Wheat Grass Supplementation Decreases Oxidative Stress in Healthy Subjects: A Comparative Study with Spirulina

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LETTER TO THE EDITOR
THE JOURNAL OF ALTERNATIVE AND COMPLEMENTARY MEDICINE
Volume 13, Number 8, 2007, pp. 789–791
© Mary Ann Liebert, Inc.
789
DOI: 10.1089/acm.2007.7137
WHEAT GRASS SUPPLEMENTATION
DECREASES OXIDATIVE STRESS IN
HEALTHY SUBJECTS: A COMPARATIVE
STUDY WITH SPIRULINA
Dear Editor,
Recent investigations have shown that the antioxidant prop-
erties of plants could be correlated with oxidative stress de-
fense in different human diseases. Supplementation using nat-
ural antioxidants as functional foods such as wheat grass
(Triticum aestivum) and Spirulina (Spirulina maxima) holds
great promise in overcoming the ill effects of oxygen toxicity.
Hanninen et al. reported that humans consuming un-
cooked vegan food called living food including wheat grass
juice showed increased levels of carotenoids, vitamins C and
E, and lowered cholesterol concentration in their sera.
1
Wheat grass juice appeared effective and safe as a single or
adjuvant treatment of distal ulcerative colitis.
2
Rauma et al.
found that consumption of uncooked vegan diet significantly
increased the intakes of energy and many nutrients without
gain in weight in Finnish rheumatoid patients.
3
Various sci-
entific groups evaluated the antioxidant activity of the Spir-
ulina and reported that it provides some antioxidant protec-
tion both in vitro and in vivo.
4–6
Since wheat grass and
Spirulina are good sources of natural antioxidants as they
contain vitamin E, -carotene, and minerals, the effect of
these functional foods on human health with respect to their
role as antioxidants was evaluated in the present study.
We conducted a randomized, double-blind, placebo-con-
trolled study with the volunteers and study team being
blinded to the type of functional food intervention the sub-
jects received. The study was conducted with 30 healthy vol-
unteers ages 18–21. The subjects were consuming food from
the same place. Their socioeconomic status was the same.
The subjects who participated in the study were medical as-
sistants in the Indian Navy. They were undergoing training
at School of Medical Assistants, Institute of Naval Medi-
cine, Mumbai, India. Naval recruits are selected for the med-
ical branch in the Navy after completion of Basic/Sea train-
ing. The School of Medical Assistants at Mumbai imparts
higher-rank professional and specialist training to medical
assistants of the Navy to handle all types of emergencies on
ship and/or on shore in war and peace. The subjects were
engaged in routine physical exercise (i.e., morning physical
therapy, morning drill, parade training, classroom instruc-
tions, afternoon muster, evening games, and night study).
Written consent was obtained from the volunteers after giv-
ing full details of protocol approved by the Institute’s Ethic
Committee.
The volunteers were divided into three groups of 10 each.
Group I took placebo (calcium gluconate), Group II took
wheat grass, and Group III took Spirulina. All three sup-
plements were given as a dry powder for 30 days, at 500
mg twice daily, in capsules that were identical in appear-
ance, before breakfast and dinner. The manufacturers of the
wheat grass and Spirulina were M/s Sanat Products Ltd.,
Delhi, India, and M/s Nutraceuticals Bio-Tech, Mumbai, In-
dia, respectively. The physical parameters were recorded be-
tween 7:00
AM
and 8:00
AM
before taking food. Various bio-
chemical tests were performed before and after 30 days of
supplementation.
Blood samples were collected in heparinized tubes at
8:00
AM
to 8:30
AM
from an antecubital vein after 12 hours
of fasting and were processed for various analytes (e.g.; re-
duced glutathione [GSH]) and malondialdehyde [MDA]),
and estimations for these were made on the same day. While
for other biochemical variables, plasma and red blood cells
were separated immediately by centrifugation at 1000g for
15 minutes and were frozen at –80°C until the assay. For
vitamin C, plasma samples were treated with 10% meta-
phosphoric acid and then they were frozen at 80°C until
the analysis was completed. MDA was estimated by using
Utley et al.’s method,
7
and GSH was assayed using an Ell-
man reagent.
8
The total antioxidant status in plasma and ery-
throcytes superoxide dismutase (EC 1.15.1.1; SOD), were
estimated using commercially available kits supplied by
Randox Laboratories (Ardmore, UK). Erythrocytes glu-
tathione reductase (EC 1.6.4.2; GR) was estimated using
Racker et al.’s method.
9
Plasma vitamin C was estimated by
a method used by Zannoni et al.
10
Statistical analysis was carried out using a paired t-test
within each group to make conclusions that there was a sig-
nificant change from baseline. Data were reported as
mean  standard error of the mean. A value of p  0.05 was
considered statistically significant.
All three groups were homogeneous with respect to age
and body–mass index (Table 1). We found that supplemen-
tation with wheat grass for 30 days (Group II) resulted in
decreased blood MDA and enhanced concentrations of

