Food Chemistry 201 (2016) 80–86

Contents lists available at ScienceDirect

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

Effect of cooking methods on nutritional quality and volatile compounds

of Chinese chestnut (Castanea mollissima Blume)
Qian Li a, Xianhe Shi a, Qiaojiao Zhao a, Yahui Cui a, Jie Ouyang a,⇑, Fang Xu b,⇑
a
Department of Food Science and Engineering, College of Biological Sciences and Technology, Beijing Key Laboratory of Forest Food Processing and Safety, Beijing Forestry
University, Beijing 100083, China
b
Analytical and Testing Center, Beijing Forestry University, Beijing 100083, China

a r t i c l e i n f o a b s t r a c t

Article history: This study aimed to evaluate the effects of different cooking methods on the content of important nutri-
Received 4 October 2015 ents and volatiles in the fruit of Chinese chestnut. The nutritional compounds, including starch, water-
Received in revised form 17 December 2015 soluble protein, free amino acids, reducing sugar, sucrose, organic acids and total flavonoids, of boiled,
Accepted 18 January 2016
roasted and fried chestnuts were significantly (P < 0.05) lower than those of fresh chestnuts after cooking,
Available online 18 January 2016
while the amylose, fat, crude protein and total polyphenol content varied slightly (P > 0.05). L-Aspartic
acid, L-glutamic acid and L-arginine were found to be the main reduced free amino acids in cooked chest-
Chemical compounds studied in this article:
nuts. The main aromatic compositions in fresh chestnuts were aldehydes and esters, while ketones, fur-
Sucrose (PubChem CID: 5988)
Fructose (PubChem CID: 5984)
fural and furan were formed in cooked chestnuts due to the Maillard reaction and degradation of
Glucose (PubChem CID: 5793) saccharides, amino acids and lipids. Principle component analysis demonstrated that roasting and frying
L-Aspartic acid (PubChem CID: 5960) had a similar effect on the nutritional composition of chestnuts, which differed from that of the boiling
L-Glutamic acid (PubChem CID: 611) process.
L-Arginine (PubChem CID: 6322) Ó 2016 Elsevier Ltd. All rights reserved.
Oxalic acid (PubChem CID: 971)
Malic acid (PubChem CID: 525)
Ascorbic acid (PubChem CID: 54670067)
Citric acid (PubChem CID: 311)
Fumaric acid (PubChem CID: 444972)

Keywords:
Chinese chestnut
Cooking method
Nutritional quality
Volatile compound

1. Introduction
Chestnuts belong to the family Fagaceae and are mainly dis-
tributed in Eastern and Southwestern Asia, Southern Europe and
North America. Among the 12 world chestnut species, the annual
fruit production of the Chinese chestnut (Castanea mollissima
Blume) is 925,000 t, compared with 108,000 t for the European
chestnut (C. sativa Miller) and 55,800 t for the North and South
American chestnut (C. dentata Borkh) (De Vasconcelos, Bennett,
Rosa, & Ferreira-Cardoso, 2010; Fernandes et al., 2011). The best
development conditions are found at altitudes above 500 m and
low winter temperatures, as in the Yan Mountain region of North-
ern China. Chestnuts are an important food resource with high
nutritional value. Fresh Chinese chestnut fruits contain 52.0%
⇑ Corresponding authors.
E-mail addresses: ouyangjie@bjfu.edu.cn (J. Ouyang), casxu@sohu.com (F. Xu).
water, 42.2% carbohydrates, 4.2% proteins and 0.7% lipids (Yang,
Pan, & Wang, 2009), while the starch, total sugar, crude protein
and fat content in Spanish chestnuts are 42.2–59.8%, 9.5–22.2%,
4.8–6.9% and 1.7–4.0% d.m. (dry matter), respectively, varying by
cultivar and region (Pereira-Lorenzo, Ramos-Cabrer, Díaz-
Hernández, Ciordia-Ara, & Ríos-Mesa, 2006). Chinese chestnut
fruits can improve the function of kidneys according to the ancient
encyclopedia of China Compendium of Materia Medica (Ben Cao
Gang Mu) of the Ming Dynasty (A.D. 1590). From the various
compositions and health studies, it is clear that chestnut fruits,
and potentially other extracts from chestnut trees, have consider-
able potential as a functional food or as food ingredients (De
Vasconcelos, Bennett, et al., 2010).
Studies in the literature mainly focused on fresh materials
(varieties) or assessed the impact of different heat treatments on
European chestnut composition and antioxidant activity
(Attanasio, Cinquanta, Albanese, & Di Matteo, 2004; Barreira,

