Food Chemistry 285 (2019) 347–354

Contents lists available at ScienceDirect

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

Tea aroma formation from six model manufacturing processes T

⁎ ⁎
Zhihui Feng, Yifan Li, Ming Li, Yijun Wang, Liang Zhang, Xiaochun Wan , Xiaogen Yang
State Key Laboratory of Tea Plant Biology and Utilization, Anhui Agricultural University, Hefei 230036, Anhui, China

A R T I C LE I N FO A B S T R A C T

Keywords: Tea aroma is determined by the nature of the plant, the production processes, and many other factors influencing
Tea types its formation and release. The objective of this study was to investigate the impact of manufacturing processes on
Tea aroma the aroma composition of tea. Fresh tea leaves from the same cultivar and growing area were selected for
Precursors producing the six types of tea: green, white, yellow, oolong, black, and dark teas. Comprehensive analysis by gas
Volatiles
chromatography mass spectrometry (GC/MS) was performed for the volatiles of tea infusion, prepared by solid-
Tea processing
Aroma character impact
phase microextraction (SPME), solid-phase extraction (SPE), and solvent assisted flavor evaporation (SAFE). A
Quantitative SPME total of 168 volatile compounds were identified. Black tea has the highest volatile concentration of 710 µg/g,
while green tea has the lowest concentration of 20 µg/g. Significantly affected by these processes, tea aroma
molecules are formed mainly from four precursor groups: carotenoids, fatty acids, glycosides, and amino acids/
sugars.

1. Introduction
As a consumer product, the most appealing attribute of tea (Camellia
sinensis (L.) Kuntze) is its sensory characteristics. Fresh tea leaves on the
plant are almost odorless. Upon plucking, they start to form aromas
mainly through enzyme assisted reactions. The purpose of the tea
manufacturing process is to induce and direct the reactions towards
generating desired aroma according to the intention of the tea maker.
Evolved over thousands of years, tea emerges on the market by rich and
intriguing finished products with wide varieties of flavor profile.
According to a survey, there are more than 200 kinds of “most re-
presentative” and distinctive commercial products of finished tea in
China market alone (Wan, 2007). Attempts have been made to classify
tea leaves in many different ways, such as by form, tenderness of tea
leaves, cultivation region, manufacturing process, production season,
and/or sensory profile. The generally accepted concept of tea classifi-
cation was proposed by Chen (Chen, 1979). Based on the processing
methods and sensory qualities of products, the six types of tea were
defined as green, yellow, dark, oolong, white, and black teas (Shitandi,
Ngure, & Mahungu, 2013). Three types are widely available on the
global markets and more often cited as non-fermented (green tea),
semi-fermented (oolong tea), and completely fermented (black tea)
(Yamanishi & Kobayashi, 1999). Although there are many different
styles within each type that vary in processing technique and tea leaf
cultivar, the same type of tea shares the similarity of key elements in
sensory quality and chemical composition.
Tea aroma formation has been studied in various tea manufacturing
processes (Ravichandran & Parthiban, 1998; Sanderson & Graham,
1973). However, the aroma profile is determined not only by the pro-
cess, but also by the tea plant cultivar, terroir, growing conditions, and
many other factors affecting flavor formation and release. In order to
study the influence of manufacturing processes on tea aroma, it is ne-
cessary to keep all other affecting factors the same. High quality teas of
specific type and style are generally made from a particular cultivar,
terroir, tenderness, and require specific processing steps. Although it is
possible to produce different tea types using the same leaves, it is very
difficult to find a tea cultivar suitable to make all six types of tea with
high quality and authentic profile. In addition, variations of processing
within any type of tea can significantly affect the sensory profile of the
final product. Thus, the scope of the study is not to produce any par-
ticular premium tea, but rather to generate samples with the typical
character of each type of tea using thoughtfully designed and simplified
processes. The objective of this study was to investigate the underlying
chemistry of the processes and to test the hypothesis that the tea pro-
cesses promote the precursor reactions selectively and thus generate the
typical aroma characters of each type of tea.

