PHARMACOGNOSTICAL STUDIES ON THE STEM BARK OF CAESALPINIA BONDUC (L.) ROXB.
HTML Full TextPHARMACOGNOSTICAL STUDIES ON THE STEM BARK OF CAESALPINIA BONDUC (L.) ROXB.
J. Shashikanth
Center of Medicinal Plants Research in Homoeopathy (CMPRH), Central Council for Research in Homeopathy (CCRH), Ministry of AYUSH, Government of India, 3/126, Indira Nagar, Emerad, The Nilgiris, Tamil Nadu, India.
ABSTRACT: Background: Caesalpinia bonduc (L.) Roxb. belonging to family Caesalpiniaceae, commonly known as ‘Kuberaksha’, is a large scandant shrub. The stem bark is useful as febrifuge, emmenagogue, anthelmintic, germicidal and anti-pyretic. Aim: The present study was conceived to standardize the stem bark of Caesalpinia bonduc to determine correct identity and purity of the drug have been undertaken besides in detection of adulteration. Materials and Methods: Pharmacognostical studies include (study of morphological characters), microscopical characters and powder microscopy and physicochemical parameters along with phytochemical, histochemical studies and fluorescence behavior were studied. Results: Transverse section of stem bark revealed the presence of outer most cork interspersed with stone cells, inner phelloderm cells with rhombic crystals and biseriate medullary rays. Tangential Longitudinal Section (T.L.S) revealed presence of uni to biseriate medullary rays, mostly biserate. Physicochemical standards showed that foreign organic matter (Nil), loss on drying (11.07%), total ash (5.45%), acid insoluble ash (0.45%), water soluble ash (1.45%), petroleum ether extractive (0.45%), chloroform (1.65%), ethanol (6.5% ), methanol (7.55%), water (11.65%). Preliminary phytochemical screening of various extracts showed the presence of alkaloids, phenolic compounds and tannins, phytosterols, flavonoids, terpenoids and carbohydrates (were found be major components), besides histo-chemical studies and fluorescence behavior is given. Conclusion: Established Pharmacognostical characters, physic-chemical and phytochemical parameters of Caesalpinia bonduc stem bark will serve as standards of drug in the crude form and also would help in distinguish the drug from its adulterants or other closely related species. The study will provide referential information for correct identification of the crude drug.
Keywords: Pharmacognostic, Stembark, Macroscopic, Microscopic Caesalpinia bonduc, Physicochemical, Phytochemical
INTRODUCTION: Caesalpinia bonduc (L.) Roxb. belonging to family Caesalpiniaceae, in Ayurveda it is known as Putikaranja, is medicinally important plant predominantly distributed in tropical and subtropical regions of Asia 1 (up to 1000 m in the Himalayas also in the plains, waste
lands and coastal areas widely distributed all over the world specially in India, Sri Lanka, Burma and Andaman-Nicobar Islands). Description: It is Large scandant or scrambling shrubs; branches armed with recurved prickles. Leaves bipinnate, pinnae 6-11 pairs, leaflets stalked, flowers yellow, or fragrant, in axillary and terminal racemes.
Pods oblong, densely armed with sharp wiry prickles, dehiscent, seeds 1 or 2, globose or ovoid, grey. It is locally known as Nata karanja (Hindi), gachha (Telugu) and possess a lot of applications in folk medicines. Review of literature: Seed and bark extracts have been used as Anthelmintic, anti-cancer, anti-malarial, anti-rheumatic and anti-pyretic agents 2, 3. The traditional practioners residing in the vicinity of Western Ghats of Karnataka are using the leaves and stem bark to cure jaundice and liver disorders 4. In Ayurveda it is used to treat febrifuge and emmenagogue, anthelmintic, germicidal, anti-pyretic, anthelmintic and syptic 5, in homoeopathyit is used for treating febrifuge, emmengoue, anthelmintic, tonic, styptic, depression, intermittent fever, headache, worm infestation, chronic fever and malaria 6. Stem bark is reported as febrifuge, anthelmintic, antimalarial, hypoglycemic, antipyretic, anti-inflammatory and antirhematic 7. Some of the chemical compounds found in stem barks of Caesalpinia bonduc are Hematoxylol, Stereochenol - A, 61-O-aceylloganic acid, 41-O- acetylloganic acid, 2 - 0 - β – D - glucosyloxy - 4 - methoxybenzeneneropanoic acid, Ethacrynic acid 8.
