J Food Sci Technol
https://doi.org/10.1007/s13197-020-04474-7
ORIGINAL ARTICLE
Analysis of flavor and taste attributes differences treated
by chemical preservatives: a case study in strawberry fruits
treated by 1-methylcyclopropene and chlorine dioxide
Xiaoying Yang1 • Ran Yan1 • Qingmin Chen2 • Maorun Fu1
Revised: 28 March 2020 / Accepted: 23 April 2020
Ó Association of Food Scientists & Technologists (India) 2020
Abstract Flavor and taste attributes of fruits varied by
different preservatives treatments. Changes in sugars,
organic acids, amino acids as well as volatiles of straw-
berries treated with 1-methylcyclopropene (1-MCP) and/or
chlorine dioxide (ClO2) were evaluated during storage
period in this study. Our results revealed that the decreases
of tartaric acid, malic acid, citric acid, titratable acidity
(TA), sucrose and soluble sugar contents were significantly
inhibited by 1-MCP ? ClO2. The fructose and glucose
contents of all groups remained stable and slightly
increased at the last period of 10 days. However, different
treatments had no influence on content of succinic acid.
Moreover, the highest sweet taste (77.37 mg 100 g-1 fresh
weight) and lowest bitter taste (3.44 mg 100 g-1 fresh
weight) free amino acids (FAA) were observed in the
strawberries treated by 1-MCP combined with ClO2 treat-
ment as compared to other treatments and control. (E)-2-
hexenal was the most abundant volatile and showed a
significant increase trend during strawberry storage. More
interestingly, ethyl butyrate, fruit-like aroma, could be
recovered in content by 1-MCP, ClO2 alone and their
Xiaoying Yang and Ran Yan ontributed equally to this work.
Electronic supplementary material The online version of this
article (https://doi.org/10.1007/s13197-020-04474-7) contains sup-
plementary material, which is available to authorized users.
& Maorun Fu
skyfmr@163.com
1
School of Food Science and Engineering, Qilu University of
Technology (Shandong Academy of Sciences), Jinan,
People’s Republic of China
2 (Chen et al. 2014). Besides sugars, organic acids and
Department of Food Science and Engineering, Shandong
Agriculture and Engineering University, Jinan, People’s
Republic of China
combination treatment. Compared with other treatments,
the significant different flavor in ClO2 treatment was
identified by principle component analysis. In addition,
methyl hexanoate and 4-methoxy-2,5-dimethylfuran-
3(2H)-one (DMMF) were the major factors that affected
the volatile organic compounds (VOCs) of strawberries
through the whole storage. Taken together, 1-MCP coupled
with ClO2 could be a complex preservative to maintain
strawberries quality by regulating the flavor and taste
attributes.
Keywords Strawberry  Flavor  1-MCP  ClO2  Vocs
Introduction
Strawberries (Fragaria 9 ananassa) are cultivated all over
the world and consumed for their nutritional value, color,
and flavor (Enomoto et al. 2018; Hajji et al. 2018; Zhang
et al. 2018). However, strawberries are perishable and have
a short shelf life as they are prone to physical and bio-
chemical damages during harvest and transportation
(Pagliarulo et al. 2016). The quality attributes of straw-
berries are based on various indices, for instance, size,
firmness, and flavor (Van de Poel et al. 2014; Vanden-
driessche et al. 2013). Among them, the flavor of fruit,
perceived by odor, taste, and mouth feel, mostly depends
on the balance between sugars, organic acids, VOCs and
FAA. Sugars and organic acids are the two key chemicals
that affect the internal quality of fruits (Goldenberg et al.
2014). The FAA contributes to the food taste by regulating
the corresponding human taste receptor differentially
FAAs, which are vital to the good taste of fruit, the vola-
tiles are the primary factor determining the unique aroma
123