Abbreviations: GC/MS, gas chromatography mass spectrometry; SPMS, solid-phase microextraction; SPE, solid-phase extraction; SAFE, solvent assisted flavor
evaporation; PCA, principal component analysis; OAV, odor activity value; ACI, aroma character impact
⁎
Corresponding authors.
E-mail addresses: yijun@ahau.edu.cn (Y. Wang), zhangliang@ahau.edu.cn (L. Zhang), xcwan@ahau.edu.cn (X. Wan), xiaogen.yang@gmail.com (X. Yang).

https://doi.org/10.1016/j.foodchem.2019.01.174
Received 27 October 2018; Received in revised form 23 January 2019; Accepted 31 January 2019
Available online 02 February 2019
0308-8146/ © 2019 Elsevier Ltd. All rights reserved.
Z. Feng et al.
Fig. 1. Illustrative diagram of the model processes for making the six types of tea: temperature treatment and main reactions during the tea processes.
2. Materials and methods
2.1. Fresh tea leaves
The tea plant cultivar, Longjing 43, was chosen as the source of
fresh tea leaves for the experiments. The tea leaves were plucked as
“one bud with three leaves” from the Shucheng 916 tea factory, com-
mercial plantation in Shucheng, Anhui Province. Longjing 43 is a na-
tional breed of clones from the Tea Research Institute of the Chinese
Academy of Agricultural Sciences, which is a state-level fine breed, with
an elegant shape, flat smooth, brilliant color, long-lasting aroma, and
refreshing taste.
2.2. Model processes
The temperature treatments during the tea processes are illustrated
in Fig. 1. Fixation was accomplished at 150 °C for 3 min using a drum
machine. Drying was performed at 90–110 °C for 5–15 min. The tea leaf
processing experiments were duplicated during the spring season of
2015 and 2016. The tea samples were stored at −80 °C immediately
after production.
2.3. Quality assessment
Color, aroma, and taste of the finished tea products made by the
processes were evaluated by a panel composed of six tea experts ac-
cording the Chinese national standard procedure for evaluating tea
leaves (GB/T 23776, 2018). Tea leaves were brewed with boiling water
for 5 min in a tea-to-water ratio of 1:50. After filtration, the tea infu-
sions were evaluated towards the quality standard of each types of tea.
2.4. Tea aroma analysis
2.4.1. Chemicals and materials
All reference compounds were purchased from commercial sources.
Benzaldehyde, (E)-2-hexenal, benzyl alcohol, phenylacetaldehyde, li-
nalool, 2-phenylethanol, geraniol, nonanoic acid, coumarin, acet-
aldehyde, methanethiol, propanal, 3-methylbutanal, 2-methylbutanal,
348
Food Chemistry 285 (2019) 347–354
1-penten-3-one, dimethyl disulfide, (E)-2-pentenal, dimethyl sulfide, 2-
methylpropanal, 2,3-butanedione, butanal, and 2-furfural were pur-
chased from Sigma-Aldrich (Australia). 2-Pentanone, 2-pentanol, 3-
pentanol, heptanoic acid, (E, Z)-2,6-nonadienal, 1-nonanol, octanoic
acid, 2,6-dimethylocta-3,7-diene-2,6-diol, geranial, n-decanoic acid,
(Z)-3-hexen-1-ol, furfuryl alcohol, isophorone, neral, naphthalene, 1,1-
diethoxyethane, pentanal, and indole were acquired from DRE
(Germany). (E)-2-Hexen-1-ol, 2-n-butylfuran, 1-ethyl-1H-pyrrole, 2-
ethyl-furan, 1-penten-3-ol, and 2-pentylfuran were obtained from
ZZStandard (Shanghai, China). Propanal, 2-methylbutan-1-ol, 1-pen-
tanol, hexanal, 2-hexanol, 3-methylbutanoic acid, 2-methylbutanoic
acid, 1-hexanol, 2-heptanone, pentanoic acid, heptanal, 1-heptanol,
phenol, hexanoic acid, octanal, 2-ethyl-1-hexanol, 1-octanol, nonanal,
maltol, methyl salicylate, 2,4-bis-(1,1-dimethylethyl)-phenol, tetra-
decanoic acid, and n-hexadecanoic acid were purchased from Aladdin
(Shanghai, China).
Methyl tert-butyl ether, pentane, sodium sulfate, and sodium
chloride were obtained from TEDIA (Fairfield, Ohio, USA). Ethyl de-
canoate and p-xylene-d10 were purchased from Sigma-Aldrich
(Australia). Linalool-d3 was purchased from CDN isotopes (Pointe-
Claire, Quebec, Canada). Methyl tert-butyl ether and pentane were
distilled prior use. SPE cartridge (LC-Si, 6 mL tube with 500 mg packing
Part#505374) was purchased from Supelco (USA).
2.4.2. Preparation of internal standard solutions
For solid-phase extraction (SPE) and solvent assisted flavor eva-
poration (SAFE): linalool-d3 (10.0 mg) were dissolved in absolute
ethanol (10.0 mL). Solid-phase microextraction (SPME) internal stan-
dard: 10 μL of p-xylene-d10 (0.05 mg/mL in EtOH) and 10 μL of lina-
lool-d3 (1 mg/mL) were diluted in 1.0 mL of pure water.
2.4.3. Preparation of stock solutions for standard addition
Solution of individual compounds in the concentration of 1 mg/mL
in absolute ethanol was prepared: acetaldehyde, methanethiol, pro-
panal, 3-methyl butanal, 2-methyl butanal, 1-pentene-3-one, dimethyl
disulfide, (E)-2-pentenal, dimethyl sulfide, 2-methyl propanal, 2,3-bu-
tanedione, butanal, 2-furfural.
Z. Feng et al.
2.4.4. Preparation of tea infusion
The amount of tea infusion prepared was according to the needs of
the specific experiments. In general, tea leaves (3.00 g) were placed in
120 mL water at 95 °C. After 3 min, the infusion was filtered through a
