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*Corresponding Author: Vanisha S Nambiar, Email: vanishanambiar@gmail.com,Contact No. 09327223374
ISSN 0976 – 3333
CASE STUDY
Available Online at
Effect of Drumstick Leaves supplementation on Hematological Indices
of young girls (16-21 years)
Vanisha S Nambiar* Shilpa Parnami, Parul Guin
*Department of Foods and Nutrition, Faculty of Family and Community Science, The Maharaja SayajiRao University of
Baroda, Vadodara, Gujarat. India
www.ijpba.info.
International Journal of Pharmaceutical & Biological Archives 2010; 1(2): 261 - 266
Received 3 May 2010; Accepted 28 July 2010
ABSTRACT
Background: Drumstick leaves (DL) with its high beta carotene content (19690 mcg/100g) along with
vitamin C from lemon juice may have a positive impact in the mobilization of stored iron and increase
hemoglobin levels of anemic subjects. Aim: To determine the effect of DL and vitamin C
supplementation on hematological indices of young girls (16-21 years). Method: Based on pallor,
100/700 girls studying in first and second year the Faculty of Family and Community Sciences were
selected and assessed for hematological Indices (Hb, Hct, PCV, MCV, MCHC) and red cell
morphology and were divided into Group A (329 RE of Beta carotene from DL rich recipes (boiled
mung/desi chana/kabuli chana) and 5.85 mg Vitamin C as lemon juice, n=21), Group B (329 RE from
the above DL recipes, n=20) and Group C (recipes without DL leaves, n=21) for 45 days respectively.
Results: Post supplementation data revealed 28.6% reduction of anemia in Group A, by followed by
5% in group B and 4.7% in group C. There was a positive change observed in red cell morphology
(normocytic normochromic) in Group A (18%) and B (2.6%) respectively. Similar results were seen for
RBC, MCV and MCHC. Group B which received drumstick leaves showed small positive change in
MCH showing a causal association between VA and iron metabolism. Conclusions: The study
emphasizes that anemia is still prevalent even in young adult women of the middle and high-income
groups of urban Vadodara (54%). It also shows a strong association between vitamin C from lemon
juice and iron and a causal association between VA available from DL and iron metabolism.
Keywords: Anemia, Vitamin A, Beta Carotene, Drumstick leaves, Young girls, Hematological
Indices.
INTRODUCTION
Women and adolescent girls are at high
risk of micronutrient malnutrition especially Iron
Deficiency Anemia and Vitamin A Deficiency.
Globally the most important cause of anemia is
believed to be iron deficiency due to inadequate
dietary intake, physiologic demands of pregnancy
& rapid growth & iron losses due to parasitic
infection. Other prevalent causes of anemia
include malaria, chronic infection and nutritional
deficiencies of vitamin A, folate & Vitamin B
12.
[1]
Vitamin C is a well known enhancer of iron
bioavailability. Several researchers have reported
relationship between Iron and vitamin A. Hodges
et al and Mejia & Arroyave
[2,3]
established an
association between iron metabolism &
hypovitaminosis A. Anemia provoked by VAD
resembles Hypochromic anemia. Vitamin A
binds with iron during digestive process and
forms a complex that acts as a chelating agent,
thus blocking the effect of hydroxyl radicals
present in phytates and polyphenols in flour and
tea / coffee respectively
[4]
. After supplementation
with Vitamin A, hematologic evidence and
measures of iron status have improved
[5, 6]
.
Vitamin A can play a role in improving
utilization of available iron in young girls.
[7]
Drumstick leaves (Moringa oliefera) is
reported to contain alkaloids, flavonoids
[8]
anthocyanins, proanthocyanidins and cinnamates
and is highly reputed in folklore and traditional
system of medicine as a remedy for variety of
ailments. Drumstick leaves (Moringa oleifera)
with a beta carotene (a precursor of vitamin A)
content of 19690mcg/100gm FW is acceptable in
Vanisha S Nambiar et al. / Effect of Drumstick Leaves supplementation on Hematological Indices of
young girls (16-21 years)
© 2010, IJPBA. All Rights Reserved. 262
IJPBA, July – August, 2010, Vol. 1, Issue,
traditional Indian diets both in fresh as well as
dry forms, has a good bioavailability in fresh and
dry forms may thus be useful in improving
utilization of iron especially from high phytate
Indian diets.
