International Journal of Food Science and Technology 2003, 38, 869875

Maintaining postharvest quality of raspberries with

natural volatile compounds
Chien Y. Wang
Produce Quality and Safety Laboratory, Plant Sciences Institute, ARS, US Department of Agriculture, Beltsville, MD 207052350, USA
(Received 30 December 2002; Accepted in revised form 27 May 2003)

Summary

The postharvest quality of raspberries (Rubus idaeus L.) was evaluated after treatment with
several natural volatile compounds and storage at 0, 10 or 20
C. At high storage
temperature (20
C), raspberries deteriorated rapidly and none of the natural volatile
compounds that were used were eective in extending storage life of the fruit. At low
storage temperature (0
C), little dierence could be discerned among the various volatile
treatments. However, at 10
C storage, raspberries treated with methyl jasmonate (MJ),
allyl isothiocyanate (AITC), tea tree oil (TTO), or absolute ethyl alcohol had less decay.
MJ- and TTO-treated fruit also maintained higher levels of sugars, organic acids and
oxygen radical absorbance capacity compared with untreated fruit. Samples treated with
acetic acid or vinegar vapour did not dier from control fruit. Measurement of oxygen and
carbon dioxide concentrations within the containers during storage revealed that none of
the treatments caused accumulation or depletion of these gases to a level that would be
harmful to raspberries. Colour measurements of the berries showed that all raspberry fruit
became darker and less red after storage, but fruit treated with MJ were found to have the
highest intensity of red colour while AITC application caused the berries to appear lighter
in colour.

Keywords

Antioxidant, methyl jasmonate, Rubus idaeus, storage.

Introduction

In addition to having a distinct pleasant aroma,

raspberries contain high antioxidant capacity that
oers signicant health benets (Wang & Lin,
2000). However, raspberries soften and deteriorate rapidly after harvest. Their storage life is
limited by fungal decay such as that caused by
grey mould rot (Botrytis cinerea Pers.), rhizopus
rot (Rhizopus stolonifer Ehrenb.) and cladosporium rot (Cladosporium herbarum Fr.) (Harvey &
Pentzer, 1960).
Several natural volatile compounds have been
reported to possess antimicrobial activity. Methyl
jasmonate (MJ) either as a vapour or as an
emulsion has been shown to reduce microbial
Correspondent: Fax: +1 301 504 5107;
1 e-mail: wangc@ba.ars.usda.gov

 2003 Blackwell Publishing Ltd

contamination of fresh-cut celery and peppers

(Buta & Moline, 1998), inhibit grey mould
infection in strawberries (Moline et al., 1997),
suppress green mould growth in grapefruit (Droby
et al., 1999) and control Botrytis rot in cut rose
owers (Meir et al., 1998). MJ also decreased the
severity of postharvest brown rot in sweet cherries
when used as a co-fumigant with thymol or
carvacrol (Tsao & Zhou, 2000). Ethanol has also
been found to have antimicrobial properties, and a
postharvest ethanol dip eliminated most of the
fungal and bacterial populations on the surface of
grapes without impairing bunch appearance or
berry rmness (Lichter et al., 2002). Ethanol
vapour also prevented scald development in apples
(Ghahramani et al., 2000; Chervin et al., 2001)
and reduced leaf blackening in the stems of the cut
ower Protea Pink Ice (Crick & McConchie,
1999). Allyl isothiocyanate (AITC) has been

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Raspberry fruit quality maintenance C. Y. Wang

