
chestnut and orange honeys from EU countries)
Kynurenic acid Honeys (32) (Botanical origins (multiflora, acacia, SEC-UV-DAD HCA; PCA Beretta et al. (2012)
chestnut, dog rose, mango and honeydew. Geographical
origins (Italy, Kosovo, Cameroon, Kenya and Brazil))
Perseitol Honeys (140) (Avocado nectar) NIRS PCR; PLS; SECV Dvash et al. (2002)
5-Hydroxymethylfurfural, citric acid, ethanol, lactic acid, phenylalanine, sucrose, Honeys (46) (Brazilian wildtype-, citrus-, eucalyptus, assa- NMR HCA; KNN; PCA; PLS-DA; SIMCA; Boffo et al. (2012)
tyrosine peixe, sugar-cane) SECV
4-(1-Hydroxy-1-methylethyl)benzoic acid, 4-(1-Hydroxy-1-methylethyl)- Honeys (305) (Linden, chestnut, orange and eucalyptus NMR 02PLS-DA Schievano et al. (2013)
cyclohexa-1,3-dienecarboxylic acid, 4-(1-Hydroxy-1-methylethyl)cyclohexa- from Italy)
1,3-dienecarboxylic acid methyl ester, 4-(1-methylethenyl)-cyclohexa-1,3-
dienecarboxylic acid, 2-quinolone, 3-oxo-α-ionone, 4-quinolone, 8-

150
hydroxylinalool, caffeine, deoxyvasicinone, dehydrovomifoliol
Acetyl-dicaffeoylglycerol, acetyl-feruloyl-caffeoylglycerol acetyl-p-coumaroyl- Propolis (13) (Germany) TLC, TLC-MS – Betrams et al. (2013)
caffeoylglycerol, acetyl-p-coumaroyl-feruloylglycerol, acetyl-di-p-
coumaroylglycerol, apigenin, caffeic acid, caffeic acid phenethyl ester, chrysin,
galangin, kaempferol, naringenin, pinocembrin, quercetin
Kaempferol-dimethylether Propolis (40) (Portugal) LC-DAD-ESI-MS(n) – Falcao et al. (2013)
4-(4′-Hydroxyphenyl)-2-butanone Arabica coffees (27) (Ethiopia, Tanzania, Guatemala) GC–MS, GC-O DA; PCA Akiyama et al. (2008)
2-Methylpyrazine, 2,3-diethylpyrazine, 2,3-dimethylpyrazine, 5-methylfurfural Arabica coffees (47) (Brazil, Colombia, Costa Rica, GC-TOF-MS PCA Risticevic, Carasek, and
Guatemala, Ethiopia, Indonesia) Pawliszyn, (2008)
(−)-Epicatechin, caffeine cinnamtannin A2, cyanidin-3-O-arabinoside, cyanidin-3- Cocoa (31) (31 different geographical origins) HPTLC – Pedan et al. (2017)
O-galactoside, proanthocyanidin B2 and C1, theobromine
1-Hexanol, (E)-2-hexenal, (E)-2-hexen-1-ol, hexanal, (Z)-3-hexenyl acetate Olive oils (36) (Southern Italian regions) GC–MS PCA Benincasa et al. (2003)
Olive oils (37) (Calabrian and Tunisian areas) GC–MS ANOVA; LDA; Wald–Wolfowitz Cavaliere et al. (2007)
test
3-Methylbutanal, ethanol tetrahydro-2-(3-pentynyloxy)-2H-pyran, hexanal Tomatoes (80) (Italian regions) GC–MS LDA; PCA; SIMCA kCV Feudo et al. (2011)
(E)-2-hexenal, (E,E)-2,4-heptadienal, ethyl hexanoate, hexyl 2-methylbutyrate, p- Apples (9) (Madeira Islands regions) GC–MS LDA; PCA; LOOCV Ferreira et al. (2009)
ethyl styrene
1-Butanol, 1-hexanol, (E)-3-hexenol, hexanal, (Z)-2-hexenal Apple juices (210) (New Zealand, South Africa, Chile) APCI-MS, GC–MS ANOVA, PCA; PLS-DA; LOOCV; Gan et al. (2014)
ECV
Citrusin D Oranges (65) (Spain, Brazil, Argentina, South Africa) UHPLC-q-TOF MS PLS; OPLS-DA; ECV Diaz et al. (2014)
4-Aminobutyrate, O-phosphocholine, acetate, asparagine, isoleucine, leucine, Cabbages (322) (Korea, China) NMR PCA; ICV Kim et al. (2013)
phenylacetate, phenylalanine, succinate, sucrose, tyrosine, valine
Diacylglycerols, γ-tocopherol, phosphatidylcholines, phosphatidylethanolamines, Hazelnuts (196) (Germany, France, Italy, Turkey, UPLC-q-TOF-MS ANOVA; Studentś t-test; PCA; Klockmann et al. (2016)
triacylglycerols Georgia) LDA; PLS-DA; SIMCA; SVM; ICV
1-Hexanol, 1-pentanol, 1-penten-3-ol, hexanal, pentanal, trimethylbenzenes Walnuts (China, Ukraine, Chile) GC–MS ANOVA Elmore, Nisyrios, and Mottram,
(2005)
2-Furaldehyde, 5-(hydroxymethyl)-2-furaldehyde, vanillin, syringaldehyde, Sherry vinegars (Aging in acacia or cherry wood barrels) GC–MS LDA; BPANN García-Parrilla et al. (2011)
coniferaldehyde
(continued on next page)
Food Chemistry 278 (2019) 144–162
 Table 3 (continued)

Biomarkers Matrix (number of samples) Methodology Statistical analysis References

S. Medina et al.

(+)-Catechin; (−)-epicatechin, cis-piceid, p-hydroxybenzoic acid, caftaric acid, Wines (28) (Czech Republic regions) HPLC ANOVA Lampir and Pavlousek (2013)
protocatechuic acid
(+)-Catechin, (−)-epicatechin, (−)-epicatechingallate, (−)-epigallocatechin, p- Wines (90) (China regions) HPLC ANOVA; CDA; ICV Sun et al. (2015)
coumaric acid, p-hydroxy benzoic acid, caffeic acid, chlorogenic acid, ferulic
acid, gallic acid, gentisic acid, epigallocatechin gallate, protocatechuic acid,
sinapic acid, syringic acid, vanillic acid
Malvidin-3-O-glucoside, malvidin-3-O-glucoside-4-vinylguaiacol, malvidin-3-(6-O- Wines (27) (Argentina regions) LC-MS PLS; LOOCV Pisano et al. (2015)
acetylglucoside)
1-Octen-3-ol, 2-ethylhexan-1-ol, 3-ethylbutanoic acid, 3-ethoxypropan-1-ol, 3,5,5- Wines (34) (Azores, Canary, Madeira Islands) GC–MS PLS-DA Perestrelo et al. (2014)
trimethylhexan-1-ol, 4-methylphenol, 4-(methylthio)-1-butanol, decanoic acid,
(E)-whiskey lactone, ethyl 2-methylbutanoate, ethyl DL-2-hydroxycaproate,
ethyl 3-hydroxybutanoate, γ-octalactone, isoamyl lactate, (Z)-3-hexenyl
butanoate
(E)-β-damascenone, α-bisabolene, α-terpineol, α-ylangene, β-bourbonene, β- Grapes (22) (Madeira Island regions) GC–MS PCA; HCA Perestrelo, Barros, Rocha, and
ionene, geraniol, hotrienol, linalool Câmara, (2014)
1
Citrate, malate, proline, threonine, trigonelline Grapes (20) (South Korea regions) H NMR PCA; PLS-DA Son et al. (2009)
1
Acetate, alanine, aspartate, choline, choline phosphate, citrate, citrulline, Lentils (85) (Italy, Canada) H NMR PCA-LDA; PLS-DA; SIMCA; MCCV Longobardi et al. (2017)
isoleucine, galactose, glucose, malate
2-Hydroxy-4-isopropylnaphthalene, 2-heptenal, 2-methylbutanal, 2- Truffles (Italy regions) PTR-TOF-MS PCA; HCPC Vita et al. (2015)
methylbutanoate, 2-methyl-1-propanol, 2-pentylfuran, 2,3-octanedione, 2,4-
dimethylfuran, acetaldehyde, acrylic acid, carveol, cyclopentyl-1-thiaethane,
diethanol sulfide, ethenone (akyl fragment), ethyl;, pyridine
2-Carene, 3-carene, α-phellandrene, α-pinene, β-pinene, γ-terpinene, camphene, Cheeses (41) (Five Slovenian PDO) GC–MS CA Tompa et al. (2013)
limonene, tricyclene
β-Caryophyllene Cheeses (35) (PDO Asiago Mountain) GC–MS — Favaro, Magno, Boaretto, Bailoni,
and Mantovani, (2005)

151
Pachymic acid Mushroom (60) (Wolfiporia extensa from five China UFLC, UV ANOVA; PLS-DA; HCA Li et al. (2016)
regions) spectroscopy
Glycerophospholipids and their oxidized lipids Saffron (44) (Spanish PDO La Mancha-Aragón) UHPLC-q-TOF-MS/ PCA; OPLS-DA; ICV Rubert et al. (2016)
MS

ANOVA: analysis of variance; APCI-MS: atmospheric-pressure chemical ionization-mass spectrometry; BPANN: back propagation artificial neural network; CA: cluster analysis; CDA: canonical discriminant analysis; CE:
capillary electrophoresis; DA: discriminant analysis; DPLS: dispersive partial least squares; ECV: external cross validation; GC–MS: gas chromatography mass spectrometry; GC-O: gas chromatography-olfactometry; HCA;
hierarchical cluster analysis; HCPC: hierarchical clustering on principal components; HPLC: high performance liquid chromatography; HPTLC: high performance thin layer chromatography; ICV: internal cross validation;
kCV: k-fold cross validation; KNN: K-nearest neighbour; LC-DAD-ESI-Ms(n): liquid chromatography- diode array detector-electrospray ionization- tandem mass spectrometry; LC-MS/MS: liquid chromatography tandem
mass spectrometry; LDA: linear discriminant analysis; LOOCV: leave-one-out cross validation; MCCV: monte carlo cross validation; NIRS: near-infrared spectroscopy; NMR: nuclear magnetic resonance; OPLS-DA:
orthogonal projections to latent structures modeling discriminant analysis; PCA: principal component analysis; PCR: principal component regressions; PLS-DA: partial least squares-discriminant analysis; PDO: protected
designation of origin PTR-TOF-MS: proton transfer reaction time of flight mass spectrometry; RMSCV: root mean square error of cross validation; SEC-UV-DAD: size exclusion chromatography ultraviolet diode array
detection; SECV: standard error of cross validation; SIMCA: soft independent modeling of class analogy; SVM: support vector machines; TLC: thin layer chromatography; UFLC: ultra-fast liquid chromatography; UHPLC-q-
TOF-MS: ultra-high performance liquid chromatography-quadrupole-time of flight-mass spectrometry; UV: ultraviolet.
Food Chemistry 278 (2019) 144–162
 S. Medina et al.

