Indian Journal of Natural Products and Resources

Vol. 3(3), September 2012 pp. 407-410

Extraction and scanning electron microscopic studies of

Curcuma angustifolia Roxb. starch
Abha Rani* and Pravin H Chawhaan
Arid Forest Research Institute, PO-Krishi Upaj Mandi, New Pali Road, Jodhpur 342 005, Rajasthan, India
Received 07 January 2011; Accepted 29 September 2012
Starch is the main carbohydrate produced by all green plants and is basically found in the seeds, fruits, tubers, roots and
stems of various plants, notably corn, potatoes, wheat and rice. Starch varies widely according to its source owing to its
granular characteristics. About 50% of starch produced is used in food industry, either as food additive or to improve
functional properties and shelf life of food products. Curcuma angustifolia Roxb., commonly known as Tikhur in Hindi,
occurs wildly in many parts of India. It is traditionally recognized as medicinal plant and also contains starch in its
rhizome. In some forest tubers extraction of starch is simple, whereas this is not always so with other tuber starches.
Maximum recovery of starch with economical extraction is thus important. Therefore, extraction of the starch in Tikhur was
done using 1% ammonium oxalate and 0.03 M ammonia solution and purification of starch was done according to the
method of Badenhuizen, 1964. The yield of starch in 0.03 M ammonia solution and 1% ammonium oxalate was obtained
38.46 and 37.64%, respectively. The granular shape and size of starch granules were recorded using Scanning Electron
Microscope. The shape of C. angustifolia starch granules were small rounded, oval to elliptical, spherical, elongated and
3.32 to 32.55 in length and 2.29 to 8.47 in width. The study demonstrated that for extraction of starch from C.
angustifolia 0.03 M ammonia solution method described is best and yields significant quantity of starch. The results would
aid the authentication and to check the adulteration of starch of this species.
Keywords: Rhizome, Starch, Starch granules, Curcuma angustifolia, Tikhur, Food product.
IPC code; Int. cl. (2011.01)A61K 36/00, C08B 30/04

Introduction
Tropical tubers contain starch as the major
component and are staple or subsidiary food for
the low income group. In Asian countries like
India, China, Thailand, Indonesia except Manihot
esculenta Crantz (Cassava) and Ipomoea batatas
Lam. (Sweet potato), other tuber crops have not
been exploited for extraction of starches. Studies on
different starches in India and elsewhere have
revealed wide diversity in starch characteristics of
tuber crops and the possibility of using these native
starches instead of chemically modified starches1-3.
In some forest tubers like cassava extraction of
starch is simple; the isolated starch is white in colour
and relatively free from other chemical impurities,
whereas this is not always so with other tuber
starches4. The settling of starch granules is often
hindered by presence of various components like
proteins, fats, mucilage's and latex, leading to
lowering of quantity of extracted starch. The long

*Correspondent author:
E-mail: abha@icfre.org, abhapravin@yahoo.co.in;
Phone: 0291-2729145(O), 0291-2721680(R), 9414071269 (Mob)

time for extraction may promote microbial growth

and also deteriorate colour and quality of starch.
Economical extraction and maximum recovery of
starch from forest tubers is thus utmost important.
Hence different chemicals were used for extraction
and improving yield of starch from tropical tubers5,6.
Starch occurs in plants as granules having a
characteristic size and shape for each plant species.
The suitability of starch for a specific use depends on
size and shape of the starch granules. In Curcuma
malabarica, C. caesia Roxb., Curcuma longa Linn.
syn. C. brog Valeton, C. amada Roxb. and other
Curcuma species except C. angustifolia Roxb. starch
yield have been estimated and granular shape and size
of the starch granules have been studied7,8. Suitability
of Curcuma angustifolia starch for pharmaceutical
excipient has also been reported9.
Curcuma angustifolia Roxb., commonly known as
Tikhur, occurs in the hilly tracts of Central India,
West Bengal, erstwhile Bombay and Madras
provenances and some of the lower Himalayan
ranges. The plant grows wild in the moist and cool
areas at altitudes of about 2000 m and is cultivated to

