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Original Article 111
J Health Res
2009,
23
(3): 111-115
QUALITY ASSESSMENT OF RHINACANTHUS NASUTUS
Tanasorn Tunsaringkarn1, Chanida Palanuvej1, Somchai Issaravanich1,
Niran Vipunngeun1, Anusorn Rungsiyothin1, Anchalee Chuthaputti2 and
Nijsiri Ruangrungsi1,3,*
1College of Public Health Sciences, Chulalongkorn University, Bangkok 10330, Thailand
2Department for Development of Thai Traditional and Alternative Medicine, Ministry of Public Health,
Nonthaburi 11000, Thailand 3Department of Pharmacognosy, Faculty of Pharmaceutical Sciences,
Chulalongkorn University, Bangkok 10330, Thailand
ABSTRACT: Dried whole plants of Rhinacanthus nasutus (L.) Kurz (Acanthaceae) were collected
from 15 Thai traditional drugstores located around four regions of Thailand. The
pharmacognostic specifications were performed according to the World Health Organization
(WHO) guidelines for herbal standardization. Microscopic investigation revealed multicellular
and glandular trichomes as well as lithocyst cells occured on both leaf surfaces. Calcium
carbonate and raphide crystal were found on the epidermal and parenchyma cell of the stem.
The powdered crude drug displayed microscopic identification of both leaves and stems. The
mean contents of foreign matter, acid insoluble ash, total ash, moisture content, ethanol-
soluble extractive, water-soluble extractive and loss on drying were 0.2, 6.5, 10.7, 8.7, 1.8, 16.9
and 9.5 % of dry weight respectively. TLC fingerprint of the methanolic extracts were
demonstrated.
Keywords: Rhinacanthus nasutus, pharmacognostic specification
INTRODUCTION: Rhinacanthus nasutus (L.) Kurz
(Acanthaceae family) is widely distributed in some
parts of the sub-continent India, China and in the
region of Southeast Asia including Thailand. R.
nasutus have been used for various illness
treatments. The literatures reported of traditional
medicine preparations from roots and leaves of
this shrub were widely advocated for prickly heat
and scurf and whole plant for ringworm, various
skin problems such as eczema1,2) as well as
pulmonary tuberculosis3). The active ingredients of
extracts have been reported for antifungal1-3), anti-
bacterial3,4), antivirus5,6), cytotoxic7), anti platelet8),
antitumor9,10) and antiproliferative11) activities. This
study aimed to examine the pharmacognostic
identification of R. nasutus crude drug distributed
among traditional drug stores in Thailand.
MATERIALS: The dried whole plants of R.
nasutus were collected from 15 Thai traditional
drugstores located at 4 regions of Thailand as
follow : Bangkok (3 stores), Chon Buri, Nakorn
Pathom, Kamphaeng Phet, Pichit, Nakorn Sawan,
Chiang Mai, Nakorn Ratchasima, Ubon Ratchathani,
Roi et, Songkhla, Trang and Nakorn Si Thammarat.
The samples were identified by one of the authors
(N.R.). The voucher and numbers of specimens
were deposited at College of Public Health Sciences,
Chulalongkorn University, Thailand.
METHODS: Size, colour and other visual
inspections were examined for macroscopic
characteristics. Transverse sections and ground
powders were inspected under microscope
(Olympus BX41) for anatomical and histological
characteristics. The foreign matter was excluded
and recorded the percentage amount. The total
and acid-insoluble ashes remained after burning
at 500-600C were determined for inorganic
components part. The loss on drying test was
performed to measure the amount of water and
volatile matters by drying the sample at 105C
until constant weight. In addition, the moisture
content was also quantitated using Azeotropic
distillation method. Extractable matters in
absolute ethanol as well as water were determined
by maceration to represent the amount of active
constituents. Chemical fingerprint was performed
by thin-layer chromatography after reflux in
methanol. The details of methodology were
described elsewhere12,13).
To whom correspondence should be addressed.
E-mail: nijsiri.r@chula.ac.th, Tel./fax. +66 2218 8158
112 Original Article
J Health Res
2009,
23
(3): 111-115
RESULTS AND DISCUSSION: R. nasutus is an
erect, branched shrub, tall up to 2 m. Its stem is
obtusely quadrangular. The leaves are opposite,
simple, ovate to lanceolate or elliptical, 3-10 cm
long and 1-5 cm wide. The young stem and leaves
are puberulent. The flowers are white with red
markings at base (Figure 1). Dried whole plants
have been used as crude drug in Thai traditional
drug stores (Figure 2). Anatomical and histological
characteristics of R. nasutus were presented (Figure
3-6). Multicellular and glandular trichomes as
well as lithocyst cells occured on both leaf
surfaces (upper and lower epidermis). The micro-
scopic characters of R. nasutus dried leaves
observed in this study were in agreement with the
previous report of fresh leaves by Phisutthanan S.
and Phisutthanan N.14). In addition to the leaves,
this study explored the stems microscopically and
found multicellular and glandular trichomes as
well. The epidermal cell containing calcium
carbonate and parenchyma cell containing
raphide crystals were also presented. The
powdered crude drug displayed microscopic
identification of both leaves and stems, for
example, stomata, trichomes, calcium carbonate,
raphide crystals, fibers, vessels and sclereids
(Figure 6).
