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Indian Journal of Natural Products and ResourcesVol. 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 andstems of various plants, notably corn, potatoes, wheat and rice. Starch varies widely according to its source owing to itsgranular characteristics. About 50% of starch produced is used in food industry, either as food additive or to improvefunctional 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 itsrhizome.  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  wasdone using 1% ammonium oxalate and 0.03 M ammonia solution and purification of starch was done according to themethod of Badenhuizen, 1964. The yield of starch in 0.03 M ammonia solution and 1% ammonium oxalate was obtained38.46 and 37.64%, respectively. The granular shape and size of starch granules were recorded using Scanning ElectronMicroscope. The shape of C. angustifolia  starch granules were small rounded, oval to elliptical, spherical, elongated and3.32 µ  to 32.55 µ  in length and 2.29 µ  to 8.47 µ  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 wouldaid 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

IntroductionTropical tubers contain starch as the major

component and are staple or subsidiary food forthe 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 oftuber crops and the possibility of using these native

starches instead of chemically modified starches1-3

.

In some forest tubers like cassava extraction ofstarch 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

time for extraction may promote microbial growthand also deteriorate colour and quality of starch.

Economical extraction and maximum recovery ofstarch 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 onsize 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. starchyield have been estimated and granular shape and size

of the starch granules have been studied7,8. Suitabilityof 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 Himalayanranges. The plant grows wild in the moist and cool

areas at altitudes of about 2000 m and is cultivated to

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

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INDIAN J NAT PROD RESOUR, SEPTEMBER 2012408

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 forthe 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 unexploitedstarch source. 

Materials and MethodsThe 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 rhizomespieces 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% ammoniumoxalate5. The suspension was then filtered through

muslin cloth and kept for sedimentation of starch.The residue retained on muslin cloth was againhomogenized 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 wasextracted 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

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 starchwas 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 roomtemperature, 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. Thesedimentation cycle was repeated 2-3 times. After

sedimentation of the starch granules, the protein-

toluene layer was discarded and the starch wasdefatted by refluxing with methanol (80%), washed

with acetone and ether, and dried under reduced

pressure. The yield of starch was then quantifiedgravimetrically.

Scanning Electron Microscopy

Starch sample was taken in a small stoppered glasstube and dehydration of starch was done serially in

30, 50, 70 and 90% ethyl alcohol for few minutes.

Afterwards absolute alcohol was added and sampleswere 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 thestarch sample was preserved in amyl acetate until

used for gold coating.

For preparation of sample for gold coating amylacetate 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 stuband 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 coatingmachine and viewed in Stereoscan Leica 410

microscope at low magnification 500 and EHT 5 KV

using SE1 detector. The size of starch granules weremeasured by point to point method and photographed

through camera attached with the SEM.

Results and DiscussionThe 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 ofPlate 1—Rhizome of Curcuma angustifolia 

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ABHA RANI & CHAWHAAN: EXTRACTION & SCANNING ELECTRON MICROSCOPY OF CURCUMA ANGUSTIFOLIA STARCH 409

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 ofC. 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 and10.34%, 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 forextraction 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 materialreleasing 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 wasfound most effective in improving the yield of starch

in aroids and yams

21,22

.The Scanning Electron Microscopy (SEM) studiesrevealed that the size and shape of starch granules

of C. angustifolia  were small, rounded, oval

to elliptical, spherical, elongated 3.32 µ-32.55 µ  in

length and 2.29 µ-8.47 µ in width (Plate 2). The sizeof the granules observed in the present investigation

is coherent with the average size of starch granules

ranging between 3-20 µm for small granules and

20-48 µm for large granules. The elliptical shape

and granular size of 14-46 µm and 1.6-4.2 µm,

respectively of starch granules were reported in

C. zedoaria  and C. malabarica23

. The SEM studiesrevealed 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 µ while in C. malabarica, C. longa and C. sylvatica 

granular size 10-40 µm were reported7.  The starchgranules size of Casimiroa edulis, Careya arborea,

 Aesculus assamica, Canna edulis, Pueraria tuberos

and Pachyrrhizus erosus (Linn.) Urban were reportedin the range of 3.6-21.6 µm, 7-24 µm, 5-40 µm,

13.05-57.63 µm and 1.6-22.8 µm and 6-35 µm,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 µm was reported19.

ConclusionThe study demonstrated that for extraction of

starch from C. angustifolia 0.03 M ammonia solution

method described is best yielding significant quantityand better quality of starch. The findings can be usedto check adulteration and to provide authentic sample

of starch of Tikhur . From the results, it can be

concluded that Tikhur could become source of starchfor commercial utilization.

AcknowledgementsWe express our sincere thank to the Director,

Tropical Forest Research Institute (TFRI), Jabalpur,

M.P. for providing facilities. We are also grateful to

Plate 2—Scanning electron micrograph of starch of Curcuma

angustifolia 

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INDIAN J NAT PROD RESOUR, SEPTEMBER 2012410

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, TFRIare gratefully acknowledged.

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