Table 4
Biomarkers candidates to detect food adulteration.
Biomarkers Matrix (number of samples) Methodology Statistical analysis References

Filbertone, lupeol Olive oil (Adulteration with hazelnut oil) GC–MS – Azadmard-Damirchi (2010)
Lactose semicarbazone, semicarbazide Milk (Adulteration with nitrogen-containing LC-MS – Abernethy and Higgs (2013)
compounds)
Linoleic acid, oleic acid, saturated fatty acids (total concentration), triacylglycerol, cholesterol Milk (Adulteration with vegetable oils and GC–MS t-test Kim et al. (2015)
contents animal fats)
Hexanal, heptanal, methyl-butanal, nonanal, pentanal Maté tea (6) (Adulteration of Ilex GC–MS – Dallago, Valduga, Luccio, Benin,
paraguariensis with other Ilex species) and Tres, (2011)
3-Oxohexadecanoic acid glycerides, arabitol, CE(22:5), Cer(d18:1/24:1), citric acid, creatinine, Meat (85) (Adulteration of beef mince with GC–MS; UHPLC-MS ANOVA; PCA; PLS-DA Trivedi et al. (2016)
decanoylcholine, ethanolamide, glucose 6-phosphate, glycine, glycyl-lysine, heptadecane, pork mince)
hexano-dibutyrin, malic acid, myo-inositol, N-carboxyethyl-gaminobutyric acid, oleic acid,
pentadecane, PG(36:4), phosphate, prostaglandin D2, pyroglutamic acid, TG (16:0/15:0/18:4),
xi-2-ethyl-1-hexanol
Caffeoylphenylalanine, p-coumaroyltyrosine Coffee (Adulteration arabica with robusta) QTOF-MS; FT-ICR PCA; PLS-DA; LOOCV Garrett et al. (2013)

152
2,5-Dimethylfuran, 4-ethyl-2-methoxyphenol, indole, maltol, phenol Coffee (Roast defects) GC–MS ANOVA; PCA Yang et al. (2016)
Catechol Propolis (22) (Adulteration with poplar HPLC-MS; 1H NMR; 13C – Huang et al. (2014)
extract) NMR
2-acetylfuran-3-glucopyranoside Honey (186) (Adulteration by rice syrup) HPLC-DAD; LC-q-TOF-MS; – Xue et al. (2013)
NMR
1
Dimethylproline Orange juice (313) (Adulteration with pulp H NMR ANOVA; LDA; PLS; Le Gall et al. (2001)
wash) SLDA; CVA
Arbutin Apple juice (132) (Adulteration with pear HPLC-PDA – Thavarajah and Low (2006)
juice)
4-O-p-Coumarylquinic acid, abscisic acid, arbutin, isorhamnetin-3-O-rutinoside Apple juice (58) (Adulteration with pear HPLC-PDA; LC-MS/MS; – Willems and Low (2018)
juice) NMR

ANOVA: analysis of variance; BPANN: back propagation artificial neural network; CVA: canonical variates analysis; FT-ICR: Fourier transform ion cyclotron resonance; GC–MS: gas chromatography mass spectrometry;
HPLC-DAD: high performance liquid chromatography-diode array detector; HPLC-PDA: high performance liquid chromatography-photodiode array; LC-MS/MS: liquid chromatography-tandem mass spectrometry; LC-q-
TOF-MS: liquid chromatography quadrupole time of flight-mass spectrometry; LDA: linear discriminant analysis; LOOCV: leave-one-out cross validation; NMR: nuclear magnetic resonance; PCA: principal component
analysis; PLS-DA: partial least squares-discriminant analysis; SLDA: stepwise linear discriminant analysis; UHPLC-MS: ultra-high performance liquid chromatography mass spectrometry.
Food Chemistry 278 (2019) 144–162
 S. Medina et al.

Table 5
Metabolites as potential biomarker compounds for the detection of food spoilage and freshness.
Biomarkers Matrix (number of Methodology Statistical analysis References
samples)

Spoilage 2-Ethyl-1-hexanol, 2-methyl-1-butanol, 3-methylbutanal, 3-methyl-1-butanol, ethanol, ethyl acetate, Sea bass (8) GC–MS PCA, Tukeýs test Parlapani et al. (2015)
ethyl isobutyrate, ethyl propionate
Dimethyl sulfide, trimethylamine Oyster GC–MS Bioelectronic – Lee et al. (2018)
nose
3-Methyl butanoic acid, acetic acid, acetoin, ethyl benzene, propyl benzene, styrene Salmon (12) GC–MS ANOVA, REML Wierda et al. (2006)
2,3-Butanediol, 3-methyl-1-butanol, acetic acid, dimethyl sulfide, ethyl acetate, isobutyl alcohol, Brown shrimp Atlantic SIFT-MS HCA, PCA, PLS; LOOCV Kuuliala, Abatih, et al.
trimethylamine cod (18) (2018)
2,3-Butanediol, 3-methyl-1-butanol, ethanol, ethyl acetate, trimethylamine Atlantic cod (31) SIFT-MS Friedmańs test Kuuliala, Al Hage, et al.
(2018)
2,4,6-Trimethypyridine, allyl methyl sulfide, dimethyl disulfide, dimethyl trisulfide, methyl Chicken (12) GC–MS – Lovestead and Bruno
thiolacetate, phenyl sulfide (2010)
2-Butanol, 3-methyl-1-butanol, acetic acid, ethanol, dimethyl disulfide, dimethyl trisulfide, proponic Chicken (168) GC–MS/FASST ANOVA, CA, HCA, PCA Miks-Krajnik et al. (2016)
acid, sulfides methanethiol
Acetone, alanine, arginine, asparagine, carbon disulfide, dimethyl sulfide, ethanol, ethyl acetate, Chicken (128) HPLC; GC–MS – Alexandrakis et al. (2012)
glucose, glutamine, glycine, hexanal, heptane, isoleucine, L-lactate, lysine, methionine, methyl

153
benzoate, methyl ethyl ketone, phenylalanine, serine, threonine, toluene, tyrosine
Acetoin, acetone, diacetyl, dimethyl sulfide ethanol Chicken (21) PTR-MS – Franke and Beauchamp
(2017)
3-Methylindole, acetoin, adenosin diphosphate, aspartic acid, betaine, butanoic acid, creatine, Beef (28) GC–MS HCA, PLS, Pearson correlation Ercolini et al. (2011)
1
glycogen, histidine, inosine monophosphate, lactate, leucine, proline; threonine H NMR
2,3,5,-Trimethyl pyrazine, 2,6-dimethyl pyrazine, cyclohexanone, ethanol 2,2 (ethoxy-ethoxy), Iberian ham (30) GC–MS ANOVA, Pearson correlation Martín et al. (2010)
tetramethyl pyrazine, trimethyl pyrazine
Ergosterol Grape (10) HPLC Pearson correlation Porep et al. (2014)
5,10-Diethoxy-2,3,7,8-tetrahydro-1H,6Hdipyrrolo[1,2-a:1′,2′-d]pyrazine, pyruvic acid, tryptophan, Bread (25) GC x GC-TOF-MS PCA, PLS-DA, Studentś test; Cheng et al. (2015)
2,3-butanediol, leucinic acid, malonic acid, Egg, (25) Jonckheere-Terpstra test; LOOCV
Nitrosotrimethylurea, putrescine, Cucumber (25)
Ethanolamine, fumaric acid, homoproline; pyroglutamic acid, threonic acid

Freshness 1-Penten-3-ol, 1-octen-3-ol, cyclopentanol, (E)-2-hexenal, (E)-2-pentenal, hexenal, octanal, (Z)-2- Salmon (12) GC–MS ANOVA, REML Wierda et al. (2006)
penten-3-ol
Carotenoids Citrus fruits (18) Raman spectroscopy – Nekvapil et al. (2018)