rapid filter paper. The tea leaves were washed with 30 mL water. The
infusion was cooled down quickly to room temperature.
2.4.5. Standard addition for headspace solid-phase microextraction
(SPME)
Headspace SPME was based on a modified method (Yang &
Peppard, 1999) and standard addition was applied for SPME for
quantitative analysis. Proper amounts of stock solutions of aroma re-
ference compounds were mixed together and diluted with water. SPME
stable flex fiber (DVB/CAR/PDMS, 50/30 µm) from Supelco (Oakville,
Canada) was used for the headspace experiments. Tea infusion (10 mL)
was mixed with 2.0 g sodium chloride and 10 µL of SPME internal
standard and reference solutions. The headspace vial was incubated for
30 min at 30 °C with shaking and SPME fiber sampling for 30 min
without shaking.
2.4.6. Aroma enrichment by solvent extraction and clean-up by solid-phase
extraction (SPE)
The SPE method was established based on the recommendations of
SPE cartridge supplier and literature (Watson & Raynie, 2014). Tea
infusion (300 mL made with 6 g of tea leaves) was fortified with in-
ternal standard solution (10 µL). The infusion was then extracted three
times with 60 mL of pentane and methyl tert-butyl ether mixture (4:1).
The organic layer was combined and dried with anhydrous sodium
sulfate, then concentrated to 2 mL by TurboVap (Biotage, Sweden),
further concentrated to 0.5 mL with a gentle nitrogen stream. After
conditioned the SPE cartridge with 6 mL solvent system of pentane and
methyl tert-butyl ether (1:1), the tea extract was loaded in the cartridge,
then eluted with 10 mL of the solvent system. The eluent was con-
centrated to 200 µL under a gentle nitrogen stream.
2.4.7. Concentration of volatiles by solvent assisted flavor evaporation
(SAFE) followed by solvent extraction
Tea infusion (150 mL made from 3 g tea leaves) was fortified with
1.00 mL of internal standard solution. The infusion was distilled at
40 °C under a vacuum of 10−3 pa using the solvent assisted flavor
evaporation (SAFE) method (Engel, Bahr, & Schieberle, 1999). The
SAFE distillate was extracted three times with 50 mL of methylene
chloride. The extract was dried over anhydrous sodium sulfate, con-
centrated to 2 mL by TurboVap, then to 200 µL using a gentle nitrogen
stream.
2.4.8. Gas chromatography–mass spectrometry (GC–MS) analysis
The headspace SPME and the aroma concentrates prepared above
were analyzed with an Agilent 6890 GC equipped with 5975B mass
selective detector using fused silica capillary columns DB-5,
30 m × 0.25 mm, 0.25 µm film thickness for SPME analysis and samples
prepared by SPE, DB-5, 60 m × 0.32 mm, 1 µm film thickness for
samples prepared by SAFE. Hydrogen was used as carrier gas with a
linear velocity of 50 cm/s. Splitless injection mode was used for SPME
analysis. Both split and splitless injection modes were used for samples
prepared by SPE. Cold on-column technique was used for samples
prepared by SAFE. GC oven temperature was programmed as follows:
For 30 m column, 40 °C (hold 2 min) to 120 °C at 4 °C/min to 260 at
30 °C/min; for 60 m column, 40 °C (hold 5 min) to 200 °C at 5 °C/min
then to 280 °C at 10 °C/min (hold 10 min). The mass selective detector
was operated in positive EI mode with a mass scan range from m/z 30 to
350 at 70 eV.
2.4.9. Identification of volatile compounds and aroma character impact
(ACI) calculation
GC–MS data processing was achieved using the following software
349
Food Chemistry 285 (2019) 347–354
programs: automated mass spectral deconvolution and identification
system (AMDIS, version 2.72, National Institute of Standards and
Technology, Gaithersburg, MD), NIST mass spectral search program
(version 2.2, National Institute of Standards and Technology), and an
in-house Microsoft Excel-based data analysis program for GC–MS data
processing, which identifies GC peaks based on both mass spectrum and
retention index match using an in-house mass spectral library and re-
tention index database. Concentrations of the compounds listed in
“stock solutions for standard addition” were quantitatively determined
by standard addition using headspace SPME method. Semi-quantitative
concentrations of individual volatile component in SAFE and SPE
concentrates were estimated relative to the internal standards by the
MSD response (total ion current). Aroma character impact (ACI), a ratio
of odor activity value (OAV) in a mixture (Yang, 2008), was calculated
for a total of 119 identified aroma molecules using the odor threshold
value determined in water by the formula:
Pi/ Ti
ACIi =
∑k Pk / Tk
where Pi is the concentration percentage and Ti is the odor threshold
value of the compound i .
2.4.10. Statistical analysis
Principal component analysis (PCA) and hierarchical cluster ana-
lysis were performed using SIMCA 13.0 (Umetrics, Sweden).
3. Results and discussion
3.1. Tea process design and tea leaf evaluation
The principle of the experiment design is to simplify the tea man-
ufacturing processes yet to ensure retaining the most important ele-