of strawberries. Approximately 360 different VOCs were
detected in strawberries including aldehydes, alcohols,
esters and ketones. (Abonyi et al. 2002; Urruty et al. 2002;
Van de Poel et al. 2014). Among the VOCs, methyl or ethyl
esters of butyric acid, hexanoic acid, 4-methoxy-2,
5-dimethylfuran-3(2H)-one (DMMF) and 4-hydroxy-2,
5-dimethylfuran-3-one (DMHF) are the major volatiles in
strawberry fruit (Zabetakis and Holden 1997; Zhang et al.
2009a). All those flavor components have a great impact on
acceptance of consumers. Consequently, it is very impor-
tant to evaluate the flavor changes in postharvest
strawberries.
1-methylcyclopropene (1-MCP), an ethylene inhibitor,
has been widely used to extend the shelf life of fruit and
vegetables (Li et al. 2016). Chlorine dioxide (ClO2), a
disinfectant, has a strong oxidation capacity at low con-
centration and wide pH range (Yan et al. 2019). When
application, ClO2 can transform to much chloride, little
chlorite and/or chlorate. And there are no lasting chemical
residues (ClO2, ClO2- and ClO3-) in fruits and vegetables.
(Praeger et al. 2016; Yan et al. 2019). It was reported that
no residues were detected in plums even after 10 min
treated by ultrasound and 40 mg L-1 ClO2 (Chen & Zhu
2011). Currently, both 1-MCP and ClO2 are used as
preservatives to retain fruit quality during storage, such as
bananas (Zhu et al.2019), mango (Razzaq et al. 2016),
longan fruit (Saengnil et al. 2014) and apples (Remorini
et al. 2015). The effectiveness of 1-MCP alone and in
combination with ClO2 on the internal quality of straw-
berries was evaluated by our pervious study (Yang et al.
2018). However, there are very limited studies about effect
of individual and combination treatment of 1-MCP and
ClO2 on the flavor of strawberries, especially on changes of
VOCs and FAA during storage period.
In this study, we evaluated the effect of 1-MCP and
ClO2 alone and the combination on the flavor attributes of
strawberries. The results revealed regulation capacity of
different treatments on strawberry flavor, and provided an
angle for development of strawberry preservation. More-
over, we think it is necessary to design a portable device
for quality judgement. The device can be equipped with a
smell sensor, responding to a special VOC in strawberries
treated by 1-MCP and/or ClO2 during storage.
Materials and methods
Fruit and chemicals
Strawberries (Fragaria 9 ananassa Duch. cv. Tianbao)
were hand-picked at commercial ripening stage from an
orchard located at Changqing District, Jinan, China
(116°780 E, 36°580 N). The fruits uniform in size, shape, and
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J Food Sci Technol
color, without any wounds or rot were transported to the
laboratory immediately. 240 fruits were randomly divided
into four groups (60 fruits for per group, 3 replicate in per
group, 20 fruits for per replicate).
The chemical standards of phenolphthalein, (NH4)2-
HPO4, NaCl, anthrone, sulfosalicylic acid, H3PO4 and
NaOH were obtained from Sinopharm Chemical Reagent
Co., Ltd (Beijing, China). H2SO4, ethyl acetate, ethanol
and acetonitrile (HPLC grade) were purchased from Fuyu
Fine Chemical Co., Ltd (Tianjin, China).
Tartaric acid, malic acid, citric acid, succinic acid,
glucose, sucrose and fructose were obtained from Yuanye
Bio-Technology Co., Ltd (Shanghai, China). L-Gly, L-Ala,
L-Cys, L-Val, L-Met, L-Glu, L-Asp, L-Thr, L-Ser, L-Ile,
L-Leu, L-Tyr, L-Phe, L-Lys, L-His, L-Arg and L-Pro were
purchased from Sigma-Aldrich (St. Louis, MO, USA).
Fruit treatments
ClO2 solid-releasing tablets were prepared following the
methods of Yang et al. (2018). In brief, the tablets were
mixed by ClO2 powder (Beijing Golden Pool Scientific Co.
Ltd, Beijing, China), b-cyclodextrin, and b-zeolite in a
ratio of 1:3:1 (w w-1). Four groups of fruit were conducted
by four different treatments for 24 h at 20 °C. 1-MCP
treatment group was placed in a sealed plastic containers
with 1 lL L-11-MCP (AgrofreshSM, USA). ClO2 treatment
group was put in a sealed plastic containers with 33 lL L-1
ClO2 (1 g tablets). Combination treatment group was car-
ried out by using 1 lL L-1 1-MCP ? 33 lL L-1 ClO2.
Control group was stored directly without any treatment.
After treatment, all groups were stored at 4 °C and 90%
relative humidity. Sample collection was conducted at 0, 2,
4, 6, 8 and 10 d. At each sampling time, six strawberries
were selected from each treatment group to determine
firmness and total soluble solids (TSS). After that, the rest
strawberries were cut into pieces, packaged with tinfoil,
frozen in liquid N2 immediately, and then stored at –80 °C
for further analysis.
Determination of firmness, TSS, titratable acidity
(TA) and soluble sugar
Fruit firmness was determined using a hand-held firmness
tester (GY-2, Tuopu Co. Ltd, Zhejiang, China). The firm-
ness was expressed in terms of newton (N). TSS was
measured with strawberry fruit juice using a brix meter
(PAL-1, Atago Co. Ltd, Tokyo, Japan). The TSS was
expressed as %. Titratable acidity was determined using
0.01 M NaOH and phenolphthalein indicator. The titrat-
able acidity was calculated using the following formula
(Hajji et al. 2018):
J Food Sci Technol
Titratable acidity (%) =V1 VNaOHV20:010:064
m  100
where, V1 is the total volume of fruit juice, V2 is the volume
of juice required for titration, VNaOH is the volume of
NaOH consumed in titration, 0.01 is the molarity of NaOH,
0.064 is the conversion factor of citric acid, and m is the
fruit mass required for titration analysis.
Soluble sugar was determined according to Langer et al.
(2018) with minor modifications. Briefly, 1.5 g strawberry
fruit from each group was crushed using mortar and pestle.
Next, 0.25 g crushed fruit was weighed and added to
10 mL distilled water. The homogenate was boiled for
30 min in boiling water and cooled to room temperature
(25 °C) for filtration. The supernatant was diluted using
25 mL distilled water. Further, 0.2 mL sample was mixed
with 0.5 mL anthrone solution (1 g anthrone dissolved in
50 mL ethyl acetate) and 5 mL of 98% H2SO4 (v v-1) in
1.8 mL distilled water. The mixture was heated for 1 min
at 100 °C and cooled to 25 °C. The absorbance of the
solution was measured at a wavelength of 630 nm using a
spectrophotometer (V-1100D, Meiouda Co. Ltd, Shanghai,
China). The soluble sugar content was expressed as %.
Extraction and determination of organic acids
The extraction and determination of organic acids (tartaric,
malic, citric, and succinic acid) were performed following
the protocols of Zheng et al. (2016) with minor modifica-
tions. Briefly, 2 g fruit from each group was grinded into
powder. The powder was homogenized with 5 mL of 80%
ethanol and incubated for 20 min at 35 °C. The homo-
genate was centrifuged twice at 10,000 9 g for 10 min at
20 °C. The supernatant was diluted to 25 mL using dis-
tilled water. Next, 2 mL of the diluted sample was evap-
orated under vacuum at 45 °C using a rotary evaporator
(RE-52AA, Yarong Co. Ltd, Shanghai, China) to remove
ethanol completely. The vacuum-dried extract was dis-
solved in 1 mL distilled water, and filtered through a syr-
inge filter (U 0.2 lm, u 13 mm).
For high performance liquid chromatography (HPLC)
analysis, 20 lL sample was injected into the ODS-C18
column (200 mm 9 4.6 mm, 5 lm) in an HPLC instru-
ment (LC-20AT, Shimadzu, Japan). The mobile phase was
50 mM (NH4)2HPO4, whose pH was adjusted to 2.7 using
H3PO4. The flow rate was 0.5 mL min-1. The levels of
individual organic acids were measured at a wavelength of
