Volume 2, Number 2, July-December 2014

EVALUATION OF MYANMAR THANAKHA (Hesperethusa

Crenulata (Roxb.) M. Roem) AND PROCESSING OF ITS

Dr. Nwe Nwe Aung

Department of Industrial Chemistry
Yadanabon University, Mandalay, Myanmar

ABSTRACT by E nergy Dispersive X -ray F luores-

cence T echnique ( EDXRF). Surface
Thanakha, g rown a bundantly
morphology of Thanakha powder was
in central Myanmar, is well-known as
also s tudied by S EM. T his research
a traditional, natural and organic skin
shows that M yanmar T hanakha i s a n
care h erbal m aterial w hich i s ad ored
anti-bacterial/ an ti-fungal. I n addition,
and widely us ed by t he M yanmar
the presence o f toxic h eavy m etals
society since cen turies ago. A ll p arts
like m ercury an d arsenic w ere not
of this pl ant are us eful as i ndigenous
found in m ost of t he T hanakha
medicine, co smetics an d perfumery.
samples a nd thus t oxic pollutants
Application of T hanakha e nhances
cannot be occurred.
beauty, a cts as pe rfume and ha s sun
protection, astringent a nd a ntiseptic
properties. Thanakha pow der, one of
the e cofriendly pr oducts, i s 100% KEYWORDS
organic s ince t he ba rks a nd r oots are Thanakha, herbal material, anti-
simply t horoughly g round. T he m ain microbial, trace elements, surface
aim o f r esearch w as t o s tudy t he morphology
characteristics of T hanakha by pr e-
liminary p hytochemical te st, a nti-
microbial activity of different solvent INTRODUCTION
extract an d p rocessing o f i ts natural
Thanakha i s w ell-known i n
products. T hanakha s amples w ere
Myanmar as a kind of make-up which
collected f rom S againg R egion and
is adored a nd w idely used by t he
Magwe R egion. D ifferent s olvent
Myanmar w omen o f al l ag es s ince
extracts from Thanakha showed anti-
over 2000 y ears ago. T he fragrance
microbial a ctivities a gainst certain
of T hanakha s timulates t he m ind t o
microorganisms. F rom t he poi nt of
be f resh and cl ear and i mparts the
view of he alth hazard, e valuation of
beauty of f ace and body and
the trace elements in Thanakha is also
smoothness of skin in the form of soft
important f actor. The c ontents of
and wet t exture by c ontinuous us e.
elements in T hanakha w ere d etermined
Most Myanmar women use branches,

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stems and roots of Thanakha plant (U
Tha H la, 1974). T hanakha, well-
known in M yanmar, belongs t o t he
family " Rutaceae" a nd g enus
"Hesperethusa ".
OBJECTIVES OF THE STUDTY
To study the characteristics of
Thanakha by pr eliminary phy to-
chemical t est, an timicrobial a ctivity
of di fferent s olvent e xtract a nd
processing of its natural products.
MATERIALS AND METHODS
Sample Collection
Fresh s amples o f Thanakha
were collected f rom S hinmataung,
Yesagyo Township, P akokku T own-
ship, Magwe R egion a nd S hwebo
Township, Sagaing Region(Figure 1).
Sample Preparation
Different E xtracts o f T hanakha
samples from S hinmataung w ere
prepared by us ing di fferent s olvents
for phytochemical a nd anti-microbial
activity. T hanakha s amples co llected
from Yesagyo T ownship, P akokku
Township, Magwe R egion a nd
Shwebo T ownship, S againg R egion
were collected , air-dried and the bark
was removed from the stem. The bark
was chipped into very small pieces of
approximately 1 c ubic c entimeter in
size a nd g round a nd pa ssed through
200 mesh, 300 m esh a nd 400 mesh
screens. T he p owder s amples w ere
stored in a n a ir-tight c ontainer f or
determining trace elements.
Volume 2, Number 2, July-December 2014
Phytochemical Analysis
Solvent extracts of Shinmataung
Thanakha w ere t ested t o r eveal the
presence of a lkaloids, g lycosides,
saponin glycosides, steroids, phenolic
compounds, flavonoids and tannin.
Anti-microbial Activity
25 g of powder from stem and
root of S hinmataung T hanakha w as
filled in a th imble a nd e xtracted
respectively w ith n -hexane, be nzene,
ethyl acet ate, ethanol an d special
boiling poi nt s olvent ( SBP) us ing a
Soxhlet extractor. A ll th e e xtracts
were s ubjected t o an ti-microbial
activity a ssay. The s ensitivity testing
of the extracts were determined using
agar-well di ffusion m ethod. The
following microorganisms were used:
Bacillus pumalis, Bacillus subtilis,
Candida albicans, Mycobacterium
species, Pseudomonas aerguinosa
and Staphylococcus aureus.
Determination of Trace
Elements
The T hanakha pow der of
various r egions w ere analyzed for
trace el ements co ntent using E nergy
Dispersive X-ray Fluorescence T ech-
nique.
SEM Studies
The microstructure of Thanakha
powder of va rious r egions were
analyzed us ing S EM ( Scanning
Electron M icroscopy) JSM -6510
JEOL, Japan.
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Volume 2, Number 2, July-December 2014