ments and producing finished teas with the basic sensory characters of
the corresponding tea type. Six model manufacturing processes ac-
cording to the commonly accepted classification (Chen, 1979) were
designed and executed by the process experts at Anhui Agricultural
University, Hefei, China. The process to make finished tea leaves starts
from leaf plucking when the enzyme mediated reactions begin. Tea
fermentation is usually referred as the atmospheric oxidation of ca-
techins (flavanols) catalyzed by the tea leaf endogenous enzymes
(Harbowy & Balentine, 1997; Engelhardt, 2010), which produces
polyphenol pigments such as theaflavins, thearubigens, and many other
oligomers and polymers. Flavanol-quinones produced in the reaction
are strong oxidizing agents which further oxidize other components
present in tea leaves leading to the formation of aroma molecules. To
create desired sensory quality, the tea making process manipulates
enzyme activities and directs the reactions by controlling temperature,
moisture content, disruption degree of the leaf tissues, and reaction
time. The temperature changes and key events of the six model pro-
cesses are illustrated in Fig. 1.
The green tea and white tea processes represent two strategies of tea
processing. After four hours of withering, the fresh tea leaves during
green tea process were heated to 150 °C for 3 min to deactivate enzymes
(fixation), followed by a rolling process for 1 h to rupture the leaf tis-
sues. The green tea leaves were finished by drying at 90–110 °C for
5–15 min to stop the reactions by lowering the moisture content. Yellow
tea and dark tea processes can be viewed as derived from the green tea
process by inserting a piling time between rolling and drying. During
the piling time, the tea leaves were kept at a relatively high moisture
content. Dark tea process maintains an even higher moisture content of
the tea leaves and longer piling time than yellow tea process. In these
processes, tea leaf endogenous enzymatic reactions are predominant
before fixation although under mild conditions and in a relatively short
time. In the piling stage of yellow and dark tea processes, non-enzy-
matic oxidation and degradation play a major role at the early period.
Z. Feng et al.
With the rapid growth of microbials, ectoenzymatic reactions also take
place (Hua et al., 2015). Furthermore, the activities of residue en-
doenzymes survived after fixation cannot be ignored. Microbial fer-
mentation plays a very significant role in the dark tea process (Zheng,
Wan, & Bao, 2015).
There are only two steps in the white tea process: prolonged 70 h of
withering followed by drying. Although the white tea process is the
simplest one among all processes, it requires skilled expertise in
maintaining proper temperature, moisture, and timing to produce high
quality products. Oolong tea and black tea processes can be regarded as
modified white tea processes by promoting the tea endoenzymatic
oxidations through rocking/rolling in the first step. Black tea process
drives the enzymatic reactions to a great extend to reach “complete
fermentation”, while the oolong tea process manages the fermentation
in a more controlled manner to achieve intended sensory profiles. In
these processes, tea leaf endoenzymes play a crucial role in the bio-
transformation of the components in fresh leaves to that of the finished
tea.
The quality of the finished tea products made by the model pro-
cesses were evaluated by an expert panel. The panelists agreed that the
tea samples of each type have recognizable typical sensory characters
(Table 1). Green tea has a fresh chest-nutty aroma and a mellow taste
while yellow tea smells more like ripe chestnuts and a cooked note.
Oolong tea has a floral aroma, with mellow and sweet notes, whereas
black tea has a fruity aroma with sweet and astringent notes. White tea
has a pleasant sweet floral and honey-like aroma, as well as a sweet
taste. Dark tea smells slightly stale and rough and tastes slightly mellow
and astringent. The color of the tea infusions reflected the oxidation
degrees of the tea polyphenols after processing. According to the expert
panel, the dark tea produced by the model process was under-fer-
mented because of the short piling time and the relative small experi-
ment scale.
3.2. Comprehensive analysis of tea volatiles
Volatile concentrates of the tea infusions were prepared prior
GC–MS analysis by three sample preparation techniques: headspace
solid-phase microextraction (HS-SPME), solvent extraction followed by
solid-phase extraction (SPE), and solvent extraction after solvent as-
sisted flavor evaporation (SAFE). The aroma enrichment methods
covered a wide range of compounds with various volatilities.
Headspace solid-phase microextraction is a sensitive method for highly
volatile compounds such as acetaldehyde, dimethyl sulfide, etc., which
have no or low recovery by the other two methods. Due to the selective
adsorption of fiber coatings, the relative concentration of compounds