http://dx.doi.org/10.1016/j.foodchem.2016.01.068
0308-8146/Ó 2016 Elsevier Ltd. All rights reserved.
 Q. Li et al. / Food Chemistry 201 (2016) 80–86
Pereira, Oliveira, & Ferreira, 2010; Morini & Maga, 1995a). The
effects at the four sequential major stages of industrial processing
(fresh, after storing for 3 months at ±0 °C and keeping the relative
humidity at 90%, after industrial peeling by flame or fire at 800–
1000 °C, after freezing in a tunnel with a CO2 flow at
65 °C) on
the nutritional content of six cultivars of C. sativa were thoroughly
evaluated, including starch, fat, energy, fiber (De Vasconcelos,
Bennett, Rosa, & Ferreira-Cardoso, 2009a), crude protein, free
amino acids, phenolic phytochemicals (De Vasconcelos, Bennett,
Rosa, & Ferreira-Cardoso, 2009b), minerals, free sugars, carotenoids
and antioxidant vitamins (De Vasconcelos et al., 2010). The vitamin
C content of fresh chestnuts varied from 400 to 693 mg/kg dry
weight between the different European cultivars, and a significant
decrease, 25–54% for the boiling process and 2–77% for the roast-
ing process, was observed (Barros, Nunes, Gonçalves, Bennett, &
Silva, 2011).
The traditional methods for cooking Chinese chestnuts include
frying them unshelled with sugar (tang chao li zi) or shelled and
cooking them with other food materials. Currently, a type of indus-
trially processed and packaged chestnut kernel has become popu-
lar in East Asia and is generally cooked via the boiling method. The
kernel of a fresh chestnut has a weak smell of fruit, which becomes
a strong flavor during cooking after thermal processing. A total of
30–33 components, including hydrocarbons, alcohols, aldehydes,
ketones, furans, pyranone and acids, were firstly identified from
the flavor extract of boiled and roasted Chinese chestnuts, respec-
tively (Morini & Maga, 1995b). Monoterpenes and derivatives of
butane, pentane, hexane and heptane were identified as important
aroma impact compounds from roasted Italian chestnuts (Krist,
Unterweger, Bandion, & Buchbauer, 2004).
The purpose of this study is to analyze the nutritional and aro-
matic components in fresh and boiled, roasted and fried chestnuts
to investigate the effects of different cooking methods on the con-
tents of proximate composition and other chemical constituents. It
is important to assess these changes so that the industrialized pro-
cessing of chestnut fruits can be optimized for nutritional and aro-
matic qualities.
2. Materials and methods
2.1. Reagents
The standards (amylose, sucrose, glucose, fructose, oxalic acid,
malic acid, ascorbic acid, citric acid and fumaric acid) were pur-
chased from Sigma–Aldrich (USA). Acetonitrile was HPLC grade
(Fisher, USA). All other chemicals, unless otherwise noted, were
of analytical grade and purchased from Sinopharm Chemical
Reagent Beijing Co., Ltd of PR China. The water was obtained from
a Milli-Q water purification system (Millipore, Belford, MA, USA).
2.2. Raw materials and sample preparation
The Chinese chestnut cultivar ‘Zaofeng’ was purchased from a
commercial market in Qianxi, Hebei province of North China. The
fruits were harvested in September of 2013 and stored at 2 °C in
the dark for one month before use. Chestnuts were divided into
four groups: group 1 had no treatment, i.e., fresh chestnuts; group
2 was boiled in 100 °C water (chestnut: water = 1:2, w/v) for
20 min, i.e., boiled chestnuts; group 3 was roasted at 200 °C for
25 min in an electric oven (T3-L383b, Guangdong Midea Kitchen
Appliance Manufacturing Co. Ltd., China), i.e., roasted chestnuts;
group 4 was fried at 240 °C for 15 min in an electric pan
(WK2102T, Guangdong Midea Kitchen Appliance Manufacturing
Co. Ltd., China), i.e., fried chestnuts. Each group of 500 g was hulled,
lyophilized, and ground using a 40-mesh, and the obtained chest-
81
nut powder was sealed in a polyethylene bag and stored at 2 °C in
the dark before analysis. Water content was measured by oven
drying at 105 °C until constant weight was achieved (AOAC
method 925.40) (AOAC, 2000). Therefore, the calculation of the
content of the chemical composition of chestnuts was based on
d.m.
2.3. Analysis of total starch and amylose
Chestnut starch was isolated by using the alkaline method as
previously reported (Correia & Beirão-da-Costa, 2012). A chestnut
sample of 500 g was immersed in 1000 mL of NaOH solution
(0.2%, w/v) for 2 h. Then, the mixture was homogenized and
filtered through a 75 lm stainless sieve to remove large particles.
The filtrate was centrifuged at 4000g for 5 min; then, the
mucilaginous layer was scraped away and the precipitate was
washed with water three times. The extracted starches were dried
for two days at 40 °C in a ventilated drying oven. Total starch
content was determined according to the AOAC method 996.11
(AOAC, 1997). Amylose content was analyzed using a K-AMYL
07/11 Amylose/Amylopectin assay kit (Megazyme International
Ireland, Ireland). Chestnut starch samples were completely dis-
persed by heating in dimethyl sulfoxide (DMSO) (Maršálková
et al., 2010; https://secure.megazyme.com/Amylose-Amylopectin-
Assay-Kit).
2.4. Analysis of crude fat, crude protein and water-soluble protein
The crude fat and nitrogen content was determined
according to AOAC official methods 920.39 and 954.01 (AOAC,
1997). Crude protein content was calculated by multiplying the
nitrogen content by 5.30 (FAO, 1986). Chestnut powder was
extracted by using distilled water three times, and the water-
soluble protein content was measured by using Coomassie Brilliant
Blue (Bradford, 1976). Bovine serum albumin (BSA) was used as the
standard.
2.5. Analysis of reducing sugar, sucrose, glucose and fructose
The fresh and fried chestnuts, 10 g of each, were shelled and
homogenized in 50 mL of Et-OH/H2O (80%, v/v). The slurry was
shaken at 70 °C for 30 min and then centrifuged at 8000g for
15 min; Et-OH was removed from supernatant via vacuum
evaporation at 60 °C. The concentrated extract was added to
0.5 mL of zinc acetate (1 M) and 0.5 mL of potassium ferrocyanide
(0.25 M) and supplemented, using deionized water, to 5 mL. The
mixture was kept at room temperature for 30 min and then cen-
trifuged at 8000g for 15 min. The supernatant was filtered using
0.2 lm Millipore Express Membrane Filters (Millipore, Belford,
MA, USA) and stored at 4 °C until analysis. The reducing sugar con-
tent was analyzed by using Fehling’s reagent titration method
(Ayoola et al., 2008). The sucrose, glucose and fructose content in
chestnuts was determined via HPLC (high performance liquid
chromatography).
HPLC was performed on a LUMTECH (Lumiere, German) system
with a refractive index detector (50D) and a Waters WAT084038
NH2 column (4.6  250 mm, 5 lm). The analyses of sucrose, glu-
cose and fructose were carried out separately. The mobile phase
was acetonitrile–water (75:25, v/v), the detection wavelength
was 285 nm, the injection volume was 20 lL and the flow rate
was 1.0 mL/min.
2.6. Energetic value