shown to have strong antimicrobial activity in

both liquid and vapour forms (Isshiki et al., 1992;
Delaquis & Sholberg, 1997; Lin et al., 2000).
AITC is a constituent of cruciferous vegetables
and is produced during the enzymatic degradation
of glucosinolate (Smolinska & Horbowicz, 1999).
The bactericidal activity of AITC against pathogens on iceberg lettuce and tomatoes has also been
demonstrated (Lin et al., 2000). The essential oil
of Melaleuca alternifolia, also known as tea tree oil
(TTO), is becoming increasingly popular, particularly as a naturally occurring antimicrobial agent
(Carson & Riley, 1995). TTO was eective in
inhibiting the storage pathogen Botrytis cinerea on
Dutch white cabbage (Brassica oleracea, var.
Capitata) (Bishop & Reagan, 1998). Its vapour
phase has shown a high level of antifungal activity
in fteen common postharvest pathogens on a
variety of crops (Bishop & Thornton, 1997).
Acetic acid (AA) or vinegar vapour was eective
in preventing conidia of brown rot, grey mould
and blue mould from germinating and causing
decay of stone fruit, strawberries and apples
(Sholberg et al., 2000). AA vapour also reduced
postharvest brown rot of apricot and plums (Liu
et al., 2002). However, no information is available
on the eects of these natural volatile compounds
on the development of fungal infection or decay in
raspberries. This study was undertaken to determine if these natural products would reduce decay
and maintain better quality of raspberries. The
physiological changes in raspberry fruit aected
by the volatile treatments were also investigated.
Materials and methods

used in this study include MJ (22.4 lL L)1),

AITC (5 lL L)1), essential oil of Melaleuca
alternifoli (TTO, 100 lL L)1), absolute ethyl
alcohol (AEA, 200 lL L)1) and AA (vinegar,
4 mL L)1). The specied volume of each volatile
compound was placed into individual small
beakers, which were subsequently placed inside
the plastic containers just before the lids were
covered. The volatile compounds were allowed to
vaporize inside the containers spontaneously at
20
C for 16 h. The containers were then transferred to the designated storage temperatures:
0, 10 and 20
C. Three containers were used for
each treatment. Control samples were handled
similarly with the exception of the volatile
treatment.
Decay evaluation
The severity of decay was evaluated after 10 days
of storage at 10
C. The degree of infection on the
fruit was rated using a scale of 1 to 5, where
1 clean with no infection, 2 trace infection,
3 mild infection, 4 moderate infection and
5 severe infection.
Measurements of O2 and CO2
To analyse the accumulation of CO2 and depletion
of O2 inside the containers during storage, 10 mL
of headspace gas were taken from each container
every other day for analysis. Carbon dioxide and
oxygen concentrations were measured with a CO2
and O2 analyser (AMETEK Applied Electrochemistry, Pittsburgh, PA, USA; model CD-3A for
CO2 and S-3A for O2).

Plant materials
Raspberries (Rubus idaeus L., cv. Heritage) were
freshly harvested at commercial maturity from an
orchard near Beltsville, MD. Fruit with uniform
size, colour, ripeness and free from defect or injury
were used in this study. The selected fruit were
randomized before being used for treatments with
various volatiles.
Treatments with natural volatile compounds
Fifty fruit were placed into 1-L polystyrene
containers with snap-on lids. Volatile compounds

Measurement of colour
Raspberry fruit colour was measured with a
Minolta colorimeter (model CR-10; Minolta
Corp., Ramsey, NJ, USA) equipped with an
8 mm measuring aperture and calibrated with a
white standard tile. Colour of the fruit was
expressed as L*, a*, b*, where L* indicates
lightness, a* and b* are chromaticity coordinates.
The coordinates a* and b* indicate colour directions: +a* is the red direction, )a* is the green
direction, +b* is the yellow direction, and )b* is
the blue direction.

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Raspberry fruit quality maintenance C. Y. Wang