Stem barks extracted and tested with different solvents were found to possess anthelmintic properties proved by analysing the mechanism of paralyzing and killing when tested against earthworms 9. Chloroform extracts of C. bonduc leaves, stem barks and isolated constituents were reported to have anti-bacterial effects. The stem barks extract exposing a minimum inhibitory concentration of 2 mg/ml against B. subtilis and P. aeruginosa 10. From the study conducted 11 for analyzing the anti-inflamatory and anti-cancer property of Caeselpinia stem barks having high levels of phenolics and flavonoids found to be influencing such inhibitory activities. Objectives: standardization of quality parametersin stem bark of Caesalpinia bonduc (L.) Roxb., identification and differentiation of raw drug from closely related or allied specis like Caesalpinia sappan L., Caesalpinia coriaria (Jacq.) Willd., Caesalpinia decapetala (Roth) Alstonand Caesalpinia crista L.
MATERIALS AND METHODS:
Pharmacognostical Studies: The fresh stem bark material of Caesalpinia bonduc was collected from Osmania University campus, Hyderabad and identified. The voucher specimens of the plant were deposited in Herbarium Hyderbadensis (HY), Osmania University, Hyderabad. The material was preserved in glycerin and alcohol mixture (1:1) for softening. Sections of stem bark were cut in transverse and longitudinal planes, 8-12 um thickness on Leica RM 2155 microtome. The sections were stained with crystal violet and basic fuchsin and mounted in Canada balsam. portion of fresh stem bark was macerated and examined for variyuos elements. Dried stem bark was ground to fine powder. A small amount of powder boiled in water, stained in safranin and mounted in glycerin and observed for microscopical characters. The photomicrographs were taken with Olympus ch-2 microscope 12.
Physico-Chemical Studies: Stem bark was dried in under shade; the dried material was mechanically powdered. These powdered materials was used for physico-chemical parameters viz., Moisture content, foreign organic matter, total ash, water soluble ash, acid insoluble ash values and extractive values were determined as per standard procedure 13,14. Histochemical studies, phytochemical constituents and Fluorescence studies have been carried out by standard methods 15-22.
RESULTS
Macroscopic: From the microscopic figures, the stem bark pieces was observed as they were thin, double or compound quilled; 1-2 mm thick, scaly; externally blackish brown with intermittent creamish brown patches; inner surface was smooth, pale brown; fracture uneven, splintery (Plate-1A & 1B).
PLATE 1: CAESALPINIA BONDUC (LINN.) ROXB. STEM BARK. (a,b) Dried Stem bark, (c) TS of stem bark showing cork and phelloderm, (d). TLS of stem bark, (e) TS of stem bark showing secondary phloem, (f) TS of stem bark showing cork enlarged. 1= Cork , 2= phellogen , 3=Phelloderm , 4= Stone Cells , 5= Medullary rays , 6= Phloem fibers , 7= Obliterated tissue , 8= Medullary rays , 9=Sieve elements, 10=Tannin , 11=Cork, 12=Rhombic crystals, 13=Stone cells , 14= Phelloderm
Microscopic: In the transverse section, the stem bark shows, outer most cork 40-45 layered; cells rectangular, tangentially elongated, measuring 17.5 - 70 μm (31.5) long and 3.5 - 21μm (7) wide; walls slightly thick, contents slightly with golden yellow coloured, interspersed with stone cells, isolated or in groups, mostly rectangular, measuring 21 - 42 μm (35) long and 7 - 17.5 μm (10.5) wide; walls thick, contents scanty. Phellogen is 1-2 layered. Phelloderm is separated into outer and inner zones, outer 9-12 layered, cells rectangular or tangentially long, measuring 21-108.5 μm (49.5) long and 7-24.5 μm (17.5) wide; interspersed with stone cells in bands, in between outer and inner zones phelloderm cells are large, banded stone cells are irregular, mostly isodiametric, measuring 10.5 – 35μm (21) in diameter; measuring 10.5 - 38.5μm (35) in diameter; walls thick, lignified, lumen small with ramified pits. Inner phelloderm is multilayered, cells varying shape and size, mostly polygonal, measuring 17.5 - 73.5 μm ( 28) long and 10.5 - 21μm (17.5) wide; walls slightly thick, contents dense with golden yellow or reddish contents and rhombic crystals; inner phelloderm is interspersed with cortical fibers.
