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ScienceDirect
Methodologies to advance the understanding of flavor
chemistry
Michele Eliza Cortazzo Menis-Henrique
Flavor is an important attribute of food and beverage quality
control; thus it has always been the focus of much research. It is
formed by the combination of taste (attributed to non-volatile
compounds), aroma (related to volatile compounds), and
chemesthetic sensations. The characteristic flavor of food is
due to its volatile compounds, thus many studies have focused
on the analysis of these compounds. All advances in flavor
chemistry are due to the development of new technologies that
facilitate and allow the analysis of volatile and non-volatile
compounds in foods or the pattern recognition, as observed in
the E-nose and E-tongue equipment. However, further studies
are required to understand the food flavor, supported by the
constant development of new technologies.
Address
School of Chemical Engineering, University of Campinas, Av. Albert
Einstein, 500, Campinas, SP, Brazil
Corresponding author:
Menis-Henrique, Michele Eliza Cortazzo (micheleh@unicamp.br)
Current Opinion in Food Science 2020, 33:131–135
This review comes from a themed issue on Food physics & materials
science
Edited by Maria Teresa Pedrosa Silva Clerici
https://doi.org/10.1016/j.cofs.2020.04.005
2214-7993/ã 2020 Elsevier Ltd. All rights reserved.
Introduction
The flavor of food is an important quality attribute driving
consumers’ choice. It is the result of the combination of
taste, aroma, and oral or chemesthetic sensations. Taste is
due to the non-volatile compounds present in food, which
characterizes the five basic tastes: sweet, salty, bitter,
sour, and umami. The aroma is related to volatile com-
pounds, which are responsible for the characteristic flavor
of food [1]. Finally, the chemesthesic sensations are
usually perceived through the stimulation of the human
trigeminal nerve endings within the mouth, nose, or eyes.
They are chemically induced sensations and involve
sensations considered irritating, which may be related
to heat (pepper and other spices) or not (mustard, radish),
tear-inducing stimuli (onion), cooling sensations (men-
thol, among others), and irritation (carbon dioxide). Still,
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sensations in this category include astringency (chemi-
cally induced tactile sensation) and the so-called metallic
taste [2]. Thus, understanding the flavor of food is quite
complex, requiring chemical and/or sensory evaluation
and sophisticated equipment.
Basic tastes
The five basic tastes (sweet, bitter, sour, salty, and
umami) are basic elements of the experience of human
flavor and important for the assessment of the nutritional
quality of food by humans and other organisms [3]. The
sour and salty taste, for example, indicate potentially
spoiled food, while both the sweet and umami tastes
are indicative of nutritious foods, and the bitter taste
serves as a warning against poison intake [4,5].
The taste is perceived by receptor cells organized in
groups, the taste buds. They are modified epithelial cells
found on the tongue and the soft palate, which have an
average lifespan of a week [2]. Sweet, bitter, and umami
tastes are mediated by G protein-coupled receptors
(GPCRs), while sour and salty tastes are modulated by
specialized membrane channels (ion channels). Thus, for
the flavor perception, there is the interaction between a
taste-activating molecule (substance) and a specialized
receptor located in the taste buds at the level of the cell
membrane. The recipient cells then translate the chemi-
cal stimulus into nerve signals to the brain, leading to
sensory perception [4].
The compounds responsible for sweet taste include nat-
ural sugars (sucrose), sugar alcohols (xylitol), dipeptides
(aspartame), sulfonylamide (saccharin), and terpenoid
glycosides (stevioside). Concerning bitter taste, plants
are the main source of human food. It is estimated that
there are more than tens of thousands of molecules
responsible for bitter taste belonging to different chemi-
cal classes such as salts, flavonoids, alkaloids, amino acids,
peptides, and lipids. Umami taste consists of typical
monosodium glutamate (MSG) found naturally in many
protein-rich foods, such as seafood, meat, cheese, and
certain fruits and vegetables. In foods, Na+ (NaCl) is the
main salty compound, but other cations, such as Ca2+, K+,
Li+, and NH4+, also have a salty taste, though only the ion
Na+ can provide a clean salty taste, without interferences.
The sour taste is characteristic of the detection of protons
and is generated by acids such as citric, tartaric, lactic, and
acetic acids. These acids are naturally found in citrus
fruits (lemon, orange, and grapefruit) and fermented
foods [6]. Studies have suggested a taste sensitivity also
Current Opinion in Food Science 2020, 33:131–135
132 Food physics & materials science