Energetic value was calculated as described by Fernandes et al.
(2011):
82 Q. Li et al. / Food Chemistry 201 (2016) 80–86
Energy ðkcal=100 gÞ ¼ ðg of protein þ g of starch þ g of sucrose
þ g of glucose þ g of fructoseÞ  4
þ ðg of lipidÞ  9
2.7. Analysis of total polyphenols and total flavonoids
Three grams of chestnut powder was reflux-extracted with
75 mL of Et-OH/H2O (70%, v/v) at 70 °C for 30 min. The extract
was filtered using 0.22 lm Millipore Express Membrane Filters
and stored at 4 °C until analysis. For the determination of total
polyphenols content, the reaction system included 1.0 mL of
sample solution, 5.0 mL of Folin–Ciocalteu reagent and 4 mL of
Na2CO3 (7.5%, w/v). The mixture was kept at room temperature
for 60 min, and then the absorbance at 765 nm was measured
(Ramanauskienė, Inkėnienė, Petrikaitė, & Briedis, 2013). The con-
centration of polyphenols was calculated according to the standard
curve, with gallic acid as the standard. The results were expressed
as mg of gallic acid equivalent (GAE)/g d.m.
To measure total flavonoid content, the reaction system con-
sisted of 2 mL of sample and 0.5 mL of NaNO2 (5%, w/v). After reac-
tion for 6 min, 0.5 mL of Al(NO3)3 (10%, w/v) was added and left to
stand for 6 min; then, 4.0 mL of NaOH (4%, w/v) was added and 70%
Et-OH/H2O (v/v) was supplemented to increase to 10 mL. The
absorbance was measured at 510 nm after reaction for 15 min
(Gao et al., 2011). The concentration of flavonoid was calculated
according to the standard curve, with rutin as the standard. The
results were expressed as mg of rutin equivalent/g d.m.
2.8. HPLC analysis of organic acids
HPLC analysis was carried out according to a previous study
(Ribeiro et al., 2007), with some modifications. 5 g of chestnut
powder was extracted with 50 mL of Et-OH/H2O (80%, v/v) at room
temperature for 20 min and then centrifuged at 8000g for 15 min.
Et-OH was removed from the supernatant by flushing with nitro-
gen. The concentrated extract was added to 0.2 mL of phosphoric
acid (1 M) and supplemented with deionized water to reach
10 mL. The solution was filtered using a 0.2 lm microfiltration
membrane and stored at 4 °C until HPLC analysis. The determina-
tion of organic acids, including oxalic acid, malic acid, ascorbic
acid, citric acid and fumaric acid, was performed on a Shimadzu
LC-2010 with a C18 column (4.6 mm  250 mm, 5 lm; Shimadzu).
The mobile phase was 0.01 M (NH4)2HPO4 (pH 2.7) and the detec-
tion wavelength was 210 nm. The injection volume was 20 lL and
the flow rate was 1.0 mL/min.
2.9. HPLC analysis of free amino acids (FAA)
The fresh and cooked chestnuts, 10 g of each, were shelled and
homogenized in 150 mL of Me-OH/H2O (80%, v/v). The slurry was
centrifuged at 8000g for 15 min, and the sediment was homoge-
nized in 150 mL of Me-OH/H2O (80%, v/v) for a second time and
centrifuged at 8000g for 15 min again. The two supernatants were
combined, vacuum concentrated and lyophilized. The obtained
powder was dissolved in 5% (w/v) trichloroacetic acid to reach a
constant volume of 25 mL, filtered using a 0.2 lm microfiltration
membrane, and stored at 4 °C until HPLC analysis. The samples
were hydrolyzed using 7.5 M HCl at 110 °C for 24 h. The concentra-
tions of amino acids were analyzed via HPLC on a Hitachi
Automatic Amino Acid Analyzer L-8900 (Hitachi, Japan) system
with aUV–VIS spectrometer and a Hitachi #2622pH column
(4.6  60 mm). The mobile phase was 0.075 M of sodium citrate
buffer (pH 3.3) and the detection wavelength was 570 nm. The
flow rate was 0.4 mL/min, the injection volume was 20 lL and
the oven temperature was set at 57 °C.
2.10. Analysis of volatile components
The analysis was carried out according to the method described
by Krist et al. (2004), with some modifications. The fresh, boiled,
roasted and fried chestnuts were each hulled and cut into particles
of 2 mm  2 mm  2 mm. The volatiles of the samples were ana-
lyzed by using a dynamic headspace sample (DHS) and automatic
thermal desorption-gas chromatography–mass spectrometric
(ATD-GC/MS) system. The adsorption tubes were pretreated in a
desorption disposer (TP-2040, Beijing BeiFenTianPu Instrument
Co. Ltd, China) to remove impurities by flushing with helium at
270 °C for 120 min. The chestnuts were put into a sampling bag
(Reynolds, 406 mm  444 mm, Richmond, VA, USA); the air in
the bag was pumped out for 40 min and then filtrated air was
pumped into the bag. The sampling bag was then sealed and left
alone for 90 min to accumulate volatiles. The volatiles were
pumped out by using an Atmosphere Sampling Instrument (QC-
1S, Beijing Municipal Institute of Labor Protection, China) for
60 min and adsorbed in an adsorption tube.
The filled-in adsorption tube was put in the ATD (Perkin Elmer
Turbo Matrix 650, USA); GC–MS analyses were performed using a
gas chromatograph and mass spectrometer (Perkin Elmer Clarus
600 Gas Chromatograph & 600 Mass Spectrometer, USA) equipped
with a DB-5MS capillary column (30 m  0.25 mm, 0.25 lm film
thickness). The first stage of desorption was carried out at 260 °C,
with helium as the carrier gas and the flow rate being 1.5 mL/min.
The desorbed volatiles were adsorbed in cold hydrazine (
25 °C),
followed by the second stage of desorption. The cold hydrazine tem-
perature increased from
25 °C to 300 °C at a rate of 40 °C/s. The
re-desorbed volatiles flowed into GC–MS through a pipe (250 °C).
Column temperature was initially kept at 40 °C for 2 min, then
increased to 200 °C at a rate of 6 °C/min, held for 5 min and finally
raised to 270 °C at 20 °C/min, holding for 5 min. Helium was used
as the carrier gas at a constant flow of 1.5 mL/min. Mass spectra
were recorded in EI mode, with a 29–500 amu scan range and
0.2 s scan time. Interface temperature was 250 °C; ion source tem-
perature was 220 °C; ionization voltage was 70 eV. Statistical anal-
yses of the data were conducted via a computer search using
digital libraries of mass spectral data (NIST2008). Retention indices
(RI) of constituents were determined using standard C8–C25
straight chain hydrocarbons (Shanghai Anpel Co. Ltd., China). The
RI references were obtained from the NIST Chemistry WebBook
http://webbook.nist.gov/chemistry/.
2.11. Statistical analysis
All experiments were carried out in triplicate, and the results
were expressed as mean values based on dry matter. For HPLC
analyses of the organic acids and FAA, the extractions were per-
formed in triplicate, and each sample was quantified in duplicate.
Means were compared using Tukey’s honestly significant differ-
ence (HSD) multiple comparison test by SPSS17.0 software (IBM
Corporation, Armonk, NY, USA). A principal component analysis
(PCA) of the data was performed using UnscramblerÒ X10.2 (CAMO
software, Oslo, Norway).
3. Results and discussion
3.1. Effect of cooking on the variation of proximate composition
Results obtained for the proximate composition of fresh, boiled,
roasted and fried chestnuts are shown in Table 1, which includes
 Q. Li et al. / Food Chemistry 201 (2016) 80–86 83