ANOVA: analysis of variance; CA: cluster analysis; FASST: fast automated scan/sim type; HCA; hierarchical cluster analysis; HPLC: high performance liquid chromatography; GC x GC-TOF-MS: two dimensional gas
chromatography time of flight mass spectrometry; GC–MS: gas chromatography mass spectrometry; LOOCV: leave-one-out cross validation; PCA: principal component analysis; PLS-DA: partial least squares-discriminant
analysis; PTR-MS: proton-transfer-reaction mass spectrometry; REML: residual maximum likelihood; SIFT-MS: selected-ion-flow-tube mass spectrometry.
Food Chemistry 278 (2019) 144–162
S. Medina et al.
Fig. 1. Potential candidates for biomarkers in food authenticity and safety.
(2007), that used headspace stir bar sorptive extraction-gas chromato-
graphy-mass spectrometry (HS-SBSE-GC–MS) to characterize Tuber
melanosporum (black truffle) and Tuber indicum (Chinese truffle) and
identified 1,2-dimethoxybenzene, 2-phenyl-2-buten-1-al and 5-methyl-
2-phenyl-2-hexenal as potential biomarkers to differentiate the selected
truffles. In another study involving GC-O and HS-SPME-GC–MS tech-
nologies, the VOCs 1-octen-3-ol, 1-octen-3-one, 3-octanol, 3-octanone
and octanal were also reported to distinguish between these species of
truffles (Culleré, Ferreira, Venturini, Marco, & Blanco, 2013). A similar
application involved the characterization of the volatile fraction of six
wild mushroom species (Clitocybe odora, Clitocybe fragrans, Hebeloma
crustuliniforme, Lepista nuda, Tricholoma fracticum and Tricholoma ter-
reum). Using HS-SPME-GC–MS followed by unsupervised analysis, as
PCA, 1-octanol, 1-pentanol, 3-octanol, 3-octanone, (E)-2-octen-1-ol,
(E)-2-octenal, hexanal, hexanol, linalool, and ρ-anisaldehyde were de-
scribed as markers of the six mushrooms species analysed. This study
corroborated the importance of using the volatile metabolites as a
powerful taxonomic tool (Malheiro et al., 2013).
On the other hand, a recent study described an untargeted lipi-
domics strategy to discriminate common (Blasco, Triticum aestivum) and
durum wheat (Odisseo, Triticum turgidum spp. durum) varieties. In this
work, alkylresorcinols, in particular 5-heptadecylresorcinol (AR 17:0),
triacylglycerols (56:2, 56:3, 56:4 and 56:5), monogalactosyl diglyceride
(36:3, 36:5 and 38:6) and digalactosyl diglyceride (34:2, 36:3 and 36:4)
were reported as potential biomarkers to differentiate common and
durum wheat varieties. This result demonstrated that untargeted lipi-
domics is a powerful tool for the detection of wheat frauds and it might
be applied in other lipid rich matrices (Righetti, et al., 2018). Olive oil
is very susceptible to adulteration, particularly with oils from lower
quality and species. Some olive oil cultivars are recognized as being of
high quality, originating more expensive olive oils and so are legally
protected by different certifications of origin. Using a high-performance
liquid chromatography-electrospray ionization coupled with tandem
mass spectrometry (HPLC-ESI-MS/MS) technique, Di Donna et al.
(2010) analysed the leaves of five cultivars of Olea europaea (Carolea,
154
Food Chemistry 278 (2019) 144–162
Cassanese, Coratina, Nocellara and Leccino) and reported that four
phenolic compounds, 2-methoxyhydroxytyrosol glucoside, dimethyl
oleuropein, hydroxytyrosol glucoside and methoxytyrosol glucoside,
were discriminant biomarkers of the selected cultivars. A similar work
using HS-SPME-GC–MS was carried out with different tomato cultivars
(Lycopersicum esculentum L., plum, campari, grape, cherry and regional)
and identified five VOCs (2-methyl-6-methyleneoctan-2-ol; 3-octanone;
4-octadecylmorpholine; 2,2,6-trimetylcyclohexanone and (Z)-methyl-3-
hexenoate) as discriminant markers (Figueira, Camara, Pereira, &
Camara, 2014). The same strategy was applied to five banana cultivars
of the Musaceae family (dwarf cavendish, prata, maçã, ouro, and pla-
tano). In this case, the esters group was the largest chemical class
identified in all cultivars and 2-methyl-2-methylpropyl butyrate, 2-
pentyl acetate; 3-methylhexyl butyrate, heptan-2-ol, isopentyl hex-
anoate, n-butyl butyrate and (Z)-2-hexenyl butyrate were identified as
metabolites that clearly classify the banana samples by cultivars
(Pontes, Pereira, & Câmara, 2012). 1H NMR also constitutes a powerful
metabolomics tool, as the work from Eisenmann, et al. (2016) showed.
Using an untargeted metabolomics approach, the authors applied
multivariate analysis to the peel and pulp extracts of 14 apple cultivars
and observed a remarkable variation in the content of sugars, poly-
phenols and acids among the selected cultivars. In turn, this allowed a
clear discrimination between the 14 apple cultivars analysed (Table 1).
Cinnamon is a common spice with multiple uses in cosmetic, food, and
pharmaceutical industries. It is nevertheless, very susceptible to adul-
teration, with many species of plants being labelled as cinnamon;
however, the “true cinnamon” refers only to the dried inner bark of
Cinnamon verum J.S. Presl. Regarding this fraud, a study carried out a
robust method to authenticate the “true cinnamon” and biomarkers as
coumarin, cinnamaldehyde, methyl cinnamate and various sesqui-
terpenes were reported to differentiate C. verum from other species
(Avula, et al., 2015). Another recent report added that several meta-
bolites are capable of distinguishing between “cinnamon” species C.
verum and C. cassia, particularly eugenol (2-methoxy-4-prop-2-en-
ylphenol), that was found mostly in C. verum (see Table 1) (Farag, et al.,
S. Medina et al.
2018). All studies referred above emphasize that genetic diversity
(plant cultivars) modulate the intrinsic composition for several com-
pounds. These putative biomarkers have different physicochemical
properties, mainly involving amino acids, phenolic compounds, sugars
and organic acids, but also VOCs from a wide diversity of chemical
families. Therefore, it is important to consider different analytical
platforms to identify biomarkers able to reliably discriminate foodstuffs
by the plant cultivar used in its production.
2.2. Production systems
The systems behind the production of different foodstuffs poten-
tially affect the levels of certain phytochemicals. This may result in the
identification of food production biomarkers, similarly to the examples
given in the previous section. In this context, several studies have as-
sayed the influence of farming, rearing or feed practices in the meta-
bolites profile of different foodstuffs (food metabolome) (Cubero-Leon,
et al., 2018). This discussion is particularly relevant in the context of
the growing consumer’s interest in organic foods, whose authentication
requires robust analytical platforms and bioinformatics tools. Re-
garding this, the influence of farming practices (organic and conven-
tional; O/C) in the context of a single nutrient or class of compounds is
still matter of debate in the scientific literature. Nevertheless, un-
targeted metabolomics approach has a great potential to certify the
authenticity of organic plant products (Llano, et al., 2018). In recent
years, several studies presented potential biomarkers to discriminate
samples from O/C farming (Table 2). Vallverdú-Queralt, Medina-
Remón, Casals-Ribes, Amat, and Lamuela-Raventós, (2011), for in-
stance, compared tomato-derived products (O/C ketchup) using high-
performance liquid chromatography-quadrupole time of flight-mass
spectrometry (HPLC-q-TOF-MS) and reported that glutamylphenylala-
nine and N-malonyltryptophan were biomarkers able to differentiate
organic ketchup from the conventional one. Also, in tomato products, a
method developed to analyse tomatidine using LC coupled to LTQ-Or-
bitrap mass spectrometry, identified higher levels of this compound in
conventional samples. Additionally, the authors of this study also used a
non-targeted metabolomics approach to identify another two metabo-
lites, dehydrotomatine and β1-tomatine, that discriminate the two types
of samples (Caprioli, Logrippo, Cahill, & James, 2014). A previous
metabolomics fingerprinting assay employing direct analysis in real
time (DART) coupled to TOF-MS for authentication of tomatoes and
peppers from O/C farming was also reported. This approach identified
several metabolites altered in both tomatoes and peppers (2-furanone,
furfural, α-ketobutyric acid, citric acid, dehydroascorbic acid, glutaric
acid, hydroxymethylfuran, itaconic acid, malic acid, pyruvic acid, and
succinic acid), showing the effect of agronomic systems on metabolome
and specific metabolic pathways, irrespective of the matrix tested
(Novotná, et al., 2012). The potential of metabolomics for the au-
thentication of organic crops has been also explored in cereals. Zörb
and co-workers, for instance, determined the levels of 52 metabolites,
including amino acids, organic acids, sugars, sugar phosphates, alco-
hols, and nucleotides in wheat (Triticum aestivum L.) grown under O/C
conditions. The authors reported that only eight of them (alanine, β-
alanine, glycerate, hydroxyglutarate, myo-inositol, panthotenic acid,
urea and valine) showed significant differences between two farming
systems (Table 2) (Zörb, Langenkämper, Betsche, Niehaus, & Barsch,
2006). Another study involving wheat grains, detected 11 metabolites
with significant differences between the O/C farming practices (Bonte,
et al., 2014). Moreover, among these metabolites, three of them, ala-
nine, myo-inositol and urea, were also identified in the work of Zörb
and co-workers (Zörb, et al., 2006) previously reported in this section.
Finally, Weesepoel, et al. (2016) also reported that protocatechuic acid
may serve as a single marker for discrimination between O/C produced
wheat. Metabolomics assays were also performed in potato samples
(Solanum tuberosum L.) grown under both O/C fertilizer systems and
most of the metabolites identified, mainly amino acids, were
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Food Chemistry 278 (2019) 144–162
statistically different between the two crops (see Table 2). This fact may
be due to N levels in the tubers, which are significantly higher in
conventional fertilization regimes when compared to the organic ones
(Shepherd, et al., 2014). The application of a comprehensive metabolite
approach using GC–MS to discriminate between O/C productions was
also applied to maize samples (Zea mays). In this case, malic acid, myo-
inositol and phosphate were the metabolites responsible for such dif-
ferentiation. However, in this work, other factors like the genotype,
farming location, and growing season may have contributed to the
different metabolite profile of maize kernels obtained (Röhlig & Engel,
2010). In contrast, the influence of these factors has been considered in
a recent untargeted metabolomics approach performed in carrot sam-
ples (Daucus carota L.). Cubero-Leon, et al. (2018) found a consistent
influence of the production systems on the crop’s metabolome of two
carrots varieties (Nerac and Namur) produced by different fertilization
approaches, in different regions of the same country, and over a period
of four years. In this case, markers related to carbohydrate metabolism
and plant defence mechanisms were identified as responsible for the
differences between O/C carrot samples (Table 2) (Cubero-Leon, et al.,
2018). The influence of the farming system has been also assayed in the
goldenberry fruits (Physalis peruviana). In a recent report, untargeted
metabolomics analysis identified two withanolides and one fatty acyl
glycoside as putative markers to differentiate O/C goldenberry fruits.
However, targeted metabolomics assays involving carotenoids and as-
corbic acid did not find statistical differences between both crops.
Therefore, untargeted metabolomics seems to be more suitable for the
authentication of organic products (Llano, et al., 2018). In addition,
other studies are based on carotenoids profiling to verify the differences
between farming systems. In this context, a previous report highlighted
the changes in carotenoids profile between O/C chicken eggs. The re-
sults of this study indicate that food fingerprinting approach is a va-
luable tool for analytical verification of the production systems of or-
ganic eggs (van Ruth, et al., 2011). Related with this, another study
showed that the volatile fraction of organic bananas was higher than
conventional ones and these augmented VOCs could be used as tenta-
tive biomarkers to authenticate organic bananas. This shows that the
method of cultivation also alters the fruit aromatic composition, pro-
viding an higher aromatic quality to the fruits grown under organic
farming, which is certainly a valuable information for producers and
consumers (Dong, Chen, Wang, Huang, & Yi, 2014). Nevertheless, ap-
proaches involving complementary analytical techniques, as the ones
combining metabolomics fingerprinting by ultra-performance liquid
chromatography high resolution mass spectrometry (UPLC-HR-MS),
volatile profile analysis by HS-SPME-GC–MS, and chemometrics
models, certainly constitute a more powerful framework for the au-
thentication of organic foodstuffs. This integrated approach involving
the characterization of both the volatile and non-volatile metabolites
was recently carried out in orange juices and allowed the differentiation
between O/C juices (Table 2) (Cuevas, Pereira-Caro, Moreno-Rojas,
Muñoz-Redondo, & Ruiz-Moreno, 2017). The phenolic compounds
content of organic products relative to their conventional counterparts
has been extensively investigated. Although this is still matter of debate
the scientific literature, it appears that organic products have a higher
