Research Article

Received: 26 November 2012 Revised: 5 March 2013 Accepted article published: 28 March 2013 Published online in Wiley Online Library: 8 May 2013

(wileyonlinelibrary.com) DOI 10.1002/jsfa.6151

Comparative studies on chemical parameters

and antioxidant properties of stipes and caps
of shiitake mushroom as affected by different
drying methods
Ning Zhang, Haixia Chen,∗ Yu Zhang, Lishuai Ma and Xufeng Xu

Abstract
BACKGROUND: Shiitake, the second most cultivated mushroom, is famous for its high nutritional value and medicinal properties.
In this study, various chemical parameters and antioxidant properties of caps and stipes of shiitake mushroom dried by different
methods (freeze-drying, shade drying and hot air drying) were comparatively investigated by spectrophotometric assays, high-
performance liquid chromatography, 1,1 -diphenyl-2-picrylhydrazyl assay, ferric reducing power assay and lipid peroxidation
inhibition assay.

RESULTS: The contents of amino acids, neutral sugar and total phenolics in stipes were higher than those in caps of shiitake,
while caps showed advantages in terms of their contents of protein and eritadenine. The chemical parameters and antioxidant
activities of shiitake were significantly affected by the drying method used.

CONCLUSION: The contents of total phenolics, amino acids and neutral sugar in stipes were higher than those in caps of shiitake,
which suggested that stipes were more nutritional than caps in some respects. Hot air drying at 50 ◦ C resulted in high total
phenolic, amino acid, uronic acid and neutral sugar contents and antioxidant activities, which could be useful for the application
of shiitake and related products in the food industry.


c 2013 Society of Chemical Industry

Keywords: shiitake; caps and stipes; drying methods; chemical parameters; antioxidant activity

INTRODUCTION activity was caused by a decrease in the phosphatidyl-

Shiitake mushroom (Lentinula edodes) has been valued as a food
and medicine for thousands of years in Asia, and China accounts for
over 70% of the world’s shiitake production.1 It is served in many
steamed and simmered dishes owing to its nutritional value and
beneficial effects on health.2 Shiitake is reported to have multiple
bioactivities, including anticancer, antidiabetic, hypotensive,
antinociceptive, anti-inflammatory, hypocholesterolaemic and
antioxidant activities.3 It is an important nutritional resource
because of its high contents of polysaccharides (especially
lentinan), protein, lectins, polysaccharide–protein complexes,
dietary fibre, flavour compounds, ergosterol, vitamins B1 , B2 and C,
essential amino acids (especially lysine and leucine) and minerals
(K, Mg, P, Zn, Fe and Cu).3 – 9
In Japan and China the chemical constituents of shiitake have
been analysed for their medicinal properties, and extracts have
been used in treating cancer. Several constituents have been iso-
lated and identified. One water-soluble polysaccharide isolated
was lentinan, which was found to have antitumour, antiviral and
antimicrobial activities.10 The hypocholesterolaemic activity of
shiitake was first reported by Kaneda et al.11 Eritadenine (2(R),3(R)-
dihydroxy-4-(9-adenyl)butyric acid) was identified as the main
hypocholesterolaemic factor of shiitake by several investigators.12
It was shown that total plasma cholesterol levels were decreased
choline (PC)/phosphatidylethanolamine (PE) ratio.13 – 16 Other
constituents such as flavonoids, lignans and phenolic acids found
in shiitake might also contribute to the antioxidant activities.17
Shiitake mushroom is a rapidly perishable commodity, and its
quality begins to deteriorate immediately after harvest. Fresh shi-
itake has to be processed to extend its shelf life for off-season use.
Mushrooms can be processed in many ways, such as drying, picking
and canning. It was reported that drying was a comparatively cheap
method for mushroom processing.18 Dried mushrooms packed in
airtight containers could have a shelf life of over 1 year.19,20 Some
studies on drying of shiitake have been reported. The polypheno-
lic contents and antioxidant activities were measured in shiitake
heated at 100 or 120 ◦ C for 15 or 30 min using an autoclave.4 The
drying characteristics were studied under conditions of different
drying temperatures (50, 55, 60 and 65 ◦ C) and different vacuum
pressures (0.1, 0.2, 0.3 and 0.4 bar).21 The edible part of shiitake
∗
Correspondence to: Haixia Chen, Tianjin Key Laboratory for Modern Drug
Delivery & High-Efficiency, School of Pharmaceutical Science and Technology,
Tianjin University, Tianjin 300072, China. E-mail: chennhxx@yahoo.com.cn
Tianjin Key Laboratory for Modern Drug Delivery & High-Efficiency, School of
Pharmaceutical Science and Technology, Tianjin University, Tianjin 300072,
3107