TABLE 1: ORGANO-LEPTIC CHARACTERS OF STEM BARK OF CAESALPINIA BONDUC (L.) ROXB.
Character | Observation |
Colour | Pale reddish brown |
Touch | Coarse |
Odour | Disagreable |
Taste | Bitter |
TABLE 2: PHYSICO-CHEMICAL STANDARDS OF STEM BARK OF CAESALPINIA BONDUC (L.) ROXB.
Parameter | Standards or content (% by weight) |
Loss on drying% w/w | 11.07% |
Foreign organic matter % w/w | Nil |
Total ash % w/w | 5.45% |
Acid insoluble ash % w/w | 0.45% |
Water soluble ash % w/w | 1.45% |
Pet. Ether % w/w | 0.45% |
CHCl3 % w/w | 1.65% |
EtOH % w/w | 6.25% |
MeOH % w/ w | 7.55% |
Water % w/ w | 11.65% |
TABLE 3: HISTOCHEMICAL COLOUR REACTIONS OF STEM BARK OF CAESALPINIA BONDUC (L.) ROXB.
Reagents | Constituents | Colour | Result |
Pinch of Phloroglucinol + Dil. HCl | Lignin | Magenta (or) pink | Present |
Aqueous ferric chloride solution | Tannins | Black | Present |
Iodine solution | Starch | Blue | Present |
Sudan-III | Oil globules | _ | _ |
Ruthenium red solution | Mucilage | _ | _ |
TABLE 4: PRELIMINARY PHYTOCHEMICAL SCREENING IN STEM BARK OFCAESALPINIA BONDUC (L.) ROXB.
S. no. | Test for (or) Constituents | Water extract | Pet. ether extract | Chloroform extract | Ethanolic extract | Methanolic extract |
1 | Alkaloids | _ | + | + | _ | _ |
2 | Phenolic compounds &Tannins | + | _ | _ | + | + |
3 | Phytosterols | _ | _ | + | + | + |
4 | Fixed oils and Fats | _ | _ | _ | _ | _ |
5 | Saponins | + | _ | _ | _ | _ |
6 | Flavanoids | _ | _ | _ | + | + |
7 | Aminoacids | _ | _ | _ | _ | _ |
8 | Terpenoids | _ | _ | + | + | + |
9 | Carbohydrates | + | _ | _ | + | + |
“+” indicates presence of constituents “_” indicates absence of constituents.
TABLE 5: FLUORESCENCE BEHAVIOUR OF STEM BARK O CAESALPINIA BONDUC (L.) ROXB.