[9-13]
With this hypothesis in mind, the present
study was planned. The broad objective was to
study the effect of Drumstick Leaves (Moringa
oleifera) with or without vitamin C rich lemon
juice supplementation along with traditionally
consumed high phytates recipe on the
hematological indices of young anemic/non
anemic girls.
MATERIALS AND METHODS
Study area: The study was conducted in the
Faculty of Family and Community Sciences of
The M. S. University of Baroda, Vadodara,
Gujarat.
Study design: Out of the 700 girls (16-19y)
screened for pallor, 100 positive subjects were
selected and divided into 3 groups. These girls
were supplemented with 30gm cooked (1 cup) of
boiled mung/desi chana/kabuli chana (phytates
201-265 mg percent), with or without freshly
blanched drumstick leaves or lemon juice for a
period of 45 days during the lunch hour.
However due to drop outs the final data is
presented for 62 subjects. Group A subjects
received freshly blanched drumstick leaves in
boiled mung/desi chana/kabuli chana along with
200 ml of lemon juice containing 329 RE of Beta
carotene from DL and 5.85 mg Vitamin C as
lemon juice, (n=21), Group B received freshly
blanched drumstick leaves in boiled mung/desi
chana/kabuli chana containing 329 RE beta
carotene (n=20) and Group C received boiled
mung/desi chana/kabuli chana recipes without
DL leaves (n=21) respectively. Pre-post
estimations for hematological indices were done
after 45 days and after 3 months to assess the
wash out effect of the supplementation.
Product Development Sensory evaluation and
standardization: Drumstick leaves obtained from
drumstick trees situated in university campus,
Vadodara, were selected as a food source of beta
carotene for supplementation. Different levels of
freshly blanched drumstick leaves were tried: 6-
25 gm per 30 gm raw weight i.e. one serving of
each recipe namely kabuli chana, desi chana and
mung. All recipes were prepared fresh and
presented to the judges for sensory evaluation.
The final test was conducted with incorporation
of 20 gms fresh DL in each recipe and was
selected for the study. Details of product
development and organoleptic evaluations are
described in our earlier paper.
[14]
Baseline data: Baseline data were collected on
the socioeconomic status, clinical examination
for anemia, anthropometry (weight, height, BMI)
and dietary intake (24-h DRM and FFQ
methods).
Biochemical Estimation: Pre-post analysis was
done on the hematological indices to assess the
impact of supplementation. Semi-automated
hematology analyzer F-620 was used for in vitro
diagnosis of Red Blood Cell (RBC/Hb),
Hemoglobin Concentration (HGB), Hematocrit
(HCT) and Mean Corpuscular Volume (MCV).
Hemoglobin was measured using Sysmax Lysing
reagent; HCT was measured electronically; The
MCV is calculated from HCT and the RBC. A
thin blood film was used to study the Red Cell
Morphology of the erythrocytes, using
Leishman’s stain.
Statistical Analysis: The data was entered and
analyzed using Micro Soft Excel. Means and
standard errors were calculated for the entire
parameters. Chi square, Paired‘t’ and
Independent‘t’ tests were performed to test the
differences between the control and experimental
groups.
RESULTS
Baseline Data: Data on family type reveal that
79% girls belonged to nuclear families and 77.3%
has an average family size of 3-6 members. The
mean age of menarche was 14 y. Anthropometric
and FFQ data revealed no significant differences
between the weight, height or BMI or in the
consumption of iron or vitamin A rich foods
among the 3 groups. Out of the total 700 girls of
FY and SY, 100 girls were selected on the basis
of pallor which accounts for 14% prevalence.
Based on hemoglobin estimations 54% were
found to be anemic (Hb<12g/dl).
Out of these 39% were mildly anemic,
14% moderately anemic and 1% severely anemic.
Almost 87% girls had lower Hematocrit (%) than
normal, 73% had lower MCHC, 49% had lower
MCH followed by 40% had lower RBC counts
and 20% had lower MCV.