plasma total antioxidant status, vitamin C, erythrocytes ac-
tivity of SOD significantly (p  0.05) from baseline (Table
2).
Blood MDA is a good marker of lipid oxidation and MDA
measurement may provide further indication of oxidative in-
jury. The present study showed that Spirulina supplementa-
tion did not alter the blood concentration of MDA signifi-
cantly, but a trend toward lower values was evident.
However, wheat grass supplementation showed significant
reductions in blood concentrations of MDA.
Plasma total antioxidant status is usually considered to
provide indications of the body’s global antioxidant status.
In the present study, plasma total antioxidant status im-
proved significantly after wheat grass supplementation. This
may be due to the fact that wheat grass supplementation sig-
nificantly increased the plasma concentration of major an-
tioxidants, vitamin C and erythrocytes activity of the an-
tioxidant enzyme, SOD. The contribution to the plasma total
antioxidant status of small amounts of other antioxidant
compounds absorbed from wheat grass may be the reason
for the increased total antioxidant status of plasma. Vitamin
C, -carotene, and -tocopherol are well-characterized an-
tioxidants in wheat grass.
Spirulina contains phenolic acids, tocopherols, and -
carotene, which are known to exhibit antioxidant proper-
ties. In our study, supplementation with Spirulina did not
bring about any significant change in the plasma total an-
tioxidant status, although a trend toward higher values was
evident.
The study highlights the potent antioxidant properties of
wheat grass in healthy subjects. Supplementation with wheat
grass provided better protection against lipid peroxidation
and thereby decreased oxidative stress as shown by a de-
creased concentration of MDA and increased concentrations
of endogenous antioxidant levels such as plasma total an-
tioxidant status and vitamin C, and improvement in activi-
ties of erythrocytes SOD. Spirulina supplementation also in-
creased plasma total antioxidant status; however, it was
statistically nonsignificant. Hence, this study showed that
wheat grass is a better antioxidant as compared to Spirulina.
LETTER TO THE EDITOR
790
T
ABLE
1. P
HYSICAL
C
HARACTERISTICS OF THE
V
OLUNTEERS
BMI
Group Age (years) Weight (kg) Height (m) (kg/m
2
)
Placebo 19.80  0.24 57.82  1.43 1.67  0.019 20.66  0.34
Wheat grass 19.70  0.21 58.70  1.68 1.70  0.017 20.30  0.34
Spirulina 19.85  0.25 58.25  1.41 1.68  0.017 20.61  0.45
Values are mean  standard error of the mean; n  10 in each group.
BMI, body–mass index.
T
ABLE
2. C
ONCENTRATIONS OF
V
ARIOUS
O
XIDATIVE AND
A
NTIOXIDANT
B
IOMARKERS IN
H
EALTHY
H
UMANS
B
EFORE AND
A
FTER
30 D
AYS OF
S
UPPLEMENTATION WITH
W
HEAT
G
RASS AND
S
PIRULINA
Placebo Wheat grass Spirulina
After 30 After 30 After 30
Initial days Initial days Initial days
Blood malondialdehyde 1.73  0.05 1.90  0.25 1.93  0.10 1.44  0.11* 1.94  0.24 1.59  0.19
(mol/L)
Total antioxidant status 0.76  0.09 0.70  0.06 0.63  0.03 0.78  0.06* 0.72  0.09 0.82  0.10
(mmol/L)
Reduced glutathione 37.55  0.95 34.04  0.82* 39.02  2.21 31.96  0.89* 38.01  1.53 34.27  1.40*
(mg/dL)
Vitamin C 1.01  0.20 0.98  0.12 0.81  0.08 1.31  0.17* 0.84  0.11 0.99  0.09
(mg/dL)
Glutathione reductase 0.27  0.03 0.24  0.01 0.26  0.02 0.28  0.03 0.26  0.02 0.25  0.02
(M NADPH
oxidized/min/mL)
Superoxide dismutase 260.77  4.50 260.58  3.67 257.18  3.70 267.67  3.90* 253.83  3.10 267.99  4.99
(U/mL)
Values are mean  standard error of the mean; n  10 in each group.
NADPH, nicotinamide adenine dinucleotide phosphate, reduced.
*p  0.05 as compared to initial.

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Radhey Shyam, M.Sc.
1
Som N. Singh, Ph.D.
1
Praveen Vats, Ph.D.
1
Vijay K. Singh, D.Pharma.
1
Rajeev Bajaj, M.D.
2
Shashi B. Singh, Ph.D.
1
Pratul K. Banerjee, Ph.D.
1
1
Defence Institute of Physiology and Allied Sciences,
Delhi, India
2
Institute of Naval Medicine, Colaba, Mumbai, India
Address reprint requests to:
Radhey Shyam, M.Sc.
Occupational Health Division
Defence Institute of Physiology and Allied Sciences,
Lucknow Road
Timarpur, Delhi-110054, India
E-mail: radheyshyam_dipas@rediffmail.com
LETTER TO THE EDITOR
791