Table 1

Different methodologies for the study of flavor in food and beverages

Product Methodology Reference

Argan oil Olfactometry using a GC-FID (Hewlett-Packard 5890 Series II, Waldbronn, Germany). [24]
Arabica coffees Olfactometry using a Shimadzu GC-2010 Plus equipped with an AOC-20 s autosampler (Shimadzu, Japan). [25
]
Compounds were separated using an HP-INNOWax column (30 m  0.25 mm  0.25 mm) (Agilent, Woodbridge,
USA), with the GC effluent split into a ratio of 1:1 between the sniffing port and FID.
Ciders Olfactometry using Shimadzu QP2010 GC–MS fitted with an OP275 olfactory detection port (ODP; GL Sciences, [20]
Tokyo, Japan). The effluent from the capillary column was split 1:3 between the MS and the ODP.
Wines Nosespace using the Proton Transfer Reaction-Mass Spectrometer (PTR-MS) instrument equipped with a Time- [26]
of-Flight (ToF) analyzer (PTR-ToF-MS 8000, Ionicon Analytik, Innsbruck, Austria).
Dark chocolates Nosespace using the (PTR-Qi ToF- MS) (Ionicon Analytik G.m.b.H., Innsbruck, Austria). [27]
Expanded snack Nosespace with the chewing simulator coupled to PTR-TOF-MS 8000 detector (Ionicon Analytik, Innsbruck, [10

]
Austria).
Black tea E-nose: five MOS sensors which are composed of commercially available gas sensors. E-tongue: five [28]
electrodes made of five different noble metals (viz. gold, iridium, palladium, platinum, and rhodium), a counter
electrode made of platinum (P H Ionics, Hyderabad, India), and an Ag/AgCl reference electrode (saturated KCL,
Gamry Instruments Inc., USA).
Apple fruits E-nose: PEN 3 E-nose (Insent Company, Japan). The sensor array contains 10 metal oxide gas sensors. E- [29
]
tongue: SA402B e-tongue (Insent Company, Japan). The taste sensor consists of five test sensors and two
reference sensors. SPME: 50/30 mm divinylbenzene/carboxen/polydi-methylsiloxane (DVB/CAR/PDMS) fiber
(Supelco, Bellefonte, USA). GC–MS: Agilent Technologies 7890A/5975C.
Golden pompano E-nose: PEN3 (Airsense Analytics GmbH, Schwerin, Germany) contained 10 metal oxide semiconductors. E- [30]
(Trachinotus tongue: equipped with seven chemical sensors (TS-5000Z, Insent Inc., Japan). SDE apparatus (Tianchang Great
blochii) fillets Wall, China) for extraction and GC-MS (Agilent 7890B series and Agilent 5975C series, respectively).

to free fatty acids in taste receptor cells, due to the
presence of a fatty acid transporter, CD36. This event
would make fat the 6th basic taste, but studies are not yet
conclusive [6,7].
Aroma
The aroma is another component of flavor, and responsi-
ble for the so-called characteristic flavor of food. During
chewing, volatile compounds are released from food and
transported from the oral cavity to the pharynx during
swallowing. With the subsequent exhalation, the volatile
compounds are swept by the air that comes from the lungs
reaching the olfactory epithelium and exhaled through
the nose. In this process, the sensory perception occurs by
the olfactory epithelium via the retronasal olfaction (from
the mouth) rather than the ortonasal olfaction (from the
nose) [8].
Although more than 1000 volatile compounds are
detected in food, only 5–10% contribute to food aroma
as metional (notes of boiled potatoes), 2-methylbutanal
(malted) and 3-methylbutanal (malted), butane-2,3-dione
(buttery), (E, E)-2,4-decadienal (fried, greasy) and fur-
aneol (caramel), followed by sotolon (seasoning), 1-
octene-3-one (mushroom), acetic acid (vinegar), acetal-
dehyde (fruity, fresh), ethyl 2-methylbutanoate (fruity)
and 3-methylbutanoate (fruity), (E)-2-nonenal (similar to
cardboard), vanillin (similar to vanilla), 2-acetyl-1-pyrro-
line (toast, popcorn), 2-methylbutanoic acid (sweaty) and
3-methylbutanoic acid (sweaty), and butanoic acid
(sweaty) [6].
In addition to the concentration and characteristics of the
volatile compounds, the composition of the food matrix,
including the presence of aqueous phase or oil phase, for
example, can also affect the intensity of perception of the
aroma. In high-fat foods, the release of volatile com-
pounds is slower when compared to low-fat and fat-free
products. First, these compounds should be released from
the oily phase of food to saliva, and then to the headspace
of the mouth, with a lower mass transfer rate in the oily
phase than in the aqueous phase or air [9,10