RESULTS AND DISCUSSION found i n skin lesions a nd leprosy. I n

addition, n -hexane ex tract o f r oot
Phytochemical analysis of solvent
bark showed s ignificant i nhibition
extracts of S hinmataung T hanakha
zones against B.substilis, S-aureus
(Table1) r evealed the p resence of
and Mycobacterium species.
alkaloids, glycosides, saponin glycosides,
According to t his s creening t est,
steroids, phenolic compounds, flavonoids
various parts of Thanakha were found
and t annins a nd a bsence of r educing
to possess anti- microbial activity.
sugars. A lkaloids, s aponins a nd
In Myanmar, T hanakha i s
tannins are t o be bi ologically a ctive.
used not only as cosmetics but also as
Due to t he pr esence of a lkaloids i n
indigenous m edicine. From t he point
Thanakha plant, it gives a slight bitter
of view of he alth hazard, i t i s be tter
taste.This a nalysis s howed t hat the
to a void t he us e of cosmetics
active pr inciple r esponsible f or the
containing toxic elements. According
anti-microbial activity was a phenolic
to t hese facts, ev aluation of t he trace
compound. F rom t he results of the
elements i n T hanakha i s a lso a n
phytochemical a nalysis, Thanakha
important f actor (Table 2, 3, 4 a nd
could be us ed to f ormulate ne w and
Figure 2, 3 ).
more pot ent anti-microbial dr ugs of
natural origin (Table 1). Trace el ements ar e indication
of t he pr esence of non -volatile
Biological s creening i s a
inorganic compounds. T he c ontents
useful t ool for s creening pharma-
of el ements i n T hanakha were
cological a ctivity. Agar well d iffusion
determined by E nergy D ispersive X-
method i s s imple, e conomical a nd
ray F luorescence Technique (EDXRF).
reproducible and is quick to perform.
According to th e EDXRF r esults in
In vitro study is the fundamental and
Table (5 ) an d i ts r elevant E DXRF
essential f or t he de velopment of t he
graph (Figure 4) , it is found that the
natural pr oduct. From t he r esults of
contents of c alcium, iron a nd potas-
Table (2), (3), (4) and Figure (2) and
sium were r elatively h igher th an
(3), a nti-microbial a ctivity a ssay of
those of t he other m inerals. T he
all the f ive extracts r evealed t hat
presence o f t oxic h eavy m etals l ike
ethyl acetate an d b enzene ex tract
mercury an d arsenic were not f ound
showed significant activity against all
in T hanakha from di fferent regions
organisms. O ther s olvent e xtracts
(Table 5 and Figure 4).
viz., n -hexane, e thanol, a nd S BP di d
not s how significant zone of By using S EM t o s tudy t he
inhibition for s ome or ganisms. surface m orphology of Thankha
However, et hanol ex tract o f 3 stem powder (Figure 5), the powder which
bark i nhibited 5 out of 6 tested had a particle s ize (200 m esh) could
organisms an d al l et hanol ex tracts of be s een t o c ontain a l ot of f ibre. F or
the w hole Thankha pl ant e xhibited the p reparation of T hanakha
against Mycobacterium which ar e products, t he particle s ize of

