Plant Foods for Human Nutrition 37:41-46 (1987)

Martinus Nijhoff/Dr W. Junk Publishers, Dordrecht - - Printed in the Netherlands

41

Changes in starch, oil, protein and amino acids in developing

seeds of okra (Abeimoschus esculentus L. Moench)
THEYMOLI BALASUBRAMANIAN & S.SADASIVAM 1
Department of Biochemistry, Tamil Nadu Agricultural University, Coimbatore--641 003, Tamil
Nadu, India(l for correspondence)
Key words: okra, seed maturation, protein, oil, starch
Abstract. The okra seeds of variety Pusa sawani were analysed for protein, non-protein nitrogen,
total free amino acids, lysine, and tryptophan from the 7th day to the 42nd day after flowering.
Starch, total sugars and oil percent were also estimated in these seeds. During the early stages of
maturation, the soluble components (non-protein nitrogen, free amino acids and total sugars) were
found in higher quantities than in the later stages. Protein, oil and starch contents increased
gradually from day 7 to day 42. The rate of accumulation of oil was found to be at the highest level
between 21-28 days after flowering, while the rate of protein deposition was greatest between days
35-42. Initiation of seed maturation seems to start 21 days after flowering in okra seeds.

Introduction

O k r a seed was investigated for its seed protein for the first time by Karakoltsides
and Constantinides [5] in 1975. Later, M a r t i n and Rhodes [7] reported on
physicochemical characteristics o f ten varieties o f okra seed. The amino acid
composition o f mature okra seeds and chemical scores o f amino acids in okra
seed meal revealed that valine, isoleucine and lysine were the limiting amino
acids [10]. Protein fractionation o f okra seed meal has also been reported [12].
Changes in the chemical composition o f whole okra fruits during m a t u r a t i o n
have been studied [4, 6]. Studies on changes in chemical constituents in seeds
during m a t u r a t i o n have not been reported, except for the report o f Bhalla et
al. [1] on accumulation and developmental changes in alkali and water soluble
proteins. Here, we report the results o f the study on changes in biochemical
c o m p o n e n t s during m a t u r a t i o n o f okra seeds.

Materials and methods

The variety, Pusa sawani (raised by the Faculty o f Horticulture, Tamil N a d u

Agricultural University) was utilised for this study. The okra flowers were
tagged on the day o f flowering and the fruits were collected at weekly intervals
f r o m day 7 till day 42 after flowering. The seeds were separated, weighed
immediately, and the fresh weight o f 100 seeds recorded. Then they were dried
at 100-110C to constant weight to determine the dry weight o f 100 seeds.

42

x--x
o

Fresh
o Dry

weight
weight_

P~" " " " ' X %

- 14.0

7.(;

/
6.0 -

5,0

Ot

- h?-..O

- I0.0

4.0 _

-8.0

e"

w
4,-

, m

3.0 --

-6.0

..

>~
L

2.0

@
I.
h

-4.0

_ 2.0

i-0

30

40

,,,

i0

20
Doys

offer

50

flowering

Fig. 1, G r a i n m a t t e r a c c u m u l a t i o n a t different stages o f m a t u r a t i o n .

The dried seeds were ground finely to 80 mesh and used for chemical analyses.
The total nitrogen content was estimated by a standard micro-kjeldahl method
and crude protein was calculated by multiplying the total N by 6.25. Nonprotein nitrogen was estimated by the microkjeldahl method in the supernatant
of TCA extracts of seed flour. The difference between total and non-protein
nitrogen was expressed as protein nitrogen. Total free amino acids and sugars
were estimated in 80% ethanol extract by the methods of Misra et al.[9] and
Dubois et al. [2] respectively. Estimations of lysine and tryptophan were carried
out in papain hydrolysates of defatted okra seed meal as per Vogel and Shimura
[13] and Friedman and Finely [3], respectively.

43
Table 1. Protein, non-protein nitrogen, total free amino acids, lysine and tryptophan contents in
okra seeds during maturation?
Days

7
14
21
28
35
42

Crude
proteinb

Non-protein
nitrogen

N x 6.25 (%)

(mg/g seed)b

2.5
2.2
3.9
6.7
11.2
18.3

1.1
0.3
0.5
0.3
0.3
0.5

Total free
amino acids

6.1
1.9
0.9
0.7
1.1
2.3

Lysine

Tryptophan

(rag/16 mg N) c
8.4
5.1
3.9
5.7
3.7
4.8

1.1
1.7
2.7
4.5
2.5
3.8

" Results are average of duplicate determination.

b Expressed on fresh weiglat basis.
In dry defatted seeds.

Oil content was estimated by extracting the oil in diethyl ether in soxhlet
apparatus for 6 h. Starch content was estimated in the defatted meal by first
hydrolysing the starch with 52% perchloric acid[8] and then estimating the
sugars by the phenol-sulphuric acid method of Dubois et al.[2].

