Origin and distribution
Walnut (Juglans regia L.) is the most widespread tree nut in
the world. The tree is commonly called as the Persian walnut, white
walnut, English walnut or common walnut. It belongs to juglandaceae
and has the scientific name Juglans regia. The walnut tree
species is native to the old world. It is native in a region
stretching from the Balkans eastward to the western Himalayan chain
(Fernandez-Lopez et al., 2000) and was cultivated in Europe as early
as 1000 BC. At present, walnut is cultivated commercially throughout
southern Europe, northern Africa, eastern Asia, the USA and western
South America. World production of whole walnut was around 1.5 × 106
t in 2008 (FAO, 2008). China is the leading world producer, followed
by the USA, Iran, Turkey, Ukraine, Romania, France and India, but
production in other countries such as Chile and Argentina has
increased rapidly in recent years (Martinez et al., 2010).
Walnut composition and nutritional value
Walnut has been used globally in human nutrition since ancient
times. The high protein and oil contents of the kernels of
Juglans regia L. (Juglandacea) make this fruit indispensable for
human nutrition. Therefore, the walnut is classified as a strategic
species for human nutrition and is included in the FAO list of
priority plants (Gandev, 2007). The seed part of the fruit (kernel)
is consumed fresh, toasted, or mixed with other confectionaries. In
the Middle East walnuts are added alone or along with almonds, date,
and raisin as a special pastry preparation called Ma'moul. Walnuts
are nutrient-rich food due to high contents of fats, proteins,
vitamins and minerals. They are also good source of flavonoids,
sterols, pectic substances, phenolic acids and related polyphenols.
The nutritional contents differs from a cultivar to another which
can be influenced by genotype, cultivator, different ecology and
different soil (Caglarirmak, 2003; Crews et al., 2005; Martinez et
al., 2010; Muradoglu et al., 2010). The major components of walnut
oil are triacylglycerols (980 g/kg oil), in which monounsaturated
fatty acids (FAs) (mainly oleic acid) and polyunsaturated FAs (PUFAs;
linoleic and α-linolenic acids) are present in high amounts in all
genotypes (Table 1). Oil contents reported by Pereira et al. (2008)
(78.83 to 82.4%) were higher than those reported by other
researchers (Savage, 2001; Muradoglu et al., 2010). In general, the
FA composition of walnut oil resembles that of soybean oil, but
walnut oil contains a greater concentration of linolenic acid. In
fact, among vegetable oils, walnut oil has one of the highest
amounts of PUFAs (up to 78% of the total FA content).
Walnuts have high amount of omega-6 and omega-3 PUFA, which are
essential dietary fatty acids. Clinical studies suggest that omega-3
PUFA have significant role in prevention of coronary heart disease
(Davis et al., 2007). Oil rich in oleic acid displays greater
oxidative stability therefore; it could be widely used as frying
oil. According to an investigation conducted by several researchers,
It was found that the average value for protein was 18.1% (Amaral et
al., 2003); Muradolu, 2005; Mitrovic et al., 1997; Muradoglu, 2010;
Savage, 2001). They are mainly composed of glutelins (about 70% of
the total seed proteins) together with lesser amounts of globulins
(18%), albumins (7%) and prolamins (5%) (Martinez et al., 2010). The
amino acid (AA) composition of walnut flour is dominated by the
acidic AA residues of aspartate and glutamate together with
relatively high levels of arginine. Walnut proteins contain all
essential AAs required for the needs of a human adult. The lysine/arginine
ratio in walnut proteins is lower than those observed in other
common vegetable proteins, and this fact has been identified as a
positive feature in the reduction of atherosclerosis development (Sza-Tao
et al., 2000; Venkatachakm and Sathe; 2006; Martinez et al., 2010).
Walnut cultivars analyzed have recorded rich mineral composition,
especially potassium, magnesium, and calcium. The minimum and
maximum macro and micro nutrient contents of walnut are presented in
Table 1 (Ravai, 1992; Payne, 1985; Souci et al., 1994; Cosmulescu et
al., 2009). Walnuts contain high levels of potassium, phosphorus and
magnesium and lower sodium. These elements play an important role
for many enzymes activity especially as cofactor.
Nutritional value of Juglans regia L.