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Thanakha pow der plays a significant
part in d etermining f ormulation
elegance, r ate o f settling, ab sence of
caking a nd f inal s tability o f the
product. Therefore, Thanakha powder
having particle size (300mesh) should
be u sed t o i ncrease t he adhesive
power and reduce the abrasion of the
skin (Figure 5).

Table 1. Phytochemical Investigation of Thanakha.

No Test Solvent Reagent Observation Result
Stem Root Stem Root
1 Alkaloids 5% Mayer’s reagent White White + +
HCl precipitate precipitate
2 Glycosides H2O 10% Lead precipitate precipitate + +
acetate solution
3 Reducing H2O Fehling’solution No No _ _
sugars precipitate precipitate
4 Tannins H2O 1% Ferric precipitate precipitate + +
chloride solution
5 Steriods Pet. Acetic Green Green + +
ether anhydride and colour colour
conc. Sulphuric
acid
6 Saponins H2O Distilled water Frothing Frothing + +

7 Flavonoids EtOH Mg ribbon & Pink Pink + +

conc. HCl colour colour
8 Phenolic H2O Ferric chloride Violet Violet + +
Group solution color color
(+) Present (-) Absent
Source: Original Research

Table 2. Microorganisms and Their Respective Code Number used for

Antimicrobial Activity.
No. Organisms Code No.
1 Bacillus pumalis IFO-12102
2 Bacillus subtilis ap-0225015
3 Candida albicans IFO-1060
4 Mycobacterium species IFO-3158
5 Pseudomonas aeruginosa IFO-3080
6 Staphylococus aureus ATCC-12877
IFO : Institute of Fermentation, Osaka, Japan
ATCC : American Type Culture Collection
Source: Development Centre for Pharmaceutical Technology, Ministry of
Industry.

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Table 3. Anti-microbial Activity of Different Solvent Extracts of Thanakha

Stem.
Organisms
Sample Solvents B.subtilis S.aureus P.aeruginosa B.pumalis C.albicans M.spscies
n-
- - - - - -
hexane
13mm 12mm 14mm 14mm 14mm 17mm
benzene
(+) (+) (+) (+) (+) (+)
ethyl 33mm 24mm 32mm 31mm 37mm 29mm
Stem
acetate (+++) (+++) (+++) (+++) (+++) (+++)
Ethyl 12mm
- - - - -
alcohol (+)
12mm
SBP - - - - -
(+)
Agar well- 10mm
10mm~14mm (+)
15mm~19mm (++)
20mm above (+++)
Source: Original Research

Table 4. Anti-microbial Activity of Different Solvent Extracts of Thanakha

Root.
Organisms
Sample Solvents B.subtilis S.aureus P.aeruginosa B.pumalis C.albicans M.spscies
n- 17mm 12mm 12mm
- - -
hexane (+) (+) (+)
15mm 15mm 14mm 12mm 14mm 15mm
benzene
(+) (+) (+) (+) (+) (+)
ethyl 19mm 25mm 20mm 20mm 20mm 23mm
Root
acetate (++) (+++) (+++) (+++) (+++) (+++)
Ethyl 12mm 14mm 14mm
- - -
alcohol (+) (+) (+)
12mm 12mm 14mm 14mm
SBP - -
(+) (+) (+) (+)
Agar well- 10mm
10mm~14mm (+)
15mm~19mm (++)
20mm above (+++)
Source: Original Research

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Table 5. EDXRF Analysis for Trace Elements in Thanakha.