Table 1
Proximate composition of fresh and cooked chestnuts.

Chestnuts Total starch Amylose Crude fat Crude Water-soluble Reducing Sucrose Glucose Fructose Energetic
(%, d.m.) in starch (%, d.m.) protein protein sugar (%, d.m.) (%, d.m.) (%, d.m.) value
(%, d.m.) (%, d.m.) (%, d.m.) (%, d.m.) (kcal/100 g)
Fresh 71.08 ± 0.12a 19.87 ± 0.52a 2.27 ± 0.06a 8.27 ± 0.75a 2.65 ± 0.31a 2.06 ± 0.04a 9.85 ± 0.87a 0.22 ± 0.03a 0.18 ± 0.02a 405 ± 43a
Boiled 57.69 ± 0.66c 18.88 ± 0.95a 2.27 ± 0.10a 8.40 ± 0.81a 0.89 ± 0.08b 1.15 ± 0.03b 5.90 ± 0.65c 0.19 ± 0.02b 0.11 ± 0.01c 354 ± 41b
Roasted 61.25 ± 0.14b 21.21 ± 0.28a 1.96 ± 0.08b 8.49 ± 0.86a 0.74 ± 0.07c 0.67 ± 0.01d 9.21 ± 0.91b 0.23 ± 0.02a 0.17 ± 0.02ab 318 ± 28c
Fried 62.91 ± 2.27b 19.65 ± 0.22a 1.42 ± 0.12c 8.44 ± 0.89a 0.77 ± 0.09c 0.80 ± 0.01c 9.26 ± 0.82b 0.18 ± 0.03a 0.16 ± 0.02b 324 ± 39c