in rats fed eritadenine in their diets. The hypocholesterolaemic China

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c 2013 Society of Chemical Industry
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consists of caps and stipes. However, because of their bad taste,
stipes are usually discarded. It is necessary to study the differences
in bioactive constituents and antioxidant activities between caps
and stipes for their better utilisation. There have been no compara-
tive studies on the bioactive constituents and antioxidant activities
of shiitake caps and stipes as affected by different drying methods.
The purpose of this study was to compare the differences in
bioactive constituents of caps and stipes of shiitake as affected by
different drying methods (freeze-drying, shade drying and hot air
drying). The antioxidant properties of shiitake extracts were also
investigated and the relationship between the constituents and
the antioxidant activity was discussed.
MATERIALS AND METHODS
Materials and chemicals
Raw shiitake were purchased from Qingguang Vegetable Base
(Tianjin, China). Phenic acid, Coomassie brilliant blue, bovine serum
albumin (BSA), carbazole, D-galacturonic acid and 1,1 -diphenyl-
2-picrylhydrazyl (DPPH) were obtained from Sigma (St Louis, MO,
USA). All other chemicals and reagents were purchased locally and
were of analytical grade.
Drying process of shiitake
After shiitake harvest, the fresh mushrooms were lightly washed
with distilled water to remove large impurities and then divided
into caps and stipes. The cap and stipe samples were further
divided into five parts each. The five parts were dried by freeze-
drying at -50 ◦ C, shade drying at 28 ◦ C and hot air drying at
50, 100 and 150 ◦ C respectively. All fresh caps (100 g each) and
stipes (100 g each) were dried until they reached constant weight
(<0.01 g difference between consecutive weighings). All drying
experiments were performed in triplicate.
Preparation of water-soluble extracts of shiitake
The dried cap and stipe samples were extracted three times with
distilled water (1:10 w/v, 2 h, 100 ◦ C). Each combined extract was
filtered through Whatman No. 1 filter paper and the filtrate was
concentrated in a rotary evaporator at 50 ◦ C under vacuum. All
extracts were freeze-dried and kept in a desiccator until analysis
of water-soluble constituents.
Determination of water-soluble constituents of shiitake
Protein content was determined by the method of Bradford22 using
BSA as standard. Briefly, 100 µL of sample solution (1 mg mL−1 )
was mixed with 5 mL of Bradford protein assay kit solution. After
shaking and incubation at room temperature, the absorbance
at 595 nm was measured. Protein contents were calculated
according to the following equation obtained from the standard
BSA graph: absorbance = 0.504 × BSA (mg mL−1 ) + 1.148 (R2
= 0.995).
Amino acid content was determined by the ninhydrin method
with slight modification.23 Briefly, the sample was diluted to 5 mL
with phosphate buffer (pH 8). Then the solution was mixed with 1
mL of hydroxide solution (5 mg mL−1 ) and heated in a boiling water
bath for 15 min. The solution was cooled to room temperature and
the absorbance at 570 nm was measured. L-Glutamate was used
as standard. Amino acid contents were calculated according to the
following equation obtained from the standard L-glutamate graph:
3108
absorbance = 25.28 × L-glutamate (mg mL−1 ) − 0.399 (R2 = 0.992).
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c 2013 Society of Chemical Industry
N Zhang et al.
Uronic acid content was determined by the carbazole/sulfuric
acid method using galacturonic acid as standard.24 Briefly, 250 µL
of sample solution (1 mg mL−1 ) was mixed with 1.5 mL of 9 g L−1
borax/sulfuric acid solution in ice-water. The mixture was heated