210 nm by the ultraviolet detector (SPD-20A). The organic
acids contents were expressed as mg per g fresh weight
(mg g-1 FW).
Extraction and analysis of sugar components
The extraction and determination of sugar components
(glucose, fructose and sucrose) were performed by the
methods of Terry et al. (2007). Briefly, 1.5 g powdered
sample was added to 40 mL of 80% (v v-1) ethanol. The
mixture was subjected to ultrasound extraction for 30 min.
After filtered, the supernatant was evaporated under vac-
uum at 50 °C using a rotary evaporator. The residue was
dissolved in 10 mL of acetonitrile and water (70:30; v
v-1). Finally, the solution was filtered through a 0.45 lm
filter (U 0.45 lm, u 13 mm).
For HPLC analysis, 20 lL sample was loaded into the
NH2 column (5 lm, 250 mm 9 4.6 mm) in an HPLC
instrument (LC-20AT, Shimadzu, Japan) equipped with
refractive index detector (RID-10A). The flow rate and
oven temperature were 0.7 mL min-1and 40 °C. The
mobile phase used for HPLC analysis was acetonitrile and
water (80:20; v v-1). The concentrations of sucrose, glu-
cose, and fructose were expressed as mg g-1 FW.
FAA analysis
FAA content was analyzed following the protocols of Song
et al. (2019) with minor modifications. Briefly, 1 g straw-
berry fruit was homogenized with 5 mL of 4% (v v-1)
sulfosalicylic acid using mortar and pestle. The homo-
genate was centrifuged at 10,000 9 g for 15 min at 4 °C.
The supernatant was diluted by fivefold using distilled
water. The diluted sample was filtered using 0.22 lm
aqueous phase membrane filters. The amino acid content in
20 lL sample was analyzed using the Hitachi model
L-8900 amino acid analyzer (Hitachi Co. Ltd, Tokyo,
Japan). The amino acid contents were expressed as mg
100 g-1 FW.
VOCs analysis
HS-SPME (headspace solid phase micro-extraction)
analysis
The VOCs were extracted by HS-SPME with a carboxen/
polydimethylsiloxane (75 lm, CAR/PDMS) fiber (Zhang
et al. 2009b). Strawberry fruit (3 g) was powdered in liquid
nitrogen. The powdered samples (3 g) were transferred into
15 mL headspace bottle, sealed and 2 g of NaCl was added
with continuous heating and agitation for 30 min. The
extraction method was same as Zhang et al. (2009b) with
slight modifications. The samples were placed into the
injection port of gas chromatography-mass spectrometer
(QP2010Plus, Shimazu, Japan) and desorbed for 3 min at
250 °C.
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GC–MS (gas chromatography–mass spectrometry)
analysis
Gas chromatography-mass spectrometer equipped with a
DB-WAX capillary column (30 m 9 0.25 mm 9 0.25
lm) was used to separate and detect the complex VOCs
from strawberries according to the procedure reported by
Zhang et al. (2015) with minor modifications. The GC oven
temperature was from 40 °C (held for 3 min) to 140 °C at
5 °C min-1 and held at 250 °C for 5 min at 10 °C min-1.
The carrier gas was Helium at a flow rate of 1.0 ml/min
with the splitless GC inlet mode. The MS spectral using
full-scan mode was obtained by electronic impact (EI)
mode at 70 eV and scan range was 35–650 m/z. The
transmission line and source temperatures were 250 °C and
200 °C, respectively. The VOCs of strawberries were
identified by comparing the retention index (RI) with mass
spectra NIST11 library in the GC–MS data system. VOCs
with matching degree over 85% were exhibited in Table 3.
Statistical analysis
All data were analyzed using SPSS 20.0 software (SPSS
Inc., Chicago, IL, USA) and Microsoft Office Excel 2016.
One-way ANOVA followed by Duncan’s test was used to
determine difference between control and treatment group.
Principal component analysis (PCA) was conducted with
Matlab R2018a software. All experiments were performed
with three replicates and the data were expressed as
mean ± standard deviation.
Results and discussion
Storage index changes of strawberries in different
treatments
The firmness changes of strawberry fruit during storage are
presented in Fig. 1a. The firmness decreased remarkably
both in treated and control group fruit with prolong storage
time. Compared with control, the firmness of treated
groups was well maintained during storage. By the end of
storage (10 d), the firmness of control fruit (1.70 N) and
ClO2 (1.76 N) were significantly lower compared with that
of 1-MCP (1.90 N) and 1-MCP ? ClO2 treatment (1.98 N)
(P \ 0.05). The results indicated that 1-MCP alone and
combined with ClO2 treatment delayed the reduction of
fruit firmness, which was agreed with previous reports,
such as strawberries (Yang et al. 2018), plum (Chen & Zhu
2011).
TSS is an important index for quality evaluation and
contributes to the sweet flavor of strawberries. The TSS
content of treated and control strawberries are shown in
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J Food Sci Technol
Fig. 1b. A sudden drop was observed in the TSS content of
control group after 4 d, while that of other treatment groups
declined less until 10 d. The decrease of TSS in fruit was
responsible for the respiration consumption as a substrate
(Petriccione et al. 2015). TSS contents in the whole storage
among different groups was 1-MCP ? ClO2 [ 1-
MCP [ ClO2 [ control group. By the end of storage (10
d), the TSS content of 1-MCP ? ClO2 group was signifi-
cantly higher than other treated and control groups
(P \ 0.05). However, Yang et al. (2018) reported that
1-MCP combined with ClO2 treatment had no effect on
TSS of strawberries compared with control.
TA variations of the treated and control fruit during
storage are shown in Fig. 1c. The TA of all three treatment
groups decreased slightly in the whole storage period
compared with control group. At 10 d, the TA content of
control group (0.19%) was lower than that of 1-MCP, ClO2
and 1-MCP ? ClO2 (0.20, 0.21 and 0.24%, respectively).
In addition, the TA content of combination treatment
showed a significant difference compared with other
groups at the end of storage (P \ 0.05). This indicated that
1-MCP ? ClO2 was able to delay the reduction of TA in
strawberry, which is associated to inhibit the respiration
consumption and microorganism growth (Hajji et al. 2018;
Tadesse & Abtew 2016).
Soluble sugar is not only the products of photosynthesis,
but also the substrates for respiration. The increase of
respiration was accompanied by the decrease of soluble
sugar, causing the deterioration of postharvest fruit and
vegetables quality (Yang et al. 2009). A decrease in the
soluble sugar content was observed in all groups during
storage period (Fig. 1d). At 4 d, the soluble sugar content
of control group (4.49%) was remarkably lower than ClO2
one (4.57%), 1-MCP group (4.77%) and 1-MCP ? ClO2
group (4.90%). This difference lasted for the end of stor-
age. This indicated that 1-MCP ? ClO2 and 1-MCP
maintained the soluble sugar content of strawberry fruit
during storage. The result was similar to the report by Xu
et al. (2018) who found that the application of 1-MCP
alone and combined with nano-packing was beneficial to
maintain the soluble sugar in pleurotus eryngii.
Effects of 1-MCP or/and ClO2 on organic acids
in strawberries during storage
Organic acids, related to fruit acidity, are important attri-
butes of flavor. Combined with sugars, organic acids have
an impact on the sensory quality of strawberry (Gündüz
and Özdemir, 2014). Organic acids accumulate during fruit
growth and development, and are consumed as substrates
in the tricarboxylic acid cycle (TCA cycle) for respiration
at maturity stage. As shown in Fig. 2, among four organic
acids (tartaric, malic, citric and succinic acid), tartaric acid
J Food Sci Technol