Values are expressed as means ± SD (n = 3). Different letters within one column represent significant difference at P < 0.05.

Table 2
Free amino acid content of raw, boiled, roasted and fried chestnuts.

Free amino acid (mg/g DW) Fresh chestnut Boiled chestnut Roasted chestnut Fried chestnut
Asp 4.23 ± 0.32a 2.12 ± 0.17b 1.82 ± 0.21c 2.07 ± 0.20b
Thr 0.10 ± 0.02a 0.02 ± 0.00b 0.03 ± 0.00b 0.04 ± 0.01b
Ser 0.15 ± 0.02a 0.05 ± 0.01b 0.05 ± 0.01b 0.06 ± 0.01b
Glu 1.29 ± 0.11a 1.09 ± 0.09b 0.97 ± 0.10c 0.96 ± 0.08c
Pro 0.14 ± 0.02a 0.04 ± 0.01b 0.05 ± 0.01b 0.05 ± 0.01b
Gly 0.09 ± 0.01a 0.05 ± 0.01c 0.05 ± 0.01c 0.07 ± 0.01b
Ala 0.15 ± 0.02a 0.11 ± 0.02b 0.12 ± 0.03b 0.10 ± 0.02b
Val 0.08 ± 0.01a 0.03 ± 0.01b 0.03 ± 0.00b 0.03 ± 0.01b
Ile 0.03 ± 0.01a 0.01 ± 0.00a 0.02 ± 0.00a 0.01 ± 0.00a
Leu 0.04 ± 0.01a 0.03 ± 0.00a 0.03 ± 0.01a 0.02 ± 0.00a
Tyr ND ND ND ND
Phe 0.11 ± 0.02a 0.02 ± 0.00b 0.02 ± 0.00b 0.02 ± 0.00b
Lys 0.04 ± 0.01a 0.02 ± 0.00ab 0.02 ± 0.00ab 0.01 ± 0.00b
His 0.11 ± 0.01a 0.03 ± 0.01b 0.03 ± 0.00b 0.04 ± 0.01b
Arg 1.11 ± 0.09a 0.11 ± 0.02b 0.11 ± 0.02b 0.11 ± 0.01b
Total 7.67 ± 0.67a 3.73 ± 0.36b 3.35 ± 0.40c 3.78 ± 0.37b

Values are expressed as means ± SD (n = 3). Different letters within one row represent significant difference at P < 0.05. ND: not detected.