content of phenolic compounds than the traditional ones. This ob-
servation was reported, for instance, for polyphenols in grapes juices
(Granato, et al., 2015), anthocyanins in blueberries var. Bluecrop
(Vaccinium corymbosum L.) (Wang, Chen, Sciarappa, Wang, & Camp,
2008), and different antioxidant compounds (phenolic compounds,
carotenoids, vitamin C, among others) in sweet bell peppers (Hallmann
& Rembiałkowska, 2012), suggesting that organically grown foods have
higher content of these compounds and this fact may have positive
effects on human health. Regarding this, an animal experiment showed
that coffee (both O/C) had beneficial effects on rat health, although
chlorogenic acids levels were higher in organic coffees than conven-
tional ones. More studies are therefore necessary to clarify whether
there is an effective higher content in phenolics in the organic products
S. Medina et al.
relatively to conventional products and if this represents a real positive
effect in human health (do Carmo Carvalho et al., 2011). Food pro-
cessing adds an extra level of complexity to the authentication problem
because its effects on food composition and eventually can mask the
discrimination of frauds. Regarding this, two fatty acids, phytanic acid
and pristanic acid, were found significantly higher concentration in
organic products from German markets like cheese, milk, cream and
butter than conventional ones. A level of 200 mg of phytanic acid per
100 g of lipids that was then suggested as the value for control for or-
ganic products and this will constitute a valuable biomarker for the
authentication of this organic product (Vetter & Schröder, 2010).
Discrimination of the rearing practices used to obtain certain food
product is certainly very relevant to the producers and consumers. The
work from Mannina, et al. (2008), used NMR to differentiate between
wild and cultured sea bass (Dicentrarchus labrax). The metabolite pro-
filing in conjunction with statistical analysis identified fatty acids
(monounsaturated fatty acids (MUFA), diunsaturated fatty acids
(DUFA), and polyunsaturated fatty acids (PUFA)) and other metabolites
as compounds responsible for the discrimination between different
rearing systems. Fumaric and malic acids, which are metabolites of
Krebs cycle, were found among the identified metabolites, suggesting
differences at the level of energy metabolism in cultured fish versus wild
type fish. Moreover, accumulation of glutamine was also reported in the
wild type fish, what can be correlated with the higher level of these
organic acids, since glutamine metabolism is closely related to Krebs
cycle (Mannina, et al., 2008). As can be seen in Table 2, these results
are in accordance with another work reporting fatty acids and free
amino acids as discriminative compounds between farmer and wild sea
bass (Fuentes, Fernández-Segovia, Serra, & Barat, 2010). Recently, the
influence of feed practices in the production of different foodstuffs has
been investigated by different researchers. Segato, et al. (2017), for
instance, were able to distinguish among three cheese production sys-
tems with milk from cows fed, pasture-based, hay-based and maize si-
lage-based, which one was used in the production of cheese from
Asiago, and Italian protected designation of origin (PDO). Using GC and
the discriminant analysis, the authors found nine fatty acids and fat-
soluble vitamins as useful biomarkers to separate the three production
systems. Moreover, high contents of conjugated linoleic acids (CLA),
antesio-C15:0 and vitamin A were enough to discriminate pasture-based
cheese (Segato, et al., 2017). Also related with the cheese production,
another study reported the discrimination of the simultaneously use of
milk originating from two different production systems (animals
feeding on pasture and feeding with concentrates or silages) to produce
the same cheese type. This allowed an effective product protection and
a corresponding increase of economic value (Povolo, Pelizzola, Ravera,
& Contarini, 2009). Similarly, another work reported the presence of
lactobacillic acid and dihydrosterculic acid (components of bacterial
membranes) in milk and dairy products from cows fed with silage.
Therefore, these acids may be used as biomarkers of silage feedings for
milk-based products (Caligiani, Nocetti, Lolli, Marseglia, & Palla,
2016). Lactobacillic acid was also detected in milk samples from cows
fed with forages containing maize silage, which is not allowed to pro-
duce milk for some PDO cheeses as Parmigiano-Reggiano, constituting
therefore a very useful biomarker to detect cheese adulteration
(Marseglia, et al., 2013). In the same context, but using volatile markers
characterized through GC–MS analysis, skatole, 3-undecanone, cuminic
alcohol, 1-butanol, and 2-methyl were able to discriminate between
animals fed concentrate and animals fed non-concentrate diets and the
terpenoid germacrene D was reported as a prospective marker of grass
feeding (Vasta et al., 2011).
2.3. Geographical/botanical origin
Ongoing consumer’s demands about greater labelling specifications
regarding the foods geographical origin have driven great research ef-
forts using advanced technologies. This has produced complex results
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Food Chemistry 278 (2019) 144–162
that required robust statistical methods in order to extract the relevant
features related with product origin and allow an eventual assessment
of the influence of the product origin on food metabolome (Klockmann
et al., 2016). As previous emphasized, the verification of the true
composition of different foodstuffs is very important to certify their
authenticity and defend their economic value. This is particularly re-
levant for products whose specificity is based on its origin and orga-
noleptic characteristics, as wines, honeys, coffees, cheeses or olive oils.
Usually, this certification is achieved through the identification of
biomarkers of different botanical species used in the production of a
given product. Honeys, for instance, were subject of extensive studies to
certify their authenticity and detect frauds and adulterated products.
Overall, honeys can be discriminated by the type of flowers (unifloral or
multifloral) that different species of bees pollinate in different geo-
graphies, resulting in almost unlimited number of honeys with different
characteristics and flavours (Beretta, Fermo, & Maffei Facino, 2012;
Boffo, Tavares, Tobias, Ferreira, & Ferreira, 2012; Spanik, Pazitna,
Siska, & Szolcsanyi, 2014). Originally, honeys were certificated by its
botanical origin based in the microscopic analysis and quantification of
the pollens used in their production (melissopalynological analysis).
Meanwhile, alternative methodologies were explored to certify the
authenticity of honeys and a huge number of studies can be found in the
literature (Pattamayutanon, et al., 2017). Here we will present selected
examples of different methodologies (HPLC, GC, NMR, NIRS) and
markers reported to differentiate specific honeys, as orange honeys
(Tette, et al., 2017), manuka honey (Lin, Loomes, Prijic, Schlothauer, &
Stephens, 2017), chestnut honeys (Beretta, et al., 2012) or even honeys
obtained with avocado nectar (Dvash, et al., 2002), as shown in Table 3.
In addition, propolis, a beehive sealing material and one of the most
heterogeneous natural cocktail built from plant sources, has been also
subjected to chemometric analysis and several studies successfully as-
signed propolis type to the corresponding plant source (Betrams,
Muller, Kunz, Kammerer, & Stinzing, 2013; Falcao et al., 2013). On the
other hand, coffee constitutes another commodity of great economic
value and so it has been extensively studied, particularly its different
aroma and flavours. Table 3 presents potential biomarkers for coffee
discrimination by geographic origin using chemometric analysis of the
volatile composition. In the same sense, wine is a matrix that has been
extensively analysed regarding the contribution of different factors to
its typicity, namely grape varieties and winemaking processes used.
Wine composition in bioactive compounds, like phenolic compounds,
constitute a reliable signature to classify wines according to their bo-
tanical and geographical origin, as exemplified in Table 3 for Chinese
red wines (Sun, et al., 2015), Czech white wines (Lampir & Pavlousek,
2013) and Argentinean wines (Pisano, Silva, & Olivieri, 2015). The
wine aroma, usually assessed using GC–MS, has a high potential for
wine discrimination, particularly in what concerns to complex wines,
allowing a deeper characterization of their aging and quality. In Table 3
are presented volatile markers that were reported to discriminate dif-
ferent aspects of Madeira wine, one of the oldest fortified wines known,
according to the cultivars used in its production (Câmara, Alves, &
Marques, 2007; Perestrelo, Silva, & Câmara, 2014). In general, it can be
easily observed that the botanicals used in food production, like food
supplements, are the main source of markers for their certification and
traceability. In this context, several classes of compounds have been
explored as botanical markers, particularly phenolics, flavonoids, or-
ganic acids, terpenes, sugars, and amino acids. Phenolic compounds, for
instance, are secondary metabolites arising from shikimate/phenyl-
propanoid pathway or ‘polyketide’ acetate/malonate pathway, or both,
and are produced by plants, often in response to abiotic and abiotic
adverse conditions (García-Parrilla, Cerezo, Tesfaye, & Troncoso,
2011). Furthermore, several health protective properties beyond their
antioxidant effect have been associated to phenolics and hundreds of
such compounds have been reported so far in different plants. This
constitutes a reliable fingerprint to discriminate different commodities
according to their geographic and botanic origin, including cocoa
S. Medina et al.
(Pedan et al., 2017), wines (Lampir & Pavlousek, 2013; Pisano, et al.,
2015), and fruits, as apples or oranges (Diaz, Pozo, Sancho, &
Hernandez, 2014; Gan, Soukoulis, & Fisk, 2014). On the other side,
terpenoids (or isoprenoids), arising from mevalonic acid (MVA, or
isoprenoid) pathway and 1-deoxy-D-xylulose 5-phosphate (DXP, or non-
mevalonate) pathway, constitute the largest class of known plant me-
tabolites, with over 40,000 structures identified. In this respect,
monoterpenoids, sesquiterpenoids diterpenoids, triterpenoids, and
carotenoids are among the most relevant terpenoids, and they have
important applications as pharmaceuticals, flavours, or fragrances.
Most of these terpenoids are volatile, conferring unique flavours and
fragrances and they may be used as biomarkers to authenticate the
origin of foodstuffs in which they are present, such as cheeses (Tompa,
Susic, Rogelj, & Pompe, 2013), honeys (Pattamayutanon, et al., 2017;
Schievano, Morelato, Facchin, & Mammi, 2013; Spanik, et al., 2014;
Stanimirova, et al., 2010), fruits (Ferreira, Perestrelo, & Camara, 2009)
or wines (Perestrelo, et al., 2014). Several volatile carbonyl compounds
are often reported as markers of different plant species and foods in
terms of geographical origin. This is the case of green leaf volatiles
(GLVs), consisting of C6 and C9 aldehydes, alcohols and their esters, as
alkyl esters, carbonyl compounds (e.g., hexanal, trans-2-hexenal) and
alcohols (e.g., 1-hexanol, 1-butanol, cis-3-hexenol) reported by Gan,
et al. (2014) to discriminate apple juices from different geographic
origins (New Zealand, South Africa and Chile; see Table 3). GLVs are
obtained from the oxylipins pathway, through the peroxidation of
PUFA and they are involved in plant defence, plant–plant interactions
and plant–insect interactions. Beyond the previous example, different
GLVs have been pointed as markers of botanical or geographical origin
in olive oils (Benincasa, et al., 2003; Cavaliere, et al., 2007) and in a
variety of fruits and vegetables, as apples and tomatoes (Ferreira,
Perestrelo, Caldeira, & Camara, 2009; Feudo, Macchione, Naccarato,
Sindona, & Tagarelli, 2011). Sugars also present huge variations among
different botanical and geographical origin, making them also excellent
biomarkers. In higher plants, sugar metabolism is essentially driven by
different invertases that hydrolyse sucrose into glucose and fructose. In
turn, these are the key molecules that plants use as nutrients, energy
sources, and signalling for growth, yield formation, and cope with stress
(Wan, Wu, Yang, Zhou, & Ruan, 2018). Broadly, plant invertases can be
classified in two types; cytoplasmic invertases (CINs) and acid in-
vertases, which in turn can be associated to the cell wall (CWIN) or exist
in a vacuolar soluble form (VIN) (Wan, et al., 2018). Thus, CINs pro-
duce sugars that are important to maintain cytosolic sugar homeostasis
for cellular functions and are more conserved in phylogenetic terms.
However, CWINs and VIN, are involved in plant response and adapta-
tion to biotic and abiotic stresses, being able to process oligosaccharides
containing both sucrose and fructose, generating a myriad of different
sugars that can be used as plant biomarkers. This is the case of complex
sugar derivatives as perseitol (a sugar alcohol) detected in honey and it
may be used to certify that honeybees have harvested avocado nectar
(Dvash, et al., 2002). Also, citrusin D, a fatty acyl glycoside of mono-
and disaccharides, was tentatively identified as optimal marker to dif-
ferentiate Valencian (Spain), Argentinean, Brazilian and South African
oranges (Diaz, et al., 2014). Furthermore, amino acids not only are the