]. In this
case, with the slower release of aroma compounds, the
consumer perceives the high-fat product as tastier than a
similar product with a lower fat content.
Analyzing the flavor of food
The study of the chemistry of food flavor is quite com-
plex. The characteristic flavor of food is a consequence of
its volatile compounds; thus several studies have focused
on the analysis of these compounds (Table 1). The
present review will discuss the methodologies frequently
used for the analysis of volatile compounds in foods and
beverages, which have contributed greatly to the under-
standing of the flavor chemistry.
Volatile compounds profile of food can be determined
using gas chromatography coupled to mass spectrometry

Current Opinion in Food Science 2020, 33:131–135 www.sciencedirect.com


(GC–MS). Ma et al. [11] evaluated the volatiles profile in
sea buckthorn (Hippophae rhamnoides L.) berries and
found 45 compounds, and alcohols, aldehydes, hydrocar-
bons, and ketones were the main responsible for the
flavor.
Some authors have also investigated the changes in the
volatiles profile during food processing and storage. Alasti
et al. [12] studied the changes in the volatiles profile of
cocoa powder during processing, aimed at determining
the ideal profile for obtaining a high-quality cocoa pow-
der. Sirtori et al. [13] evaluated the evolution of the
volatile compounds in Tuscan ham during the curing
period, which allowed the separation between the sam-
ples from 0 to 6 months from those from 12 to 18 months,
using 26 identified volatile compounds.
In addition to knowing the volatiles profile of food, the
compounds that effectively contribute to the food flavor
should also be assessed, once not all of the volatile
compounds identified have a significant contribution to
the products’ aroma. In that context, the gas-chromatog-
raphy olfactometry (GC-O) method can be used. It con-
sists of the sensory evaluation of volatile compounds in
chromatographic effluents that leave the column, that is,
the volatiles separated by the chromatographic column
are diverted to a deactivated column rather than the
detector, thus reaching the evaluator’s nose [14]. In the
GC-O-MS equipment, the column flow is divided, with a
part going to the mass spectrometer to identify the
compounds, and a part going to a deactivated column,
for sensory evaluation. Currently, the most accepted and
used technique in GC-O is the OSME [15,16], in which
individuals evaluate the chromatographic effluent using
the time-intensity analysis, in which the evaluator records
the duration of each olfactory stimulus on an intensity
scale, and describes the quality of the perceived odor.
Then, an aromagram is obtained from the data, with high
peaks and/or larger areas, thus indicating the significant
odorous compounds [1].
However, although olfactometry has contributed to a
major advance in flavor chemistry, it still does not provide
the perception of aroma in real time, that is, during food
consumption. This online assessment was made possible
with snosespace analysis, which uses a proton transfer
reaction mass spectrometer to analyze the volatile com-
pounds released from food during chewing [17].
For this analysis, a plastic accessory is placed in the eva-
luator’s nostrils to capture the volatile compounds that are
exhaled during food consumption. These compounds are
then conducted through a heated transfer line until reach-
ing the mass spectrometer [18]. Two devices can monitor
the compounds in real time quickly and with high sensitiv-
ity: the atmospheric-pressure chemical ionization mass
spectrometry (APCI-MS) and the proton-transfer-reaction
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Understanding of flavor chemistry Menis-Henrique 133
mass spectrometry (PTR-MS) [8]. In this equipment, the
ionization of volatile compounds occurs by proton transfer
reaction (PTR) with hydronium ion (H3O+) as a reagent. It
is a smoother chemical ionization when compared to elec-
tron impact ionization. The proton transfer reaction occurs
when the proton affinity of the volatile compound is greater
than that of the hydronium ion and is represented by the
following equation, where R represents the volatile
compound:
H3O+ + R ! H2O + RH+
The reaction can occur at atmospheric pressure (APCI-
MS), which is the simplest process. In this process, the
formation of hydronium ions has a limited control, and
water clusters (H2O)nH+ are also formed, impairing the
efficiency of proton transfer while in PTR-MS, the hydro-
nium ion is formed separately and introduced to react
with the analyte after a cleaning process, which ensures
the inclusion of H3O+ ions only [14]. The proton transfer
reaction occurs at reduced pressure in a drift tube, and the
ions are analyzed by a quadrupole or time-of-flight (TOF)