Analysis of sugars and organic acids

Two grams of raspberry fruit tissue were homogenized with a Polytron homogenizer (Brinkmann Instruments, Westbury, NY, USA) in
imidazole buer (20 mm, pH 7.0). The extracts
were centrifuged and the supernatants were dried
in vacuo in vials that were used during derivalization. Procedures described by Li & Schuhmann
(1980) were modied for the derivatization of
sugars and organic acids. A known amount of bphenyl-d-glucopyranoside was included in all
samples as an internal standard. One millilitre
Trisil reagent (Pierce, Rockford, IL, USA) was
mixed vigorously with each sample and then heated
at 75
C for 30 min. After silylation, 1 lL of each
derivatized sample was injected into a HewlettPackard 5890 gas chromatograph (Hewlett-Packard, Palo Alto, CA, USA) equipped with a ame
ionization detector and a 25 m crosslinked
methyl silicon gum capillary column (HewlettPackard, HP-1; 0.2 mm i.d., 0.33 lm lm
thickness) and using helium as carrier gas.
Temperatures were as follows: injector 250
C,
detector 275
C and column temperature was
programmed to increase from 100 to 250
C at
10
C min)1, then hold constant at 250
C for
23 min. Organic acids were analysed after extraction with imidazole buer (20 mm, pH 7.0) and
purication with a Baker-10 solid phase extraction system. Supernatants from the extract were
passed through quaternary amine columns, which
were previously conditioned with hexane and
methanol. The samples were then eluted from the
columns with 0.1 n HCl. The eluates were concentrated to dryness in vacuo in derivatized vials.
Procedures of derivatization and chromatography
for organic acids were the same as those for
sugars except that column temperature was held
at 180
C for 3 min, then increased to 250
C at
10
C min)1 and held at 250
C for 12 min. The
sugars and organic acids were quantied by
comparison with derivatized standards.
Oxygen radical absorbance capacity (ORAC) assay
Five grams of raspberry fruit tissue were extracted
in 45 mL phosphate buer (75 mm, pH 7.0) using
a homogenizer and then centrifuged at 20 000 g
at 4
C for 30 min. The supernatant was used for
 2003 Blackwell Publishing Ltd

the ORAC assay after suitable dilution with

phosphate buer. The ORAC assay procedure
was modied from a method previously described
by Cao et al. (1993). This assay measures the eect
of antioxidant components in fruit extracts on the
decline in R-phycoerythrin (R-PE) uorescence
induced by a peroxyl radical generator, 2,2-azobis
(2-amidinopropane) dihydrochloride (AAPH).
The reaction mixture contained 1.7 mL of 75 mm
phosphate buer (pH 7.0), 100 lL of R-PE
(3.4 mg L)1), 100 lL of 320 mm AAPH and
100 lL of sample. Phosphate buer was used as
a blank and 1 lm of Trolox (a water-soluble
a-tocopherol analogue) was used as a standard
during each run. The nal volume of 2 mL was
used in a 10 mm wide uorometer cuvette. R-PE,
phosphate buer and samples were pre-incubated
at 37
C for 15 min. The reaction was started by
the addition of AAPH. Fluorescence was measured and recorded every 5 min using emission at
570 nm and excitation at 540 nm in a Shimadzu
RF-Mini 150 Recording Fluorometer (Columbia,
MD, USA) until the uorescence of the last
reading declined to <5% of the rst reading.
This usually took approximately 70 min. One
blank, one standard, and a maximum of ten
samples were analysed at the same time. Each
sample was repeated three times. The ORAC value
refers to the net protection area under the
quenching curve of R-PE in the presence of an
antioxidant. The nal results (ORAC values) were
calculated and expressed using Trolox equivalents
per gram fresh weight basis.
Results and discussion

Effect of volatile compounds on decay

Storage temperature had a profound eect on the
postharvest life of raspberry fruit. Temperature
also aected the response of raspberries to volatile
treatments. At high storage temperature (20
C),
fruit deteriorated rapidly and none of the volatile
treatments were eective in extending the storage
life of the fruit. In contrast, low storage temperature (0
C) nullied the eect of volatiles and little
dierence could be discerned among the various
volatile treatments (data not shown). Apparently,
the eect of low temperature overshadowed the
eect of volatile compounds on the quality of

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Raspberry fruit quality maintenance C. Y. Wang

raspberries. However, at 10
C storage, several

natural volatile compounds were found to be
benecial in retarding decay and maintaining
quality of raspberry fruit (Table 1). The most
eective compounds were found to be MJ and
AITC, followed by AEA and TTO. MJ has been
shown to induce the synthesis and expression of
some stress proteins, such as heat shock proteins
and pathogenesis-related proteins, which lead to
increased resistance to pathogens and decreased
incidence of decay (Ding et al., 2001, 2002). AITC
is a natural component in plant tissue and has
been used as a food additive, for example AITC is
used as a major avouring constituent of wasabi
(Japanese green mustard) and is the key compound responsible for its antimicrobial activity
(Delaquis & Mazz, 1995; Lin et al., 2000). AITC
isolated from cruciferous vegetables has been
reported to provide chemoprotection against cancer (Zhang & Talalay, 1994; Talalay & Zhang,
1996). AA was not eective in reducing mould
growth on raspberries at the concentration that we

