PHCOG J.

ORIGINAL ARTICLE

Pharmacognostic Standardization of the Leaves

and Root Bark of Caesalpinia benthamiana
Dickson RA1*, Annan K2, Komlaga, G1
1
Department of Pharmacognosy, 2Department of Herbal Medicine, Faculty of Pharmacy and Pharmaceutical Sciences,
Kwame Nkrumah University Science & Technology, Kumasi

ABSTRACT

Caesalpinia benthamiana (Baill.) Herend. and Zarucchi (Mezoneuron benthamianum Baill.) (Caesalpiniaceae) has been
traditionally used in management of erectile dysfunction, dysentery, urethral discharges, skin diseases and wounds.
Despite a long tradition of use in the treatment of various ailments, no systematic pharmacognostic standardization
work has been carried out on C. benthamiana. One major obstacle in the systematic exploration of the plant may be the
non-availability of authentic plant material. In the present investigation, various pharmacognostic standards for the
plant have been generated including the macro and micro morphological studies, one of the WHO accepted parameters
FORÒIDENTIµCATIONÒOFÒMEDICINALÒPLANTS by way of establishing the salient diagnostic characters and constants. This was
carried out on the leaves and root bark of C. benthamiana.
The leaves were bipinnate, oblong, with entire margin. The apex was obtuse, and possessed a symmetric base and
reticulates venation. The terminal leaves were however obovate in shape. Actinocytic and paracytic types of stomata were
observed. Trichomes were uniseriate and unicellular and epidermal cells were found to be wavy. Surface data analysis
revealed the stomatal index to be 1.69% to 11.11% for the upper surface and 16.94% to 28.52% for the lower surface. Vein-
islet number was recorded as 12.5 to 16.5 whilst the veinlet termination number was 22.25 to 35 with palisade ratio
ranging between 11.25 and 13.75. The water- soluble extractive value was 9.2% and 3.7% for the leaves and root bark
respectively. Whilst the alcohol-soluble extractive value for the leaves was found to be 6.7% and 2.6% for the root bark.
The total ash value determinations were observed to be 5.6% for the leaves and 7.9% for the root bark. The result of this
STUDYÒMAYÒBEÒUSEFULÒINÒSETTINGÒDIAGNOSTICÒINDICESÒFORÒTHEÒIDENTIµCATIONÒANDÒPREPARATIONÒOFÒAÒMONOGRAPHÒFORÒTHEÒPLANT
Key words: Caesalpinia benthamiana, Pharmacognostic studies, Quantitative microscopy and Extractive values

INTRODUCTION An infusion of the dried root is drunk or used as a bath

for general malaise in Senegal.[4] The aqueous decoction
Caesalpinia benthamiana (Baill.) Herend. and Zarucchi (Mezoneuron of the roots is used in traditional medicine as aphrodisiacs
benthamianum Baill.) (Caesalpiniaceae),[1] is an African tropical and the vasorelaxant properties have been reported.[5]
shrub found mostly in the secondary forest zones.
The anti-diarrhoeal effects of the aqueous extract of the
The roots of C. benthamiana are considered to be an effective plant have also been studied. The results obtained showed
dysentery remedy in Ghana.[2] The powdered roots are used that the plant possessed anti-diarrhoeal activity due to its
mixed with shea butter or palm kernel oil to treat skin inhibitory effects on gastrointestinal propulsion and
diseases and wounds in Ghana.[3] A decoction of the root, intestinal uid accumulation.[6] The analgesic, antipyretic
bark, and leaves is used in Guinea for urethral discharges.[2] and anti-in ammatory effects of the aqueous extract of
the plant have been also been evaluated in mice, rats and
*Address for correspondence: rabbits. The data obtained show that C. benthamiana root
RA. Dickson, Department of Pharmacognosy, bark extract possesses analgesic and antipyretic activities
Faculty of Pharmacy and Pharmaceutical Sciences, but lacked anti-in ammatory properties.[6] In a phytochemical
College of Health Sciences, Kwame Nkrumah University
of Science and Technology, Kumasi-Ghana.
investigation conducted by Dickson et al.,[7] the bioactivity-
Tel.: +233-(0)3220-60366 guided fractionation of the light petroleum extract of the
E-mail: [email protected] root bark of the plant led to the isolation of two novel
cassane diterpenoids, designated as benthaminin 1 and 2.
DOI: 10.5530/pj.2011.24.6
A third compound, a deoxy form of caesaldekarin C (also