Fig. 1 Effect of 1-MCP and/or

ClO2 on storage index of
strawberries during storage:
a Firmness; b Total soluble
solid (TSS);
(c) Titratable acidity (TA);
(d) Soluble sugar. Each
treatment involved the three
replicates. And the standard
deviations of the averages were
represented by bars. Different
uppercase letters indicate
significant difference within a
group at a same day, different
lowercase letters indicate
significant difference between
groups (P \ 0.05)

was the predominant organic acid in strawberries, followed
by malic, citric and succinic acid. In previous study, citric
acid was found to be the predominant organic acid (Hwang
et al. 2019). This may be explained by different strawberry
varieties (‘Seolhyang, Jan-ghee, and Maehyang’) used in
two studies.
As shown in Fig. 2a, the content of tartaric acid showed
a decrease tendency during fruit storage. However, a small
increase of tartaric acid in control was observed at 4 d,
followed by a decrease until the end of storage. While the
content of tartaric acid in treatment groups increased
slightly at 6 d of the storage and then reduced. This indi-
cated that the treatments delayed the reduction of tartaric
acid during storage period. At day 8, there was a significant
difference between treated and control fruit (P \ 0.05).
The content of tartaric acid in strawberries was ranged as
follows: 1-MCP ? ClO2 group [ 1-MCP group [ ClO2
group [ control group. Additionally, the tartaric acid
content of 1-MCP ? ClO2 group was 16.95 mg g-1 FW,
which was at least twice than that of control group
(7.79 mg g-1 FW) at 8 d. This indicated that the combi-
nation treatment prevented the degradation of tartaric acid,
maintaining the acidity of strawberry. The changes of
malic acid and citric acid contents were similar to those of
tartaric acid shown in Fig. 2b and c. At 6 d, malic and citric
acid content of 1-MCP ? ClO2 and 1-MCP group were
higher than control and ClO2 treatment group, though no
significant difference was found between 1-MCP and
1-MCP ? ClO2 group (P [ 0.05). However, at day 8,
1-MCP ? ClO2 was beneficial to maintain the content of
malic and citric acid in strawberries compared with control
and other treatments (P \ 0.05). The succinic acid content

123

Fig. 2 Effect of 1-MCP or/and ClO2 on organic acid metabolism of
strawberries during storage: a Tartaric acid; b Malic acid; c Citric
acid; d Succinic acid. Each treatment involved the three replicates.
And the standard deviations of the averages were represented by bars
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J Food Sci Technol
of strawberries was unaffected by 1-MCP, ClO2 alone and
their combination during whole storage (Fig. 2d). Hajji
et al. (2018) noted that organic acids could be used for the
growth of microorganisms. Especially, fungal infection
accelerates the losses of citric acid. However, ClO2, as a
disinfectant, can be more effective in compensating for the
deficiency of 1-MCP in inhibiting the microbial growth.
Thus, the combination of 1-MCP ? ClO2 maintained
content of tartaric, malic and citric acid in strawberries by
interact between two preservatives (Yang et al. 2018).
Effects of 1-MCP or/and ClO2 on sugar
in strawberries during storage
Sugars, responsible for sweet taste, are closely linked to
consumer acceptance (Jouquand et al. 2008). As the most
abundant soluble components in strawberries, the contents
of fructose, glucose and sucrose were detected by HPLC
during fruit storage (Fig. 3). The fructose and glucose
contents of all groups remained stable and slightly
increased at the last period of 10 days, while sucrose
content declined obviously during storage period. The
reason was that sucrose gradually transformed into
monosaccharides (fructose and glucose), taking part in
some physiological and biochemical reactions during fruit
storage (Caleb et al. 2016). As shown in Fig. 3a, the
fructose of 1-MCP ? ClO2 group was the highest among
all groups by the end of storage (day 10), and there was no
significant difference between ClO2 and control groups
(P [ 0.05). The difference in glucose content of treated
and control fruit started from day 8 (Fig. 3b). The glucose
content of strawberries treated by 1-MCP combined with
ClO2 was dramatically higher than control group at the
10th day. For sucrose, no remarkable difference was noted
between 1-MCP and 1-MCP ? ClO2 group at day 8, but
those sucrose content were significantly higher than ClO2
and control group (P \ 0.05) (Fig. 3c). The results were
consistent with previous report which discovered that
Japanese pear treated by 1-MCP inhibited the loss of
sucrose during cold storage (Itai and Tanahashi 2008).
FAA in strawberries under different treatments
during storage
The FAA contents of fresh (0 d) and stored strawberries (10
d) in control, 1-MCP, ClO2 alone and their combination
treatment group are presented in Table 1. Only 15 types of
FAA were determined in our study, whereas Pro and Met
were also detected in strawberry fruit (Zhang et al. 2014).
This may be due to the differences in the exaction method
and the variety of strawberries used in the two studies. In
comparison to fresh fruit (144.28 mg 100 g-1 FW), the
total FAA contents showed a decrease tendency at the end
J Food Sci Technol
Fig. 3 Effect of 1-MCP, ClO2 alone and the combination on organic
acid metabolism of strawberries during storage: a Fructose; b Glucose;
c Sucrose. Each treatment involved the three replicates. And the
standard deviations of the averages were represented by bars
of storage no matter in control or treated groups. Among
stored samples, the lowest FAA content was seen in the
ClO2 treatment group (91.75 mg 100 g-1 FW), while the
higher contents of 1-MCP ? ClO2, 1-MCP and control
reached 108.58, 105.74 and 102.81 mg 100 g-1 FW,
respectively. The most abundant FAAs were Thr and Ser,
accounting for 46–59% of the total FAA content, followed
by Asp, Glu and Ala. The contents of Ala and Phe in fresh
fruit were 17.53 and 3.62 mg 100 g-1 FW respectively,
which were approximately three and four-fold higher than
the stored samples.
The taste-active amino acids contributing to umami,
sweet, bitter, and none taste were detected in strawberries
after treatments (Xu et al. 2019). Umami taste amino acids
are Asp and Glu; sweet taste amino acids are Thr, Ser, Gly,
Ala and Pro; bitter taste amino acids are Val, Met, Ile, Leu,
Phe, His, and Arg and tasteless amino acids are Cys, Tyr,
and Lys (Table 2). All varieties of taste-active amino acids
exhibited a decreased tendency after storage. The sweet
and umami taste amino acids were the main taste-active
amino acids and they contributed over 90% to the whole
FAA contents. In addition, the combination treatment had
the highest sweet taste amino acids (77.37 mg 100 g-1
FW) and the lowest bitter taste amino acids (3.44 mg
100 g-1 FW) when compared to other treatment and con-
trol groups during storage period. This indicated that
1-MCP ? ClO2 was able to improve the quality of straw-
berries by regulating the flavor and taste attributes.
VOCs affected by 1-MCP or/and ClO2
in strawberries during storage
Table 3 shows the results of aroma compounds detected by
GC–MS in fresh and stored fruit with or without treatment.
A total of 54 VOCs was tested. Among them, the number
of VOCs both in 1-MCP ? ClO2 and 1-MCP group was
45, followed by 44 and 42 in ClO2 and control group
respectively at end of storage (10 d). And the lowest
number of VOCs (40) was detected in fresh fruit at 0 d.
Aldehydes were the most abundant compounds in
postharvest strawberries, which contributed 32.59, 84.53,
86.63, 66.27 and 75.87% in fresh, control, 1-MCP, ClO2
and 1-MCP ? ClO2 groups, respectively. However, the
content of ketones and acids was very low.
The most abundant volatile aldehydes in postharvest
strawberry were hexanal and (E)-2-hexenal, which impart
grass aroma in fruit (Song et al. 2019). The contents of
hexanal and (E)-2-hexenal in the control and other treat-
ment groups were significantly higher when compared to
those in fresh fruit (P \ 0.05). The content of hexanal in
1-MCP ? ClO2 was higher than other treatments and
control by the end of the storage. It was reported that
hexanal was effective in inhibiting the growth of spoilage
microbes (Seethapathy et al. 2016). Thus, the combination
treatment of 1-MCP and ClO2 was able to improve defense
system by inducing hexanal to delay senescence and pre-
vent quality deterioration in strawberries. Earlier studies
reported that (E)-2-hexenal synthesis was induced upon
exposure to high temperature, pest and disease infection to
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 J Food Sci Technol