in boiling water for 10 min, chilled rapidly to room temperature
and then added to 50 µL of carbazole/alcohol solution. After
incubation, heating in boiling water for 15 min and then chilling
to room temperature, the absorbance at 525 nm was measured.
Uronic acid contents were calculated according to the following
equation obtained from the standard galacturonic acid graph:
absorbance = 4.734 × galacturonic acid (mg mL−1 ) + 0.000
(R2 = 0.999).
Neutral sugar content was determined by the phenol/sulfuric
acid method using D-glucose as standard.25 Briefly, 200 µL of
sample solution (1 mg mL−1 ) was mixed with 200 µL of phenol.
The mixture was incubated and added to 1 mL of concentrated
sulfuric acid. After incubation and chilling to room temperature,
the absorbance at 490 nm was measured. Neutral sugar contents
were calculated according to the following equation obtained
from the standard D-glucose graph: absorbance = 5.975 ×
D-glucose (mg mL−1 ) + 0.037 (R2 = 0.995).
Preparation of ethanolic extracts of shiitake
The dried shiitake caps and stipes were extracted three times
with 800 mL L−1 ethanol (1:12 w/v, 78 ◦ C). Each combined
extract was filtered through Whatman No. 1 filter paper and
the filtrate was concentrated in a rotary evaporator at 45 ◦ C
under vacuum. The concentrated liquid was freeze-dried to
obtain the ethanolic extracts. All extracts were kept at 4 ◦ C until
analysis of phenolics and eritadenine and assay of antioxidant
activities.
Determination of phenolics and eritadenine in shiitake
Phenolic content, expressed as gallic acid equivalent, was
determined by the method of Singleton and Rossi26 with slight
modification. A 3 mg aliquot of each ethanolic extract was added
to 3 mL of 700 mL L−1 ethanol. Then 500 µL of sample solution was
mixed with 500 µL of Folin–Ciocalteu’s phenol reagent (Sigma).
After 3 min, 500 µL of saturated 350 g L−1 Na2 CO3 solution was
added and the mixture was made up to 5 mL by adding distilled
water. The reaction was kept in the dark for 60 min, after which
the absorbance at 725 nm was read. The standard curve was as
follows: absorbance = 17.42 × gallic acid (mg mL−1 ) − 0.046 (R2
= 0.998).
Eritadenine content was determined by high-performance
liquid chromatography (HPLC) (LC-10AT, Shimadzu, Kyoto, Japan)
on an ODS-C18 reverse phase column (5 µm, 250 mm × 4.6
mm i.d.; YMC Co. Ltd, Kyoto, Japan) equipped with an analytical
KJ0-4282 C18 guard cartridge system (Phenomenex, Torrance, CA,
USA). A 3 mg aliquot of each ethanolic extract was added to 3 mL of
50 mL L−1 methanol. The solution was filtered through a 0.22 µm
hydrophobic membrane. The mobile phase was methanol/water
(5:95 v/v) at a flow rate of 1 mL min−1 . The eluate was monitored by
UV detection at 260 nm. All data were analysed using LCsolution
software (Shimadzu). An eritadenine standard was provided by
our laboratory and its chemical structure was confirmed by com-
paring UV, IR, ESI-MS and 1 H NMR measurements with literature
data. The purity of the standard was checked by HPLC at four
wavelengths (210, 240, 254 and 280 nm) and found to be above
98%. The standard curve was as follows: Y = 2 × 106 X + 13726
(R2 = 0.995).
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Antioxidant activities of ethanolic extracts of shiitake
Scavenging effect on DPPH radicals
The scavenging effect of the ethanolic extracts on DPPH radicals
was measured as described previously.27 Briefly, 2.9 mL of 0.1
mmol L−1 DPPH solution was added to 0.1 mL of ethanol solution
containing different amounts of shiitake ethanolic extract (0.4,
1.2, 2, 3.2, 4 mg). The absorbance of the solution at 517 nm
was measured using a UV–visible spectrophotometer (UV-2450,
Shimadzu). The DPPH radical-scavenging rate of the sample was
calculated as

