Effect of different levels of pulse proteins (Bengal gram and lentil) on calcium and phosphorus balance and calcium and phosphorus contents of different tissues and serum


Khan, A.; Chakrabarti, C.H.

Indian Journal of Nutrition and Dietetics 15(11): 371-376

1978


The effect of different levels of pulse proteins was investigated on phosphorus and calcium metabolism. The rats fed 18 per cent casein (control diet) showed high calcium balance than the rats fed 18 per cent and 12 per cent pulse protein. There was no significant change in phosphorus balance in rats fed 18 per cent casein diet when compared with the rats fed 18 per cent and 12 per cent pulse protein diet. The calcium content of serum in the rats fed 12 per cent and 18 per cent pulse protein diet have also been found significantly lowered than in the rats fed 18 per cent casein (control diet). The decrease in contents of calcium serum, calcium balance and percentage of calcium retention in the rats fed 18 per cent and 12 per cent protein diet than the 18 per cent casein diet (control diet) has been attributed to insufficient supply of essential amino acids like tryptophan, lysine, methionine and threonine.

The
Ind.
J.
Nutr.
Dieter.,
(1978),
15,
371.
EFFECT
OF
DIFFERENT
LEVELS
OF
PULSE
PROTEINS
(BENGAL
GRAM
AND
LENTIL)
ON
CALCIUM
AND
PHOSPHORUS
BALANCE
AND
CALCIUM
AND
PHOSPHORUS
CONTENTS
OF
DIFFERENT
TISSUES
AND
SERUM
AQUEEL
KHAN
AND
C.
H.
CHAKRABARTI
(
Department
of
Biochemistry,
Nagpur
University,
Mahatma
Gandhi
Marg,
Nagpur-440
010
)
(
Received
8th
September,
1978)
Introduction
Many
workers
have
claimed
that
there
was
an
association
of
protein
and
calcium
in
the
diet,
calcium
absorption
and
utilization
being
better
with
high
protein
intakes.
The
beneficial
response
to
higher
protein
intake
has
been
found
to
be
very
much
in
case
of
phosphorus
than
in
the
case
of
calcium.'
Sherman
eta!'
found
in
rats
that
the
total
body
calcium
was
raised
during
growth
by
increased
protein
in
the
diet
but
this
difference
disappeared
as
the
animals
grew
older.
Based
on
their
earlier
observations
that
the
solubility
of
insoluble
calcium
salts
is
increased
in
solution
of
aminoacids,
Mc
Cance
etal'
found
that
an
increased
protein
intake
results
in
a
better
calcium
absorption
and
retention.
The
addition
of
one
of
several
amino
acids
like
L-lysine,
L-arginine,
L-tryptophan,
L-
leucine,
L-histidine,
L-metbionine
and
L-isoleucine
significantly
enhanced
calcium
absorption
in
rats'.
Raven
eta!'
showed
that
the
lysine
had
the
most
marked
effect
in
the
above
series.
Also
the
Ca/P
ratio
has
effect
on
calcium
utilization.°
Most
of
the
people
in
India
are
vegetarians
and
live
on
pulse
proteins
as
the
sole
source
of
proteins.
In
view
of
the
fact
that
the
pulse
proteins
are
deficient
in
lysine,
tryptophan,
threonine
and
methio-
nine%
it
was
thought
worthwhile
to
investigate
their
effect
on
the
absorption
of
calcium
and
phosphorus
in
comparison
with
the
casein
which
is
a
complete
biological
protein.
Experimental
The
experiment
was
performed
for
each
pulse
on
twenty
four
male
albino
rats
of
Wistar
strain
weighing
between
60
to
90
grams.
The
animals
were
divided
in
three
groups
of
eight
each.
The
animals
in
group
I
were
fed
18
per
cent
casein
diet
and
were
treated
as
control.