enriched by SPME have significant deviation from the volatile compo-
sition in the headspace. Therefore, a standard addition was applied for
the quantitative analysis of headspace SPME. Both volatile concentrates
prepared by SPE and SAFE had a slight difference in odor profile al-
though they smelled similar to the tea infusion. SAFE concentrate was
Table 1
Quality assessment of tea infusions of the experimental samples.
Tea Infusion
Green Tea Yellow Tea
Color green with yellow tint, yellow green, bright
brighter
Aroma clean, fresh, chest-nutty Chest-nutty, clean, slightly
sweet and cooked, fresh
Taste mellowish mellowish
Food Chemistry 285 (2019) 347–354
free of non-volatiles and suitable for GC cold on-column injection. Split
and splitless injections were employed for SPE concentrates and
achieved better sensitivity for less volatile compounds. All procedures
were repeated at least three times for each sample.
A total of 168 volatile compounds were identified in the tea infu-
sions. The total volatile concentration varies widely among the tea
samples from the six types of processing (Fig. 2). Processes for oolong,
white, and black teas with predominant tea leaf endogenous enzyme-
mediated oxidations produced aroma at concentrations ranging from 91
to 710 µg/g. Yellow and dark tea processes formed a lower aroma
concentration at the level of 55–81 µg/g, while aroma from green tea
process had the lowest concentration of 20 µg/g. It is interesting to note
that the largest number of aroma compounds was detected in the oo-
long tea infusion although its overall aroma concentration is sig-
nificantly lower than that of white and black teas. Top ten compounds
with the highest concentration for each tea type are listed in Table 2a.
3.3. Aroma formation in the manufacturing processes
Fresh tea leaves contain only a minute amount of aroma compounds
in free form which is stored in the glands of the plant. The majority of
aroma molecules in finished tea leaves is generated from their pre-
cursors during processing (Zheng, Li, Xiang, & Liang, 2016; Yang,
Baldermann, & Watanabe, 2013). Ho et al. conducted a thorough re-
view and summarized the formation mechanism of aroma molecules
during the tea manufacturing process (Ho, Zheng, & Li, 2015). The
authors concluded that the major aroma molecules are produced from
four main groups of precursors: carotenoids, lipids, glycosides, and
amino acids/carbohydrates.
During tea processing, the oxidation of flavanols induced by ca-
techol oxidase generates strong oxidizing quinones (Wang & Ho, 2009)
which lead to Strecker degradation of amino acids and the oxidation of
carotenoids. Non-enzymatic oxidations can also occur for the de-
gradation of carotenoids, amino acids, and carbohydrates. Glycoside
hydrolases hydrolyze the glycosidic linkage of glycosides and release
the free aglycon. Oxidation of fatty acids can be induced by lipox-
ygenase or initiated by free radicals. Thus, the important reaction
pathways for aroma formation during tea processing can be coarsely
grouped into three types: a) Maillard reaction and degradation of amino
acids and hydrocarbons, b) enzyme-mediated oxidation and hydrolysis,
as well as c) non-enzymatic degradation and oxidation. We categorized
precursor molecules into five groups: i) carotenoids, ii) fatty acids, iii)
glycosides, iv) amino acids/carbohydrates, and v) other precursors in-
cluding isoprenoids, polyphenols, and unknowns.
In order to understand the aroma formation in the tea processes, the
volatile molecules identified in this study were assigned to their pre-
cursor groups based on literature (Ho et al., 2015; Ravichandran &
Parthiban, 1998; Sanderson & Graham, 1973; Yamanishi & Kobayashi,
1999; Yang et al., 2013; Zheng et al., 2016; Zheng et al., 2015). Fig. 3a
shows the volatile concentration by tea types and precursor groups and
Dark Tea Oolong Tea White Tea Black Tea
darker orange red, orange yellow, orange yellow, bright orange red, bright
cloudy, brighter brighter
slightly stale and rough floral, slightly sweet floral, honey-like fruity, pure
fermented
slight mellow, astringent thicker body, mellow, thicker body, sweet mellow, thicker
mellowish slightly astringent body
350
Z. Feng et al.
Fig. 2. Overall aroma concentration (upper, µg/g tea leaves relative to internal standard) and the number of volatile compounds (lower) identified in the six tea
samples.
along with a PCA plot of the tea types and a cluster diagram of the
precursors. Black and white teas have the highest volatile concentration
from each precursor groups. Both are very different in composition and
form two distinctive groups in the PCA plot. The precursor groups fatty
acids and glycosides produce more or less comparable volatile con-
centration for the teas and show a close similarity in the cluster dia-
gram. From carotenoids and other groups, a lower volatile concentra-
tion was formed. The amino acids/carbohydrates group is more distant