Value per 100 g
Vitamins (USDA, 2010)
(Muradoglu et al., 2010)
Unsaturated fatty acids
Palmitoleic acid C16:1
Oleic acid C18:1
Gadoleic acid C20:1
Linoliec acid C18:2
Lineliec acid C18:3
Saturated fatty acid
Myristic acid C14:0
Palmitic acid C16:0
Stearci acid C18:0
Archidic acid C20:0
leaves have been used mostly in worldwide traditional medicines as
antimicrobial, antihelmintic, astringent, keratolytic,
antidiarrhoeal, hypoglycaemic, depurative, tonic, carminative, and
for the treatment of sinusitis, cold and stomach ache (Girzu et al.,
1998; Mouhajir et al., 2001; Vaidyaratnam, 2005). In Turkish folk
medicine, fresh leaves applied on the naked body or forehead to
reduce fever or on swelled joint to alleviate the rheumatic pain
(Fujita et al., 1995; Yesilada, 2002).
kernel of J. regia has been used for the treatment of
inflammatory bowel disease in Iranian traditional medicine
(Kim et al., 2006).
In Palestine, it is
used for treatment of diabetes and asthma (Jaradat, 2005; Kaileh et
al., 2007) and to treat prostate and vascular disturbance (Spaccarotella
et al., 2008). The plant is used as a topical remedy for dermal
inflammation and excessive perspiration of the hands and feet. It is
also a common home remedy for the treatment of chronic eczema and
scrofula. The leaves of this plant is used topically to treat scalp
itching and dandruff, sunburn and superficial burns as well as an
adjunctive emollient in skin disorders (Gruenwald et al., 2001;
Robbers et al., 1999; Ali-Shtayeh and Abu Ghdeib, 1999; Blumenthal,
2000; Baytop, 1999). It also has high anti-atherogenic potential and
a remarkable osteoblastic activity that adds to the beneficial
effect of a walnut enriched diet on cardioprotection and bone loss (Papoutsi
et al., 2008). The bark, branches and exocarp of the immature green
fruit of this medicinal plant have been used to treat gastric, liver
and lung cancer a long time in China (Liu et al., 2004; Baytop,
1999). It is used by traditional healer in northeastern region of
Mexico to protect against liver damage (Torres-gonzalea et al.,
2011). The bark is used as miswaks for teeth cleaning (Ibrar et al.,
In Nepal the bark paste is useful in arthritis, skin diseases,
toothache, and hair growth. Seed coat is used for healing wounds
(Kunwar and Adhikari, 2005).
The shell of Juglans regia is used in Calabria folk medicine
to heal malaria (Tagarelli et al., 2010).
Hot and cold solvent and aqueous extract of leaves,
barks, fruits and green husks of J. regia
from different countries revealed broad spectrum
antibacterial activity against gram-positive and
gram-negative bacteria viz. Bacillus cereus,
Bacillus subtilis, Staphylococcus aureus,
Pseudomonas aeruginosa, Escherichia coli,
Klebsiella pneumoniae, Staphylococcus
epidermidis,Micrococcus luteus, Salmonella
typhimurium, Enterococcus faecalis, Bacillus
thuringiensis, Protomonas extroquens, and
Proteus sp. using agar streak method, disc diffusion
method and microplate alalmar blue assay (Deshpande
et al., 2011; Poyrazolu et al., 2010; Pereira
et al., 2008; Oliveira et al., 2008; Pereira et al.,
2007; Qa’dan et al., 2005ab; Citoglu and Altanlar, 2003;
Upadhyay et al., 2010b). The antimicrobial
activity against gram-negative bacteria were
selective since not all the fruit extract of J.
regia cultivator inhibited the growth of
Pseudomonas aeruginosa and E. coli. cv.
Lara inhibited the growth of K. pneumoniae
(MIC of 100 mg/mL), cv. Mayette inhibited the
development of P. aeruginosa and E. coli
with minimum inhibitory concentrations (MICs) of 50
and 10 mg/mL, respectively, and cv. Mellanaise
inhibited the growth of E. coli and K.
pneumoniae at concentration of 100 mg/mL
(Ali-Shtayah et al., 1999).Mexican
aqueous bark and leaves extract exhibited no
antimycobacterial activity. Only the hexane and
methanol extract showed antimycobacterial activity
with MIC of 100 and 125 mg/ml, respectively using
Soxhlet extractor (Cruz-Vega et al., 2008). Over
45% of Iranian clinical isolates of
Helicobacter pylori strain were
inhibited by J. regia aqueous and equal
mixture of methanol, diethyl ether and petroleum
benzene extract (Nariman et al., 2004). In a recent
was shown to potently inhibit
the three key
Helicobacter pylori, cystathionine γ-synthase
(HpCGS), malonyl-CoAacyl carrier protein
transacylase (HpFabD), and β-hydroxyacyl-ACP
with the half maximal inhibitory concentration (IC50)
values of 7.0±0.7, 20±1, and 30±4 μmol/L,
respectively. Therefore, HpCGS, HpFabD, and HpFabZ
are considered to be the potential targets of
juglone (Kong et al., 2008).