Table 1 Free amino acids contents of fresh strawberry fruits (0 d) and stored samples (10 d)
Name Samples

mg/100 g FW Fresh Control 1-MCP ClO2 1-MCP ? ClO2

Asp 12.63 ± 0.73a 12.25 ± 0.59a 10.66 ± 0.30bc 9.98 ± 0.20c 11.67 ± 0.02ab
a ab a b
Thr 32.73 ± 3.32 26.19 ± 1.63 33.41 ± 5.07 18.83 ± 5.11 31.14 ± 1.28a
Ser 48.83 ± 8.02a 35.13 ± 2.48b 36.28 ± 1.29b 41.04 ± 2.02ab 39.72 ± 0.69ab
a ab b b
Glu 18.41 ± 3.57 14.93 ± 0.46 13.66 ± 0.24 12.16 ± 0.61 14.29 ± 0.35ab
a b b b
Gly 0.72 ± 0.11 0.29 ± 0.02 0.36 ± 0.00 0.36 ± 0.10 0.37 ± 0.00b
a b b b
Ala 17.53 ± 4.76 5.48 ± 0.75 6.22 ± 1.03 3.78 ± 1.45 6.14 ± 0.39b
a ab b c
Cys 1.36 ± 0.18 1.23 ± 0.05 1.10 ± 0.03 0.41 ± 0.02 1.13 ± 0.00ab
Val 1.94 ± 0.12a 1.66 ± 0.36a 1.45 ± 0.21a 1.74 ± 0.21a 1.68 ± 0.05a
a b d c cd
Ile 1.17 ± 0.18 0.84 ± 0.09 0.22 ± 0.07 0.54 ± 0.01 0.40 ± 0.03
b a bc cd d
Leu 0.72 ± 0.05 0.88 ± 0.06 0.61 ± 0.02 0.52 ± 0.07 0.44 ± 0.01
Tyr 1.76 ± 0.04a 1.30 ± 0.06b 0.46 ± 0.00d 0.58 ± 0.02c 0.56 ± 0.05c
Phe 3.62 ± 0.25a 0.91 ± 0.18b 0.21 ± 0.02c 0.17 ± 0.02c 0.21 ± 0.01c
a b c c
Lys 0.79 ± 0.07 0.28 ± 0.10 0.11 ± 0.00 0.13 ± 0.00 0.11 ± 0.00c
His 1.59 ± 0.35a 0.96 ± 0.14b 0.39 ± 0.00c 1.12 ± 0.08b 0.43 ± 0.03c
ab ab a bc
Arg 0.45 ± 0.09 0.47 ± 0.07 0.59 ± 0.03 0.38 ± 0.05 0.29 ± 0.03c
a ab ab b
Total 144.28 ± 11.48 102.81 ± 2.44 105.74 ± 2.44 91.75 ± 6.75 108.58 ± 0.66ab
Data presented were based on three replicates and expressed as mean ± standard error. Different lowercase letters in the same row indicate a
notable difference among groups (P \ 0.05)

Table 2 Free amino acids varieties and contents in fresh strawberry fruit (0 d) and stored samples (10 d)
Varieties (mg/100 g) Samples
Fresh Control 1-MCP ClO2 1-MCP ? ClO2