backbone of proteins, but also of other molecules that plant use for
different functions. Many of these molecules constitute botanical mar-
kers, while other metabolites accumulate in response to certain abiotic
factors and they could be considered geographical markers. For in-
stance, synephrine, an aromatic amine formed in citrus by a pathway
involving tyramine and N-methyltyramine, which in turn derive from
the amino acid tyrosine, has been recently reported as a potential
biomarker for orange honey authenticity (Tette, et al., 2017). Likewise,
kynurenic acid, arising from the L-tryptophan metabolism, was reported
as a marker of chestnut honey and was found in honey varieties of
arboreal origin, but it was absent in honeys obtained from herbal
flowers. This distribution may be explained as a plant defence me-
chanism against fungal pathogens and herbivorous parasites, ever-
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Food Chemistry 278 (2019) 144–162
present on wild trees (Beretta, et al., 2012).
2.4. Food adulteration
Nowadays, food adulteration represents a major concern because
foods are often contaminated with additives to increase bulk, disguise
spoilage and enhance the benefits. This challenge requires a continuous
effort to improve the analytical power to uncover sophisticated pro-
ducts adulterations, particularly when food safety, rather than food
quality, is a real issue. In this context, olive oil, due to its commercial
value, is the product more often adulterated with other cheaper oils
from different species. Hazelnut oil is the oil most often used to adul-
terate olive oil inasmuch as it has a similar composition of triacylgly-
cerols, fatty acids, and sterols (Azadmard-Damirchi, 2010). This fact
makes it very difficult to detect olive oil adulterated with hazelnut oil,
but lupeol and filbertone, which are specific to hazelnut oil, could as-
sess the fraud down to an adulteration level of 2% and 10%, respec-
tively. Lupeol is a pentacyclic triterpene with promising anticancer
activity and filbertone, (2E)-5-methyl-2-hepten-4-one, also known as
hazelnut ketone, is an unsaturated ketone often described as a highly
potent plant flavour molecule (Azadmard-Damirchi, 2010) (Table 4).
The adulteration of olive oil with cheaper olive oils from other regions
and quality is also very frequent. However, a chemometric analysis is
reliable enough to obtain markers for the detection of these adultera-
tions, as is shown in Table 3 with Italian olive oils (Benincasa, et al.,
2003; Cavaliere, et al., 2007). Milk is the second product more often
adulterated. This is usually achieved with the addition of vegetable
proteins and fats (Kim, Kim, & Park, 2015), milk from different species,
and an addition of whey and watering are more and less harmless to
human health. However, there are milk adulterations which are not
safe, causing adverse health effects, as urea (Abernethy & Higgs, 2013),
formalin, detergents, ammonium sulfate, boric acid, caustic soda,
benzoic acid, salicylic acid, hydrogen peroxide, sugars and melamine.
Nevertheless, these adulterations, unlike the less harmful to human
health referred above, are easily detected. Overall, lipids diversity
constitutes a good marker of adulteration of different foodstuffs,
namely milk and derivatives. The lipid fraction of milk, for instance,
contains mono-, di- and triglycerides, glycolipids, cholesterol and re-
spective esters derivatives, free fatty acids, phospholipids and sphin-
golipids, and the presence, diversity and amount of each of these group
of lipids forms a specific lipid profile that allows an easy detection of
adulteration (Kim, et al., 2015). Many studies are proposing strategies
to certify meat composition, a concern that is related not only with
meat quality but also with meat safety. This is the case, for example of
the adulteration of mince with meat from other species. In this context,
it is noteworthy to refer the integrated approach followed by (Trivedi
et al., 2016) that underwent a metabolite profiling and lipidomics ap-
proach to detect the adulteration of beef with pork meat. By combining
data from GC–MS and UHPLC-MS analyses using chemometric tools,
these authors reported over 20 metabolites from different classes that
allowed the detection of beef meat adulterated with pork meat. Fur-
thermore, pathways involved in this process may be researched to de-
tect significant differences in glutathione, inositol and sphingolipid
metabolism that justify the differences reported between the two types
of meat studied (Table 4). The fraudulent addiction of a juice of lesser
economic value to a more expensive one, known as juice-to-juice de-
basing, is often detected. In this regard, a recent study reported that
arbutin (4-hydroxyphenyl-β-D-glucopyranoside) was a specific marker
for apple juice debasing with pear juice (Thavarajah & Low, 2006).
More recently, another work informed that 4-O-p-coumarylquinic acid,
isorhamnetin-3-O-rutinoside and ABA were able to demonstrate the
debasing procedure (Willems & Low, 2018). Thus, complex sugars as
arbutin and isorhamnetin-3-O-rutinoside can be also reliable markers of
food fraud because they are specifically produced by some species that
often are not the ones used to obtain the original product.
S. Medina et al.
2.5. Spoilage and freshness markers
Food spoilage is a complex process to which many factors, as tem-
perature, moisture, pH level or oxygen pressure, contribute. It has a
crucial importance in food safety as many metabolites produced during
spoilage are toxic to humans (Kuuliala, Al Hage, et al., 2018). There-
fore, the identification of key markers of food spoilage is very relevant.
Likewise, food freshness certainly corresponds to the consumer pre-
ference and simultaneously ensures food safety, at least in what con-
cerns to the toxic effects caused by spoilage. Thus, rapid monitoring of
food deterioration is considered essential, and apart from the tradi-
tional chemical spoilage markers, such as total volatile bases nitrogen
(TVB-N) and trimethylamine nitrogen (TMA-N), research for new
markers able to monitor food spoilage processes is essential. In this
context, Parlapani, Haroutounian, Nychas, and Boziaris, (2015) used
SPME-GC–MS to analyse sea bass storage and reported changes in
specific VOCs, mainly alcohols, aldehydes, and esters, which could be
used as markers of deterioration (Table 5). In the same way, a recent
study studied freshness reduction in oyster (Crassostrea gigas) by the
detection of dimethyl sulfide by GC–MS upon 4 days of harvesting and
trimethylamine upon 2 days of harvesting, in this case using a bioe-
lectronics nose. Hence, these biomarkers have a powerful potential to
monitor seafood spoilage for real-time and on-site analysis, thus con-
tributing to prevent seafood-borne diseases (Lee, et al., 2018). Other
study carried out in salmon samples (Oncorhynchus tshawytscha) iden-
tified VOCs as biomarker candidates to supervise salmon spoilage, as
well as certain alcohols and aldehydes able to reveal salmon freshness
(Table 5) (Wierda, Fletcher, Xu, & Dufour, 2006). Two recent and
promising studies carried out in Atlantic cod (Gadus morhua) were
performed using selected-ion-flow-tube mass spectrometry (SIFT-MS).
This approach eliminated the need of sample preparation procedures
while allowing fast and sensitive analysis of the VOCs profile over
storage time. Fish packaging and storage conditions affected the evo-
lution of the VOCs profile and should be considered in the selection of
spoilage indicators, which included VOCs as 2,3-butanediol; 3-methyl-
1-butanol; ethyl acetate and trimethylamine. Thus, the evolution of
these spoilage markers in conjunction with microbial growth assays and
sensory rejection could be used for the development of intelligent
packaging applications (Kuuliala, Abatih, et al., 2018; Kuuliala, Al
Hage, et al., 2018). Chicken meat is widely appreciated across the globe
and so the identification of volatile markers to certify its degree of
conservation is certainly highly relevant. In this context, Lovestead and
Bruno (2010) developed an headspace analysis using cryo-adsorption
with a porous layer open tubular plot (PLOT) column to separate and
detect the VOCs emitted by the sample by GC–MS. Then, the authors
applied this methodology to identify markers of chicken spoilage in
breast tissue samples. In another recent study performed with chicken
breast, a PCA analysis unveils some details about the spoilage process.
According to the results obtain, breast chicken spoilage seems to occur
in two steps or, alternatively, involve two different pathways in which
the glycolytic and proteolytic reactions occur simultaneously, but at
different rates. This mechanism is supported by the observation of a
first increase in alcohols and free fatty acids levels (primary spoilage
markers), followed by a rise in the sulfide contents (secondary spoilage
markers) caused by the nitrogenous compounds and amino acids cat-
abolism, mainly cysteine and methionine (Table 5). Overall, ethanol
was the predominant VOCs in all spoiled samples, a fact that may result
from the metabolic activity of lactic acid bacteria, as Pseudomonas spp.,
often present in spoilage food. In turn, 3-methyl-1-butanol and acetic
acid are usually generated by activity of Brochothrix thermosphacta and
lactic acid bacteria. Together, these three VOCs could be considered as
helpful indicators to evaluate the deterioration of aerobically stored
chicken meat (Miks-Krajnik, Yoon, Ukuku, & Yuk, 2016). A third study
on chicken breast using HPLC and GC–MS reported amino acids and
VOCs as putative biomarkers of chicken spoilage prior to organoleptic
changes in this foodstuff. A relevant aspect of this study is that certain
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Food Chemistry 278 (2019) 144–162
metabolites, mainly sulfur compounds, were detected just after 4 days
of storage, while microbial populations were still below the accepted
sensory level of spoilage. This fact is very significant and it may in-
crease our knowledge for the early detection of spoilage process
(Alexandrakis, Brunton, Downey, & Scannell, 2012). More recently,
proton-transfer-reaction mass spectrometry (PTR-MS), an on-line tech-
nique that enables the detection of compounds in real-time, has been
also applied to study chicken spoilage, identifying most the volatile
biomarkers previously referred, as acetone, dimethyl sulfide and
ethanol (Franke & Beauchamp, 2017). The combination of advanced
analytical platforms, as GC–MS and 1H NMR, to analyse the same
sample, allows a deeper and comprehensive analysis, but is simulta-
neously very important to identify specific metabolites. In this context,
several markers able to monitor the progress of the spoilage process
were detected in beef meat. Among them, butanoic acid and acetoin (3-
hydroxy-2-butanone) were related to the growth of different microbial
groups. Butanoic acid, commonly associated with meat spoilage, could
be derived from microbial consumption of free amino acids via the
Stickland reaction. In addition, the accumulation of acetoin has been
associated not to spoiled meat, but rather to “not fresh” product. Re-
frigeration is a common process of meat storage, but this procedure
causes the dehydration of the meat surface and stimulates the pro-
duction of betaine, another relevant marker to monitor the deteriora-
tion process in beef that was identified in the study (Ercolini, et al.,
2011). Still related with meat products, a work carried out in Iberian
ham identified many VOCs correlated with the deterioration of this
product, particularly pyrazines, ethers and carbonyls (Table 5) (Martín
et al., 2010). Furthermore, it has been also reported that fungi infec-
tions, particularly the ones caused by Botrytis cinerea and different
species of Penicillium and Aspergillus destroy food flavours, affecting
very negatively food quality and safety. Regarding this, ergosterol was
recently reported as a specific marker for the evaluation of fungi in-
fection and phytosanitary status of grapes (Porep, Walter, Kortekamp, &
Carle, 2014). Another study was carried out in different food matrices
(bread, chicken egg and cucumber), using an untargeted metabolic
profiling approach based on two-dimensional GC-TOF-MS, allowed the
differential identification of 13 metabolites, including amino acids,
biogenic amines, and organic acids. The major metabolic changes in the
spoilage process of the target foods were closely related to some me-
tabolic pathways, as amino acids metabolism, Krebs cycle and ammonia
recycling (Cheng et al., 2015). One of the most recent studies about
food freshness performed in three different citrus varieties (C. clem-
entina, C. tangerina and C. reticulata), reported that the Raman signal
due to the carotenoids from fruit peel was a good indicator of citrus
freshness, therefore introducing an objective criterion to assess citrus
freshness. These results could have a strong impact in food industries
and consumer satisfaction (Nekvapil, et al., 2018).
3. Factors affecting the robustness of food fingerprints
There are many factors affecting the analysis of food fingerprints
and the robustness of the models obtained to certify food authenticity.
One of the main bottlenecks is the critical lack of standardization in the
experimental layouts used. The sample size we find with this review, for
instance, presents a great heterogeneity, ranging from five to 374
samples (Tables 1–5). This fact certainly represents a major problem in
the comparison of different studies for authentication purposes. Al-