from the other four precursor groups. Due to the high volatile con-
centration of black and white teas, the other four tea types are lumped
together in the PCA plot. To reveal the compositional differences of the
tea types, relative concentration was used for PCA and cluster analysis
(Fig. 3b). The results show that all tea samples do have their own
characteristic volatile composition and the differences in precursor re-
actions. Except for white tea, oxidation/degradation of fatty acids
generated the largest amount of volatile compounds for the other five
tea types. Hydrolysis of glycosides, which formed the highest con-
centration of volatile compounds for white tea, was the second im-
portant precursor reactions. Maillard reactions generated a substantial
amount of volatile compounds for green and yellow teas. Although
carotenoids and other precursors formed the lowest volatiles con-
centrations, it does not mean they are unimportant. Potent compounds,
even in trace amounts, can have significant impact to the overall aroma
character.
Contribution of an aroma molecule to the sensory profile of a pro-
duct can be approximated by its odor activity value (OAV), which is
expressed as the quotient of the concentration of the molecule over its
odor threshold value. To obtain OAV, the absolute concentration of the
odorant is necessary. More often in practice, however, the relative
concentration of aroma compounds is more important than the accurate
quantitative measurement of individual compounds, because it is the
ratio of aroma molecules determining the overall flavor profile.
Therefore, the relative value of OAV, expressed as aroma character
impact (ACI), is more useful to compare aroma contribution of in-
dividual components in a mixture (Yang, 2008). ACI is based on relative
concentration and therefore the determination of absolute concentra-
tion of individual compounds is unnecessary. Although ACI value is by
no means an accurate measurement of aroma contribution of a com-
pound, it can be considered an estimated impact rank of odor active
compounds.
The top ten compounds with highest aroma character impact value
351
Food Chemistry 285 (2019) 347–354
for each types of tea are listed in Table 2a. Four aroma molecules in the
top ten list are common for all six types of tea: β-ionone (warm, woody,
fruity), 2-pentylfuran (fruity, green, earthy, beany), hexanal (grassy,
green), and (E, Z)-2,6-nonadienal (cucumber-like, violet leaf). Com-
paring Table 2a with Table 2b, it should be mentioned that only 7 out of
34 compounds in the top ten for each type of tea are in common, i.e.,
dimethyl sulfide, hexanal, (E, E)-2,4-heptadienal, (E)-2-hexenal, phe-
nylacetaldehyde, indole, and linalool. Thus, high concentrations do not
necessarily mean high impact to the overall odor profile.
A heatmap was constructed based on the ACI value of the aroma
compounds for each types of tea. In the horizontal axis, the ACI value
was sorted by the results of a hierarchical cluster analysis of the aroma
compounds. In the vertical axis, the types of tea were arranged by the
cluster grouping. A bar chart of the precursor groups was aligned with
the heatmap by the aroma compounds (Fig. 4). Aroma contributions of
individual compounds for each types of tea are illustrated in the ACI
heatmap and corresponding precursor assignment. As expected, black,
white, and oolong teas through endoenzyme-predominated processes
form a cluster in the ACI composition. Green and yellow teas are si-
milar. The aroma of dark tea is produced mainly from ectoenzymatic
reactions which is distantly similar with the cluster of endoenzymatic
processes. Cluster analysis for the aroma components indicates that the
key odorants have a close similarity among the six types of tea. For each
type, the aroma character-determining molecules are a small group of
identified compounds: the top ten compounds with the highest ACI
factors contribute 86% (black tea) – 96% (green tea) to the overall
aroma (Table 2b). This also agrees with the analysis that characteristic
ratios of aroma are composed only about 3 to 40 key odorants for each
food (Dunkel et al., 2014). Nevertheless, the compounds with small
value of ACI are not necessarily non-contributors. They could play an
important role collectively in supporting the aroma character by further
modulating and shaping the overall aroma profile. The ACI heatmap
associated with the precursor bar-map shows that aroma components
with significant ACI value are almost always associated with one or
more of the four precursor groups carotenoids, fatty acids, glycosides,
and amino acids/carbohydrates. There are two interesting clusters
which are composed with a single compound. The first one is dimethyl
sulfide which is the most important contributor for green, yellow, and
oolong teas. The second one is β-ionone which has the highest ACI
value for dark, white, and black teas (Table 2b).
Concentration of aroma compounds in the finished tea leaves and
Z. Feng et al. Food Chemistry 285 (2019) 347–354