Pharmacognosy Journal | August 2011 | Vol 3 | Issue 24 31

 Dickson, et al.: Pharmacognostic Standardization of the Leaves and Root Bark of Caesalpinia benthamiana

referred to as methyl vouacapenate) which had previously Microscopic evaluation

been isolated from Caesalpinia major, Caesalpinia bonducella, Leaves were cut into smaller sizes and cleared in chloral
Vouacapoua americana and Vouacapoua macropetala, was also hydrate, mounted with glycerin and observed under a
isolated, together with beta-sitosterol and stigmastenone. compound microscope. The presence /absence of the
The antibacterial and antioxidant activities of these cassane following were observed: epidermal cells, stomata (type
diterpenoids have also been assessed.[7] The resistance and distribution) and epidermal hairs (types of trichomes
modifying activities of extracts from this plant on standard and distribution). The transverse section through the fresh
antibiotics against Staphylococcus aureus have also been assessed petiole of the leaf was also examined.
by the same group of researchers. A 4-fold potentiation
of the activity of nor oxacin was observed for the ethanol Preliminary phytochemical investigation
extract, whilst the petroleum spirit extract resulted in a The leaf and root bark powders of the plant were separately
2-fold potentiation.[8] subjected to Soxhlet extraction using 70% ethanol. Chemical
tests were employed in the preliminary phytochemical
Despite a long tradition of use in the treatment of various screening for various secondary metabolites such as tannins
ailments, C. benthamiana has not been explored properly by (Ferric chloride test), cardiac glycosides (Keller-Killiani and
way of establishment of standards in the identi cation and Kedde tests), alkaloids (Mayer’s; Dragendorff ’s; Wagner’s
quality control of this plant. The cassane-type diterpenoids and 1% picric acid reagents), Saponin glycosides (frothing
possessing antimicrobial, antioxidant and wound healing and haemolysis tests), anthracene derivatives (Borntrager’s
properties isolated from the plant could serve as leads in test for combined and free Anthraquinones) and
the search for new biomolecules as drugs. The need to Cyanogenetic glycosides (sodium picrate paper test).[10,11,12,13]
standardize this plant can therefore not be overemphasized.
It is also worthwhile to note that some drugs of plant origin Quantitative investigations
in conventional medical practice are not pure compounds Quantitative leaf microscopy to determine palisade ratio,
but direct extracts or plant materials that have been prepared stomata number, stomata index, vein- islet number and
appropriately and standardized. The use of Artemisia annua, veinlet termination number were carried out on cleared
Digitalis and Senna leaves are a few examples. The sections of the leave. Other parameters determined for the
establishment of the pharmacognostic pro le of the leaves powdered leaves were moisture content, total ash, acid -
and root bark of C. benthamiana will assist in standardization, insoluble ash, water - soluble ash, alcohol (90% ethanol)
which can guarantee quality, purity and identi cation of and water soluble extractive values.[14] The moisture content
samples to ensure that only the authentic plant is explored of the powdered leaves and root bark of the plant was also
properly for its traditional claims. determined using the Dean-Stark apparatus.[9]

Determination of total ash

MATERIALS AND METHODS 2 g of the powdered leaves and root bark of C. benthamiana
were weighed separately in a pre-weighed ash-less lter
The fresh leaves and roots of the plant was collected from paper and incinerated at 400°C for about 3-4 min or until
the Ayeduase in the Ashanti Region of Ghana and the vapours completely ceased. The temperature was
authenticated by Mr. Henry Sam of the Department of gradually reduced to come to normal and then the ash was
Herbal Medicine, Kwame Nkrumah University of Science collected and weighed.
and Technology, Kumasi, Ghana, where voucher specimens
were deposited with the numbers RADCB 01 and RADCB11 Determination of alcohol soluble extractive
respectively for the leaves and roots of the plant. Accurately weighed powder (10 g) of both leaves and root
bark were taken separately and macerated with 100 ml of
Organoleptic evaluation 95% alcohol for 24 hours. The contents were frequently
In the organoleptic evaluation, various sensory parameters shaken during the rst 6 hours and allowed to remain for
of the plant material such as colour, odour, taste and texture another 18 hours. After 24 hours, the extract was ltered
were investigated.[9] and 20 ml of the ltrate was evaporated to dryness. The
extract was dried at 105°C to a constant weight.
Macroscopic evaluation
The following macroscopic characters for the fresh leaves Determination of water soluble extractive
were noted: the type of leaves, its arrangement, colour, Water soluble extractive value was determined using the
shape, length and width of the leaves, size and surfaces, procedure described for alcohol soluble extractive, except
venation, presence or absence of petiole, the apex, margin, that chloroform water was used for maceration in this
base, lamina, texture.[9,10] instance.