Almost 29% girls had abnormal morphology
which included 16% Microcytic Hypochromic,
8% Normocytic Hypochromic and 5%
Macrocytic. When the Hb levels were compared
with the red cell morphology (RCM) it was seen
that a 14% of anemic girls had a normal RCM
indicating that they were in initial stages of
Vanisha S Nambiar et al. / Effect of Drumstick Leaves supplementation on Hematological Indices of young
girls (16-21 years)
© 2010, IJPBA. All Rights Reserved. 263
IJPBA, July – August, 2010, Vol. 1, Issue 3
IDA which can be easily reverted to normal by
appropriate food based approaches.
IMPACT OF DRUMSTICK LEAVES
SUPPLEMENTATION: The mean intake after
supplementation showed an increase in the total
energy intake (201, 149 and 133 Kcal); protein
intake (7.6, 7.55 and 6.98 gms); Beta Carotene
intake (3980.5, 3960.5 and 42.5 mcg); vitamin C
(51.85, 46 and 2 mg) and iron intake (1.5, 1.49
and 1.35 mg) of all the three groups Group A, B
and C respectively. The change in hemoglobin
levels of subjects after supplementation is shown
in (Table 1).
Table 1 Changes in Hemoglobin Levels after Supplementation of 45 days (PI) and after a gap of 3 months (PII)
Mean H Levels
A (n=21)
B (n=20)
C (n=21)
Total (n=62)
Initial
10.67±0.31
11.09±0.21
10.92±0.253
10.893±1.22
PI
11.33±0.37
11.18±0.31
11.29±0.27
11.26±0.188
PII
11.58±0.36
11.22±0.28
11.59±0.43
11.46±0.21
Difference (PI-I) (PII-I)
(+0.66) (+0.91)
(+0.09) (+0.13)
(+0.37) (+0.67)
(+0.36) (+0.57)
Paired ‘t’
2.462*
0.305
1.478
2.345*
* p < 0.05 Hb = haemoglobin
There is significant difference seen which is
mainly contributed by Group A which was
supplemented by Drumstick leaves recipes and
lemon juice, Group B and C do not show any
significant change though there is a little change
in their final hemoglobin levels. Post
supplementation data revealed 28.6% reduction of
anemia in Group A by followed by 5% in group B
and 4.7% in group C (Table 2).
Table 2 Comparison of prevalence of anemia after 45 days of supplementation
Hemoglobin level (g/dl)
Group A
%
Group B
%
Group C
%
19.04 (4)
Initial
≥ 12
25 (5) 23.8 (5)
< 12
80.95 (17)
75 (15)
76.1 (16)
(Mild) 10-11.9
76.47 (13)
87(13)
75 (12)
(Moderate) 7-9.9
17.64 (3)
13 (2)
25 (4)
(Severe) <7
5.88 (1)
-
-
48 ↓ (10)
Final
≥ 12
30 (6) 28.57 (6)
<12
52.38 ↑ (11)
70 (14)
71.42 (14)
(Mild) 10-11.9
55 (6)
71 (10)
86 (12)
(Moderate) 7-9.9
45 (5)
29 (4)
14 (2)
(Severe) <7
-
-
*Figures in Parenthesis indicate number of subjects (A=Drumstick Leaves+Lemon Juice, B= Drumstick Leaves, C=Plain
Recipe)
Table 3 Red cell morphology of subjects after supplementation of 45 days (pi) and after a gap of 3 months (pii)
Red Cell
Morphology
A
B
C
Initial
PI
PII
Initial
PI
PII
Initial
PI
PII
Normal
Normocytic
Normochromic
57.1 12) 75 (16) 76.47 (13) 65 (13) 66.6(14) 73.3 (11) 66.6(14) 66.6 (14) 83.3 (10)
Abnormal
Macrocytic
4.76(1)
-
-
10 (2)
-
-
-
-
-
Normocytic
Hypochromic
19 (4) - - 5(1) - - 4.76(1) 47.6 (1) -
Microcytic
Hypochromic
19(4) 25(5) 23.6 (4) 20(4) 33.3(6) 26.6(4) 28.5(6) 28.5(6) 16.6 (2)
*Figures in Parenthesis indicate number of subject (A=Drumstick Leaves+Lemon Juice, B= Drumstick Leaves, C=Plain
Recipe)
These results highlight the fact that
vitamin C as well as beta carotene rich
supplements have brought about a positive change
in the anemic girls. In Group B there was shift of
5% these results are encouraging and support the
fact that vitamin A helps in the metabolism of iron
as this group received 329 RE as beta carotene
supplement. Considering the fact that both the
pulses contained enough inhibitors of iron as
phytic acid (201-265 mg percent,
[15]
even a 5%
increase in Hb after 45 days of supplementation is
a positive result and emphasizes that vitamin A
from drumstick leaves would have helped in
preventing inhibitory effects of phytates. In Group
C, there was shift of 4.7% which received plain
recipes.