total starch, amylose, crude fat, crude protein, water-soluble pro-
tein, reducing sugar, sucrose, glucose, fructose and calculated ener-
getic value. The total starch content in fresh chestnuts was 71.08%
d.m., higher than that (53.8%) of Chinese chestnuts (Liu, Wang,
Chang, & Wang, 2014) and (48.74–53.89%) of Portuguese chestnuts
(De Vasconcelos, Bennett, et al., 2010). After cooking, the total
starch content significantly (P < 0.01) decreased in boiled
(57.69%), roasted (61.25%) and fried (62.91%) chestnuts, which
may be attributed to the starch degradation at high temperature
(Bryce & Greenwood, 1963). Meanwhile, the starch content in
boiled chestnuts was lower than that of other cooked chestnuts
because part of the water-soluble starch was dissolved in water
during boiling. The ratios of amylose/total starch in fresh, boiled,
roasted and fried chestnuts were 19.87%, 18.88%, 20.21% and
19.65%, respectively, exhibiting no difference (P > 0.05). In previous
studies, high temperature treatment had the effect of lowering
starch content; the starch content decreased from 53.83% to
51.19% d.m. in chestnut fruits (C. sativa) after industrial peeling
by flame or fire at 800–1000 °C (De Vasconcelos et al., 2009a )
and reduced from 58.3% to 56.2% and 53.9% d.m. after drying at
40 and 60 °C, respectively (Attanasio et al., 2004). Meanwhile, the
ratio of amylose also increased from 32.9% to 43.3% and 57.8% after
drying at 40 and 60 °C, respectively. The fresh chestnut granules
appeared to be round or oval and were changed to being shapeless,
with their surfaces being quite rough after drying (Attanasio et al.,
2004).
The crude fat content was 2.27% d.m. in fresh chestnuts, which
was in accordance with the previously reported 2.1–2.4% (Liu et al.,
2014) and 1.91–4.39% (De Vasconcelos, Bennett, et al., 2010). After
cooking, the crude fat content became 2.27%, 1.96% and 1.42% in
boiled, roasted and fried chestnuts, respectively. It seems that boil-
ing had no effect on chestnut fat, but roasting and frying could
lower crude fat content by decomposing fat at a high temperature
(Das, Babylatha, Pavithra, & Khatoon, 2013). Similar results were
observed by Gonçalves et al. (2010), who found crude fat content
in fresh and roasted chestnuts of 3.20% and 3.08% d.m.,
respectively. Künsch et al. (2001) also found that total fatty acids
decreased after roasting in Switzerland native chestnuts
(C. sativa Mill).
Crude protein in fresh chestnuts was 8.27% d.m., which was
similar to the 8.5% of Liu et al. (2014), and increased slightly after
boiling (8.40%), roasting (8.49%) and frying (8.44%). The crude
protein content increased from 48.9–49.1 to 50.2–53.9 mg/g d.m.
after industrial peeling by fire (De Vasconcelos et al., 2009b ).
Gonçalves et al. (2010) noted that the cooking processes signifi-
cantly (P < 0.0001) affected the primary and secondary metabolites
of chestnuts, with the protein content in roasted and boiled chest-
nuts being 67.1 and 62.8 mg/g d.m., respectively, varying from that
of fresh chestnuts (65.1 mg/g). Water-soluble protein, mainly pep-
tides and hydrophilic proteins, decreased significantly (P < 0.01)
when subject to the Maillard reaction during cooking.
It’s well known that the Maillard reaction is the main formation
mechanism of aromatic components in thermal processed nuts;
thus, the analysis of the variation of FAA in fresh and cooked chest-
nuts is very important. The content of fresh chestnuts with regard
to 14 types of FAA was analyzed (Table 2), with a total amount of
7.67 mg/g d.m., which mainly included L-aspartic acid (4.23 mg/g),
L-glutamic acid (1.29 mg/g) and L-arginine (1.11 mg/g). Similarly,
the free amino acid profiles in Portugal chestnuts were dominated
by L-aspartic acid (0.70–1.41% d.m.), followed by L-glutamic acid
(0.61–1.03%), leucine (0.40–0.74%), L-alanine (0.45–0.74%) and
L-arginine (0.22–1.16%) (Borges, Gonçalves, de Carvalho, Correia,
& Silva, 2008). The total FAA decreased by 50.7% to 3.78 mg/g after
frying, while in boiled and roasted chestnuts, it decreased by 51.4%
and 56.3%, respectively. The sum of decreasing amounts of
L-aspartic acid, L-glutamic acid and L-arginine accounted for
84 Q. Li et al. / Food Chemistry 201 (2016) 80–86

Table 3
Total polyphenols, total flavonoid and organic acids of fresh and cooked chestnuts (mg/g d.m.).