S. no. | Powder+Reagents Used | Light source | colour |
1 | Powder as such | Visible | Brown |
Short UV(254 nm) | Brown | ||
Long UV (366 nm) | No | ||
2 | Powder + 50% HNO3 | Visible | Light green |
Short UV(254 nm) | Green | ||
Long UV (366 nm) | Leaf green | ||
3 | Powder + 50%H2SO4 | Visible | Pae green |
Short UV(254 nm) | Light greeen | ||
Long UV (366 nm) | Light greeen | ||
4 | Powder + 50% KOH | Visible | Brown |
Short UV(254 nm) | Brown | ||
Long UV (366 nm) | No | ||
5 | Powder + 1N HCl | VISIBLE | Light orange |
Short UV(254 nm) | Misty vale | ||
Long UV (366 nm) | English elms | ||
6 | Powder + 1N NaOH IN Ethanol | Visible | Thar desert |
Short UV(254 nm) | Leaf green | ||
Long UV (366 nm) | Milk toffee | ||
7 | Powder +1N NaOH IN methanol | Visible | Casablanca |
Short UV(254 nm) | Leaf green | ||
Long UV (366 nm) | English elms | ||
8 | Powder + Pet.ether | VISIBLE | Milky white |
Short UV(254 nm) | White | ||
Long UV (366 nm) | White | ||
9 | Powder + Methanol | Visible | Thar desert |
Short UV(254 nm) | Light green | ||
Long UV (366 nm) | Sandal wood | ||
10 | Powder + Acetone | Visible | Sporty yellow |
Short UV(254 nm) | Leaf green | ||
Long UV (366 nm) | Wild yellow |
Secondary phloem is scanty consists of phloem parenchyma, sieve tubes accompanied with companion cells and medullary rays. Phloem parenchyma cells isodiametric, 10.5 - 38.5μm (35) in diameter; walls thick, contents slightly dense with light red or bluish colour matter and resinous mass. Medullary rays straight, uni to biseriate, 8-10 cells high, ray cells rectangular, radially elongated, measuring 21 - 52.5 μm (42) long and 7 -21μm (17.5) wide; walls thin, contents scanty
In tangential longitudinal section structure of the medullary rays, phloem fibres and sieve elements were seen.
Medullary rays are straight and exclusively uni to biseriate. The rays cells are rectangular, thin walled, compact, contents are scanty. The rays are uniformly homocellular consisting with only one type cells. [Plate-1 (c, d, e & f) and Plate 2 (a, b, c, d, e & f)].
TABLE 6: POWDER TREATED WITH DIFFERENT CHEMICAL REAGENTS OF STEM BARK OF CAESALPINIA BONDUC (L.) ROXB.
S. no. | Powder + Reagent | Colour |
1 | Powder as such | Pale brown |
2 | Powder + con.Sulfuric acid | Pink |
3 | Powder + Con.HCl | Pale brown |
4 | Powder + Con. Nitric acid | Pale yellowish with fumes |
5 | Powder + Acetic acid | Pale brown |
6 | Powder + 10% NaOH | Black |
7 | Powder + Ammonia | Dark brown |
8 | Powder + Acetone | Light brown |
9 | Powder + Ethyl acetate | Colourless |
10 | Powder + 1N HCl | Colourless |
Powdered Microscopy: On microscopic examination of stem bark powder shows cork consisting of polygonal cells in surface and rectangular cells in sectional view; prismatic crystals of calcium oxalates; lignified stone cells, isolated or grouped fibers; crystal fibers, radially cut medullary rays; abundant starch grains. Plate-3(1, 2, 3, 4, 5 & 6).
PLATE 2: CAESALPINIA BONDUC (LINN.) ROXB. STEM BARK. (a) TS of stem bark showing cork enlarged, (b) TS of stem bark showing starch grains, (c,d) TS of stem bark showing stone cells enlarged, (e) TS of stem bark showing secondary phloem enlarged, (f) TS of stem bark showing meddullary rays enlarged, 1= Stratified cork layers, 2= Phellogen, 3= Rhombic crystals, 4= Phloem fibers, 5= Starch grains, 6= Stone cells, 7= Prismatic crystal, 8= Stone cells, 9= Starch grains, 10= Phloem parenchyma, 11= Obliterated tissue, 12= Medullary rays, 13= Sieve elements, 14= Medullary rays, 15= Starch grains.
PLATE-3: POWDERED ELEMENTS OF CAESALPINIA BONDUC (L.) ROXB. 1= Parenchyma , 2= Crystals and fibers, 3 =Stone cells, 4 =Cork, 5 =Medullary ray, 6= Crystal fibers.
Organoleptic Characters: By observing the organoleptic properties the colour of the bark was found to be Pale reddish brown and was coarser in touch, bitter in taste with a disagreeable odour Table 2.