though an optimal samples number may be difficult to establish, this
number should be very large (Novotná, et al., 2012), preferentially
higher than the number of variables identified so the statistical models
can produce reliable classification models. Regarding the quality of the
samples sets, the sampling period is also an important factor. In fact, in
previous studies, year-to-year variations in the samples set have been
reported for several food matrices, as wheat, potato, honey or orange
(Diaz, et al., 2014; Shepherd, et al., 2014; Stanimirova, et al., 2010;
Weesepoel, et al., 2016). Overall, most studies have investigated the
S. Medina et al.
differences on the chemical composition among two years of produc-
tion, with the exception of two works, involving carrot samples
(Cubero-Leon, et al., 2018) and wines samples (Perestrelo, et al., 2014),
where the differences among four years of production were in-
vestigated. Nonetheless, the seasonal variability can also produce dif-
ferences in the fingerprints in the same year. Souard, et al. (2018), for
instance, reported that most of the coffee samples harvested in No-
vember had lower intensities for most of the metabolites than the
samples harvested in other periods. In other studies, however, there
were not described differences among wine samples of different years of
production, suggesting that, at least in these cases, the influence of the
genetic background may be higher than the environmental conditions
fluctuations (Câmara, et al., 2007; Righetti, et al., 2018). Considering
the high variability of the results reported, more efforts are needed to
understand the conjugated effects of genetics and seasonal climate
variations on the chemical composition of food (Longobardi, et al.,
2017). Still related with this heterogeneity in the results, the influence
of other aspects as natural contaminations should be considered. For
example, in honeys from diverse botanical origins, overlapping flow-
ering periods of different plants may occur, resulting in a cross con-
tamination of the samples (Schievano, et al., 2013). Besides, storage
conditions before analysis must be standardized because samples are
often stored frozen, refrigerated, kept at room temperature or even on
ice, and these variations may modulate metabolite levels.
In this review, hundreds of metabolites presented in Tables 1–5 are
described as putative biomarkers and many of them have been char-
acterized according to specific authentication issues. In addition,
identical molecules have been described among different studies in the
same matrix. This is the case of β-damascenone that was found as
discriminant metabolite in wine samples of different cultivars in two
studies (Câmara, et al., 2007; Culleré, et al., 2008). Also, three VOCs, 1-
hexanol, 2-methylbutyl acetate, and hexyl acetate, were reported in
different works as molecules able to distinguish different varieties/
cultivars of apple juices, with an assignment success rate of 100% in
classification and prediction test (with internal and external validation
set for both studies - inter-laboratory reproducibility) (Gan, et al., 2014;
Guo, et al., 2012). In contrast, the amino acids alanine and γ-amino-
butyric acid have been described in three reports as discriminant
markers of different rice cultivars/varieties (Danial, et al., 2013; Frank,
et al., 2012; Hu, et al., 2014). The sample preparation, extraction, and
storage (−18, −20 and −80 °C) were quite different among these
three works, so the detection of these specific amino acids was largely
independent of experimental design. Regarding the farming systems,
three metabolites, alanine, myo-inositol, and urea, were described as
significant metabolites for the discrimination between O/C wheat
samples in two different studies using GC–MS (Bonte, et al., 2014; Zörb,
et al., 2006). Similarly, lactobacillic acid was established as a molecular
marker for cheese authentication by two different studies using GC–MS
and confirmed by NMR (Caligiani, et al., 2016; Marseglia, et al., 2013).
Another two works allowed wine differentiation according to geo-
graphic origin from Czech Republic (Lampir & Pavlousek, 2013) and
China regions (Sun, et al., 2015) with identical molecules as dis-
criminant markers ((+)-catechin, (−)-epicatechin, and protocatechuic
acid). Although, more evidences are necessary, these metabolites are
strong candidates to authenticate PDO wines.
Related to food adulteration issue, arbutin is a molecular marker
able to detect juice-to-juice debasing between apple and pear juices
described in two articles (Thavarajah & Low, 2006; Willems & Low,
2018). Concerning to spoilage biomarkers, several reports have de-
tected dimethyl sulfide and trimethylamine in different foods, as oyster,
cod and chicken (only dimethyl sulfide) (Kuuliala, Abatih, et al., 2018;
Lee, et al., 2018; Lovestead & Bruno, 2010). Particularly for chicken
samples, it is very important to note that dimethyl disulfide, dimethyl
trisulfide, acetone and dimethyl sulfide ethanol have been reported in
several works (Alexandrakis, et al., 2012; Franke & Beauchamp, 2017;
Lovestead & Bruno, 2010; Miks-Krajnik, et al., 2016). Moreover,
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Food Chemistry 278 (2019) 144–162
acetoin, lactate and threonine have also been identified as spoilage
markers in both chicken and beef meat (Alexandrakis, et al., 2012;
Ercolini, et al., 2011; Franke & Beauchamp, 2017). These metabolites
may be very useful as spoilage markers because they have been de-
tected in several foodstuffs, but they should be quantitatively evaluated
to know their sensory contribution to the “gone-off” odour
(Alexandrakis, et al., 2012). Furthermore, several metabolites have
been detected in the same food matrix using different technologies. For
instance, caffeine, caffeoylquinic acid and sucrose have been reported
as significant metabolites to differentiate coffee varieties by 13C NMR
(Wei, et al., 2012) and MS (FT-ICR-MS and LC-q-TOF-MS) (Garrett,
et al., 2013; Souard, et al., 2018). Another example is the detection of
cinnamaldehyde in cinnamon by both DART-q-TOF-MS and NMR
technologies (Avula, et al., 2015; Farag, et al., 2018). These technolo-
gies combined with chemometrics tools have a powerful potential for
food authentication with traceability and certification purposes. How-
ever, the selection and quality of statistical classification models are
essential for the robustness of the biomarkers identification and dis-
crimination. In this context, the implementation of model validation
through, for example, the leave-one-out cross validation (LOOCV, is a
particular case of leave-p-out cross-validation with p = 1) is required
not only to estimate the robustness of statistical analysis, but also to
prevent overfitting (Farag, et al., 2018; Gan, et al., 2014; Garrett, et al.,
2013). Furthermore, the models should allow a reliable sample dis-
crimination with a correct classification (∼100%) upon external vali-
dation (Gan, et al., 2014). It is noteworthy that the choice of statistical
models may influence the outcomes obtained. In this context, in a re-
cent work carried out with two varieties of carrot samples during four
production years, PCA was not able to discriminate between O/C car-
rots, but it was a successful model for the classification based on the
production year. OPLS-DA, in turn, was able to distinguish among
farming systems (Cubero-Leon, et al., 2018). In another study carried
out in grape juices (biodynamic, conventional and organic) from Brazil
and Europe, PLS-DA allowed the classification between Brazilian and
European grape juices (intraregional models with a 92% of efficiency),
kNN algorithm differentiated between European conventional and or-
ganic/biodynamic samples and SIMCA model enabled the discrimina-
tion among juices under different farming systems. However, neither
ANOVA nor HCA had significant/clear statistical results (Granato, et al.,
2015). More recently, the same author, in a detailed review about
chemometrics methods applied to food authentication, declared that
HCA is a highly arbitrary model and it should not be used with food
authenticity purposes. It is therefore a very difficult task to select the
“best” statistical model because of all of them present negative and
positive characteristics (Granato et al., 2018). This is another major gap
in food authentication protocols that, together with the lack of super-
vision from government‘s authorities and the weak communications
between consumers and producers, is making food authenticity certi-
fication a demanding and very challenging process to food handlers and
food standardization authorities.
4. Conclusions and future perspectives
Biochemical changes in foodstuffs caused by environmental, genetic
diversity or human activity result in specific variations that can be used
as markers of those products. In this context, compounds as amino
acids, phenolic compounds, organic acids and VOCs of different che-
mical classes, are the potential biomarkers for food authentication most
often reported. As these compounds possess different physicochemical
properties, several analytical platforms must be employed to obtain
reliable methodologies to detect and quantify biomarkers able to certify
foodstuffs origin, variety or production system, as well as to detect food
adulteration or spoilage/freshness indicators. Regarding this, strategies
envisaging the characterization of the volatile and non-volatile frac-
tions of food products samples, followed by robust data fusion and
processing algorithms involving advanced statistical analysis are
S. Medina et al.
becoming crucial to cope with the complexity of foodstuffs authenti-
cation. In this review, several promising biomarkers for food authen-
ticity have been pointed, but most of them still require deeper and more
extensive validation procedures. To accelerate this process, we antici-
pate that more technologies facilitating routine analysis will become
available for rapid monitoring of food authentication. In turn, in-
creasingly sophisticated untargeted methods will improve the ability to
detect the minor differences between genuine and fraudulent foods and
beverages.
Moreover, better analytical performances will allow not only con-
firming the existence of a given biomarker, but also accurately quan-
tifying it, a feature that will be very relevant in the definition of spoi-
lage markers. Additionally, PTR, SIFT and NMR outputs will certainly
result in future non-destructive analytical techniques for on-site and
real-time food authentication, to which “user-friendly” chemometrics
approaches and authentication biomarkers databases need to be gen-
erated. Future generations of consumers will certainly be even more
informed, demanding for food integrity (food traceability, safety,
quality, and authenticity), assuring that food should be not only heal-
thier and safer, but also be obtained through sustainable procedures
that respect animal welfare and are environment-friendly.
Acknowledgments
This work was supported by FCT-Fundação para a Ciência e a
Tecnologia (project PEst-OE/QUI/UI0674/2013, CQM, Portuguese
Government funds) and by ARDITI-Agência Regional para o
Desenvolvimento da Investigação Tecnologia e Inovação through the
project M1420-01-0145-FEDER-000005 – Centro de Química da
Madeira – CQM+ (Madeira 14-20). SM was supported by the Post-
Doctoral fellowship granted by ARDITI – CQM+ project (ARDITI-
CQM/2017/008-PDG). JAMP and PS were supported by project
M1420- 09-5369-FSE-000001 through the Post-Doctoral and PhD fel-
lowships, respectively, while RP was supported by FCT Post-Doctoral
grant (SFRH/BPD/97387/2013).
Conflicts of interest
The authors declare no conflict of interest.
References
Abernethy, G., & Higgs, K. (2013). Lactose semicarbazone as a marker for semicarbazide
adulteration in milk. Journal of Chromatography A, 1295, 152–155.
Akiyama, M., Murakami, K., Hirano, Y., Ikeda, M., Iwatsuki, K., Wada, A., ... Iwabuchi, H.
(2008). Characterization of headspace aroma compounds of freshly brewed arabica
coffees and studies on a characteristic aroma compound of Ethiopian coffee. Journal
of Food Science, 73(5), C335–346.
Alexandrakis, D., Brunton, N. P., Downey, G., & Scannell, A. G. (2012). Identification of
spoilage marker metabolites in Irish chicken breast muscle using HPLC, GC–MS
coupled with SPME and traditional chemical techniques. Food and Bioprocess
Technology, 5(5), 1917–1923.
Ali, K., Maltese, F., Zyprian, E., Rex, M., Choi, Y. H., & Verpoorte, R. (2009). NMR
Metabolic Fingerprinting Based Identification of Grapevine Metabolites Associated
with Downy Mildew Resistance. Journal of Agricultural and Food Chemistry, 57(20),
9599–9606.
Arbona, V., Iglesias, D. J., & Gómez-Cadenas, A. (2015). Non-targeted metabolite pro-
filing of citrus juices as a tool for variety discrimination and metabolite flow analysis.
BMC Plant Biology, 15(1), 38.
Avula, B., Smillie, T. J., Wang, Y.-H., Zweigenbaum, J., & Khan, I. A. (2015).
Authentication of true cinnamon (Cinnamon verum) utilising direct analysis in real
time (DART)-QToF-MS. Food Additives & Contaminants: Part A, 32(1), 1–8.
Azadmard-Damirchi, S. (2010). Review of the use of phytosterols as a detection tool for
adulteration of olive oil with hazelnut oil. Food Additives & Contaminants: Part A,
27(1), 1–10.
Beitlich, N., Koelling-Speer, I., Oelschlaegel, S., & Speer, K. (2014). Differentiation of
manuka honey from kanuka honey and from jelly bush honey using HS-SPME-GC/MS
and UHPLC-PDA-MS/MS. Journal of Agricultural and Food Chemistry, 62(27),
6435–6444.
Benincasa, C., De Nino, A., Lombardo, N., Perri, E., Sindona, G., & Tagarelli, A. (2003).