Umami 31.04 ± 4.30a 27.18 ± 0.13b 24.33 ± 0.55b 22.14 ± 0.81b 25.96 ± 0.37b
a b b b
Sweet 99.82 ± 16.21 67.09 ± 0.02 76.27 ± 5.33 64.01 ± 5.78 77.37 ± 0.96b
a ab c bc
Bitter 9.57 ± 0.85 5.72 ± 0.71 3.47 ± 0.26 4.48 ± 0.28 3.44 ± 0.02c
a b c d
None 3.92 ± 0.21 2.82 ± 0.02 1.67 ± 0.03 1.12 ± 0.04 1.80 ± 0.05c
Data presented were based on three replicates of twelve fruit and expressed as mean ± standard error. Different lowercase letters in the same row
indicate a significant difference among groups (P \ 0.05)

repair damaged protein, which could improve the abilities
of resisting various stress in fruit (Blée 2002). In our study,
the most abundant VOC in strawberries was (E)-2-hexenal.
Higher content of (E)-2-hexenal was observed in control
(57.92%) group compared with treatments. Especially, the
content of (E)-2-hexenal in ClO2 (37.95%) and
1-MCP ? ClO2 (47.46%) group was lower than control
and 1-MCP (58.72%) group. The reason may be that ClO2
is used as a fungicide to kill external microbes, reducing
the content of (E)-2-hexenal as a defense factor and
maintaining the quality of strawberry.
Esters, originated from the oxidation of fatty acids
(Reineccius, 2006), are considered as one of the most
considerable volatile components that impart fruity and
floral aroma to strawberries. 12 volatile esters were
detected in the 1-MCP, ClO2 and combination treatment,
while 10 volatile esters were determined in control and
fresh fruit. The content of total volatile esters in the three
treatments was significantly higher than that in control
(P \ 0.05). However, we did not observe a significant
difference in the content of total volatile esters between the
three treatment groups (P [ 0.05). This indicated that the
single or combination treatment of 1-MCP and ClO2
improved the content of esters during storage and aided in
retaining the flavor of strawberries. Methyl hexanoate,
which has sweet flavor similar to pineapple, was the pre-
dominant ester in fruit samples. The methyl hexanoate
contents in fresh fruit was 6.80%, while for the control
group, 1-MCP, ClO2, and combination treatment groups, it
was 3.94, 4.19, 7.02 and 5.62%, respectively. Ethyl

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Table 3 Volatile compounds detected by GC–MS in fresh fruit (0 d) and stored samples (10 d)
Number Compound Retention Concentration % (mean ± SD)
time
(min) Fresh Control 1-MCP ClO2 1-
MCP ? ClO2