INDIAN J NAT PROD RESOUR, SEPTEMBER 2012

408

a small extent. The plant is known to yield volatile oil

from leaves and possess antimicrobial properties10,11.
The rhizomes of C. angustifolia contain starch and
is reported to be used by tribals of India for
the preparation of milk puddings. Since, starch of
Tikhur has received little attention, despite their
potential application in food and textile industries.
The present investigation discusses the yield
determination vis-a-vis scanning electron microscopic
studies of starch granules of this hitherto unexploited
starch source.
Materials and Methods
The rhizomes of C. angustifolia were collected
from Non Wood Forest Produce Garden, Tropical
Forest Research Institute, Jabalpur, Madhya Pradesh,
India (Plate 1).
Extraction of starch

After washing and peeling of the thin skin,

rhizomes were cut into small pieces and subsequently
passed through the extraction procedures by two
different methods. In first method the cut rhizomes
pieces of 0.1 cm were dipped overnight in 0.1%
solution of sodium bisulphite12. The samples were
then homogenized in a laboratory mixer and
extraction of the starch was done in 1% ammonium
oxalate5. The suspension was then filtered through
muslin cloth and kept for sedimentation of starch.
The residue retained on muslin cloth was again
homogenized in a laboratory mixer and the extraction
of starch was done in ammonium oxalate. The process
was repeated 3-4 times or till all the starch was
extracted from the material.
In second method the sample was homogenized
directly in a laboratory mixer and extraction of
starch was done in 0.03 M ammonia6. The filtrate
so obtained was collected separately and kept for

Plate 1Rhizome of Curcuma angustifolia

sedimentation. The residue retained on muslin cloth

was again homogenized in a laboratory mixer and the
extraction of starch was done in 0.03 M ammonia.
The process was repeated 3-4 times till all the starch
was extracted from the material. Afterward the
supernatant layer was decanted from the sediment
starch layer and the sediment was collected.
The sediment starch of both procedures was
collected separately and purified5. The sediment
was then treated with saline suspension at room
temperature, followed by shaking the saline
suspension with toluene (0.1volume) to denature and
to ensure removal of residual cytoplasmic proteins
without causing degradation of starch. The
sedimentation cycle was repeated 2-3 times. After
sedimentation of the starch granules, the proteintoluene layer was discarded and the starch was
defatted by refluxing with methanol (80%), washed
with acetone and ether, and dried under reduced
pressure. The yield of starch was then quantified
gravimetrically.
Scanning Electron Microscopy

Starch sample was taken in a small stoppered glass

tube and dehydration of starch was done serially in
30, 50, 70 and 90% ethyl alcohol for few minutes.
Afterwards absolute alcohol was added and samples
were kept for few hours. The solvent was then
decanted and absolute alcohol and amyl acetate (1:1)
ratio was added and kept for 24 hours. After
decanting the alcohol and amyl acetate mixture the
starch sample was preserved in amyl acetate until
used for gold coating.
For preparation of sample for gold coating amyl
acetate was decanted from the glass tube and starch
was kept for air drying for few minutes. Afterwards
gold coating tape was pasted on the circular stub
and starch granules were sprinkled on the stub.
The prepared sample stub was coated with gold
for 30-45 seconds by using gold coating machine.
The stub was then removed from gold coating
machine and viewed in Stereoscan Leica 410
microscope at low magnification 500 and EHT 5 KV
using SE1 detector. The size of starch granules were
measured by point to point method and photographed
through camera attached with the SEM.
Results and Discussion
The yield of C. angustifolia starch in 0.03 M
ammonia solution was 38.46% whereas it was 37.64%
in 1% ammonium oxalate solution after removal of