  
inhibition (%) = Ablank − Asample /Ablank × 100
where Asample and Ablank are the UV absorbances of the sample
and blank respectively.
Ferric reducing power
The ferric reducing power of the ethanolic extracts was determined
as described previously.28 Briefly, 0.1 mL of ethanol solution
containing different amounts of shiitake ethanolic extract (0.4,
1.2, 2, 3.2, 4 mg) was mixed with 2.7 mL of 0.2 mol L−1 phosphate-
buffered saline (PBS, pH 6.6) and 2 mL of 10 g L−1 potassium
ferricyanide. The mixture was incubated at 50 ◦ C for 20 min. A 2.5 gL
aliquot of 100 g L−1 trichloroacetic acid was added to the mixture,
which was then centrifuged at 1000 × g for 10 min. The upper layer
(2.5 mL) was mixed with 2.5 mL of distilled water and 0.5 mL of 1 g
L−1 FeCl3 . The absorbance of the solution at 700 nm was measured
using a UV–visible spectrophotometer (UV-2450, Shimadzu).
Inhibitory effect on lipid peroxidation induced by Fe2+ /ascorbate
The inhibition of lipid peroxidation was assayed as described
previously.29 The reaction mixture contained 2.5 mL of liver
homogenate, 1.4 mL of 0.2 mmol L−1 PBS (pH 7.4), 2.5 mL of
10 µmol L−1 FeSO4 ·7H2 O and 2.5 mL of 100 µmol L−1 vitamin C.
Different amounts of shiitake ethanolic extract were added to the
reaction mixture and incubated at 37 ◦ C for 30 min, followed by
centrifugation at 3000 × g for 10 min. After the addition of 1 mL of
thiobarbituric acid reagent to the supernatant, the reaction tube
was placed in a boiling water bath for 15 min. The absorbance
of the solution at 530 nm was measured using a UV–visible
spectrophotometer (UV-2450, Shimadzu). The lipid peroxidation
inhibition rate of the sample was calculated as

  
inhibition (%) = Ablank − Asample /Ablank × 100
where Asample and Ablank are the UV absorbances of the sample
and blank respectively.
Statistical analysis
Values were expressed as mean ± standard deviation (SD) of three
replicates. Differences in means were analysed by one-way analysis
of variance (ANOVA) and Student’s t test using SPSS Version 16.0 for
Windows (SPSS Inc., Chicago, IL, USA). The statistical significance
of mean differences was based on a P value of <0.05.
RESULTS AND DISCUSSION
Effects of different drying methods on water-soluble
constituents of caps and stipes of shiitake
The effects of different drying methods (freeze-drying, shade
drying and hot air drying) on caps and stipes of shiitake were
evaluated. It was found that the drying method and drying
temperature affected the constituents significantly (P < 0.01)
(Table 1). The chemical parameters of caps and stipes dried at
temperatures from the lowest (-50 ◦ C) to the highest (150 ◦ C)
presented different values for each constituent.
The protein contents in caps and stipes ranged from 2.85 to
14.52 g kg−1 dry matter. The highest protein contents were found
in freeze-dried shiitake samples (14.52 g kg−1 in caps and 11.46 g
kg−1 in stipes), while the lowest were found in samples subjected
to shade drying at 28 ◦ C (5.85 g kg−1 in caps and 2.85 g kg−1 in
stipes). This might be due to enzymatic decomposition of protein
during shade drying. The protein contents in caps were higher than
those in stipes of all shiitake samples dried by different methods,
but there was no significant difference between caps and stipes
when samples were hot air dried at 150 ◦ C. The loss of protein
might be due to denaturation or changes in solubility of protein at
high temperature. The protein contents in shiitake samples were
lower than those found in other mushrooms such as Armillaria
mellea, Calocybe gambosa, Clitocybe odora and Coprinus comatus,
which ranged from 15.46 to 17.33 g kg−1 .30 The difference in
protein content might be due to the different species and sources
of mushroom samples.
The contents of free amino acids in caps and stipes ranged from
7.59 to 66.01 g kg−1 dry matter depending on the drying method.
The amino acid content in stipes subjected to hot air drying at 50
◦
C was highest (66.01 g kg−1 ), while the lowest amino acid content
(7.59 g kg−1 ) was found in freeze-dried caps. These results are in

Table 1. Contents of water-soluble constituents (g kg−1 dry matter) in caps and stipes of shiitake as affected by different drying methods

Drying method Temperature (◦ C) Part Protein Amino acids Uronic acid Neutral sugar