used (Table 1). It is possible that a dierent dose

or dierent type of vinegar might result in more
potent inhibition of the organisms causing decay
(Sholberg et al., 2000).
Changes of O2 and CO2 in the containers
Changes of O2 and CO2 concentrations inside the
containers were monitored every other day to
ascertain that O2 concentrations did not deplete
to a suboxidation level and CO2 concentrations
did not accumulate to injurious levels. Raspberries are relatively tolerant to high CO2. The
recommended controlled atmosphere storage conditions for raspberry fruit are 1520% CO2 and
510% O2 (Kader, 2001). Levels of O2 and CO2
in the headspace of the containers in all treatments did not deviate signicantly from ambient
atmosphere during the 10 days of storage at
10
C (Table 2). Oxygen concentrations uctuated between 20.3 and 18.2% during all treatments, but never dropped below 18.2%. Carbon
dioxide concentrations ranged between 0.5 and
2.4% and never increased to more than 2.4%.
Heritage raspberry fruit is nonclimacteric (Perkins-Veazie & Nonnecke, 1992), therefore, the
fruit does not have a burst of CO2 output during
its postharvest life. This helps to explain why the
CO2 concentration did not accumulate to a high
level inside the containers. Normally, CO2 of
more than 20% and O2 levels <5% are required
in order to cause injury such as discoloration
and/or o-avour (Agar & Streif, 1996; Kader,
2001). The levels of oxygen and carbon dioxide
inside the containers during all of the treatments
in our study remained in the safe range throughout the experiment.

Table 1 Effect of various volatile compounds on the severity

of decay in raspberry fruit after 10 days of storage at 10
C
Volatile compound

Concentration (lL L)1)

Scorea

Methyl jasmonate
Allyl isothiocyanate
Tea tree oil
Absolute ethyl alcohol
Acetic acid
Control

22.4
5.0
100.0
200.0
4000.0

1.0
1.0
1.7
1.2
3.0
3.3

0.0b
0.0
0.3
0.3
0.0
0.5

Scoring represents a visual rating of decay severity on

raspberry fruit using a scale of 1 to 5 with 1 no infection,
2 trace, 3 slight, 4 moderate and 5 severe infection.
b
Mean value s.e.

Table 2 Oxygen and carbon dioxide concentrations (%) inside containers of raspberries treated with methyl jasmonate (MJ),
allyl isothiocyanate (AITC), tea tree oil (TTO), absolute ethyl alcohol (AEA), or acetic acid (AA) during storage at 10
C
Control

MJ

AITC

TTO

AEA

AA

Days

O2

CO2

O2

CO2

O2

CO2

O2

CO2

O2

CO2

O2

CO2

0
2
4
6
8
10

20.32
20.21
20.03
19.36
19.94
20.18

0.55
0.98
1.67
2.28
2.06
1.89

20.14
20.08
19.54
19.69
19.32
18.21

0.86
1.25
2.01
1.97
2.35
2.36

20.25
20.12
19.62
19.83
19.51
18.47

0.68
1.19
1.82
2.14
2.26
2.18

20.12
20.03
19.89
19.21
19.18
20.06

0.57
1.01
1.35
1.88
2.06
1.97

20.06
19.63
19.42
19.04
20.01
19.21

0.74
0.96
1.03
1.76
2.14
2.08

20.28
20.01
19.68
19.13
20.07
20.26

0.52
0.91
1.42
2.01
1.83
1.32

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Raspberry fruit quality maintenance C. Y. Wang

Table 3 Effect of various volatile compounds on fruit colour

of raspberries after 10 days of storage at 10
C
Color measurement
Treatment
3 At harvest
After storage
Control
Methyl jasmonate
Allyl isothiocyanate
Tea tree oil
Absolute ethyl alcohol
Acetic acid
a

L*
37.4a

31.2c
34.2b
35.8ab
28.9d
30.5cd
29.4d

a*

b*

32.8a

18.3a

25.9c
30.7ab
24.8cd
28.7b
25.3c
22.7d

10.6c
11.5bc
13.8b
7.2d
9.6c
9.8c

Mean separation by Duncans multiple range test, P 0.05.