Table 2
Top ten compounds (number in bold denotes the top ten for each tea type).
a) Compounds with the highest concentration (µg/g tea leaves relative to internal standard) in each types of tea

Compound Green Yellow Dark Oolong White Black Precursor

(E)-2-Hexenal 0.04 0.00 0.07 6.2 24.6 113 Fatty Acids

Hexanal 1.0 1.7 2.4 9.9 34.7 48.7 Fatty Acids
n-Hexadecanoic acid 3.6 22.2 4.9 1.11 0.60 Fatty Acids
Hexanoic acid 0.12 0.25 0.80 1.78 16.1 16.53 Fatty Acids
(E,E)-2,4-Heptadienal 0.23 3.1 0.48 2.1 7.83 9.29 Fatty Acids
Stearic acid 0.6 2.4 3.9 5.4 6.09 6.83 Fatty Acids
(E,Z)-2,4-Heptadienal 0.21 2.0 3.4 1.17 3.76 3.17 Fatty Acids
2-Butanone 1.1 2.4 0.79 0.48 1.81 3.28 Fatty Acids
(Z)-3-Hexen-1-ol 0.06 0.73 0.02 1.06 30.2 51.0 Fatty Acids, Glycosides
(E)-Linalool oxide (furanoid) 0.15 0.59 4.9 2.1 58.6 67.6 Glycosides
Benzyl alcohol 0.10 0.34 0.07 3.3 18.4 38.6 Glycosides
(Z)-Linalool oxide (furanoid) 0.36 1.53 0.70 29.2 28.5 Glycosides
(E)-Linalool Oxide (pyranoid) 0.35 0.55 3.0 2.8 7.69 18.2 Glycosides
(Z)-Linalool oxide (pyranoid) 0.07 0.80 3.5 1.03 9.49 4.30 Glycosides
2-Phenylethanol 0.32 1.26 0.08 6.9 31.8 62.0 Glycosides, AA/Carb.1
Linalool 0.34 1.11 0.26 1.74 49.4 26.2 Glycosides, Carotenoids
Coumarin 1.6 5.1 0.19 1.84 2.10 6.85 Glycosides, Others
Phenylacetaldehyde 0.26 0.45 1.70 3.1 19.8 50.1 AA/Carb.1
1,1-Diethoxyethane 2.2 5.7 2.3 1.65 3.40 6.13 AA/Carb.1
Dimethyl sulfide 2.0 1.9 0.02 1.66 1.23 1.51 AA/Carb.1
Indole 1.1 1.71 0.11 0.75 0.10 0.35 AA/Carb.1
Dihydroactinidolide 0.9 4.7 8.5 1.75 9.78 10.78 Carotenoids
4-Vinylphenol 0.6 1.41 4.9 1.17 2.80 3.86 Others
2,4-Bis-[1,1-dimethylethyl]-phenol 0.6 0.57 0.65 0.66 1.03 0.00 Others
Overall Concentration 20 55 81 91 470 710
Top ten concentration% 60% 60% 73% 51% 67% 71%

b) Compounds with the highest ACI factor in each types of tea

Compound Green Yellow Dark Oolong White Black Precursor Cluster2 Odor Description