Esters
1 Methyl acetate 3.56 Nd 0.67 ± 0.02b 1.03 ± 0.33b 2.22 ± 1.28a 1.46 ± 0.03ab
a c c ab
2 Ethyl acetate 4.12 3.82 ± 0.01 0.05 ± 0.01 0.64 ± 0.85 3.10 ± 0.21 2.55 ± 0.18b
a b b b
3 Methyl butyrate 5.46 3.60 ± 1.33 0.72 ± 0.06 0.93 ± 0.14 1.22 ± 0.24 1.51 ± 0.01b
c a
4 Ethyl butyrate 6.36 Nd Nd 0.13 ± 0.00 0.53 ± 0.01 0.39 ± 0.08b
a a
5 3,6-Octadecadiynoic acid, 6.70 0.64 ± 0.37 Nd 0.20 ± 0.00 Nd 0.18 ± 0.11a
methyl ester
6 Methyl hexanoate 9.67 6.80 ± 1.69a 3.94 ± 0.40a 4.19 ± 2.60a 7.02 ± 2.94a 5.62 ± 0.02a
b b a
7 Ethyl hexanoate 10.80 0.32 ± 0.03 0.26 ± 0.00 Nd 0.91 ± 0.00 Nd
8 Hexyl acetate 11.78 1.94 ± 0.49a 0.21 ± 0.04b 0.33 ± 0.11b 0.25 ± 0.05b 0.20 ± 0.08b
9 2-Hexen-1-ol, acetate 13.34 0.75 ± 0.38a 0.45 ± 0.02bc 0.70 ± 0.31ab 0.22 ± 0.18c 0.05 ± 0.03bc
10 3-Methylheptyl acetate 14.58 0.85 ± 0.05a 0.35 ± 0.03b 1.13 ± 0.15a 0.77 ± 0.03ab 1.06 ± 0.02a
11 Methyl octanoate 14.73 0.34 ± 0.07a 0.31 ± 0.00a 0.12 ± 0.05b 0.19 ± 0.04b 0.15 ± 0.03b
a b a b
12 Heptane formate 16.28 0.38 ± 0.33 0.06 ± 0.00 0.23 ± 0.14 0.12 ± 0.01 0.09 ± 0.00b
a ab
13 Octyl caprylate 18.18 Nd Nd 0.14 ± 0.02 0.13 ± 0.07 0.08 ± 0.01a
Total 21.59 ± 5.13a 7.02 ± 1.82c 9.77 ± 0.37b 16.68 ± 5.00ab 13.34 ± 0.06ab
Aldehydes
14 Hexanal 7.31 9.36 ± 1.99b 24.24 ± 1.20a 23.48 ± 3.46a 25.42 ± 3.69a 25.87 ± 0.98a
a a b
15 cis-3-hexenal 8.64 Nd 0.64 ± 0.01 0.52 ± 0.06 0.18 ± 0.04 0.38 ± 0.04ab
c a a b
16 (E)-2-Hexenal 10.48 17.15 ± 3.54 57.92 ± 9.87 58.72 ± 7.77 37.95 ± 2.43 47.46 ± 0.52ab
17 Octanal 12.23 0.89 ± 0.11a 0.22 ± 0.02b 0.72 ± 0.23a 0.31 ± 0.08b 0.24 ± 0.05b
a b b
18 (Z)-2-heptenal 13.14 1.85 ± 0.31 Nd 0.79 ± 0.07 0.72 ± 0.12 0.64 ± 0.06b
a b a b
19 Nonanal 14.86 1.29 ± 0.24 0.63 ± 0.11 1.31 ± 0.23 0.80 ± 0.04 0.44 ± 0.19b
a a a
20 Hexa-2,4-dienal 15.17 Nd 0.16 ± 0.04 0.17 ± 0.11 0.14 ± 0.02 0.15 ± 0.01a
a b b b
21 (E)-2-Octenal 15.77 0.59 ± 0.01 0.29 ± 0.08 0.32 ± 0.02 0.25 ± 0.03 0.24 ± 0.01b
22 2,4-Dimethyl 1,4-pentadiene 16.68 0.28 ± 0.00a 0.16 ± 0.01b 0.11 ± 0.03b 0.10 ± 0.02b 0.11 ± 0.01b
23 Benzaldehyde 18.10 0.79 ± 0.12a 0.13 ± 0.05b 0.09 ± 0.03b 0.05 ± 0.01b 0.04 ± 0.00b
24 (Z)-2-Nonenal 18.33 0.22 ± 0.02a 0.09 ± 0.03b 0.19 ± 0.02a 0.19 ± 0.03a 0.16 ± 0.03a
a a a
25 (E, Z)-2,6-Nonadienal 19.53 Nd 0.03 ± 0.01 0.06 ± 0.01 0.07 ± 0.01 0.06 ± 0.01a
b a a
26 (Z)-2-Decenal 20.83 Nd 0.02 ± 0.00 0.15 ± 0.05 0.09 ± 0.03 0.08 ± 0.02ab
27 á-methyl-benzenepropanal 23.92 0.17 ± 0.04 Nd Nd Nd Nd
c a a b
Total 32.59 ± 1.07 84.53 ± 2.42 86.63 ± 0.26 66.27 ± 5.32 75.87 ± 0.09ab
Alcohols
28 trans-1,2-Cyclopentanediol 8.31 Nd 0.08 ± 0.03 Nd Nd Nd
29 2-Hexyn-1-ol 10.07 Nd 0.55 ± 0.29a 0.40 ± 0.04a 0.29 ± 0.00b 0.48 ± 0.12a
a b b
30 1-Pentanol 11.14 1.66 ± 0.29 Nd 0.63 ± 0.08 0.53 ± 0.00 0.54 ± 0.14b
31 2,4-Hexadien-1-ol 11.50 Nd 0.16 ± 0.04ab 0.1 ± 0.00b 0.21 ± 0.00a Nd
a a
32 (Z)-2-Penten-1-ol 12.94 Nd 0.12 ± 0.09 0.13 ± 0.01 0.12 ± 0.01a 0.09 ± 0.00a
a b b b
33 1-Hexanol 13.73 1.86 ± 0.14 0.25 ± 0.08 0.31 ± 0.06 0.28 ± 0.08 0.30 ± 0.17b
a b b b
34 2-Nonen-1-ol 15.07 0.71 ± 0.16 0.38 ± 0.23 0.30 ± 0.16 0.24 ± 0.01 0.14 ± 0.11b
a b b
35 1-Octen-3-ol 16.18 1.32 ± 0.12 0.21 ± 0.03 Nd 0.35 ± 0.09 0.28 ± 0.03b
36 2-ethyl-1-hexanol 17.14 29.65 ± 1.70a 2.65 ± 0.29b 3.74 ± 0.20b 2.35 ± 0.23b 2.19 ± 0.20b
37 2-Decen-1-ol 17.43 0.27 ± 0.11a 0.07 ± 0.01b 0.10 ± 0.01b 0.08 ± 0.01b 0.06 ± 0.01b
38 3,7-dimethyl-1,6-octadien-3-ol 18.54 11.88 ± 0.80a 2.69 ± 0.30b 3.40 ± 2.09b 2.92 ± 0.36b 2.81 ± 0.20b
a b a ab
39 1-Octanol 18.75 0.60 ± 0.06 0.18 ± 0.07 0.62 ± 0.14 0.37 ± 0.03 0.28 ± 0.09b
a b b b
40 2-Octen-1-ol 20.12 0.29 ± 0.00 0.06 ± 0.02 0.07 ± 0.01 0.08 ± 0.01 0.06 ± 0.01b

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Table 3 continued
Number Compound Retention Concentration % (mean ± SD)
time
(min) Fresh Control 1-MCP ClO2 1-
MCP ? ClO2

41 1-Undecanol 21.10 Nd Nd 0.05 ± 0.04a Nd 0.04 ± 0.00a

42 à-Ethyl-à-methylbenzyl alcohol 23.45 Nd Nd 0.04 ± 0.01 Nd Nd
43 Benzyl alcohol 25.51 0.11 ± 0.02 Nd Nd Nd Nd
44 3,7,11-trimethyl-1,6,10- 27.63 0.08 ± 0.00bc 0.06 ± 0.01c 0.12 ± 0.03a 0.10 ± 0.00ab 0.11 ± 0.00ab
dodecatrien-3-ol
Total 48.43 ± 3.44a 7.46 ± 0.79c 10.01 ± 0.07b 7.92 ± 0.07c 7.37 ± 0.28c
Ketones
45 1-Octen-3-one 12.53 0.33 ± 0.09a 0.12 ± 0.00b 0.13 ± 0.02b 0.11 ± 0.00b 0.10 ± 0.00b
46 6-methyl-5-hepten-2-one 13.45 0.40 ± 0.12a 0.07 ± 0.02b 0.12 ± 0.00b 0.17 ± 0.00b 0.10 ± 0.00b
47 4-methoxy-2,5-dimethylfuran- 19.67 0.43 ± 0.13c 1.97 ± 0.06b 3.81 ± 0.18a 2.90 ± 0.31ab 2.87 ± 0.32ab
3(2H)-one
48 Butyrolactone 20.59 0.66 ± 0.08a 0.14 ± 0.01b 0.11 ± 0.04b 0.09 ± 0.04b 0.07 ± 0.01b
a
49 4-hydroxy-2,5-dimethylfuran-3- 33.24 Nd 0.09 ± 0.00 Nd Nd 0.07 ± 0.00a
one
Total 1.82 ± 0.22c 2.39 ± 0.05b 4.17 ± 0.02a 3.27 ± 0.47ab 3.21 ± 0.19ab
Acids
50 Acetic acid 16.53 0.26 ± 0.20 Nd Nd Nd Nd
51 2-methyl-butanoic acid 21.53 0.17 ± 0.13a 0.03 ± 0.00b 0.06 ± 0.01b 0.04 ± 0.01b 0.04 ± 0.00b
52 Pentanoic acid 25.06 0.64 ± 0.27a 0.17 ± 0.00b 0.15 ± 0.07b 0.16 ± 0.02b 0.11 ± 0.03b
53 Heptanoic acid 26.60 0.23 ± 0.07 Nd Nd Nd Nd
54 Octanoic acid 27.91 0.18 ± 0.06 Nd Nd Nd Nd
Total 1.48 ± 0.56a 0.20 ± 0.01b 0.21 ± 0.03b 0.20 ± 0.04b 0.15 ± 0.02b
Data shown were the means of three replicates for each treatment and expressed as mean ± standard error. ‘Nd’ is not found; different lowercase
letters within a row indicate significant differences among treatments (P \ 0.05)