Table 1 showed the quality characters of R.
nasutus crude drug. The contents of foreign
matter, acid-insoluble ash, total ash, moisture
content and loss on drying were found to be 0.2
0.2, 6.5 2.1, 10.7 2.6, 8.7 1.2 and 9.5
0.9 % of dry weight respectively whereas the
determination of ethanol- and water- extractive
values were found to be 1.8 1.2 and 16.9
3.8 % of dry weight respectively. From these data,
the requirements of the specification could be set
up (in integer) using the criteria of either
mean+10% or mean-10%15) (Table 2). Phisut-
thanan S. and Phisutthanan N.14) reported the
contents of ethanol-, water-extractives and loss on
drying, of R. nasutus fresh leaves from 3 sources
in the Northern provinces of Thailand. Their
results were 5.6, 4.6 and 5.6 % of ethanol-
extractive, 29.0, 27.7 and 26.1 % of water-
extractives and 23.3, 13.4 and 13.0 % of loss on
drying. The three content values of this study
were likely lower than the earlier report especially
ethanol-extractive value14). This was under-
standable for the fresh samples collected from the
wilds versus the dried samples obtained from
drugstores.
Figure 1 Twig of R. nasutus with flowers
Table 1 The quality characters of R. nasutus crude drug
Specification Mean ± SD* Min – Max*
Foreign matter 0.20 ± 0.15 0.06 - 0.74
Acid-insoluble ash 6.53 ± 2.10 3.52 – 11.98
Total ash 10.68 ± 2.62 6.46 – 15.03
Moisture content 8.74 ± 1.15 6.00 - 10.80
Volatile oil content Not found Not found
Loss on drying 9.53 ± 0.90 6.82 - 10.98
Ethanol- extractive 1.84 ± 1.18 0.30 - 5.50
Water- extractive 16.90 ± 3.80 9.40 - 27.08
*% of dry weight
Table 2 The requirement specification of R. nasutus
crude drug
Specification*
Foreign matter Not more than 1 %
Acid-insoluble ash Not more than 7 %
Total ash Not more than 12 %
Moisture content Not more than 10 %
Loss on drying Not more than 11 %
Ethanol- extractive Not less than 1 %
Water- extractive Not less than 15 %
*% of dry weight
Original Article 113
J Health Res
2009,
23
(3): 111-115
1
2
3
4
6
5
7 8
9
10
0.01 mm
0.03 mm
1. multicellular trichome
2. glandular trichome
3. epidermal cell containing
calcium carbonate
4. epidermis layer
5. collenchymas
6. chlorenchyma
7. phloem tissue
8. xylem vessel
9. parenchyma
10. parenchyma containing
raphide crystals
Figure 3 Microscopic character of the transverse section of R. nasutus stem
1
2
3
7
8
9
10
11
6
4
5
6
1. glandular trichome
2. multicellular trichome
3. upper epidermis
4. palisade cell
5. spongy cell
6. collenchymas
7. stomata
8. xylem vessel
9. phloem tissue
10. parenchyma
11. lower epidermis
Figure 4 Microscopic character of the transverse section of R. nasutus midrib
114 Original Article
J Health Res
2009,
23
(3): 111-115
ab
cys cys
gl
gl
Figure 5 Microscopic character of the surface views of R. nasutus lamina
a. lower epidermis with stomata, lithocyst cells (cys) and glandular trichome (gl)
b. upper epidermis with lithocyst cells (cys) and glandular trichome (gl)
Figure 6 Microscopic characters of powdered R. nasutus crude drug
1. sclerenchymatous sclereids vessels
2. parenchyma in longitudinal view
3. parenchyma containing crystals
4. parenchyma containing calcium carbonate
5. calcium carbonate
6. raphide crystals
7. fragment of pitted vessels
8. fragment of reticulated
9. fragment of spiral vessels
10. stomata
11. glandular trichomes
12. multicellular trichomes
13. fragment of fiber
Original Article 114 A
J Health Res
2009,
23
(3): 111-115
Figure 2 R. nasutus dried whole plants
Figure 7 TLC fingerprint of methanolic extract of R. nasutus dried whole plants
Solvent system:
Toluene : Ethyl acetate 75 : 25
Developing distance:
20 cm
Detections:
I = detection under UV light 366 nm
II = detection under UV light 254 nm
III = detection with vanillin-sulfuric acid
IV = none
2cm
Original Article 115
J Health Res
2009,
23
(3): 111-115
TLC fingerprints of methanolic extracts were
shown in Figure 7. This study used Toluene :
Ethyl acetate (TE, 75 : 25) as the solvent system
instead of Chloroform : Methanol (CM, 19 : 1) in
the earlier study14). It was found that TLC
fingerprints by TE system showed less spots than
CM.
CONCLUSION: The pharmacognostic specifi-
cation of R. nasutus crude drug was firstly
illustrated. However, the specific chemical
constituents such as Rhinacanthin series should
be further investigated.
ACKNOWLEGEMENTS: Thanks are due to the
Department for Development of Thai Traditional
and Alternative Medicine, Ministry of Public
Health for partial financial support and College of
Public Health Sciences, Chulalongkorn University
for providing facilities.
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