The
animals
of
groups
II
and
III
were
fed
pulse
protein
(Bengal
Gram
and
Lentil)
having
18
per
cent
and
12
per
cent
protein
levels
respectively.
The
composition
of
the
experimental
diet
is
shown
in
Table
I.
The
experiment
was
performed
for
two
weeks.
The
animals
were
kept
in
ordinary
cages
for
first
nine
days
for
adjusting
them
to
the
experimental
diet.
On
10th
day
the
animals
were
trans-
fered
to
the
metabolic
cages.
The
animals
were
allowed
to
consume
distilled
water
and
fed
experimental
diet.
The
record
of
the
daily
diet
consumption
was
kept.
Every
care
was
taken
to
avoid
coprophagy
in
the
metabolic
cage
since
the
coprophagy
influences
the
mineral
diges-
tion'.
The
collection
of
excreta
was
started
from
1
lth
day.
Urine
was
collected
under
toluene,
diluted
and
estimation
of
calcium
and
phosphorus
was
carried
out
daily.
372
AQUEEL
KHAN
AND
C.
H.
CHAKRABARTI
TABLE
I.
Composition
of
Experimental
Diets
Constituents
18
per
cent
protein
level
1
2
3
12
per
cent
protein
level
4
5
Casein
18.0
0.0
0.0
0.0
0.0
Bengal
Gram
(Cicer
arietinum)
0.0
82.5
0.0
55.0
0.0
Lential
(Lens
esculents)
0.0
0.0
75.0
0.0
50.0
Arrowroot
66.0
1.5
9.0
29.0
34.0
Ground
nut
oil
9.0 9.0
9.0
9.0
9.0
Salt
mixture'
4.0
4.0
4.0 4.0
4.0
Cod
liver
oil
2.0
2.0
2.0
2.0
2.0
Vit.
mixtures
1.0
1.0
1.0
1.0
1.0
1.
Hawk
-
Oser's
salt
mixture.
2.
Vitamin
mixture
:
One
gram
of
vitaminised
sucrose
contains
:
Thiamine
H
Cl
500
Itg
;
Riboflavin
250
µg
;
Pyridoxine
200
PI
;
Calcium
pantothenate
1000
l
ag;
Niacin
1000
µg;
Cyanocobalamine
1µg:
Folic
acid
100
µg;
Choline
chloride
40U
g;
Alphatocopherol
acetate
2.2
µg;
Menadion
sodium
bisulphate
100
µg
;
Biotin
10
µg;
Vit.
A
1100
U.
P.
S.
units
;
Vit.
D.
200
I.
U.
;
Inositol
10
m
made
upto
1
g
with
sucrose.
Feces
was
collected
every
day
and
preserved
by
adding
a
few
drops
of
formaline
and
was
then
refrigerated.
The
feces
collection
of
five
days
was
pooled,
dried
in
oven,
powdered,
weighed
and
aliquot
of
it
was
then
ashed
in
electricfurnace
at
600°
C
for
12
hours.
At
the
end
of
the
experiment
the
rats
were
anaesthesized
with
intraperitoneal
dose
of
Nembutal
(25mg/kg
body
weight).
The
blood
was
collected
and
serum
was
separated.
The
kidney
and
liver
were
weighed,
dried
in
oven,
then
ashed
in
electric
furnance
at
600°C
for
12
hours.
The
ashes
of
tissues,
feces
and
diets
were
dissolved
in
very
dilute
HC1
and
calcium
was
determined
by
Cressolphthalein
comp-
lexone
method°.
The
aliquots
of
serum
and
diluted
urine
were
taken
directly
for
the
estimation
of
calcium.
The
aliquots
of
dissolved
solution
of
ashes
were
taken
and
the
volume
was
made
to
5.5m1
with
redis-
tilled
water
and
to
this
was
added
one
ml
of
one
per
cent
Quionoline
solution
in
alcohol
(w/v).
Then
2.5m1
of
ammonia
buffer
of
PH:
10.5
(prepared
by
dissolving
0.24
g
of
R.