Assay of aroma active components of virgin olive oils from southern Italian regions
by SPME-GC/ion trap mass spectrometry. Journal of Agricultural and Food Chemistry,
51(3), 733–741.
160
Food Chemistry 278 (2019) 144–162
Beretta, G., Fermo, P., & Maffei Facino, R. (2012). Simple and rapid simultaneous pro-
filing of minor components of honey by size exclusion chromatography (SEC) coupled
to ultraviolet diode array detection (UV-DAD), combined with chemometric methods.
Journal of Pharmaceutical and Biomedical Analysis, 58, 193–199.
Betrams, J., Muller, M., Kunz, N., Kammerer, D., & Stinzing, F. (2013). Phenolic com-
pounds as marker compounds for botanical origin determination of German propolis
samples based on TLC and TLC-MS. Journal of Applied Botany and Food Quality, 86,
143–153.
Boffo, E. F., Tavares, L. A., Tobias, A. C. T., Ferreira, M. M. C., & Ferreira, A. G. (2012).
Identification of components of Brazilian honey by 1H NMR and classification of its
botanical origin by chemometric methods. LWT – Food Science and Technology, 49(1),
55–63.
Bonte, A., Neuweger, H., Goesmann, A., Thonar, C., Mäder, P., Langenkämper, G., &
Niehaus, K. (2014). Metabolite profiling on wheat grain to enable a distinction of
samples from organic and conventional farming systems. Journal of the Science of Food
and Agriculture, 94(13), 2605–2612.
Caligiani, A., Nocetti, M., Lolli, V., Marseglia, A., & Palla, G. (2016). Development of a
quantitative GC-MS method for the detection of cyclopropane fatty acids in cheese as
new molecular markers for parmigiano Reggiano authentication. Journal of
Agricultural and Food Chemistry, 64(20), 4158–4164.
Câmara, J. S., Alves, M. A., & Marques, J. C. (2007). Classification of Boal, Malvazia,
Sercial and Verdelho wines based on terpenoid patterns. Food Chemistry, 101(2),
475–484.
Caprioli, G., Logrippo, S., Cahill, M. G., & James, K. J. (2014). High-performance liquid
chromatography LTQ-Orbitrap mass spectrometry method for tomatidine and non-
target metabolites quantification in organic and normal tomatoes. International
Journal of Food Sciences and Nutrition, 65(8), 942–947.
Cavaliere, B., De Nino, A., Hayet, F., Lazez, A., Macchione, B., Moncef, C., ... Tagarelli, A.
(2007). A metabolomic approach to the evaluation of the origin of extra virgin olive
oil: A convenient statistical treatment of mass spectrometric analytical data. Journal
of Agricultural and Food Chemistry, 55(4), 1454–1462.
Cheng, J., Gao, R., Li, H., Wu, S., Fang, J., Ma, K., ... Dong, F. (2015). Evaluating potential
markers of spoilage foods using a metabolic profiling approach. Food Analytical
Methods, 8(5), 1141–1149.
Cubero-Leon, E., De Rudder, O., & Maquet, A. (2018). Metabolomics for organic food
authentication: Results from a long-term field study in carrots. Food Chemistry, 239,
760–770.
Cuevas, F. J., Pereira-Caro, G., Moreno-Rojas, J. M., Muñoz-Redondo, J. M., & Ruiz-
Moreno, M. J. (2017). Assessment of premium organic orange juices authenticity
using HPLC-HR-MS and HS-SPME-GC-MS combining data fusion and chemometrics.
Food Control, 82, 203–211.
Culleré, L., Escudero, A., Pérez-Trujillo, J. P., Cacho, J., & Ferreira, V. (2008). 2-Methyl-3-
(methyldithio)furan: A new odorant identified in different monovarietal red wines
from the Canary Islands and aromatic profile of these wines. Journal of Food
Composition and Analysis, 21(8), 708–715.
Culleré, L., Ferreira, V., Venturini, M. E., Marco, P., & Blanco, D. (2013). Potential aro-
matic compounds as markers to differentiate between Tuber melanosporum and Tuber
indicum truffles. Food Chemistry, 141(1), 105–110.
Dallago, R. M., Valduga, A. T., Luccio, M. D., Benin, S., & Tres, M. V. (2011). Analysis of
volatile compounds of Ilex paraguariensis A. St. - Hil. and its main adulterating species
Ilex theizans Mart. ex Reissek and Ilex dumosa Reissek. Ciência e Agrotecnologia, 35,
1166–1171.
Danial, A. D., Neoh, B. K., Ersad, M. A., Ruzlan, N., Win, W., Musa, N. A., ... Yusof, H. M.
(2013). Differentiation of rice varieties using metabolite markers. Journal of
Agricultural Technology, 9(3), 601–610.
Di Donna, L., Mazzotti, F., Naccarato, A., Salerno, R., Tagarelli, A., Taverna, D., &
Sindona, G. (2010). Secondary metabolites of Olea europaea leaves as markers for the
discrimination of cultivars and cultivation zones by multivariate analysis. Food
Chemistry, 121(2), 492–496.
Diaz, R., Pozo, O. J., Sancho, J. V., & Hernandez, F. (2014). Metabolomic approaches for
orange origin discrimination by ultra-high performance liquid chromatography
coupled to quadrupole time-of-flight mass spectrometry. Food Chemistry, 157, 84–93.
do Carmo Carvalho, D., Brigagão, M. R. P. L., dos Santos, M. H., de Paula, F. B. A., Giusti-
Paiva, A., & Azevedo, L. (2011). Organic and conventional Coffea arabica L.: A
comparative study of the chemical composition and physiological, biochemical and
toxicological effects in wistar rats. Plant Foods for Human Nutrition, 66(2), 114–121.
Dong, T., Chen, X. J., Wang, M., Huang, Y. H., & Yi, G. J. (2014). Comparison of volatile
aroma compounds in Dwarf Cavendish banana (Musa spp. AAA) grown under organic
or traditional cultivation. The Journal of Horticultural Science and Biotechnology, 89(4),
441–447.
Dvash, L., Afik, O., Shafir, S., Schaffer, A., Yeselson, Y., Dag, A., & Landau, S. (2002).
Determination by near-infrared spectroscopy of perseitol used as a marker for the
botanical origin of avocado (Persea americana Mill.) honey. Journal of Agricultural and
Food Chemistry, 50(19), 5283–5287.
Eisenmann, P., Ehlers, M., Weinert, C., Tzvetkova, P., Silber, M., Rist, M., ... Muhle-Goll,
C. (2016). Untargeted NMR spectroscopic analysis of the metabolic variety of new
apple cultivars. Metabolites, 6(3), 29.
Elmore, J. S., Nisyrios, I., & Mottram, D. S. (2005). Analysis of the headspace aroma
compounds of walnuts (Juglans regia L.). Flavour and Fragrance Journal, 20(5),
501–506.
Ercolini, D., Ferrocino, I., Nasi, A., Ndagijimana, M., Vernocchi, P., La Storia, A., ...
Villani, F. (2011). Monitoring of microbial metabolites and bacterial diversity in beef
stored under different packaging conditions. Applied and Environmental Microbiology,
77(20), 7372–7381.
Falcao, S. I., Vale, N., Gomes, P., Domingues, M. R., Freire, C., Cardoso, S. M., & Vilas-
Boas, M. (2013). Phenolic profiling of Portuguese propolis by LC-MS spectrometry:
S. Medina et al.
Uncommon propolis rich in flavonoid glycosides. Phytochemical Analysis, 24(4),
309–318.
Farag, M. A., Handoussa, H., Fekry, M. I., & Wessjohann, L. A. (2016). Metabolite pro-
filing in 18 Saudi date palm fruit cultivars and their antioxidant potential via UPLC-
qTOF-MS and multivariate data analyses. Food & Function, 7(2), 1077–1086.
Farag, M. A., Labib, R. M., Noleto, C., Porzel, A., & Wessjohann, L. A. (2018). NMR ap-
proach for the authentication of 10 cinnamon spice accessions analyzed via chemo-
metric tools. LWT – Food Science and Technology, 90, 491–498.
Farag, M. A., Mohsen, M., Heinke, R., & Wessjohann, L. A. (2014). Metabolomic finger-
prints of 21 date palm fruit varieties from Egypt using UPLC/PDA/ESI–qTOF-MS and
GC–MS analyzed by chemometrics. Food Research International, 64, 218–226.
Favaro, G., Magno, F., Boaretto, A., Bailoni, L., & Mantovani, R. (2005). Traceability of
Asiago mountain cheese: A rapid, low-cost analytical procedure for its identification
based on solid-phase microextraction. Journal of Dairy Science, 88(10), 3426–3434.
Ferreira, L., Perestrelo, R., Caldeira, M., & Camara, J. S. (2009). Characterization of vo-
latile substances in apples from Rosaceae family by headspace solid-phase micro-
extraction followed by GC-qMS. Journal of Separation Science, 32(11), 1875–1888.
Ferreira, L., Perestrelo, R., & Camara, J. (2009). Comparative analysis of the volatile
fraction from Annona cherimola Mill. cultivars by solid-phase microextraction and gas
chromatography–quadrupole mass spectrometry detection. Talanta, 77(3),
1087–1096.
Feudo, G. L., Macchione, B., Naccarato, A., Sindona, G., & Tagarelli, A. (2011). The vo-
latile fraction profiling of fresh tomatoes and triple concentrate tomato pastes as
parameter for the determination of geographical origin. Food Research International,
44(3), 781–788.
Figueira, J., Camara, H., Pereira, J., & Camara, J. S. (2014). Evaluation of volatile me-
tabolites as markers in Lycopersicon esculentum L. cultivars discrimination by multi-
variate analysis of headspace solid phase microextraction and mass spectrometry
data. Food Chemistry, 145, 653–663.
Frank, T., Reichardt, B., Shu, Q., & Engel, K.-H. (2012). Metabolite profiling of colored
rice (Oryza sativa L.) grains. Journal of Cereal Science, 55(2), 112–119.
Franke, C., & Beauchamp, J. (2017). Real-time detection of volatiles released during meat
spoilage: A case study of modified atmosphere-packaged chicken breast fillets in-
oculated with Br. thermosphacta. Food Analytical Methods, 10(2), 310–319.
Fuentes, A., Fernández-Segovia, I., Serra, J. A., & Barat, J. M. (2010). Comparison of wild
and cultured sea bass (Dicentrarchus labrax) quality. Food Chemistry, 119(4),
1514–1518.
Gan, H. H., Soukoulis, C., & Fisk, I. (2014). Atmospheric pressure chemical ionisation
mass spectrometry analysis linked with chemometrics for food classification - a case
study: Geographical provenance and cultivar classification of monovarietal clarified
apple juices. Food Chemistry, 146, 149–156.
García-Parrilla, M. C., Cerezo, A. B., Tesfaye, W., & Troncoso, A. M. (2011). Phenolic
compounds as markers for the authentication of sherry vinegars: A foresight for high
quality vinegars characterization. ACS Symposium Series, 1081, 201–213.
Garrett, R., Schmidt, E. M., Pereira, L. F. P., Kitzberger, C. S., Scholz, M. B. S., Eberlin, M.
N., & Rezende, C. M. (2013). Discrimination of arabica coffee cultivars by electro-
spray ionization Fourier transform ion cyclotron resonance mass spectrometry and
chemometrics. LWT – Food Science and Technology, 50(2), 496–502.
Granato, D., Koot, A., Schnitzler, E., & van Ruth, S. M. (2015). Authentication of geo-
graphical origin and crop system of grape juices by phenolic compounds and anti-
oxidant activity using chemometrics. Journal of Food Science, 80(3), C584–C593.
Granato, D., Putnik, P., Kovačević, D. B., Santos, J. S., Calado, V., Rocha, R. S., ...
Pomerantsev, A. (2018). Trends in chemometrics: Food authentication, microbiology,
and effects of processing. Comprehensive Reviews in Food Science and Food Safety,
17(3), 663–677.
Guo, J., Yue, T., & Yuan, Y. (2012). Feature selection and recognition from nonspecific
volatile profiles for discrimination of apple juices according to variety and geo-
graphical origin. Journal of Food Science, 77(10), C1090–C1096.
Hallmann, E., & Rembiałkowska, E. (2012). Characterisation of antioxidant compounds in
sweet bell pepper (Capsicum annuum L.) under organic and conventional growing
systems. Journal of the Science of Food and Agriculture, 92(12), 2409–2415.
Hu, C., Shi, J., Quan, S., Cui, B., Kleessen, S., Nikoloski, Z., ... Lin, H. (2014). Metabolic
variation between japonica and indica rice cultivars as revealed by non-targeted
metabolomics. Scientific Reports, 4, 5067.
Huang, S., Zhang, C. P., Li, G. Q., Sun, Y. Y., Wang, K., & Hu, F. L. (2014). Identification of
catechol as a new marker for detecting propolis adulteration. Molecules, 19(7),
10208–10217.
Jumhawan, U., Putri, S. P., Yusianto, Marwani, E., Bamba, T., & Fukusaki, E. (2013).
Selection of discriminant markers for authentication of Asian palm civet coffee (Kopi
Luwak): A metabolomics approach. Journal of Agricultural and Food Chemistry, 61(33),
7994–8001.
Khalil, M. N., Fekry, M. I., & Farag, M. A. (2017). Metabolome based volatiles profiling in
13 date palm fruit varieties from Egypt via SPME GC–MS and chemometrics. Food
Chemistry, 217, 171–181.
Kim, J., Jung, Y., Song, B., Bong, Y. S., Ryu, D. H., Lee, K. S., & Hwang, G. S. (2013).
Discrimination of cabbage (Brassica rapa ssp. pekinensis) cultivars grown in different
geographical areas using (1)H NMR-based metabolomics. Food Chemistry, 137(1–4),
68–75.
Kim, J. M., Kim, H. J., & Park, J. M. (2015). Determination of milk fat adulteration with
vegetable oils and animal fats by gas chromatographic analysis. Journal of Food
Science, 80(9), C1945–1951.
Klockmann, Reiner, E., Bachmann, R., Hackl, T., & Fischer, M. (2016). Food finger-
printing: Metabolomic approaches for geographical origin discrimination of
Hazelnuts (Corylus avellana) by UPLC-QTOF-MS. Journal of Agricultural and Food
Chemistry, 64(48), 9253–9262.
Kuuliala, L., Abatih, E., Ioannidis, A. G., Vanderroost, M., De Meulenaer, B., Ragaert, P., &
161
Food Chemistry 278 (2019) 144–162
Devlieghere, F. (2018). Multivariate statistical analysis for the identification of po-
tential seafood spoilage indicators. Food Control, 84, 49–60.
Kuuliala, L., Al Hage, Y., Ioannidis, A. G., Sader, M., Kerckhof, F. M., Vanderroost, M., ...
Devlieghere, F. (2018). Microbiological, chemical and sensory spoilage analysis of
raw Atlantic cod (Gadus morhua) stored under modified atmospheres. Food
Microbiology, 70, 232–244.
Lampir, L., & Pavlousek, P. (2013). Influence of locality on content of phenolic com-
pounds in white wines. Czech Journal of Food Science, 31(6), 619–626.
Le Gall, G., Puaud, M., & Colquhoun, I. J. (2001). Discrimination between orange juice
and pulp wash by 1H nuclear magnetic resonance spectroscopy: Identification of
marker compounds. Journal of Agricultural and Food Chemistry, 49(2), 580–588.
Lee, K. M., Son, M., Kang, J. H., Kim, D., Hong, S., Park, T. H., ... Choi, S. S. (2018). A