butyrate, fruit-like aroma, was detected only in the three
treatment groups during storage period. Zhang et al.
(2009b) found that ethyl butyrate in strawberries samples
could not be extracted at low temperature and detected by
GC–MS. This indicated that individual or combination
treatment of 1-MCP and ClO2 could recover the content of
ethyl butyrate during strawberries storage.
Five volatile ketones were detected in control group and
combination treatment: 1-octen-3-one, 6-methyl-5-hepten-2-
one, DMMF, butyrolactone and DMHF. However, only four
volatile ketones (except DMHF) were detected in fresh fruit,
1-MCP, and ClO2 treatment groups. DMHF was first detected
in pineapple. DMHF has poor stability in strawberries and
easily convert into DMMF. DMHF-glucoside, DMHF-mal-
onyl glucoside, DMHF and DMMF comprise four derivative
forms of DMHF. And the former two are not VOCs, but they
can be changed into DMHF under certain conditions, which
imparts sweet flavor to the strawberries (Zabetakis et al.
1999). The most abundant volatile ketone in strawberries was
DMMF, accounting for 24% and 67–91% of the total volatile
ketone content in fresh fruit (0 d) and stored samples (10 d),
respectively. In addition, the DMMF content of strawberries
increased during storage period, while the content of other
ketone compounds decreased. The increase of DMMF with
caramel aroma is a symbol of fruit maturity (Zabetakis et al.
1999). The content of total ketones was 1.82 and 2.39% in
fresh and control fruit, which was lower than that of 1-MCP
(4.17%), ClO2 (3.27%) and 1-MCP ? ClO2 (3.21%). All
those indicated that 1-MCP and ClO2 single or combination
treatment could maintain the DMMF content during storage
period and enhance the flavor of strawberries.
17 types of volatile alcohols were determined in
strawberries. Alcohol compounds showed a significant
decreasing tendency in stored control fruit (10 d) compared
with fresh strawberries (P \ 0.05). However, trans-1,2-
cyclopentanediol was identified only in the control group,
and benzyl alcohol only in fresh fruit. 2-ethyl-1-hexanol
and 3,7-dimethyl-1,6- octadien-3-ol were the most abun-
dant volatile alcohols, whose content reduced by approxi-
mately 10% and 34% respectively after storage. 3,7,11-
trimethyl-1,6,10-dodecatrien-3-ol presented rose and lilies-
like aroma and the content of it in treatment groups were
significantly higher than that in control group (P \ 0.05).

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There were 5 volatile acids detected in fresh fruit,
however, only 2 in control, 1-MCP, ClO2 alone and the
combination group. From 0 to 10 d, the content of
2-methyl-butanoic acid and pentanoic acid declined obvi-
ously (P \ 0.05). However, no significant difference was
seen in the content of pentanoic acid and 2-methyl-bu-
tanoic acid between treatments and control by the end of
storage (P [ 0.05).
Principle compound analysis (PCA) of strawberries
volatiles during storage
In this study, although 54 VOCs were totally detected in
the strawberries, methyl or ethyl esters of butyric acid,
hexanoic acid, DMMF and DMHF are seemed as the most
necessary contributors to the aroma of strawberries.
Supplementary Fig. S. 1 show the correlation between
VOCs analyzed by PCA on 15 samples and 12 variables.
The samples overlap or close to each other had similar
flavor. As shown in Supplementary Fig. S. 1A (scores plot),
PC-1 and PC-2 accounted for 67.32% and 19.77% of the
total variations respectively. The combination treatment
exhibited aroma similar to those of fresh fruit, control
group, and 1-MCP treatment group. However, ClO2 treat-
ment group exhibited distinctly different flavor. As shown
in Supplementary Fig. S. 1B (loadings plot), methyl acet-
ate, ethyl acetate, methyl butyrate, methyl hexanoate and
DMMF contributed towards PC-1, while ethyl hexanoate
contributed towards PC-2. Methyl acetate, ethyl acetate,
methyl butyrate and DMMF were highly positive corre-
lated. Similarly, methyl hexanoate had a positive correla-
tion with DMMF. However, other components were
negatively correlated with DMMF. Moreover, DMMF and
methyl hexanoate were the major VOCs in strawberries.
Conclusion
In summary, 1-MCP ? ClO2 could maintain the content of
sugar and organic acids (except succinate), retain the
highest sweet taste and lowest bitter taste amino acids as
well as total FAA, and regulate the contents of alcohols,
ketones and aldehydes, which are important flavor attri-
butes for strawberry. PCA indicated that the ClO2 treat-
ment group exhibited distinct flavor in comparison to those
of fresh fruit, control, 1-MCP and the combination. Methyl
acetate, ethyl acetate, and methyl butyrate were positively
correlated with methyl hexanoate and DMMF. Moreover,
methyl hexanoate and DMMF were the major factors
affecting the aroma of strawberries. Overall,
1-MCP ? ClO2 maintained the quality of strawberries by
regulating flavor and taste. Portable smell sensor can be
designed to detect ethyl butyrate in strawberries subjected
to individual or combination treatment of 1-MCP and ClO2
for evaluating the flavor attributes of strawberries.
Acknowledgements This research project was financially supported
by the National Science Foundation of China (Project No. 31701973)
and the Youth Science and Technology Innovation Team of Shandong
Province in China (Project No. 2019KJF01).
Complaince with ethical standards
Conflict of interest The authors declare no competing financial
interest.
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