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 Dickson, et al.: Pharmacognostic Standardization of the Leaves and Root Bark of Caesalpinia benthamiana

RESULTS DISCUSSION

Macroscopically, the plant possess compound leaves which The standardization of a crude drug is an integral part
are bipinnate, alternate in arrangement, apex and base are of establishing its correct identity.[17] Caesalpinia benthamiana
obtuse, margin is entire, venation is reticulate, shape oblong is employed in ethnomedicine in the management of
with terminal leaves being obovate. Full length of leaves various disease states without standardization. The
33-38 cm, Six (6) pairs of pinnae, with length of each quantitative determination of some pharmacognostic
pinnae being 6.5-7.5 cm and four (4) pairs of lea ets on parameters is useful for setting standards for crude drugs.
each pinnae. The average size of individual leaf is 2.0- [18]
The vein islet, and vein termination numbers and the
5.0 cm ± 0.5 (length) and 1.5-3 cm ± 0.3 (width). Fresh other parameters determined in the quantitative microscopy,
leaves are green in colour, odourless with a gritty texture. are relatively constant for plants and can be used to
It is petiolated with the length of petiole between 4-5 cm, differentiate between closely related species.[19] Also, the
shape of petiole cylindrical and brownish-green in colour. physical constant evaluation of the crude drugs is an
important parameter which is a valuable tool in detecting
The leave arrangement may be similar to that of C. spinosa adulteration or improper handling of crude drugs. The
but whereas the leaves of C. spinosa lack petioles those of moisture content of the powered drugs may be said to
C. benthamiana are petiolated.[15] be high since the BP stipulates an allowable value of not
more than 10%. Any value beyond this encourages
The length of the leaves of C. benthamiana falls between microbial growth and subsequent deterioration of the
33 to 38 cm which is within those of C. pulcherrima which stored powdered drugs.
are also between 20 cm and 40 cm but are longer than
those of C. gilliesii which falls between 10 cm and 15 cm Equally important in the evaluation of crude drugs, is
long.[16] the ash value and acid-insoluble ash value determination.
The total ash is particularly signi cant in the evaluation
The leaves of C. benthamiana have two types of stomatal of purity of the crude drugs, i.e. the presence or absence
arrangements and these are the actinocytic and paracytic of foreign inorganic matter such as metallic salts and/
types. The types of trichomes observed were uniseriate or silica.[20] The macro - and micro – morphological
clothing hairs and unicellular hairs. The epidermal cells features of the leaf described, distinguishes it from other
were found to be wavy. members of the genera. For example whereas the leaves
of Caesalpinia spinosa lack petioles those of Caesalpinia
Micromorphological features revealed that anticlinal walls benthamiana are petiolated.[15] The length of the leaves of
are thin and wavy. The type of stomata revealed actinocytic Caesalpinia benthamiana falls between 33 to 38 cm which
arrangement and these were few as compared to the paracytic is within the range for those of Caesalpinia pulcherrima
ones which were more. Uniseriate covering trichomes are which are also between 20 cm and 40 cm but the leaves
present on both surfaces. It has Isobilateral leaf arrangement. of Caesalpinia gilliesii are shorter, falling within the range
The midrib bundle is surrounded by a zone of pericyclic of 10 cm and 15 cm.[16]
bres possessing double layered parenchymatous cells. The
roots are brown in colour and the texture is gritty. Vein By and large, the pharmacognostic constants including
islet number was determined to be 12.5 to 16.5, veinlet extractive values, ash values and the phytochemical pro le
termination number ranging from 22.5 to 35, stomatal index of Caesalpinea benthamiana obtained for the leaves and root
of 1.69% to 11.11% for the upper surface and 16.94% to bark of this plant, the diagnostic microscopic features and
28.52% for the lower surface and palisade ratio of 11.25 the numerical standards reported in this work could be
to 13.75. Water- soluble extractive values were 9.2% and useful for the compilation of a suitable monograph for the
3.7% for the leaves and roots respectively. The alcohol- proper identi cation and use of this plant.
soluble extractive value of 6.7% was obtained for the leaves
and 2.6% for the roots. Also, the total ash values for both
leaves and roots were 5.6% and 7.9%. Phytochemical CONCLUSION
evaluation revealed the presence of true tannins,
pseudotannins and terpenoids mainly for both leaves and These parameters which are being reported for the rst
roots. These secondary plant metabolites are known to time, could serve as useful information in preparing a
possess various pharmacological effects and may be monograph of the plant which can be locally incorporated
responsible for the various actions of C. benthamiana. See into the Ghana Herbal Pharmacopoeia which may ultimately
Tables 1 to 6 numerical and quantitative values as well as serve as a signi cant addition to international herbal
morphological descriptions. pharmacopoeias. Any crude drug which is claimed to be

Pharmacognosy Journal | August 2011 | Vol 3 | Issue 24 33

 Dickson, et al.: Pharmacognostic Standardization of the Leaves and Root Bark of Caesalpinia benthamiana

Caesalpinea benthamiana but whose characters signi cantly 9. Evans WC. Trease and Evans Pharmacognosy. WB Saunders Ltd.
London. 2006; pp 32, 33, 95-99, 512, 547.
deviate from the accepted standards above may be considered
10. Wallis, T. E. Textbook of Pharmacognosy. Published by SK Jain. 1985;
to be contaminated, adulterated, substandard or fake. 572-575.
11. Brain KR, Turner TO. Practical evaluation of phytopharmaceuticals.
Wright - Scientechnica, Bristol. 1st Ed. 1975; 144.
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