Chestnuts Total polyphenols Total flavonoids Malic acid Citric acid Oxalic acid Ascorbic acid Fumaric acid
a a a a a a
Fresh 2.24 ± 0.05 2.62 ± 0.13 1.2593 ± 0.1024 0.8569 ± 0.0832 0.1226 ± 0.0111 0.0475 ± 0.0051 0.0207 ± 0.0031a
Boiled 2.03 ± 0.02b 2.12 ± 0.09b 1.1355 ± 0.0988a 0.6941 ± 0.0456b 0.1011 ± 0.0095c 0.0281 ± 0.0032b 0.0032 ± 0.0006c
Roasted 2.26 ± 0.01a 2.25 ± 0.11a 0.7979 ± 0.0803b 0.6669 ± 0.0733b 0.1077 ± 0.0080c 0.0136 ± 0.0014d 0.0069 ± 0.0007b
Fried 2.08 ± 0.05b 2.13 ± 0.08b 0.7979 ± 0.0752b 0.2806 ± 0.0410c 0.1124 ± 0.0092b 0.0157 ± 0.0011c 0.0066 ± 0.0007b

Values are expressed as means ± SD (n = 3). Different letters within one column represent significant difference at P < 0.05.

84.0–89.7% of the total FAA loss, which indicated that they had a
close relationship with aroma formation in chestnuts.
The reducing sugar content in fresh chestnuts was 2.06% d.m.
and decreased to 1.15%, 0.67% and 0.80% in boiled, roasted and
fried chestnuts, respectively. The concentrations of sucrose, glu-
cose and fructose in varieties of chestnut fruits from Tenerife
(Spain) were between 31.10–99.40, 0.25–1.90 and 0.25–1.53 g/kg
d.m., respectively (Hernández Suárez, Rodriguez Galdón, Rios
Mesa, Diaz Romero, & Rodriguez Rodriguez, 2012). In the present
study, the sucrose content in Chinese chestnuts was 9.85% d.m.
and decreased significantly (P < 0.05) after thermal processing,
while the glucose and fructose content changed slightly. The vari-
ation of reducing sugar, sucrose, glucose and fructose in cooked
chestnuts is mainly due to four processes: the hydrolysis of starch
to oligosaccharide and monosaccharide, decomposition of sucrose
to glucose and fructose (Bernárdez, De la Montaña Miguélez, &
Queijeiro, 2004), the caramelization and degradation of sugars,
and the Maillard reaction. The sucrose content in fresh Italian
chestnuts (29.7%) decreased significantly (P < 0.05) to 22.3% after
drying at 60 °C, while fructose (1.9%) and glucose (1.4%) content
remained unchanged under the same drying condition (Attanasio
et al., 2004).
The energetic value of fresh chestnuts was 405 kcal/100 g d.m.,
which decreased to 318–354 kcal/100 g d.m. after cooking owing
to the degradation of the proximate composition, especially starch,
which is the main energy source (approximately 70%) in chestnuts.
The energetic value of Portuguese chestnuts (C. sativa) was
402 kcal/100 g d.m., which changed to 396 kcal/100 g d.m. after
30-day storage (Fernandes et al., 2011).
3.2. Effect of cooking on the variation of total polyphenols, total
flavonoids and organic acids
The total polyphenol content in fresh chestnuts was 2.24 mg/g
d.m., which remained unchanged after roasting (2.26 mg/g) and
decreased to 2.03 and 2.08 mg/g after boiling and frying, respec-
tively (Table 3). In the previous studies, total polyphenols
remained unchanged (P > 0.05) after boiling but increased signifi-
cantly (P < 0.05) after roasting (Gonçalves et al., 2010) and
increased significantly (P < 0.05) after industrial peeling by flame
(De Vasconcelos et al., 2009b ). Polyphenols may transfer from
the chestnut shell to the kernel, by which the content of polyphe-
nols in a chestnut kernel is increased (Gonçalves et al., 2010). On
the other hand, polyphenols decompose during cooking, which
leads to a decrease. The content of total flavonoids decreased from
2.62 to 2.12, 2.25 and 2.13 mg/g d.m. after boiling, roasting and fry-
ing, respectively. All five tested organic acids, including malic acid,
citric acid, oxalic acid, ascorbic acid and fumaric acid, decreased
significantly (P < 0.01) after cooking. The total content of the above
organic acids in fresh chestnut was 2.307 mg/g d.m., which
decreased by 50.6% after frying, while only being reduced by 15%
after boiling. Gonçalves et al. (2010) found that citric acid

Fig. 1. Principal component analysis score plot for the classification of fresh, boiled, roasted and fried chestnuts. The vertical and horizontal lines show the 95% confidence
interval.
 Q. Li et al. / Food Chemistry 201 (2016) 80–86 85

Table 4
Comparison of volatile components of raw, boiled, fried and roasted chestnuts.