G.
Ammonium
chloride
in
five
per
cent
ammonia
(v/v)
)
was
added.
To
this
was
added
one
ml
of
0.1
per
cent
0-Cresolphtha-
lein
complexone
solution
(prepared
by
adding
28m1
of
ammonia
buffer
to
100
mg
of
0-Cresolphthalein
complexone
and
diluted
to
100
ml
with
redistilled
water).
The
tubes
were
shaken'
thoroughly
after
the
addition
of
each
reagent.
The
standards
of
CaC1,
were
run
simultaneously
and
treated
as
above.
The
readings
were
taken
after
five
minutes
at
565
mµ.
Phosphorus
in
the
ashes
of
tissues,
feces
and
diets
was
estimated
by
dissolving
them
in
10
per
cent
TCA.
The
solutions
were
filtered
and
phosphorus
was
estimated
by
Fiske
&
Subbarow
method."
In
urine
and
serum,
the
total
phosphorus"
was
estimated
by
heating
with
concentrated
H,S0,,
completing
the
oxidation
with
hydrogen
peroxide
(100
volume)
added
drop
by
drop.
Then
distilled
water
was
added,
boiled
for
two
to
three
minutes,
cooled
and
made
upto
known
volume
with
water
and
aliquot
was
taken
for
determination
of
phosphorus
by
Fiske
&
Subbarow's
method.
EFFECT
OF
DIFFERENT
LEVELS
OF
PULSE
PROTEINS
(BENGAL
GRAM
AND
LENTIL)
373
ON
CALCIUM
AND
PHOSPHORUS
BALANCE
AND
CALCIUM
AND
PHOSPHORUS
CONTENTS
OF
DIFFERENT
TISSUES
AND
SERUM
TABLE
II.
The
Effect
of
Different
Levels
of
Pulse
Proteins
(Bengal
gram)
on
Calcium
and
Phosphorus
Balance
and
Calcium
and
Phosphorus
Contents
of
Different
Tissues
and
Serum
CALCIUM
Diet
-s
4
e
1..3.
ri
4.1
011
<
g
Element
intake
mg
Average
excretion
of
Element
Balance
mg
13
°
Average
content
of
element
in
tissues
and
serum
Urine
mg
Feces
mg
Total
mg
Serum
mg,
100m1
Liver
mg/
100g
Kidney
m
100g
g/
18%
Casein
diet
12.5±
153.9±
0.640±
46.0± 46.6±
a107.2+
69.6
*12.0+
132.4±
79.4+
(control
diet)
1.4
18.3
.0074
6.0
5.9
16.4
029
3.5
4.0
18%
Pulse
11.7±
119.9+
0.823±
36.6± 37.4±
b82.2±
68.5
"11.1+
122.8±
75.5±
Protein
diet
1.7
17.7
.0076
4A
3.9
15.2
0.25
2.6
2.5
12%
Pulse
11.9+
111.7±
0.573±
54.3±
54.9+
a56.7+
50.7
***11.0±
88.8±
27.5±
Protein
diet
1.7
12.7
.0102
2.5
2.5
15.5
0.14
3.7
2.7
PHOSPHORUS
18%
Casein
diet
12.5±
56.2+
0.071±
8.6+
8.7±
a47.5±
85.1
*6.1+
179.7±
67.3±
(control
diet)
1.4
6.4
.0037
2.3 2.3
6.7
1.06
5.6
3.3
18%
Pulse
11.7±
52.5±
0.133±
8.3±
8.4+
b44.0±
84.2
**5.4+
139.9±
90.8±
Protein
diet
1.7
7.8
.0047
1.4
1.4
8.5
0.63
4.5
3.3
12%
Pulse
11.9±
51.1±
0.131±
7.9±
8.0±
43.1+
78.9
***5.0+
72.5±
27.0±
Protein
diet
1.7
7.4
.0042
1.4
1.4
8.1
1.25
3.5
1.1
All
values
are
given
as
mean±standard
deviation.