triangle study of human, instrument and bioelectronic nose for non-destructive sen-
sing of seafood freshness. Scientific Reports, 8(1), 547.
Li, Y., Zhang, J., Jin, H., Liu, H., & Wang, Y. (2016). Ultraviolet spectroscopy combined
with ultra-fast liquid chromatography and multivariate statistical analysis for quality
assessment of wild Wolfiporia extensa from different geographical origins.
Spectrochimica Acta Part A: Molecular and Biomolecular Spectroscopy, 165, 61–68.
Lin, B., Loomes, K. M., Prijic, G., Schlothauer, R., & Stephens, J. M. (2017). Lepteridine as
a unique fluorescent marker for the authentication of manuka honey. Food Chemistry,
225, 175–180.
Llano, S. M., Muñoz-Jiménez, A. M., Jiménez-Cartagena, C., Londoño-Londoño, J., &
Medina, S. (2018). Untargeted metabolomics reveals specific withanolides and fatty
acyl glycoside as tentative metabolites to differentiate organic and conventional
Physalis peruviana fruits. Food Chemistry, 244, 120–127.
Longobardi, F., Innamorato, V., Di Gioia, A., Ventrella, A., Lippolis, V., Logrieco, A. F., ...
Agostiano, A. (2017). Geographical origin discrimination of lentils (Lens culinaris
Medik.) using (1)H NMR fingerprinting and multivariate statistical analyses. Food
Chemistry, 237, 743–748.
Lovestead, T. M., & Bruno, T. J. (2010). Detection of poultry spoilage markers from
headspace analysis with cryoadsorption on a short alumina PLOT column. Food
Chemistry, 121(4), 1274–1282.
Magdas, D. A., Guyon, F., Feher, I., & Pinzaru, S. C. (2018). Wine discrimination based on
chemometric analysis of untargeted markers using FT-Raman spectroscopy. Food
Control, 85, 385–391.
Malheiro, R., Guedes de Pinho, P., Soares, S., da Silva, César, Ferreira, A., & Baptista, P.
(2013). Volatile biomarkers for wild mushrooms species discrimination. Food
Research International, 54(1), 186–194.
Mannina, L., Sobolev, A. P., Capitani, D., Iaffaldano, N., Rosato, M. P., Ragni, P., ...
Coppola, R. (2008). NMR metabolic profiling of organic and aqueous sea bass ex-
tracts: Implications in the discrimination of wild and cultured sea bass. Talanta,
77(1), 433–444.
Marseglia, A., Caligiani, A., Comino, L., Righi, F., Quarantelli, A., & Palla, G. (2013).
Cyclopropyl and omega-cyclohexyl fatty acids as quality markers of cow milk and
cheese. Food Chemistry, 140(4), 711–716.
Martín, A., Benito, M. J., Aranda, E., Ruiz-Moyano, S., Córdoba, J. J., & Córdoba, M. G.
(2010). Characterization by volatile compounds of microbial deep spoilage in iberian
dry-cured ham. Journal of Food Science, 75(6), 1–10.
Miks-Krajnik, M., Yoon, Y. J., Ukuku, D. O., & Yuk, H. G. (2016). Identification and
quantification of volatile chemical spoilage indexes associated with bacterial growth
dynamics in aerobically stored chicken. Journal of Food Science, 81(8), M2006–2014.
Monakhova, Y. B., Ruge, W., Kuballa, T., Ilse, M., Winkelmann, O., Diehl, B., ...
Lachenmeier, D. W. (2015). Rapid approach to identify the presence of Arabica and
Robusta species in coffee using 1H NMR spectroscopy. Food Chemistry, 182, 178–184.
Nekvapil, F., Brezestean, I., Barchewitz, D., Glamuzina, B., Chiş, V., & Cintă Pinzaru, S.
(2018). Citrus fruits freshness assessment using Raman spectroscopy. Food Chemistry,
242, 560–567.
Novotná, H., Kmiecik, O., Gałązka, M., Krtková, V., Hurajová, A., Schulzová, V., ...
Hajšlová, J. (2012). Metabolomic fingerprinting employing DART-TOFMS for au-
thentication of tomatoes and peppers from organic and conventional farming. Food
Additives & Contaminants: Part A, 29(9), 1335–1346.
Oliveira, J. M., Faria, M., Sá, F., Barros, F., & Araújo, I. M. (2006). C 6-alcohols as varietal
markers for assessment of wine origin. Analytica Chimica Acta, 563(1), 300–309.
Parlapani, F. F., Haroutounian, S. A., Nychas, G.-J. E., & Boziaris, I. S. (2015).
Microbiological spoilage and volatiles production of gutted European sea bass stored
under air and commercial modified atmosphere package at 2 ͦC. Food Microbiology,
50, 44–53.
Pattamayutanon, P., Angeli, S., Thakeow, P., Abraham, J., Disayathanoowat, T., &
Chantawannakul, P. (2017). Volatile organic compounds of Thai honeys produced
from several floral sources by different honey bee species. PLoS One, 12(2),
e0172099.
Pedan, V., Weber, C., Do, T., Fischer, N., Reich, E., & Rohn, S. (2017). HPTLC fingerprint
profile analysis of cocoa proanthocyanidins depending on origin and genotype. Food
Chemistry.
Perestrelo, R., Barros, A. S., Rocha, S. M., & Câmara, J. S. (2014). Establishment of the
varietal profile of Vitis vinifera L. grape varieties from different geographical regions
based on HS-SPME/GC-qMS combined with chemometric tools. Microchemical
Journal, 116, 107–117.
Perestrelo, R., Silva, C., & Câmara, J. S. (2014). A useful approach for the differentiation
of wines according to geographical origin based on global volatile patterns. Journal of
Separation Science, 37(15), 1974–1981.
Pisano, P. L., Silva, M. F., & Olivieri, A. C. (2015). Anthocyanins as markers for the
classification of Argentinean wines according to botanical and geographical origin.
Chemometric modeling of liquid chromatography-mass spectrometry data. Food
Chemistry, 175, 174–180.
Pontes, M., Pereira, J., & Câmara, J. S. (2012). Dynamic headspace solid-phase
S. Medina et al.
microextraction combined with one-dimensional gas chromatography–mass spec-
trometry as a powerful tool to differentiate banana cultivars based on their volatile
metabolite profile. Food Chemistry, 134(4), 2509–2520.
Porep, J. U., Walter, R., Kortekamp, A., & Carle, R. (2014). Ergosterol as an objective
indicator for grape rot and fungal biomass in grapes. Food Control, 37, 77–84.
Porto-Figueira, P., Freitas, A., Cruz, C. J., Figueira, J., & Câmara, J. S. (2015). Profiling of
passion fruit volatiles: An effective tool to discriminate between species and varieties.
Food Research International, 77, 408–418.
Povolo, M., Pelizzola, V., Ravera, D., & Contarini, G. (2009). Significance of the non-
volatile minor compounds of the neutral lipid fraction as markers of the origin of
dairy products. Journal of Agricultural and Food Chemistry, 57(16), 7387–7394.
Righetti, L., Rubert, J., Galaverna, G., Hurkova, K., Dall'Asta, C., Hajslova, J., & Stranska-
Zachariasova, M. (2018). A novel approach based on untargeted lipidomics reveals
differences in the lipid pattern among durum and common wheat. Food Chemistry,
240, 775–783.
Risticevic, S., Carasek, E., & Pawliszyn, J. (2008). Headspace solid-phase microextraction-
gas chromatographic-time-of-flight mass spectrometric methodology for geographical
origin verification of coffee. Analytica Chimica Acta, 617(1–2), 72–84.
Röhlig, R. M., & Engel, K.-H. (2010). Influence of the input system (Conventional versus
Organic Farming) on metabolite profiles of maize (Zea mays) Kernels. Journal of
Agricultural and Food Chemistry, 58(5), 3022–3030.
Rubert, J., Lacina, O., Zachariasova, M., & Hajslova, J. (2016). Saffron authentication
based on liquid chromatography high resolution tandem mass spectrometry and
multivariate data analysis. Food Chemistry, 204, 201–209.
Schievano, E., Morelato, E., Facchin, C., & Mammi, S. (2013). Characterization of markers
of botanical origin and other compounds extracted from unifloral honeys. Journal of
Agricultural and Food Chemistry, 61(8), 1747–1755.
Segato, S., Galaverna, G., Contiero, B., Berzaghi, P., Caligiani, A., Marseglia, A., & Cozzi,
G. (2017). Identification of lipid biomarkers to discriminate between the different
production systems for asiago PDO cheese. Journal of Agricultural and Food Chemistry,
65(45), 9887–9892.
Shepherd, L. V. T., Hackett, C. A., Alexander, C. J., Sungurtas, J. A., Pont, S. D. A.,
Stewart, D., ... Davies, H. V. (2014). Effect of agricultural production systems on the
potato metabolome. Metabolomics, 10(2), 212–224.
Son, H.-S., Hwang, G.-S., Kim, K. M., Ahn, H.-J., Park, W.-M., Van Den Berg, F., ... Lee, C.-
H. (2009). Metabolomic studies on geographical grapes and their wines using 1H
NMR analysis coupled with multivariate statistics. Journal of Agricultural and Food
Chemistry, 57(4), 1481–1490.
Souard, F., Delporte, C., Stoffelen, P., Thévenot, E. A., Noret, N., Dauvergne, B., ...
Stévigny, C. (2018). Metabolomics fingerprint of coffee species determined by un-
targeted-profiling study using LC-HRMS. Food Chemistry, 245, 603–612.
Spanik, I., Pazitna, A., Siska, P., & Szolcsanyi, P. (2014). The determination of botanical
origin of honeys based on enantiomer distribution of chiral volatile organic com-
pounds. Food Chemistry, 158, 497–503.
Splivallo, R., Bossi, S., Maffei, M., & Bonfante, P. (2007). Discrimination of truffle fruiting
body versus mycelial aromas by stir bar sorptive extraction. Phytochemistry, 68(20),
2584–2598.
Stanimirova, I., Üstün, B., Cajka, T., Riddelova, K., Hajslova, J., Buydens, L. M. C., &
Walczak, B. (2010). Tracing the geographical origin of honeys based on volatile
compounds profiles assessment using pattern recognition techniques. Food Chemistry,
118(1), 171–176.
Sun, X., Li, L., Ma, T., Liu, X., Huang, W., & Zhan, J. (2015). Profiles of phenolic acids and
flavan-3-ols for select chinese red wines: A comparison and differentiation according
to geographic origin and grape variety. Journal of Food Science, 80(10).
Tette, P. A., Guidi, L. R., Bastos, E. M., Fernandes, C., & Gloria, M. B. (2017). Synephrine –
A potential biomarker for orange honey authenticity. Food Chemistry, 229, 527–533.
Thavarajah, P., & Low, N. H. (2006). Adulteration of apple with pear juice: Emphasis on
major carbohydrates, proline, and arbutin. Journal of Agricultural and Food Chemistry,
54(13), 4861–4867.
162
Food Chemistry 278 (2019) 144–162
Tompa, G., Susic, R., Rogelj, I., & Pompe, M. (2013). Cryotrap/SPME/GC/MS method for
profiling of monoterpenes in cheese and their clustering according to geographic
origin. Acta Chimica Slovenica, 60(3), 595–603.
Trivedi, D. K., Hollywood, K. A., Rattray, N. J., Ward, H., Trivedi, D. K., Greenwood, J., ...
Goodacre, R. (2016). Meat, the metabolites: An integrated metabolite profiling and
lipidomics approach for the detection of the adulteration of beef with pork. Analyst,
141(7), 2155–2164.
Vallverdú-Queralt, A., Medina-Remón, A., Casals-Ribes, I., Amat, M., & Lamuela-
Raventós, R. M. (2011). A Metabolomic Approach Differentiates between
Conventional and Organic Ketchups. Journal of Agricultural and Food Chemistry,
59(21), 11703–11710.
van Ruth, S., Alewijn, M., Rogers, K., Newton-Smith, E., Tena, N., Bollen, M., & Koot, A.
(2011). Authentication of organic and conventional eggs by carotenoid profiling.
Food Chemistry, 126(3), 1299–1305.
Vasta, V., Luciano, G., Dimauro, C., Rohrle, F., Priolo, A., Monahan, F. J., & Moloney, A.
P. (2011). The volatile profile of longissimus dorsi muscle of heifers fed pasture,
pasture silage or cereal concentrate: Implication for dietary discrimination. Meat
Science, 87(3), 282–289.
Vetter, W., & Schröder, M. (2010). Concentrations of phytanic acid and pristanic acid are
higher in organic than in conventional dairy products from the German market. Food
Chemistry, 119(2), 746–752.
Vita, F., Taiti, C., Pompeiano, A., Bazihizina, N., Lucarotti, V., Mancuso, S., & Alpi, A.
(2015). Volatile organic compounds in truffle (Tuber magnatum Pico): Comparison of
samples from different regions of Italy and from different seasons. Scientific Reports, 5,
12629.
Wan, H., Wu, L., Yang, Y., Zhou, G., & Ruan, Y. L. (2018). Evolution of sucrose meta-
bolism: The dichotomy of invertases and beyond. Trends in Plant Science, 23(2),
163–177.
Wang, S. Y., Chen, C.-T., Sciarappa, W., Wang, C. Y., & Camp, M. J. (2008). Fruit quality,
antioxidant capacity, and flavonoid content of organically and conventionally grown
blueberries. Journal of Agricultural and Food Chemistry, 56(14), 5788–5794.
Weesepoel, Y., Heenan, S., Boerrigter-Eenling, R., Venderink, T., Blokland, M., & van
Ruth, S. (2016). Protocatechuic acid levels discriminate between organic and con-
ventional wheat from Denmark. CHIMIA International Journal for Chemistry, 70(5),
360–363.
Wei, F., Furihata, K., Koda, M., Hu, F., Kato, R., Miyakawa, T., & Tanokura, M. (2012).
13C NMR-based metabolomics for the classification of green coffee beans according
to variety and origin. Journal of Agricultural and Food Chemistry, 60(40),
10118–10125.
Wierda, R. L., Fletcher, G., Xu, L., & Dufour, J. P. (2006). Analysis of volatile compounds
as spoilage indicators in fresh king salmon (Oncorhynchus tshawytscha) during storage
using SPME-GC-MS. Journal of Agricultural and Food Chemistry, 54(22), 8480–8490.
Willems, J. L., & Low, N. H. (2018). Structural identification of compounds for use in the
detection of juice-to-juice debasing between apple and pear juices. Food Chemistry,
241, 346–352.
Xue, X., Wang, Q., Li, Y., Wu, L., Chen, L., Zhao, J., & Liu, F. (2013). 2-acetylfuran-3-
glucopyranoside as a novel marker for the detection of honey adulterated with rice
syrup. Journal of Agricultural and Food Chemistry, 61(31), 7488–7493.
Yahagi, T., Masada, S., Oshima, N., Suzuki, R., Matsufuji, H., Takahashi, Y., ...
Hakamatsuka, T. (2016). Determination and Identification of a specific marker
compound for discriminating shrub chaste tree fruit from agnus castus fruit based on
LC/MS metabolic analysis. Chemical and Pharmaceutical Bulletin, 64(4), 305–310.
Yang, N., Liu, C., Liu, X., Degn, T. K., Munchow, M., & Fisk, I. (2016). Determination of
volatile marker compounds of common coffee roast defects. Food Chemistry, 211,
206–214.
Zörb, C., Langenkämper, G., Betsche, T., Niehaus, K., & Barsch, A. (2006). Metabolite
profiling of wheat grains (Triticum aestivum L.) from organic and conventional agri-
culture. Journal of Agricultural and Food Chemistry, 54(21), 8301–8306.