ABHA RANI & CHAWHAAN: EXTRACTION & SCANNING ELECTRON MICROSCOPY OF CURCUMA ANGUSTIFOLIA STARCH

the impurities. The extraction of starch in both the

extraction medium resulted almost similar quantity of
starch. In both the procedures feasible quantity of
native starch was obtained. Repeated washing of
C. angustifolia starch with saline solution and then
with saline solution with toluene yielded low protein
starch. In different forest species, viz. Aesculus
assamica Griff., Amorphophallus campanulatus
Blume ex Decne, Canna edulis Ker-Gawl., Careya
arborea Roxb., Casimiroa edulis Llave & Lex.,
Shorea robusta Gaertn.f., Stephania glabra Miers.,
Pueraria thomsonii Benth., P. tuberosa DC. and
Quercus leucotrichophora A. Camus starch yield is
reported as 12.10, 34.03, 60.3, 50.30, 26.30, 15.60,
18.05, 10, 34.20 and 54.70% (on dry basis),
respectively12-18. The starch yield in C. malabarica
K.C. Velayudhan, V.A. Amalraj & V.K. Muralidharan,
C. caesia Roxb., C. longa Linn., C. amada Roxb.,
C. aromatica Salisb., C. zedoaria Rosc., C. aeruginosa
Roxb., C. haritha J.K. Mangaly & M. Sabu,
C. raktakanta J.K. Mangaly & M. Sabu and
C. sylvatica Valeton were reported to be 21.40, 19.86,
18.00, 10.22, 15.00, 14.06, 14.10, 14.32, 14.20 and
10.34%, respectively7. In C. amada, Pachyrhizus ahipa
(Wedd.) Parodi (yam bean) and Ceropegia bulbosa
Roxb. var. bulbosa starch yield of 45, 56-58 and
82% (dry weight basis), respectively was reported8,19,20.
The 0.03 M ammonia solution was used for
extraction of starch from tropical forest tubers,
viz. Manihot esculenta Crantz (Cassava), Dioscorea
alata Linn., D. esculenta Burkill, D. rotundata
Poir., Ipomoea batatas Lam. (sweet patato) and
Xanthosoma sagittifolius (Linn.) Schott6. Ammonia
acts by complexing with the mucilaginous material
releasing the starch granules. Thus use of ammonium
solution enables faster settling of starch in less
viscous slurry. The short residence time also prevents
microbiological damage of the starch4. Ammonia was
found most effective in improving the yield of starch
in aroids and yams21,22.
The Scanning Electron Microscopy (SEM) studies
revealed that the size and shape of starch granules
of C. angustifolia were small, rounded, oval
to elliptical, spherical, elongated 3.32 -32.55 in
length and 2.29 -8.47 in width (Plate 2). The size
of the granules observed in the present investigation
is coherent with the average size of starch granules
ranging between 3-20 m for small granules and
20-48 m for large granules. The elliptical shape
and granular size of 14-46 m and 1.6-4.2 m,

409

Plate 2Scanning electron micrograph of starch of Curcuma

angustifolia

respectively of starch granules were reported in

C. zedoaria and C. malabarica23. The SEM studies
revealed the variation in shape of granules, with
round, elliptic, irregular and polygonal. Similar
studies have also been reported in C. amada8. The
shape of starch granules of Stephania glabra and
Pueraria thomsonii were reported spherical and
polygonal12. In C. zedoaria, C. raktakanta, C. caesia,
C. aeruginosa and C. aromatica granular size 6-25
while in C. malabarica, C. longa and C. sylvatica
granular size 10-40 m were reported7. The starch
granules size of Casimiroa edulis, Careya arborea,
Aesculus assamica, Canna edulis, Pueraria tuberos
and Pachyrrhizus erosus (Linn.) Urban were reported
in the range of 3.6-21.6 m, 7-24 m, 5-40 m,
13.05-57.63 m and 1.6-22.8 m and 6-35 m,
respectively4,13,14,16. Granular shape of starch of
C. angustifolia resembles to the starch of Canna
edulis and granule size resembles to the starch
of Amorphophallus paeoniifolius (Dennst.) Nicolson
(Elephant foot yam) and Pachyrrhizus erosus
(Yam bean). In Pachyrrhizus ahipa geometric form
starch granule with size of 5-35 m was reported19.
Conclusion
The study demonstrated that for extraction of
starch from C. angustifolia 0.03 M ammonia solution
method described is best yielding significant quantity
and better quality of starch. The findings can be used
to check adulteration and to provide authentic sample
of starch of Tikhur. From the results, it can be
concluded that Tikhur could become source of starch
for commercial utilization.
Acknowledgements
We express our sincere thank to the Director,
Tropical Forest Research Institute (TFRI), Jabalpur,
M.P. for providing facilities. We are also grateful to

410

INDIAN J NAT PROD RESOUR, SEPTEMBER 2012

Indian Council of Forestry Research & Education,

Dehra Dun for financial assistance. Help rendered by
Shri S. K. Choubey RA II and Miss Sushma Marawi
RA II of Non Wood Forest Produce Division, TFRI
are gratefully acknowledged.
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