mass detector. The PTR-MS presents greater sensitivity
when compared to the APCI-MS, and allows measuring
the volatile compounds at very low levels, reaching parts
per trillion for some compounds.
Most studies on flavor are focused on volatile compounds,
and the non-volatile compounds responsible for basic
tastes have also been studied after the advancement of
instrumental analysis. Intelmann et al. [19] detected many
types of iso-a-acids, a-acids, and b-acids in beer using
liquid chromatography-mass spectrometry (LC–MS).
High-performance liquid chromatography (HPLC) has
also been used to quantify carbohydrates (glucose, fruc-
tose, and glycerol) and organic acids in fermented apple
juice [20].
Electronic nose and electronic tongue
The techniques previously discussed allow analyzing the
flavor of food and beverages by determining the volatile
or non-volatile compounds. Such techniques are expen-
sive and require specific equipment and specialized labor.
Thus, other instruments for measuring flavor have been
developed, such as the electronic nose and the electronic
tongue (E-nose and E-tongue, respectively), as shown in
Table 1. Such instruments are composed of a set of
sensors connected to a computer with an artificial intelli-
gence system to recognize patterns of molecules, trying to
imitate the human nose and the tongue [21].
In the human nose, signals from the olfactory system are
transmitted to the brain and then interpreted as an
odorous sensation based on previous experiences. Like-
wise, the electronic nose uses sensors to capture signals
that are sent to software that will play the brain, being
able to estimate the volatile and complex gas
Current Opinion in Food Science 2020, 33:131–135
134 Food physics & materials science
combinations without separately identifying all the chem-
ical compounds in the mixture [21,22]. Electronic nose
sensors can be of the following types: metal oxide semi-
conductors, organic polymer conductor, piezoelectric
crystal sensors (QCM-Quartz Crystal Microbalance and
SAW-Surface Acoustic Wave) and metal-oxide field-
effect transistor (Mosfet) sensors. The sensors convert
the chemical interactions into analog electrical responses
(change in electrical resistance, frequency, or conductiv-
ity). When exposed to a change in the atmosphere, the
electrical signals generated by the sensors are transmitted
to a computer for processing and interpretation, which is
performed by software that compares the spectrum with
patterns of known compounds stored in the computer’s
memory [21–23]. Currently, most commercial electronic
noses employ the Artificial Neural Network (ANN),
which works like a human brain through a learning
process, that is, it learns and stores the results in memory
from the analysis of patterns for use in future determina-
tions. Thus, the most carefully collected data, the more
accurate the instrument [21].
The electronic tongue works similarly to the electronic
nose. It uses specific sensors to detect the markers and a
pattern recognition system, capable of recognizing solu-
ble non-volatile molecules and identifying the five basic
tastes (salty, sweet, sour, bitter, and umami) and recog-
nizing the product under analysis. The methods used to
distinguish taste in liquid samples include electrochemi-
cal (potentiometry and voltammetry) and optical meth-
ods, detection of mass changes, amperometry, and con-
ductometry [22].
However, e-nose and e-tongue cannot safely identify the
quality or intensity of odor or taste of multicomponent
samples. Some practical limitations of this equipment
include the interference of odorless gas mixtures, such
as water vapor in the reading of gas sensors, contamination
of sensors and oscillation due to repeated stimuli, inade-
quate sensor’s sensitivity, noise amplification as the
matrix increases, and inadequate selectivity [23]. Thus,
such equipment requires constant research and develop-
ment to improve its performance.
Conclusion
Great advances have been made toward understanding
the flavor chemistry using new technologies and sophis-
ticated equipment. Today, it is possible to determine not
only the compounds responsible for the flavor profile of
food but also those that effectively contribute to the
aroma and those released from food and beverages in
real time, that is, during consumption. In addition, instru-
ments such as the electronic nose and electronic tongue
have been developed and/or improved to replace the
human nose and tongue in routine analysis. In this
way, new technologies open space for new research to
better understand the flavor of foods.
Current Opinion in Food Science 2020, 33:131–135
Conflict of interest statement
The author declares no conflicts of interest.
References and recommended reading
Papers of particular interest, published within the period of review,
have been highlighted as:

of special interest


of outstanding interest
1. Franco MRB: Aroma e Sabor de Alimentos: Temas Atuais. Livraria
Varela; 2003.
2. Lawless HT, Heymann H: Sensory Avaluation of Food: Principles
and Practices. Springer; 2010.
3. Beauchamp GK: Basic taste: a perceptual concept. J Agric
Food Chem 2019, 67:13860-13869.
4. Spaggiari G, Di Pizio A, Cozzini P: Sweet, umami and bitter taste
receptors: state of the art of in silico molecular modeling
approaches. Trends Food Sci Technol 2020, 96:21-29.
5. Sacchi R, Caporaso N, Squadrilli GA, Paduano A, Ambrosino ML,
Cavella S, Genovese A: Sensory profile, biophenolic and volatile
compounds of an artisanal ice cream (‘gelato’) functionalised
using extra virgin olive oil. Int J Gastron Food Sci 2019,
18:100173.
6. Guichard E, Salles C, Morzel M, LeBon AM: Flavour: From Food to
Perception. Wiley Blackwell; 2017.
7. Liu D, Costanzo A, Evans MDM, Archer NS, Nowson C, Duesing K,
Keast R: Expression of the candidate fat taste receptors in
human fungiform papillae and the association with fat taste
function. Br J Nutr 2018, 120:64-73.
8. Roberts DD, Pollien P, Yeretzian C, Lindinger C: Nosespace
analysis with Proton-Tranfer-Reaction Mass Spectrometry:
intra- and interpersonal variability. In Handbook of Flavor
Chracterization: Chemistry, and Physiology Series. Edited by
Deibler KD, Delwiche J. Marcel Dekker; 2004:151-162.
9. Roos KB: How lipids influence food flavor. Overview
Outstanding Symposia in Food Science & Technology 1997, vol.
5160-61.
10. Menis-Henrique MEC, Janzantti NS, Andriot I, Sémon E,


Berdeaux O, Schlich P, Conti-Silva AC: Cheese-flavored
expanded snacks with low lipid content: oil effects on the in
vitro release of butyric acid and on the duration of the
dominant sensations of the products. LWT- Food Sci Technol
2019, 105:30-36
This study investigated the butyric acid release via aroma and odor in
cheese-flavored expanded snacks with different lipid content using the
nosespace technique. In the snacks with sunflower oil, the release was
supposedly slower compared to the control snack without sunflower oil.
11. Ma X, Yang W, Marsol-Vall A, Laaksonen O, Yang B: Analysis of
flavour compounds and prediction of sensory properties in
sea buckthorn (Hippophaë rhamnoides L.) berries. Int J Food
Sci Technol 2019, 55:1705-1715.
12. Alasti FM, Asefi N, Maleki R, SeiiedlouHeris SS: Investigating the
flavor compounds in the cocoa powder production process.
Food Sci Nutr 2019, 7:3892-3901.
13. Sirtori F, Dimauro C, Bozzi R, Aquilani C, Franci O, Calamai L,
Pezzati A, Pugliese C: Evolution of volatile compounds and
physical, chemical and sensory characteristics of Toscano
PDO ham from fresh to dry-cured product. Eur Food Res
Technol 2019, 246:409-424.
14. Taylor AJ, Linforth RST: Food Flavor Technology. Blackwell
Publishing Ltd.; 2010.
15. McDaniel MR, Miranda-Lopez BT, Watson M, Libbey LM: Pinot
noir aroma: a sensory/gas chromatographic approach. In
Flavors and Off-Flavors. Edited by Charalambous G. Elsevier
Science Publishers; 1990:23-36.
www.sciencedirect.com