Data
were
analysed
statistically
using
Student's
t-test.
Calcium
Balance:
The
values
are
significantly
different
at
b
and
c
from
a.
b
=
P<
.01;
c
=
P<.001
Serum
:
The
values
are
significantly
different
at
**
and
***
from
corresponding
*
(P
<.001)
Phosphorus
Balance:
The
values
are
not
significantly
different
at
b
and
c
from
corresponding
a.
b
=
P>.05;
c
=
P>.05
Serum
:
The
values
are
not
significantly
different
at
••
and
***
from
corresponding
*.
**-
P>.05;
•••
=
P>.05
Results
and
Discussion
Data
collected
in
Tables
II
and
III
indicate
that
animals
receiving
18
per
cent
casein
diet
(control
diet)
shows
higher
calcium
balance
than
the
animals
receiving
pulse
protein
at
18
per
cent
level
and
12
per
cent
level
and
the
difference
between
them
are
statistically
significant.
This
difference
is
more
statistically
significant
in
12
per
cent
diet
than
18
per
cent
diet
when
compared
with
control
diet
(18
per
cent
P<
0.01,
12
per
cent
P<
0.001).
The
values
of
calcium
content
of
serum
in
the
rats
fed
12
per
cent
and
18
per
cent
pulse
proteins
are
lowered
than
the
18
per
cent
casein
diet
(control
diet)
and
the
difference
is
statistically
signi-
374
AQUEEL
KHAN
AND
C.
H.
CHAKRABARTI
TABLE
III.
The
Effect
of
Different
Levels
of
Pulse
Proteins
(Lentil)
on
Calcium
and
Phosphorus
Balance
and
Calcium
and
Phosphorus
Contents
of
Different
Tissues
and
Serum
CALCIUM
Diet
0
0
Ad
bo
c*
ba
0
Element
Average
excretion
of
element
Balance
mB
Average
content
of
element
in
tissue
and
serum
°
intake
mg
Urine
g
Feces
mg
Total
mg
Serum
mg/
100m1
Liver
100
mg,
g
Kidney
100g
mgl
18%
Casein
diet
12.6±
154.6±
0.650±
46.3± 46.9±
'107.6+
69.5
*12.5±
125.0+
79.8+
(control
diet)
1.4
17.5
.0056
5.6
5.5
19.9
0.35
4.4
3.4
18%
Pulse
11.0±
120.2±
0.798+
36.3±
37.1+
b83.0±
69.0
**11.5+
120.0±
75.6+
Protein
diet
1.1
12.4
.0058
4.3
4.3
8.2
0.20
3.4
2.6
12%
Pulse
11.1+
113.6+
0.610+
57.5+
58.1+
c55.4+
48.7
***10.9+
85.4±
62.6+
Protein
diet
0.73
6.6
.0057
3.3
3.3
6.8
0.29
2.7
2.9
PHOSPHORUS
18%
Casein
diet
12.6±
56.6±
0.104±
8.2+
8.3±
a48.2±
85.1
*5.9+
181.2+
81.2+
(control
diet)
1.4
6.4
.0047
2.1
1.9
6.7
0.74
6.0
2.2
18%
Pulse
11.0+
50.0+
0.245±
7.6±
7.8+
b42.1+
84.2
**5.0+
145.2±
75.6+
Protein
diet
1.1
2.0
.0032
1.7
2.6
3.6
0.60
4.8
3.1
12%
Pulse
11.1+
48.4+
0.321+
9.9±
10.2±
08.2+
78.9
***4.8+
75.0±
58.1+
Protein
diet
0.73
1.5
.0050
1.5
1.4
3.1
0.31
4.5
2.8
All
values
are
given
as
mean±
standard
deviation.
Data
were
analysed
statistically
using
Student's
t-test.