The antibacterial activity of Jordanian J. regia
leaves extract to acne developing organism revealed
that 12.5% S. epidermidis isolates were
resistant to the leaf extract where as all
Propionibacterium acnes isolates were
sensitive even to 10% of the extract (Qa'dan et al.,
fruits, leaves and bark aqueous and solvents extract
exhibited antifungal activity against wide range of
fungi using disc diffusion method, agar dilution
method, agar streak dilution and Raddish method.
Pereira et al. (2008) reported that all the walnut
varieties exhibited antifungal activity against
Candida albicans and Cryptococcus neoformans
when soxhleted with light petroleum ether (b.p.
40-60°C). The higher inhibition was observed with cv.
Lara extract (MIC of 1 mg/mL). However, C.
albicans and C. neoformans were only
resistant to cv Mallanaise extract. Cold extraction
of fruit, leaves and bark inhibited the growth of
Microsporum canis, Trichophyton
mentagrophytes, and Trichophyton violaceum
(Ali-Shtayah et al., 1999). On the other hand, the
aqueous extract of green husks showed no antifungal
activity against C. albicans and C.
neoformans (Oliveira et al., 2008). Methanol,
acetone, chloroform and ethyl acetate bark extract
revealed antifungal activity against A. niger,
Alternaria alternata, Trihoderma viresn,
fusarium solani, Pichia guiliermondii,
Pichia jadinii and all Candida speicestested
(Upadhyay et al., 2010c;
Ahmad et al., 1973).
Mei-zhi et al. (2007) reported that 95% ethanol and
ethyl acetate leaves extract of J. regia,
inhibited tobacco mosaic virus (TMV). The methanol
extract of J. regia inhibited Sindbis
virus at a minimum concentration of 1.5 µg/ml (Mouhajir
et al., 2001).
The antioxidant potential of ethyl acetate, butanol,
meta-nol, ether and aqueous methanol extract of
walnut kernels, husks and leaves were measured by
different methods such as reducing power, scavenging
activity on 2,2-diphenyl-1-picrylhydrazyl (DPPH)
radicals and lipid oxidation inhibition by
β-carotene linoleate system. All the extracts showed
strong antioxidant activity (Qamar and Sultana,
2011; Carvalho et al., 2010; Abbasi et al., 2010;
Rahimipanah et al., 2010; Zhang et al., 2009b;
Almeida et al., 2008; Oliveira et al., 2008; Pereira
et al., 2008; Pereira et al., 2007; Fukuda et al.,
2003). Bullo et al. (2010) reported a decrease in
the antioxidant burden observed in enzymatic and
non-enzymatic antioxidant systems after the
consumption of a whole-walnut or a walnut-skin diet
in C57BL/6 mice. The same author also reported that
consumption of walnuts and walnut skins have
no deleterious effect on low-density lipoprotein (LDL)
oxidizing capability, despite their higher contents
of omega-6 PUFAs. Several phenolic compounds
isolated from J. regia such as pyrogallol, p-hydroxybenzoic
acid, vanillic acid, ethyl gallate, protocatechuic
acid, gallic acid, 3,4,8,9,10-pentahydroxydibenzo
pyran-6-one, tannins, glansrins, adenosine, adenine,
etc, could provide a chemical basis for some of the
health benefits claimed for J. regia in foods
and folk medicine (Zhang et al., 2009a;
Fukuda et al., 2003).