Fruit colour as affected by volatile treatments

After 10 days of storage at 10
C, the values of all
chromaticity coordinates decreased (Table 3).
However, fruit treated with MJ and TTO were
found to have the highest a* values as compared
with those of other treatments, indicating that
raspberries treated with MJ and TTO retained
more red colour than fruit in other treatments.
The retention or promotion of fruit colour by MJ
treatment has also been reported in other fruits
including apples (Fan et al., 1998) and mangoes
(Gonzalez-Aguilar et al., 2001). Fruit treated with
AITC retained the highest L* values while those
treated with TTO had the lowest L* values.
Consequently, AITC-treated fruit looked brighter
and TTO-treated fruit were the darkest among all
of the fruit. A high b* value (toward yellow
direction) also contributed to a lighter colour
appearance for the AITC-treated fruit and a low

Table 4 Sugar, organic acid and

oxygen radical absorbance capacity (ORAC) values in raspberries
at harvest and after 10 days of
storage at 1
C as affected by
treatments with methyl jasmonate
(MJ), allyl isothiocyanate (AITC),
tea tree oil (TTO), absolute ethyl
alcohol (AEA) or acetic acid (AA)

Sugar and organic acid levels as affected by

volatile treatments
Citric acid is the predominant organic acid in
raspberries (Table 4). The citric acid content in
raspberry fruit is more than ten times higher than
malic acid at harvest. Both citric and malic acids
decreased after 10 days of storage at 10
C. The
decrease was especially prominent in malic acid,
which declined more than 50% during storage.
Treatments with MJ, AITC and TTO tended to
maintain higher levels of organic acids than other
treatments after storage. Major sugars in raspberries were fructose, glucose and sucrose (Table 4).
Sugar content in raspberry fruit did not change
much during storage after harvest. However, fruit
treated with MJ and TTO had higher sugar
content while those treated with AEA and AA
had lower sugar contents after 10 days of storage
at 10
C.
ORAC values in fruit treated with various
volatile compounds
Antioxidant activity, measured as ORAC, in the
control fruit increased slightly after 10 days of

Sugars (g kg)1
fresh wt.)

At harvest
After storage
Control
MJ
AITC
TTO
AEA
AA
a

 2003 Blackwell Publishing Ltd

b* value (toward blue direction) added to the

darkness of the TTO treated fruit. It is common
for raspberry fruit to become darker and more
blue after storage (Robbins & Moore, 1990).
However, raspberries with a lighter red colour
and less blue colour, such as fruit from controlled
atmosphere storage (Haner et al., 2002), are
considered to be more attractive to consumers.

Organic acids
(g kg)1 fresh wt.)

ORAC

Fructose

Glucose

Sucrose

Malic

Citric

(lmol TE g)1)

33.2bca

21.4b

6.8c

1.22a

13.21a

18.6bc

31.6cd
36.3a
33.6bc
35.7ab
31.4cd
28.7d

18.7c
25.2a
20.8bc
23.9a
19.1bc
18.5c

7.2bc
8.3ab
6.8c
8.8a
6.3c
6.6c

0.51d
0.94b
0.82bc
0.91b
0.75c
0.56d

8.56d
11.63b
10.17bc
10.32bc
8.73cd
7.92d

20.2b
22.8a
19.6bc
22.3a
19.1bc
17.7c

Mean separation by Duncans multiple range test, P 0.05.

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Raspberry fruit quality maintenance C. Y. Wang

storage at 10
C (Table 4). Fruit treated with MJ

and TTO had even higher ORAC values while
fruit treated with AA had lower ORAC values as
compared with the control. Fruit treated with MJ
and TTO also had high a* values and fruit treated
with AA had low a* values (Table 3), indicating
that there may be an association between ORAC
values and red color. ORAC values have been
shown to closely correlate to anthocyanin content
in fruit (Prior et al., 1998; Kalt et al., 1999; Wang
& Lin, 2000). As high antioxidant activity oers
potential health benets, treatments with MJ and
TTO may be benecial for human health.
Taken together, several natural volatile compounds have shown promise for protection of
raspberries from microbial infections and for
maintenance of the quality of raspberry fruit.
Further study is warranted to investigate such
factors as toxicity, practicality and economical
feasibility.
Acknowledgments

The author would like to thank Hilarine Repace

and David Spaulding for technical assistance.
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