Freeze- drying Caps 14.52 ± 0.06a 7.59 ± 0.07a 14.73 ± 0.86a 18.78 ± 4.28a
-50
Stipes 11.46 ± 1.31b 14.68 ± 0.36c 11.98 ± 0.75c 25.92 ± 10.56a
Shade drying Caps 5.85 ± 1.29b 21.62 ± 2.29c 7.86 ± 0.60c 28.43 ± 5.59a
28
Stipes 2.85 ± 0.54c 30.51 ± 0.98c 17.84 ± 0.62c 57.25 ± 8.34b
Hot air drying Caps 6.82 ± 2.49a 14.05 ± 0.52c 12.08 ± 0.64b 56.55 ± 4.75b
50
Stipes 3.30 ± 0.16c 66.01 ± 5.81c 31.96 ± 0.97c 123.97 ± 26.34b
Caps 11.06 ± 0.11b 37.09 ± 1.67a 13.84 ± 0.98a 56.59 ± 8.42c
100
Stipes 9.37 ± 0.43c 24.77 ± 0.35c 12.37 ± 0.26b 43.70 ± 11.82b
Caps 8.82 ± 0.00c 15.10 ± 0.30c 13.86 ± 0.85a 88.10 ± 4.56c
150
Stipes 9.24 ± 0.19c 21.25 ± 0.94c 8.6 ± 0.13c 49.90 ± 2.19b

Values are mean ± SD of three replicate measurements. Means with different letters in the same column are significantly different at either P < 0.05
(adjacent letters) or P < 0.01 (non-adjacent letters).
3109

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 www.soci.org N Zhang et al.

agreement with those for protein contents, suggesting that high
temperature could promote proteolysis during the drying process.
The amino acid contents in stipes were higher than those in caps
when samples were freeze-dried, shade dried or hot air dried at
50 ◦ C. The total amino acid contents in fruiting bodies of other
mushrooms measured by Lee and Yun31 were 80.1, 68.6 and 56.2
g kg−1 for Agrocybe chaxingu, Pleurotus ostreatus and Flammulina
velutipes respectively. The amino acid contents in those
mushrooms were close to the amino acid contents found in shiitake
stipes hot air dried at 50 ◦ C, suggesting that stipes of shiitake could
be as nutritional as other mushrooms after suitable drying.
Different drying methods resulted in significant variation in the
contents of uronic acid and neutral sugar in shiitake caps and
stipes (P < 0.01). Uronic acid contents ranged from 7.86 to 31.96 g
kg−1 dry matter, while neutral sugar contents ranged from 18.78
to 123.97 g kg−1 dry matter. Both neutral sugar and uronic acid
contents were increased by hot air drying at 50 and 150 ◦ C. The
content of uronic acid was increased 2.7-fold in stipes and 0.8-fold
in caps and the content of neutral sugar was increased 3.0-fold
in caps (to 56.55 g kg−1 ) and 4.8-fold in stipes (to 123.97 g kg−1 )
when shiitake was dried at 50 ◦ C. One possible reason for the
increase in sugar content and decrease in protein content might
be the release of amino acids from proteins after denaturation,
which could react with sugars to produce melanoidines via the
Maillard reaction.32,33
Effects of different drying methods on contents of total
phenolics and eritadenine in caps and stipes of shiitake
The contents of total phenolics and eritadenine in caps and stipes
after different drying methods are shown in Table 2. Total phenolic
contents ranged from 1.88 to 5.83 g kg−1 dry matter when shiitake
of shiitake mushroom could increase its health-beneficial effects.
Total phenolic contents of 8.51, 10.89 and 32 g kg−1 in shiitake
were determined by other authors.34,35 The difference between
the present results and those of other studies might be due to the
different sources of shiitake samples.
Eritadenine is one of the main constituents of shiitake with
hypocholesterolaemic activity.11 In this study the effects of
different drying methods on the eritadenine content in shiitake
were determined by HPLC according to the method of Enman
et al.16 Compared with freeze-dried samples, the contents of
eritadenine in caps and stipes were reduced by about 50% in
both shade-dried and hot air-dried samples. With an increase in
drying temperature, the content of eritadenine was decreased.
The contents of eritadenine in caps were higher than those in
stipes. The highest contents of eritadenine were 16.61 and 14.25
g kg−1 in freeze-dried caps and stipes respectively. Enman et al.16
reported eritadenine contents ranging from 3.17 to 6.33 g kg−1
in dried shiitake. The difference might be due to the different
types, sources and processing procedures of shiitake samples. The
results of the present study suggested that eritadenine was stable
during the freeze-drying process, while it might react with other
components during the high-temperature drying process and
thus affect the bioactivities. Eritadenine and its derivatives were
also synthesised by Okumura et al.36 and their bioactivities were
compared. The most active derivatives were carboxylic acid esters
with short-chain monohydroxy alcohols. Therefore the drying
(a) 100
Scavenging effects (%)