2-Pentylfuran 0.01 0.03 0.30 0.08 0.14 0.12 Fatty Acids 3 fruity, green, earthy, beany
(E,E)-2,4-Nonadienal 0.04 0.000 0.10 0.17 0.03 Fatty Acids 3 fatty, green-vegatable, cucumber
Hexanal 0.02 0.02 0.04 0.08 0.09 0.12 Fatty Acids 3 grassy, green
(E,Z)-2,6-Nonadienal 0.02 0.05 0.06 0.09 0.10 0.10 Fatty Acids 3 cucumber-like, violet leaf
(E,E)-2,4-Decadienal 0.03 0.10 0.04 0.03 0.04 Fatty Acids 4 fatty, oily, green, chicken-like
(E)-2-Hexenal 0.000 0.000 0.000 0.014 0.017 0.07 Fatty Acids 4 green-fruity, pungent vegetable-like
Decanal 0.005 0.05 0.03 0.03 0.02 0.030 Fatty Acids 4 sweet, aldehydic, waxy, floral
(E,E)-2,4-Heptadienal 0.004 0.04 0.007 0.016 0.019 0.020 Fatty Acids 4 fatty, green, oily, aldehydic, vegetable
Nonanal 0.03 0.04 0.02 0.02 0.014 0.03 Fatty Acids 4 fatty-floral, waxy
Heptanal 0.01 0.010 0.02 0.013 0.007 0.017 Fatty Acids 5 Potato-like, Fatty, Rancid-like
Octanal 0.01 0.012 0.007 0.010 0.005 0.008 Fatty Acids 5 aldehydic, waxy, citrusy, green, fatty
Linalool 0.001 0.003 0.001 0.003 0.02 0.011 Glycosides, Carotenoids 5 floral, woody, citrusy
Dimethyl sulfide 0.68 0.43 0.004 0.23 0.05 0.06 AA/Carb.1 1 radish, sharp, green, cabbagelike
Phenylacetaldehyde 0.005 0.006 0.02 0.03 0.05 0.11 AA/Carb.1 4 honey-like, floral, sweet
Indole 0.01 0.008 0.001 0.002 0.000 0.000 AA/Carb.1 5 Animal-like, floral
β-ionone 0.16 0.21 0.32 0.19 0.22 0.17 Carotenoids 2 warm, woody, fruity
Limonene 0.01 0.004 0.02 0.000 0.000 0.003 Others 5 fresh, light sweet, citrusy
Sum of top ten ACI 0.96 0.92 0.93 0.91 0.90 0.86
1
Amino acids/carbohydrates.
2
Cluster group number (see Fig. 4).

infusions is determined by their formation, chemical transformation,
and evaporation rates during the process. It reflects the various reaction
intensity of the tea processes by comparing the final concentrations vs.
processing time. In the processes of green, yellow, and dark teas, there
appears to be a close association between concentration and processing
time (Table 2a). The concentrations of phenylacetaldehyde and β-Io-
none increase slowly with the processing time from green tea to dark
tea. Many compounds have the same increasing trends. Conversely,
dimethyl sulfide, indole and a few other compounds decreased their
concentration with processing time, indicating that these compounds
formed at the early stage and the evaporation and/or chemical trans-
formation rate is higher than their formation. Black tea process is short
but generates the highest concentration of many aroma compounds due
to the optimal endoenzymatic reaction conditions. It should also be
noted that the white tea process has a faster aroma formation rate than
that of yellow tea and dark tea within the processing time. This
indicates that the tea endoenzymes greatly accelerated the reactions
although under mild conditions in terms of temperature, moisture, and
the least bruised leaves. White tea is the least processed tea and often
referred as “non-fermented” or “slightly fermented” (Jiang, 2009). Al-
though tea fermentation can catalyze the reactions of aroma precursors,
it is not the predominant pathway to form aroma compounds in white
tea process. Our experiment results indicate large amount of volatile
compounds was generated mainly from the oxidation of fatty acids and
the hydrolysis of glycosides during the white tea process (Table 2a).
4. Conclusions
The model tea manufacturing processes produced finished tea
leaves with the typical sensory profile of the commonly known six types
of tea using the same fresh tea leaves. The character impact aroma
molecules are mainly formed from four groups of precursors:

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Fig. 3. Volatile composition of six types of tea by precursor groups.

glycosides, fatty acids, carotenoids, as well as amino acids and carbo- Acknowledgement
hydrates. Oxidation, hydrolysis, and degradation reactions of the pre-
cursor molecules are greatly accelerated by the tea endogenous en- We sincerely thank Shihui Fang for his crucial technical support in
zymes. Thus, fast formation rate leading to high aroma concentration the tea process design and experiment execution.
was observed in the processes of black tea and white tea, in which
enzyme mediated oxidations are predominant. Maillard reactions plays
Funding
an important role in aroma generation in green tea and yellow tea
processes, while oxidations of fatty acids in dark tea and oolong tea
This work was financially supported by the Agriculture Research
processes, and hydrolysis of glycosides and degradation of fatty acids in
System of China (CARS-19), the State Key Laboratory of Tea Plant
white tea and black tea processes.
Biology and Utilization, China, and Anhui Agricultural University,

Fig. 4. ACI heatmap and precursor groups.

353
Z. Feng et al.
China.
Declaration of interests
The authors declare that they have no known competing financial
interests or personal relationships that could have appeared to influ-
ence the work reported in this paper.
Appendix A. Supplementary data
List of compounds identified in the six types of tea, their CAS
numbers, concentrations, GC retention indices, and precursors.
Supplementary data to this article can be found online at https://doi.
org/10.1016/j.foodchem.2019.01.174.
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