Analysis Shimataung (%) Shwebo (%) Pakokku (%)
Ca 83.185 47.510 72.521
Fe 6.872 39.660 18.336
K 4.508 2.512 3.734
Sr 4.360 1.939 5.409
Cu 1.075 1.737 -
Mn - 0.941 -
Pb - - -
Zr - - -
Ti - 3.612 -
Zn - 2.088 -
As - - -
Hg - - -
Source: Original Research

Figure 1. Shinmataung Thanakha (Hesperethusa Crenulata (Roxb.)M. Roem).

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Figure 2. Anti-microbial Activity of Different Solvent Extracts from

Shinmataung Thanakha (Stem).

Source: Original Research

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Figure 3. Anti-microbial Activity of Different Solvent Extracts from

Shinmataung Thanakha (Root).

Source: Original Research

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Figure 4. EDXRF Graph of Thanakha (a)Shinmataung (b) Pakokku (c)

Shwebo.

Source: Original Research

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Figure 5. SEM Analysis of (a) Shinmataung (Heartwood Powder), (b)

Shwebo (Stem Bark- Powder), (c) Pakokku (Stem Bark Powder), (d)
Shinmataung (Stem Bark Powder), (e) Thanakha Powder from Paste (by
machine) (f) Thanakha Powder from Paste (by manual,)(g) Thanakha powder
(+200 mesh) ,(h) Thanakha powder (+300 mesh) and (i) Thanakha powder (-
300 mesh).

Source: Original Research

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CONCLUSION
Phytochemical a nalysis o f all
the s olvent extracts r evealed the
presence of a lkaloids, g lycosides,
saponin glycosides, steroids, phenolic
group, flavonoids a nd t annins a nd
absence of reducing sugars. Different
solvent e xtracts f rom Shinmataung
Thanakha s howed anti-microbial
activities against certain microorganisms.
The p resence o f t oxic h eavy m etals
like m ercury an d arsenic w ere not
found i n Thanakha f rom different
regions.
In t he pr eparation of finished
Thanakha pr oducts, i t w as not iced
that the particle size and fibre content
are of i mportant f actors concerning
especially t he T hanakha cr eam. The
stability a nd t exture of Thanakha
cream depend on the particle size and
fibre c ontent of T hanakha powder.
Using T hanakha he artwood onl y is
economical b ut it g ives impaired
quality of Thanakha Products.
In c onclusion, t he c onsumers
receive n atural p roducts o f s pecified
best quality by using Thanakha which
has va luable pr operties f or a nyone a t
any age.
ACKNOWLEDGEMENT
The author w ould l ike t o
express s incere t hanks to D r Khin
Maung Oo, R ector, D r. M aung
Maung N aing, P ro-rector a nd D r Y i
Yi M yint, P rofessor a nd H ead,
Industrial C hemistry D epartment,
Yadanabon U niversity f or t heir
permission to review and s ubmit t his
118
research ar ticle. The au thor would
like t o ex press t he m ost sincere
gratitude to Dr Kyaw Htin, Part-time
Professor, Retired, Industrial Chemis-
try D epartment, University of Yangon
and Dr M in M yint, P rofessor a nd
Head, R etired, I ndustrial C hemistry
Department, D agon U niversity, for
their c lose supervision a nd suggestions
during t he c ourse of t his research
work.
The au thor a lso t hanks the
Universities R esearch C entre (Yangon)
and t he D evelopment C entre for
Pharmaceutical Technology f or t he
assistance in p roviding f acilities to
conduct this research work.
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Burmese Medicinal Plants, (1980),
Agriculture Corp, Yangon
Dissertation, Industrial Chemistry
Department, University of
Yangon.
Nwe Nwe Aung (2006), “
Production of Traditional
Myanmar Thanakha as
Cosmetic”,PhD
Pearson, D., (1962), "The Chemical
Analysis of Foods" Churchill
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Tha Hla, U; (1974),
“Myanmar Thanakha”, Sarpai
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