80
were dried by the three different methods at temperatures from the
lowest (−50 ◦ C) to the highest (150 ◦ C). The total phenolic contents
60
in caps increased as the drying temperature was increased, while
the total phenolic contents in stipes were higher than those in
40
caps at all drying temperatures. The highest total phenolic content
in caps (3.68 g kg−1 ) was found after hot air drying at 50 ◦ C,
20
while the highest total phenolic content in stipes (5.83 g kg−1 )
was found after hot air drying at 100 ◦ C. The results were in 0
agreement with Choi et al.,4 who reported that heat treatment 0 10 20 30 40
-1
Concentration (mg mL )
-50°C drying 28°C drying 50°C drying
Table 2. Contents of total phenolics and eritadenine (g kg−1
dry
matter) in caps and stipes of shiitake as affected by different drying 100°C drying 150°C drying
methods
(b) 100
Temperature
(◦ C)
Scavenging effects (%)

Drying method Part Total phenolics Eritadenine 80

Freeze- drying Caps 1.88 ± 0.01a 16.61 ± 0.05a

-50 60
Stipes 5.26 ± 1.15b 14.25 ± 0.92b
Shade drying Caps 2.65 ± 0.05c 8.35 ± 0.22c 40
28
Stipes 3.97 ± 0.04c 6.92 ± 0.18c
Hot-air drying Caps 3.68 ± 0.13c 8.35 ± 0.32c 20
50
Stipes 5.01 ± 0.74b 6.74 ± 0.26c
Caps 3.20 ± 0.12c 8.25 ± 0.05c 0
100
Stipes 5.83 ± 0.21c 6.89 ± 0.33c 0 10 20 30 40
Caps 3.15 ± 0.02c 8.06 ± 0.26c -1
Concentration (mg mL )
150
Stipes 5.01 ± 0.16c 7.78 ± 0.08c
-50°C drying 28°C drying 50°C drying
Values are mean ± SD of three replicate measurements. Means with 100°C drying 150°C drying
different letters in the same column are significantly different at either
P < 0.05 (adjacent letters) or P < 0.01 (non-adjacent letters). Figure 1. Scavenging effects of differently dried shiitake samples on DPPH
3110

radicals: a, caps; b, stipes.

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method might result in changes in composition and thus affect (a) 100
the bioactivities of shiitake mushroom.
80

Inhibitory effects (%)

Antioxidant activities of shiitake as affected by different
drying methods 60

As the second most cultivated and popular mushroom, shiitake

40
has been reported to have antioxidant potential in several
studies.27,34 Tsai et al.37 found that ethanolic extracts of shiitake
20
were more effective than aqueous extracts. In the present study
the effects of different drying methods on the antioxidant activities 0
of ethanolic extracts of caps and stipes of shiitake were compared 0 10 20 30 40
for the first time. The ethanolic extracts of different shiitake -1
Concentration (mg mL )
samples were assayed for their scavenging activity on DPPH
radicals, their ferric reducing power and their inhibitory effect -50°C drying 28°C drying 50°C drying
on lipid peroxidation induced by Fe2+ /ascorbate. Concentration- 100°C drying 150°C drying
dependent effects were shown by all samples in the three
assays (Figs 1–3). (b) 100
Stipes showed the highest antioxidant activity (IC50 = 9.98 mg
mL−1 ) by DPPH assay (Fig. 1) and the highest antioxidant activity 80

Inhibitory effects (%)