Compounds RI (cal) RI (lit) Percentages

Fresh chestnut Boiled chestnut Roasted chestnut Fried chestnut
Ethyl acetate – 610 92.46 96.40 94.82 –
Spiro[2,4]hepta-4,6-diene – – – 0.43 – –
Hexanal 811 802 1.54 0.77 1.26 14.18
Dihydro-2-methyl-3(2H)-furanone 820 810 – – 0.04 –
Butyl acetate 825 810 1.20 0.85 0.97 –
Furfural 842 830 – – 0.37 36.58
Ethyl benzene 873 849 0.67 0.17 0.59 0.28
3-Methyl-1-butanol acetate 892 876 0.34 0.10 0.10 1.35
3-Heptanone 901 894 – 0.04 – 8.87
2-Hydroxy-2-cyclopenten-1-one 908 926 – – 0.85 1.42
4-Hydroxy-2-butanone 936 – – – – 6.76
3-Carene 942 1007 – – – 1.15
1R-a-pinene 944 937 0.33 0.14 0.24 –
1-(Methylencyclopropyl)-ethanol 965 – – – – 0.56
Benzaldehyde 971 964 0.55 0.16 0.22 3.32
(2-Hexenoic acid, methyl ester) 972 – – – – 0.30
6-Methyl-5-hepten-2-one 1002 – 0.18 0.11 0.05 4.67
2-Pentyl-furan 1005 991 – – – 0.73
Octanal 1021 1000 0.36 0.14 0.09 3.42
Acetophenone 1083 1062 0.11 0.06 0.02 1.41
Nonanal 1122 1102 1.81 0.50 0.26 10.73
Dodecane 1217 1200 0.17 0.05 0.06 0.94
Decanal 1223 1206 0.28 0.08 0.06 3.33
Total 100 100 100 100

RI (lit) from the NIST Chemistry WebBook (http://webbook.nist.gov/chemistry/).

increased after boiling and roasting, while malic acid decreased.
The content of ascorbic acid decreased by 33% and 37% after boiling
and roasting, respectively (Barros et al., 2011).
To establish the relationship between the different variables in
fresh and cooked chestnuts, PCA was applied to total starch, amy-
lose, crude fat, crude protein, water-soluble protein, reducing
sugar, sucrose, glucose, fructose, total polyphenols, total flavonoids
and organic acids (Fig. 1). An eigenvalue of 100% was achieved
using two PCs (PC1 = 99%, PC2 = 1%). Fresh chestnuts were clearly
distinguished from boiled, roasted and fried chestnuts, while
roasted chestnuts were similar to fried chestnuts because of simi-
lar cooking conditions.
3.3. Volatile components in fresh and cooked chestnuts
The main volatiles in fresh chestnuts were esters and aldehydes,
which mainly included ethyl acetate (92.46%), nonanal (1.81%),
hexanal (1.54%), butyl acetate (1.20%), and benzaldehyde (0.55%),
13 compounds in total (Table 4). The primary volatiles in fried
chestnuts were furfural (36.58%), hexanal (14.18%), nonanal
(10.73%), 3-heptanone (8.87%), and 4-hydroxy-2-butanone
(6.76%). Ethyl acetate (96.40%), butyl acetate (0.85%), hexanal
(0.77%), nonanal (0.50%) and spiro[2,4]hepta-4,6-diene (0.43%)
were found in boiled chestnuts; ethyl acetate (94.82%), hexanal
(1.26%), butyl acetate (0.97%) and 2-hydroxy-2-cyclopenten-
1-one (0.85%) were found in roasted chestnuts. There was still a
large amount of esters and aldehydes in the thermal processed
chestnuts; moreover, ketones, furfural and furan were found. The
aromatic components of cooked chestnuts mainly come from the
degradation of saccharides, protein and lipids, caramelization of
saccharides, and Maillard reaction between reducing sugar and
amino acids (Morini & Maga, 1995b). Furfural, 3-heptanone,
2-hydroxy-2-cyclopenten-1-one, 4-hydroxy-2-butanone, 3-carene,
1-(methylencyclopropyl)-ethanol, 2-hexenoic acid methyl ester
and 2-pentyl-furan were observed in fried chestnuts but not in
fresh chestnuts, which means that they were formed during ther-
mal processing. Comparing the obtained results with previous
studies, hexanal, 4-hydroxy-2-butanone, and decanal were also
identified in thermal processed Chinese chestnuts (Morini &
Maga, 1995b); furfural (6.3%) and benzaldehyde (7.2%) were also
found to be main components in roasted Italian chestnuts (Krist
et al., 2004).
4. Conclusions
After thermal processing, the proximate composition, including
starch, fat, water-soluble protein, reducing sugar, L-aspartic acid, L-
glutamic acid, L-arginine, sucrose and other nutritional com-
pounds, decreased significantly, which led to a decrease in nutri-
tional value. However, the decrease in reducing sugar and free
amino acids made a great contribution to the flavor formation.
The main volatile components in cooked chestnuts were ketones,
furfural and furan, in addition to the esters and aldehydes that
originated in fresh chestnuts.
Acknowledgements
The authors are thankful for the support of the Forestry Indus-
try Research Special Funds for Public Welfare Projects (No.
201204401) from the Ministry of Forestry of the People’s Republic
of China and the Fundamental Research Funds for the Central
Universities (2015ZCQ-SW-04).
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