Table 3 reveals that the trend of RBC,
MCV and MCHC remain same after
supplementation. In case of PCV only in group C,
normal PCV shifted to abnormal. In case of MCH
Vanisha S Nambiar et al. / Effect of Drumstick Leaves supplementation on Hematological Indices of young
girls (16-21 years)
© 2010, IJPBA. All Rights Reserved. 265
IJPBA, July – August, 2010, Vol. 1, Issue 3
in group B, girls shifted from abnormal MCH to
normal. In group A, a positive change is noted
with the Normocytic normochromic NN shifts
from 57% to 75% after supplementation. In Group
B NN shifts from 65 to 66.6 and in Group C, there
was no change seen after supplementation, but
after a gap of 3 months a better condition was
seen as most of them started taking iron tablets
realizing their anemic condition. Though the
increase of normal subjects in group B is not as
high as that in Group A, the results are
encouraging and support the fact that vitamin A
helps in the metabolism of iron.
CONCLUSIONS
Freshly blanched drumstick leaves showed
a mild positive relationship in the improvement of
anemia. The study also emphasizes that anemia is
still prevalent even in young adult women of the
middle and high-income groups of urban
Vadodara (54%). It also shows a strong
association between vitamin C from lemon juice
and iron and a causal association between VA
available from DL and iron metabolism.
DISCUSSION
The beta carotene from drumstick leaves
though theoretically high, may not have produced
equivalent amounts of vitamin A activity. The
release of carotenoids from the food matrices,
their solubilization in the gut and their absorption
into intestinal mucosal cells are critically
important processes.
[16]
Thus the study could
have given better results if dehydrated DDL
tablets with increased dosage, would have been
used as we did to study their impact on
hyperlipidemic patients.
[17]
The main effect of
vitamin A is to maintain adequate levels of iron in
plasma to supply the different body tissues
including the bone marrow with proper amounts
of this essential mineral which may be the
mechanism by which the hemopoeitic tissue
becomes flavored with more available iron
Although deficiency of iron in the diet is regarded
as the most important factor in the etiology of
nutritional anemia certain human and animal
studies have shown that supplementation of iron
with vitamin A may increase the iron status as
measured by hematological indices like
hemoglobin and hematocrit.
[18]
A significant
association of serum retinol with hematocrit,
serum iron and serum ferritin has been reported by
[7]
in a cross sectional study of children in north
east Thailand. Supplementation with vitamin A
alone has significantly increased hemoglobin,
hematocrit, plasma ferritin in children,
[19-20]
and
pregnant women.
[21-22]
The presence of vitamin
A increased iron
absorption up to twofold for rice, 0.8-fold for
wheat and 1.4-fold for corn. Beta -carotene
increased absorption more
than threefold for rice
and 1.8-fold for wheat and corn, suggesting
that
both compounds prevented the inhibitory effect of
phytates
on iron absorption.
[23]
The supplement received by Group B was
663.4 RE, obtained from Beta-carotene, which is
the most abundant provitamin A in foods.
Approximately 10-50% of the total -carotene
consumed is absorbed in
the gastrointestinal tract
and within the intestinal wall is partially
converted
into vitamin A. Βeta carotene from the drumstick
leaves is also not toxic, so it is considered a safe
source of vitamin A, thus need to be promoted not
only as a source of vitamin A to combat VAD but
also to prevent IDA in the communities. Also the
low cost and the easy availability, fast growth and
ease of cultivation enhance its uses. The Social
Forestry Schemes of India also promotes this tree
plantation by providing free samplings. The
results of the present study have highlighted
another importance of these leaves besides the
ones already recorded.
[24-25]
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ACKNOWLEDGEMENT
The authors are grateful to all the subjects for their
cooperation and support. The study was supported
by UGC-DSA Phase 1 of the Department of
Foods and Nutrition. This work was presented as a
poster at the MN Forum (2007) Istanbul. Turkey.