Calcium
Balance:
The
values
are
significantly
different
at
b
and
a
from
corresponding
a.
b
P<.01;
c
P<.001
Serum
:
The
values
are
significantly
different
at
I
,
*
and
***
from
corresponding
*
(P<.001)
Phosphorus
Balance:
The
value
is
not
significantly
different
at
b
from
corresponding
a
(P>.05)
The
value
is
significantly
different
at
c
from
corresponding
a
(P<.01)
Serum
:
The
values
are
significantly
different
at
and
***
from
corresponding
*
"
=
F<.0
01
;
***
P<.01.
ficant.
The
percentage
of
calcium
and
phosphorus
retention
(Tables
II
and
III)
is
more
or
less
same
in
animals
fed
18
per
cent
casein
diet
(control
diet)
and
pulse
protein
diet
at
18
per
cent
level.
The
percentage
re-
tention
of
calcium
and
phosphorus
is
found
to
be
lowered
in
animals
fed
12
per
cent
diet.
This
is
likely
that
the
casein
being
a
complete
protein
supplies
all
the
amino
acids
which
help
in
better
absorption
of
calcium.
These
results
are
in
agreement
with
the
observation
of
Desikachar
and
Subramanian
that
for
sufficient
calcium
utilization,
not
only
should
a
sufficient
intake
of
protein
be
insured
but
also
that
the
protein
should
contain
all
the
essential
amino
acids
in
adequate
amount.
EFFECT
OF
DIFFERENT
LEVELS
OF
PULSE
PROTEINS
(BENGAL
GRAM
AND
LENTIL)
375
ON
CALCIUM
AND
PHOSPHORUS
BALANCE
AND
CALCIUM
AND
PHOSPHORUS
CONTENTS
OF
DIFFERENT
TISSUES
AND
SERUM
There
is
a
very
little
change
in
the
phos-
phorus
balance
of
animals
receiving
pulse
protein
at
18
per
cent
and
12
per
cent
level
when
compared
with
the
animals
receiving
18
per
cent
casein
(control
diet).
This
change
is
not
statistically
significant.
The
Tables
II
and
HI
show
that
the
percent-
age
of
phosphorus
retention
is
higher
than
the
calcium
retention
in
all
the
cases
which
shows
that
the
beneficial
response
to
higher
protein
intake
is
very
much
in
case
of
phos-
phorus
than
in
case
of
calcium.
The
low
calcium
balance
and
less
percentage
of
calcium
and
phosphorus
retention
in
albino
rats
fed
pulse
protein
at
12
per
cent
protein
level
than
the
18
per
cent
casein
control
diet
may
be
attributed
to
the
insufficient
supply
of
essential
amino
acids
like
trypto-
phan,
lysine,
methionine
and
threonine
which
are
playing
important
role
in
absorption.
Summary
The
effect
of
different
levels
of
pulse
proteins
was
investigated
on
phosphorus
and
calcium
metabolism.
The
rats
fed
18
per
cent
casein
(control
diet)
showed
high
calcium
balance
than
the
rats
fed
18
per
cent
and
12
per
cent
pulse
protein.
There
was
no
significant
change
in
phosphorus
balance
in
rats
fed
18
per
cent
casein
diet
when
compared
with
the
rats
fed
18
per
cent
and
12
per
cent
pulse
protein
diet.
The
calcium
content
of
serum
in
the
rats
fed
12
per
cent
and
18
per
cent
pulse
protein
diet
have
also
been
found
signi-
ficantly
lowered
than
in
the
rats
fed
18
per
cent
casein
(control
diet).
The
decrease
in
contents
of
calcium
serum,
calcium
balance
and
percentage
of
calcium
reten-
tion
in
the
rats
fed
18
per
cent
and
12
per
cent
protein
diet
than
the
18
per
cent
casein
diet
(control
diet)
has
been
attributed
to
insufficient
supply
of
essential
amino
acids
like
tryptophan,
lysine,
methionine
and
threonine.
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