16. Miranda-Lopez R, Libbey BT, Watson BT, McDaniel MR:
Identification of additional odor-active compounds in pinot
noir wines. Am J Enol Viticult 1992, 1:90-92.
17. Blake RS, Monks PS, Ellis AM: Proton-transfer reaction mass
spectrometry. Chem Rev 2009, 109:861-896.
18. Quéré JLL, Gierczynski I, Sémon E: An atmospheric pressure
chemical ionization-ion-trap mass spectrometer for the on-
line analysis of volatile compounds in foods: a tool for linking
aroma release to aroma perception. J Mass Spectrom 2014,
49:918-928.
19. Intelmann D, Haseleu G, Hofmann T: LC-MS/MS quantitation of
hop-derived bitter compounds in beer using the ECHO
technique. J Agric Food Chem 2009, 57:1172-1182.
20. Wei j, Zhang Y, Qiu Y, Guod H, Ju H, Wang Y, Yuan Y, Yue T:
Chemical composition, sensorial properties, and aroma-
active compounds of ciders fermented with Hanseniaspora
osmophila and Torulaspora quercuum in co- and sequential
fermentations. Food Chem 2020, 306:125623.
21. Lisboa HM, Page T, Guy C: Odor management: fundamentals of
Electronic Nose. Eng Sanit Ambient 2009, 1:9-18.
22. Patel HK, Patel PH, Patel H: Innovative application electronic
nose and electronic tongue techniques for food quality
estimation. Int J Res Eng Technol 2019, 2:318-323.
23. Nagle HT, Schiffman SS: Electronic taste and smell: the case for
performance standards. Proc IEEE 2018, 9:1471-1478.
24. Matthäus B, Bonte A, Sinning B, Charrouf Z: Aroma-relevant
volatile compounds as markers for the sensory quality of
argan oil. Eur J Lipid Sci Technol 2019, 121:1900279.
25. Pua A, Lau H, Liu SQ, Tan LP, Goh RMV, Lassabliere B, Leong KC,

Sun J, Cornuz M, Yu B: Improved detection of key odourants in
Arabica coffee using gas chromatography-olfactometry in
www.sciencedirect.com
Understanding of flavor chemistry Menis-Henrique 135
combination with low energy electron ionisation gas
chromatography-quadrupole time-of-flight mass
spectrometry. Food Chem 2020, 302:125370
This study analyzed the volatile compounds responsible for the aroma of
coffee of four Arabica coffees (Brazil, Colombia, Ethiopia, and Guatemala)
using chromatography-olfactometry in combination with low energy
electron ionization gas chromatography-quadrupole time-of-flight mass
spectrometry. Key odourants were identified by olfactometry analysis.
26. Arvisenet G, Ballester J, Ayed C, Sémon E, Andriot I, Le Quere JL,
Guichard E: Effect of sugar and acid composition, aroma
release, and assessment conditions on aroma enhancement
by taste in model wines. Food Qual Prefer 2019, 71:172-180.
27. Acierno V, Liu N, Alewijn M, Stieger M, Van Rutha SM: Which
cocoa bean traits persist when eating chocolate? Real-time
nosespace analysis by PTR-QiToF-MS. Talanta 2019, 195:676-
682.
28. Banerjee MB, Roy RB, Tudu B, Bandyopadhyay R,
Bhattacharyya N: Black tea classification employing feature
fusion of E-Nose and E-Tongue responses. J Food Eng 2019,
244:55-63.
29. Zhu D, Ren X, Wei L, Cao X, Ge Y, Liu H, Li J: Collaborative

analysis on difference of apple fruits flavour using electronic
nose and electronic tongue. Sci Hortic 2020, 260:108879
Electronic nose and electronic tongue was used to evaluate the quality of
five popular apple varieties coupled with gas chromatography-mass
spectrometry. Differences were observed in the acidic, salty, and umami
tastes among the five apple varieties by electronic language and 45 vola-
tile compounds were identified for five apple varieties.
30. Zhang J, Cao J, Pei Z, Wei P, Xiang D, Cao X, Shen X, Li C: Volatile
flavour components and the mechanisms underlying their
production in golden pompano (Trachinotus blochii) fillets
subjected to different drying methods: a comparative study
using an electronic nose, an electronic tongue and SDE-GC-
MS. Food Res 2019, 123:217-225.
Current Opinion in Food Science 2020, 33:131–135