(IC50 = 3.56 mg mL−1 ) by lipid peroxidation assay (Fig. 3) when
dried at 100 ◦ C (P < 0.01). The extraction efficiency of stipes was 60
lower than that of caps, being minimum at a drying temperature
of 100 ◦ C. The second highest antioxidant activity by DPPH assay 40

was shown by stipes dried at 150 ◦ C, while the second highest

20
antioxidant activity by lipid peroxidation assay was shown by
stipes dried at 50 ◦ C. The ferric reducing power (Fig. 2) showed
0
the same trend as the antioxidant activity determined by lipid 0 10 20 30 40
-1
Concentration (mg mL )
(a) 2
-50°C drying 28°C drying 50°C drying
100°C drying 150°C drying
Absorbance at 700nm

1.6

Figure 3. Inhibitory effects of differently dried shiitake samples on lipid

1.2 peroxidation induced by Fe2+ /ascorbate: a, caps; b, stipes.

0.8

peroxidation assay. Compared with caps, stipes showed better

0.4
scavenging of DPPH radicals, especially after drying at 100 and
150 ◦ C (P < 0.01). These results are in agreement with other
0
0 10 20 30 40 studies4,34,38 in which heat treatment was found to influence the
Concentration (mg mL-1)
antioxidant activities of samples. However, most studies showed
that freeze-drying was the most suitable method to preserve
-50°C drying 28°C drying 50°C drying chemical compounds. In the present study the improvement in
100°C drying 150°C drying antioxidant activities at high drying temperature might be due
to cell disruption, whereby more antioxidant compounds were
(b) 2
extracted from shiitake. On the other hand, there is a possibility
1.6
that new antioxidant compounds occurred when shiitake samples
Absorbance at 700nm

were dried at high temperature, which will need further study.

1.2

0.8
Correlation of phenolic and eritadenine contents with
antioxidant activities of shiitake
0.4 Phenolics are among the main antioxidant components present in
natural resources, and their correlation with antioxidant activities
0 has been discussed in many reports. In this study, there were
0 10 20 30 40 good correlations between phenolic contents in caps of shiitake
Concentration (mg mL-1) and antioxidant activities measured by DPPH assay (R2 = 0.935)
and lipid peroxidation assay (R2 = 0.911). The different drying
-50°C drying 28°C drying 50°C drying
methods and drying temperatures resulted in clear differences in
100°C drying 150°C drying
total phenolic contents and antioxidant activities. The antioxidant
Figure 2. Ferric reducing power of differently dried shiitake samples: a, activities of extracts of whole fruiting bodies of shiitake were also
3111

caps; b, stipes. found to correlate strongly with total phenolic contents in previous

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 www.soci.org
studies.34,39 Therefore phenolics are one of the main antioxidant
components in shiitake.
Eritadenine has been reported to have hepatoprotective
activity.3 However, in the present study, there was no clear
correlation between eritadenine contents and antioxidant
activities in dried shiitake samples (R2 = 0.081). Eritadenine might
not be the main antioxidant constituent in vitro in shiitake. The
changes in eritadenine content in different dried samples might
correlate with other activities such as hepatoprotective activity,
but this will need to be investigated further.
CONCLUSIONS
Three drying methods, freeze-drying, shade drying and hot air
drying, were applied to freshly harvested shiitake mushroom
samples. The chemical parameters and antioxidant properties of
shiitake stipes and caps were comparatively studied for the first
time. It was shown that the contents of total phenolics, amino
acids and neutral sugar in stipes were higher than those in caps of
shiitake, while caps showed advantages in terms of their contents
of protein and eritadenine, which suggested that stipes were more
nutritional than caps in some respects. Freeze-drying was benficial
for the stability of eritadenine and protein in shiitake, while hot
air drying at 50 ◦ C was beneficial for the stability or creation
of total phenolics, amino acids, uronic acid and neutral sugar.
The antioxidant activities of shiitake were affected by the drying
method used, and samples dried at high temperature showed
better antioxidant activities. The results of this study could be
useful for the application of shiitake and related products in the
food industry.
ACKNOWLEDGEMENTS
We are grateful for the financial support of this research from the
National High Technology Research and Development Program
(‘863’ Program) of China (grant no. SS2013AA100207) and from
the Ministry of Science and Technology of the People’s Republic
of China (grant no. 2012BAD33B08).
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