from Rewa, Madhya Pradesh, India. It was authenticated by Dr. A K S Rawat, and matched with the voucher specimen and deposited in the institute's herbarium ...
Journal of Medicinal and Aromatic Plant Sciences 33 (1) (2011) 59-63
Pharmacognostic and phytochemical evaluation of Cordia macleodii VIKAS KUMAR, SHASHI SHANKAR TIWARI, ACHUTA NAND SHUKLA, SHARAD SRIVASTAVA AND A. K. S. RAWAT* Pharmacognosy and Ethnopharmacology Division, National Botanical Research Institute (CSIR), Lucknow226001, India. etc [6, 10]. Although the plant is important, there is no pharmacognostic and phytochemical study on this species so far. Hence this study has been undertaken to develop quality standards of its bark which will be utilized by researchers for exploring the possibility for herbal drug industry [2, 3].
ABSTRACT Cordia macleodii (Griff.) Hook. f. & Thoms. (FamilyBoraginaceae), a rare and endangered medicinal plant, is commonly known as ‘Dahipalas’ or ‘Dahiman’. It is distributed in Chota Nagpur, Central India, Konkan, North Kanara and Deccan, and is widely used against various disorders in Indian system of medicine. The HPTLC, physicochemical, phytochemical, fluorescence study, morphological and histological parameters presented in this paper may be helpful to establish the authenticity of C. macleodii bark and may possibly help to differentiate the drug from its other allied species.
MATERIALS AND METHODS Plant materials The plant sample was collected in December 2009 from Rewa, Madhya Pradesh, India. It was authenticated by Dr. A K S Rawat, and matched with the voucher specimen and deposited in the institute’s herbarium no. (262532). Fresh bark was preserved in 70% ethyl alcohol for histological studies. Microtome sections were cut and stained with safranin and fast green and photographed with Olympus CX 31 camera (Johansen, 1940) [1, 3].
Key words: Bark, Cordia macleodii, HPTLC, pharmacognosy, phytochemical study.
INTRODUCTION Cordia macleodii (Griff.) Hook.f. & Thomas. (Boraginaceae), is an evergreen, polygamous medium sized, deciduous tree 8-10 m high, with white hairy branches. It is a rare and endangered medicinal plant species, locally known as ‘Dahiman’ or ‘Dahipalas’ and found in moist and dry mixed deciduous forests [5, 9]. In India, it is distributed in Bihar hills, Madhya Pradesh, Chattishgarh, Karnataka, and Deccan. The recorded information is crosschecked and verified with discussion of other tribal communities. The bark of this plant has been reported to be useful in cases of jaundice, and wound healing. The tribal people believe that if the cattle rub their body with the tree, the wound is quickly healed. They also believe that it is a deity’s tree therefore they do not use its wood for making doorframes, plough
Physico-chemical and phytochemical studies like, total ash, acid insoluble ash, sugars, starch and tannins were calculated from the shade dried and powdered material (Peach and Tracy, 1955; Anonymous, 2007; Anonymous, 1984). HPTLC studies A densitometric HPTLC analysis was also performed for the development of characteristic fingerprint profile, which may be used as markers for quality evaluation and standardization of the drug. Plates were developed to a distance of 80 mm, with toluene: ethyl acetate: formic acid (80: 20: 5 v/v) as mobile phase in a Camag glass twin-trough chamber (20 cm × 10 cm) previously saturated with mobile phase vapour
*Author for correspondence; E mail: pharmacognosy1@ rediffmail.com
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Plate 1. Macro and microscopic characters of the stem bark of Cordia macleodii CK, Cork; CC, Cork cambium; PD, Phelloderm; SP, Secondary phloem; RHY, Rhytidoma; MR, Medullary rays; PF, Phloem fibre; ST, Stone cell; RC, Resin cell; SG, Starch grain.
60
for 20 min:Room temperature was 38°C.After removal of plates from chamber completely dried in Hot oven at temperature (110°C) for 15 min and -sitosterol (Rf 0.59) was simultaneously quantified by using CAMAG TLC Scanner model-3 equipped with Wincats version [ 3.2.1 ] software.
1.6 cm, oblong, or much shorter. Fruit acutely conical till nearly ripe, persistent calyx broadly funnel-shaped or subcompanulate. Macroscopic characteristics of
bark
RESULTS AND DISCUSSION
Dried mature bark greyish brown in colour, curved to channeled, 8-15 cm in length, 1-2 mm thick, outer surface rough. Inner surface fibrous, whitish in colour when fresh and turning to brown on drying. Texture coarse, fracture splintery, fibrous. No characteristic taste and odour.
A brief taxonomic description of the plant
Microscopic characteristics of bark
A polygamous medium sized tree about 12 m in height with greyish brown bark. Leaves alternate, cordate-ovate, obtuse, 3-5 nerved, permanently tomentose beneath, scarcely subopposite; mature 12.5 cm in diameter, petiole 2.5-5 cm, inflorescence corymbs, short tomentose, calyx about 1.2 cm, tubularclavate, densely tomentose, ribbed upwards or much smaller, not (or obscurely) ribbed. Corolla- lobes about
Transverse section of bark shows outer most 812 layered cork, tangentially elongated and radially arranged. Cork cambium distinct 2-3 cell broad followed by secondary cortex or phelloderm. Phelloderm parenchymatous and embedded with small groups of stone cells, prismatic crystals of calcium oxalate, tannin containing cells and groups of fibres. Secondary phloem is wide consist of sieve tubes,
The following scan condition were applied slit width- 5 x 0.45 nm, wavelength (max -600 nm), Absorption –Reflection scan mode.