DISCUSSION: Earlier studies conducted in Caesalpinia bonduc (L.) Roxb. stem barks lack microscopic and physic-chemical analysis. Pharmacognostic characters of herbal drugs play an important role since particular- macroscopic features are unique for each plants. Macro and microscopic evaluation is simple and cost effective. The microscopic and macroscopic studies of the herb should be the first and fundamental step to authenticate the botanical source. Stem bark pieces are thin, double quilled or compound quilled. External bark blackish brown and scaly while inner surface is smooth. Fracture is splintery. Cork in T.S consists of rectangular cells with golden yellowish content and interspersed with stone cells. Phelloderm is wide interspersed with many bands of stone cells.
Secondary phloem is scanty with phloem parenchyma cells encrusted with resinous masses. Medullary rays are biseriate with rectangular, radially long cells. The Powder microscopic features and organoleptic characters presented (loc.cit.) are of diagnostic significance.
Physicochemical parameters revealed absence of foreign organic matter indicating purity of drug. Standards of loss on drying 11.07% is indicating to avoid the microbial contamination, total ash, acid insoluble ash, water soluble ash values found to be 5.45%, 0.45%, 1.45% respectively and extractive values in various solvents are ( Petroleum ether extractive value 0.45%, chloroform extractive value 1.65% , ethanol extractive value 6.25%, methanol extractive value 7.55% , water extractive value 11.65%) in acceptable range. Phytochemicals have been used for the treatment and prevention of various health ailments from time immemorial. The results of preliminary phytochemical screening of various extracts reveals presence of alkaloids, phenolic compounds and tannins, phytosterols, flavonoids, terpenoids and carbohydrates helps in establishing genuinity and efficacy of the drug. The preliminary phytochemical screening shows that Caesalpinia bonduc stem bark contains different chemical constituents which indicate the presence of both polar and non-polar compounds in the drug Table 1-7.
From the result of this study it has been found that the dried whole plant of Caesalpinia bonduc stem bark can be easily identified macro-microscopically and phytochemically. The study sets specific morpho-anatomical and preliminary phytochemical studies, histochemical colour reactions, powder treated with different chemical reactions and fluorescence behavior of stem bark which are helpful in standardization and efficacy of the drug.
CONCLUSION: The present study on pharmacognostical characters of Caeselpinia bonduc L. stem bark provides useful information in regards to its correct identity and helps to differentiate from the closely related other species of Ceasalpinia genus. Standardization of herbal medicine is a key importance in establishing its correct identity, safety, efficacy and purity of drugs besides checking of the adulteration as well. Botanical authentication, physic-chemical and phytochemical parameters will give an idea about the quality of drug. The macroscopic, microscopic, Physico-chemical and phytochemical analysis are the confirmatory test for standardization and quality control, which may be helpful in authentication of crude drug and checking the adulteration of raw materials as well.
ACKNOWLEDGEMENTS: Author is grateful to Director General, CCRH (Ministry of Ayush) for constant encouragement, thankful to Head, Department of Botany, Osmania University for providing all the necessary facilities and also thankful to UGC (BSR- RFSMS) for financial assistance.
CONFLICT OF INTEREST: Nil
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How to cite this article:
Shashikanth J: Pharmacognostical studies on the stem bark of Caesalpinia bonduc (L.) Roxb.. Int J Pharmacognosy 2023; 10(9): 502-08. doi link: http://dx.doi.org/10.13040/IJPSR.0975-8232.IJP.10(9).502-08.
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J. Shashikanth
Center of Medicinal Plants Research in Homoeopathy (CMPRH), Central Council for Research in Homeopathy (CCRH), Ministry of AYUSH, Government of India, 3/126, Indira Nagar, Emerad, The Nilgiris, Tamil Nadu, India.
jaleshashikanth2000@gmail.com
13 September 2023
24 September 2023
29 September 2023
10.13040/IJPSR.0975-8232.IJP.10(9).502-08
30 September 2023