Fukuda et al. (2004) demonstrated a strong
inhibitory activity of walnut polyphenols and the
polyphenolic components like Casuarictin,
tellimagradin II and Tellimagradin I on different
enzymes like glycosidase, sucrose, maltase and
amylase. In addition to the above findings,
researchers also noticed that walnut polyphenol-rich
fraction has triglyceride lowering effect and urine
peroxide lowering effect in genetically inherited
Type II diabetes mellitus (db/db) mice
at the dose of 200mg/kg/day. The consumption of
walnut leaf pelletsin alloxan induced diabetic rats at
the dose of 185 mg/kg reduced fasting blood sugar
significantly and the histomorphometric study of
pancreas showed a sign of regeneration of β-cells in
the treated group (Jelodar et al., 2007). J.
regia leaves methanolic extract at dose of
250 mg/kg decreases the postprandinal plasma blood
glucose levels in both short and long term models.
The plant extract significantly inhibited α-glucosidase
activity in vitro for both maltase and
sucrase enzymes and showed no changes in the insulin
and glut-4 genes expression. The author attributed
the inhibitory action of the plant extract to gallic
acid and caffeoylquinic acid in the leaves (Teimori
et al., 2010).
Kale et al. (2011) reported that stem park of
J. regia acetone extract exhibited significant
activity at all dilution tested when compared to the
Albendazole standard against Eicinia feotida.
The benzene, methanol and ethanol soxhlet extracts
of J. regia stem bark on adult Indian
earthworm, Pheretima posthuma exhibited
significant anthelmintic activity as
comparable to that of standard drug Piperazine
citrate (Upadhyay et al., 2010a).The
95% ethanol, petroleum ether and ethyl acetate
extract of green walnut hull have obvious
anti-feeding effect on armyworm and the small
vegetable-moth. The research group indicated that
anti-feeding rate, death rates as well as growth
inhibition rate of armyworm
have correspondingly changed in dose dependant
manner (Me-zhi et al., 2006).
The ethanolic extracts of J. regia leaves
exhibited potent anti-inflammatory activity as
potent as indomethacin against carrageenan-induced
hind paw edema model in mice without inducing any
gastric damage (Erdemoglu et al., 2003). Mokhtari et
al. (2008) stated that the alcohol extract of walnut
leavesin dose of 1.5 mg/kg caused a
significant nociception decrease in acute phase of
formalin test where as the aqueous (2.87 and 1.64
g/kg) and ethanolic (2.044 and 1.17 g/kg) extracts
of leaves showed antinociceptive activity in
hotplate test suggesting a promising analgesic and
anti-inflammatory agents against diseases such as
rheumatoid arthritis. On the basis of Qamar and
Sultana (2011)result, a protective role of methanolic
J. regia extract against CSE-induced acute
lung toxicity in Wistar rats was suggested. The
extract significantly decreased the levels of
Lactate dehydrogenase (LDH), total cell count, total
protein and increased the glutathione (GSH) level in
bronchoalveolar lavage fluid. It also significantly
restored the levels of Glutathione reductase (GR),
catalase and reduced the xanthine oxidase (XO)
activity in lung tissue.
The macerated hexane extract of J. regia
fruit produced significant antidepressant activity
at both doses of 100 and 150 mg/kg body weight when
compared with standard drug fluoxetine on male
Wistar rats. The antidepressant activity was
evaluated by forced swimming and tail suspension
test (Rath and Pradhan, 2009).
Ozer et al. (2007) suggested that gel formulation
containing ellagic acid and plant leaves extract of
J. regia is effective in treating uneven skin
pigmentation. The ethanolic leaves extract could be
suggested as new sources of skin-whitening agents.
Aitani and Shimoda (2005) reported that melanin
formation was inhibited at concentration 1 to 30 μg/ml
in Pre-cultured B16 melanoma cells incubated with
medium containing walnut polyphenols and their
result indicated that walnut polyphenols is more
superior to the popular skin-lightening agent,
ascorbic acid and arbutin upon data comparison.
Orally fed Walnut polyphenols prepared from the
kernelpellicle demonstrated a dose dependent
lowering effect in glutamyl oxaloacetic transaminase
(GOT) and glutamyl pyruvic transaminase (GPT) in
carbon tetrachloride (CCl4) induced liver
damage in mice model after a single oral
administration (200 g/kg). Result indicated that
walnut polyphenols is more superior to Curcumin, a
commonly used hepatoprotective agent. The effect of
each active component of in vitro evaluation
of walnut polyphenols on CCl4-induced
cytotoxicity in primary cultured rat hepatocytes
I, casuarictin, tellimagrandin II, and rugosin C
(Figure 1) are inhibitory on CCl4-induced
cytotoxicity in primary cultured rat hepatocytes
however, tellimagrandin I of walnut polyphenols is
believed to be the most important active
compound responsible for hepatoprotective
effect (Hiroshi et al., 2008).