Table 1. Fluorescence analysis of the bark powder of C. macleodii S. no.
Treatment
Daylight
UV light (254mm)
1.
Powder as such
Creamish brown
Light greenish brown
2.
Powder + 1N aq. NaOH
Dark orangish brown
Dark greenish black
3.
Powder + 1N Alc. NaOH
Dark yellowish brown
Greenish black
4.
Powder + 1N HCl
Yellowish brown
Fluorescent green
5.
Powder + NH3
Yellowish Green
Dark green
6.
Powder + I2
Light Yellow
Greenish Black
7.
Powder + FeCl3
Blackish Green
Black
8.
Powder + Acetic acid
Yellowish Brown
Greenish Black
9.
Powder + 1N HNO3
Yellowish orange
Blackish Green
Table 2. Phytochemical screening of the successive solvent extractives of C. macleodii bark Extractive
Triterpenoides
Saponins
Flavonoids
Tannins
Reducing sugar
Glycosides
Alkaloids
Hexane
+
-
-
-
-
-
-
Chloroform
+
-
-
-
-
-
-
Acetone
-
-
-
-
+
-
-
Methanol
-
-
-
-
+
-
-
Water
-
+
-
+
+
+
+
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Physico-chemical and fluorescence studies
SF
On physico-chemical evaluation it contains total ash 10.55%; acid insoluble ash 0.43%; hexane soluble extractive 0.75%; alcohol soluble extractive 3.80%; water soluble extractive 26.80%; sugar 1.89%; starch 9.32%; and tannins 0.05%. Successive solvent extraction through soxhlet apparatus yielded hexane soluble extractive 6.20%; chloroform soluble extractive 3.42%; acetone soluble extractive 2.46%; methanol soluble extractive 13.48% and water soluble extractive 14.34% respectively.
STD
β- Sitosterol (STD)
Sample
Figure 1. HPTLC profile of C. macleodii bark under 600 nm along with -Sitosterol (STD)
Figure 3. HPTLC densitometric scan (at 600 nm) of marker compound-â sitosterol β- Sitosterol
Figure 2. HPTLC chromatogram of Cordia macleodii bark sample (2) with reference compound (1)
companion cells, phloem parenchyma and tangentially running bands of thick walled lignified fibers interrupted by medullary rays. The primary phloem is also wide and is transversed by multiseriate medullary rays which get broader towards peripheral regions. Medullary rays heterogenous and 4-6 cells broad and 18-24 cell height.
Figure 4. HPTLC densitometric scan (at 600 nm) of C. macleodii bark sample
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Air-dried barks were powdered using a homogenizer and bark powder was considered as drug. The bark powder in different solvents was examined under ordinary day light and in UV-light (254nm). The fluorescence was determined according to the methods of Chase and Pratt, 1949) [4].
ACKNOWLEDGEMENT
From the above studies, bark the can easily be identified on the basis of macroscopic features for e.g. dry bark is grayish brown in colour, curved to channeled in shape fracture splintery fibrous. On microscopic examination prismatic crystal, stone cells, heterogenous, multiseriate medullary rays, resin cells are observed in the bark.
1.
Anonymous. 1984. Official methods of Analysis (AOAC) 4th Edn. Association of Official Chemists, Inc. U.S.A.
2.
Anonymous. 2000. A Dictionary of India Raw materials and Industrial Products. The Wealth of India. pp. 78-84.
3.
Anonymous. 2007. The Ayurvedic Pharmacopoeia. New Delhi; Govt. of India Publication.
4.
Chase CR, Pratt R. 1949. Fluorescence of powdered vegetable drugs with particular reference to development of a system of identification. Am Pham Assoc 38: 324-331.
5.
Chopra RN, Nayer SL, Chopra IC. 1956. Glossary of Indian Medicinal Plants, CSIR, New Delhi. pp. 77.
6.
Evans WC. 2005. Trease and Evans Pharmacognosy, Saunders an Imprint of Elsevier. pp. 41-47.
7.
ICH harmonized tripartite guidelines validation of analytical procedures; text and methodology, Q2 (R1). 2005. International Conference on Harmonization (ICH), Geneva.
8.
Johansen DA. 1940 Plant Microtechnique, MC Graw Hill Book Co. New York London. pp. 182.
9.
Kirtikar KR, Basu BD. 1935. Indian Medicinal Plants, Allahabad: Vol. III.
The authors are thankful to Director, NBRI for providing all the facilities and guidance to conduct this research work. REFERENCES
Physico-chemical values viz. total ash, acid insoluble ash, hexane soluble extractive, alcohol soluble extractive, water soluble extractive are observed. The total ash, acid insoluble ashes are considered to be an important and useful parameter for detecting the presence of inorganic substance like silicate ion. Similarly the hexane, alcohol and water soluble extractives are indicators of the total solvent soluble component, sugar, starch and tannin was also determined. Successive solvent extractions were carried through soxhlet apparatus and are indicators of the total soluble component in different polarity viz. Hexane, Chloroform, Acetone, Ethanol and Water respectively. On quantitative HPTLC analysis methanol extract yield 0.447% of -sitosterol as marker compound. CONCLUSION As there is no detailed pharmacognostic and anatomical work on record of this traditionally much valued drug the present work was taken up with a view to lay down standards which will contribute significantly to quality control and authentication of this medicinally useful plant. It will also open a new vista for exploitation of this species for the development of herbal formulation(s).
10. Rastogi RP, Mehrotra BN. 1999. Compendium of Indian Medicinal Plants Central Drug Research Institute Lucknow, National Institute of Science Communication, CSIR, New Delhi. Vol.1. 11.
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