Results and discussion

The fresh weight and dry weight of one hundred seeds at different stages of
maturation are given in Fig 1. Fresh weight of 100 seeds was only 1.9 g on the
seventh day after flowering. From day 7 to day 14, a ten-fold increase in weight
was observed and it kept on increasing to 14.5 g in 21-day-old seeds. After 21
days, fresh weight was found to decrease. The maturation phase of okra seeds
appears to start 21 days or later after flowering, since a decline in the fresh
weight of seeds was reported to indicate initiation of seed maturation[11].
However, the dry weight of 100 seeds kept increasing throughout development
until day 35, afterwhich there was no change.
Crude protein was found to be 2.5% on day 7 (Table 1) and 2.2% in 14 day
old seeds. From 14 days onwards, it was found to increase steadily. The initial
decrease in crude protein might be due to the slight increase in the moisture
content. Non protein nitrogen content was found to be the highest (1.1 mg per g)
on day 7 and as the seeds matured it decreased to 0.34).5 mgperg.
The contribution of non-protein nitrogen to total nitrogen was 31% on day
7 (Fig 2). On 14 and 21 day after flowering only 10.6-10.1% of the total nitrogen
was contributed by NPN.After 21 days, the NPN contribution decreased to
2,t%. During the same period (from days 7 to 42), protein nitogen was found
to increase gradually from 2.6 mg to 26.9 mg per g fresh seed. The rate of protein
deposition was greater between the 35th and 42nd days after flowering. The
synthesis and accumulation of protein during maturation may explain the
decrease in the NPN contribution to total nitrogen [11] as the seeds mature. This

44
o,

o NPN

x--x

Per cent

Protein

30

30

X
l

/
/
Y

I
I
/

Z
0

20

I
I

O..
Z

I
I

to.
o

.4.,.
e-

U 10_

""'~0

!
X

_10

O
I.

I@

/ / X ~ o .
x--x

- ~-""~" 0
m

io

20
Days

30
offer

I
40
flowering

50

Fig. 2. Protein nitrogen and percent N P N to total N in okra seeds during maturation.

is augmented by the fact that a higher content of free amino acids was noticed
on day 7 than on days 14 to 42. Lysine, one of the limiting amino acids was
present in higher amounts in young seeds than in mature seeds. Conversely,
tryptophan content was found to be higher in mature seeds than in young seeds.
These changes in individual amino acids are suggestive of an active amino acid
metabolism in the maturing seed even during the last few days of maturation.
Another important constituent of okra seed, per cent ether extractable oil,
was found to be very low on day 7 (Table 2). From day 7 onwards, oil
accumulation was observed till day 42. The rate of increase was found to be very

45
Table 2. Changes in starch, total sugars and oiP in maturing okra seeds.

Days

Starchb
(%)

Total sugarsb
(%)

Oilb
(%)

7
14
2l
28
35
42

1.2
1.5
2.8
3.3
5.8
8.0

1.7
0.4
0.6
2.0
2.1
4.1

0.4
1.3
1.9
5.6
8.2
15.6

a Values are average of duplicate determinations.

b Values are expressed on fresh weight basis.

high from days 21 to 42. A three fold increase in oil content was seen by day 28.
An increase in dry matter accumulation and the onset of maturation on day 21
may explain the higher rate of oil deposition during this period.
The starch content of the seeds was also found to increase from day 7 onwards
to 42 days (Table 2). As starch content increased, total sugar content was found
to decrease until day 21. This is probably due to the high demand on sugars
during the initial starch accumulation process. However, after 21 days, the sugar
was found to increase like that of the starch.

Summary
I n i t i a t i o n o f seed m a t u r a t i o n seems to start 21 days after flowering since fresh
weight of seeds starts to decrease from that day onwards. C r u d e p r o t e i n c o n t e n t
of the seeds increased from 14 days onwards, t h o u g h a slight decrease was
observed d u r i n g the first 14 days after flowering. D u r i n g the early stages of
m a t u r a t i o n , soluble c o m p o n e n t s (like n o n - p r o t e i n n i t r o g e n a n d sugars) were
higher t h a n d u r i n g the latter stages. Oil a n d starch a c c u m u l a t i o n were observed
only d u r i n g the latter part o f m a t u r a t i o n .

References
1. Bhalla PL, Singh MB, Malik CP (1983) Changes in the nature of proteins during okra seed
development. Trop Plant Sci Res 1:189-190
2. Dubois M,Gilles KA, Hamilton JK, Rebers PA, Smith F (1956) Colorimetric method for
determination of sugars and related substances. Anal Chem 28:35(~356
3. Friedman M, Finely JW (1971). Methods of tryptophan analysis. J Agric Food Chem
19:626-631
4. Gopalakrishna Rao KP, Sulladmath UV (1977) Changes in certain chemical constituents
associated with maturation of okra. Veg Sci 4:37-42
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Chem 23:1204-1207
6. Longe OG, Fetuga BL, Akenova ME (1982) Changes in the composition and carbohydrate
constituents of okra A. esculentus with age. Food Chem 8:27-32

46
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