The same author,
Hiroshi et al. (2006) reported that 50% EtOH extract
from endocarps of walnuts on mice liver injury
models induced by carbon tetrachloride at the dose
of 100 and 200 mg/kg significantly suppressed GOT
and GPT deviations. Polyphenolic constituents,
tellimagrandins I and II, rugosin C and casuarictin
were found to be principal constituents with
hepatoprotective activity against oxidative damage.
Oral administration of a polyphenol-rich extract
(WP) from walnuts (100 and 200 mg/kg) in high fat
diet fed mice significantly reduced liver weight and
serum triglycerides (TG) where as hepatic
β-oxidation in cytosol, including peroxisome, was
enhanced by WP (50-200 mg/kg). A polyphenol-rich
extract was found to possess hypotriglyceridemic
activity via enhancement of peroxisomal fatty acid
β-oxidation in the liver. These results suggest that
tellimagrandin I is involved in the
hypotriglyceridemic mechanism (Shimoda et al.,
Juglone has been reported to inhibit intestinal
carcinogenesis induced by azoxymethane in rats and
might be a promising chemopreventive agent in human
intestinal neoplasia (Sugie et al., 1998). Juglone
was also proven to be a potent cytotoxic agent in
vitro in human tumor cell lines, including human
colon carcinoma (HCT-15) cells, human leukemia
(HL-60) cells and doxorubicin-resistant human
leukemia (HL-60R) cells (Kamei et al., 1998; Segura-Aguilaretal,
1992). In a recent study, Juglone inhibited the
growth and induce apoptosis of sarcoma and 180
SGC-7901 cells in vivo. The mechanism is
mediated by the activation of the mitochondrial
death pathway, which requires the generation of
reactive oxygen species (ROS), down-regulation of
Bcl-2 protein expression and up-regulation of Bax
protein expression (Ji et al., 2011). Walnut
methanolic extracts obtained from J. regia
seed, green husk and leaf showed concentration
dependent growth inhibition against human renal
cancer cell lines A-498, 769-P and the colon cancer
cell line Caco-2. Concerning A-498 renal cancer
cells, all extracts exhibited similar growth
inhibition activity (IC50 values between
0.226 and 0.291 mg/mL), while 769-P renal and Caco-2
colon cancer cells, walnut leaf extract showed a
higher antiproliferative efficiency (IC50
values of 0.352 and 0.229 mg/mL, respectively) than
green husk or seed extracts (Carvalho et al., 2010).
The tested dried fine powder of J. regia
light petroleum seed extract in cancer induced in
Swiss albino mice with the help of
7,12-Dimethylbenz(a)anthracene (DMBA) and croton oil
showed the petroleum extract was significant in
reducing the cancer cells (Kumudhavalli et al.,
Other medicinal uses
Willis et al. (2009) examined the effects of walnut
diet on motor and cognitive ability in aged rats for
8 weeks. The three treated groups (2, 6 and 9%)
revealed that the 2% walnut diet improved
performance on rod walking, while the 6% walnut diet
improved performance on the medium plank walk; the
higher dose of the 9% walnut diet impaired reference
memory, however the researcher attributed this to
the number of polyphenolic compounds that could be
negatively effecting reference memory at a higher
dose. A 2004 study by the NYS Institute for Basic
Research in Developmental Disabilities (OMRDD)
revealed that methanolic extract of walnut was able
to inhibit and defibrillize fibrillar amyloid β-
protein (the principal component of amyloid plaques
in the brains of patients with Alzheimer's). It is
proposed that polyphenolic compounds present in
walnuts may be responsible for its anti-amyloidogenic
activity (Chauhan et al., 2004). Similarly, it was
found that two of its major components in walnuts,
gallic and ellagic acid, act as "dual-inhibitors" of
the enzyme acetylcholinesterase which, in
association with amyloid inhibits protein
aggregation, and inhibit the site of
acetylcholinesterase responsible for the breakdown
of acetylcholine. These results suggest that walnuts
may reduce the risk or delay the onset of
Alzheimer's disease by maintaining amyloid-protein
in the soluble form and prevent the break down of
acetylcholine (Society for Neuroscience, 2007).
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