PHARMACOGNOSTICAL EVALUATION AND PHYTOCHEMICAL ANALYSIS OF DELONIX REGIA RAFIN. STEM BARKHTML Full Text
PHARMACOGNOSTICAL EVALUATION AND PHYTOCHEMICAL ANALYSIS OF DELONIX REGIA RAFIN. STEM BARK
Sumitra Singh * and Sonia
Department of Pharmaceutical Sciences, Guru Jambheshwer University of Science and Technology, Hisar - 125001, Haryana, India.
ABSTRACT: Delonix regia Rafin. belonging to family Fabaceae and subfamily Caesalpinioideae is flowering plant native to Madagascar and East Africa. Delonix regia reported to have anti-diarrhoeal, anti-inflammatory, antioxidant, hepatoprotective and antimicrobial activity. The present study was carried out to establish the pharmacognostical studies, physico-chemical parameters along with preliminary phytochemical screening of petroleum ether, chloroform, ethylacetate, acetone, methanol and aqueous extracts of Delonix regia Rafin. stem bark. The macroscopical and microscopical characteristics of drug powder were studied. The transverse section of stem bark indicated the arrangement of various cells in cork, cortex, phelloderm and pith region. The preliminary phytochemical screening of various extracts revealed the presence of carbohydrate, protein, glycosides, flavonoids, sterols, phenolic and tannin compounds. The physico-chemical parameters such as total ash, acid insoluble ash, water insoluble ash and sulphated ash (8, 2.005, 3 and 1.4 %w/w respectively), loss on drying (45 %w/w) extractive values, foaming index, swelling index and fluorescence analysis of stem bark powder were studied. These studies will be helpful to establish standards for quality, purity and sample identification of Delonix regia Rafin.
Antioxidant, Extractive values, Total ash, Phytochemical and flavonoids
INTRODUCTION: Delonix is a genus of flowering plants in the pea family, Fabaceae and subfamily Caesalpinioideae. The name of the genus is derived from the Greek words ‘delos’ meaning ‘evident,’ and ‘onyx’ meaning "claw," that refers to the petal 1. Delonix, a genus consists of two species growing in India Delonix elata and Delonix regia 2.
Delonix regia Rafin. with an impressive range of medicinal and biological properties, has been used in the folk medicine systems of several civilizations like for the treatment of constipation, inflammation, arthritis, hemiplagia, leucorrhoea and rheumatism 3, 4. Delonix regia Rafin. flower were used in dysmenorrhoea, as antibacterial, anti-inflammatory, broad spectrum antibacterial, analgesic, antimicrobial and antifungal 5, 6, 7, 8, 9.
The flowers have been used as traditional herbal remedies for gynecological disorders and they are also used as tablet binder. Seeds of Delonix regia contain flavonoids are used as wound healing agent in households 10, 11, 12. For the standardization and quality assurance purpose, the following three attributes must be verified: authenticity, purity and assays. Hence, in this work we make an attempt for the standardization of Delonix regia Rafin. stem bark by carrying out its pharmacognostical studies, physico-chemical parameters and preliminary phytochemical screening.
MATERIALS AND METHODS:
Plant Material: The sample of Delonix regia Rafin. was collected from Pehowa, District Kurukshetra Haryana in month of August, 2011. The herbarium of this plant was identified by Dr. H.B. Singh, Chief Scientist & Head, Raw Material Herbarium and Museum (RHMD), National Institute of Science Communication & Information Resources (NISCIR) New Delhi, Vide Reference no. NISCAIR/RHMD/Consult/-2011-12/1834/134 Dated - Sep 28, 2011. The bark of plant was selected for the present study. A voucher specimen of the same has been retained in the Department of Pharmaceutical Sciences, Guru Jambheshwar University of Science & Technology, Hisar for future reference. Shade dried and pulverized bark of the plant was used for the various studies.
Chemical and Reagent: All the chemical and solvents used for the study were of analytical grade and all methods were taken from official methods.
Macroscopical Characters: The fresh and dried stem bark of Delonix regia were studied for their macroscopical characters such as colour, odour, taste, shape, size and texture.
Microscopical Characters: Microscopical study of Delonix regia stem bark in entire form and in powdered form was performed for histological evaluation. Thin transverse sections soaked in water were cut and stained with phloroglucinol and hydrochloric acid and observed under compound microscope. Photography was done by using Carl Zeiss Primostar trinocular microimaging GmbH Hingen, Germany microscope with canon photomicrograph unit13.
Histochemical Colour Reactions: The histochemical colour reactions on the transverse section of Delonix regia stem bark were performed according to standard procedures reported 14, 15, 16. The colour tests were performed for the identification of the major cell components.
Physicochemical Parameters: The physic-chemical parameters such as percentage of total ash, acid-insoluble, water soluble and sulphated ash, loss on drying, extractive values, foaming index, swelling index, crude fiber content, hemolytic activity, foreign organic matter and bitterness value were determined according to official methods for quality control of medicinal plant 17, 18, 19, 20, 21, 22.
Fluorescence Analysis: A small quantity of dried and finely powdered of Delonix regia stem bark was placed on grease free clean microscopic slide and added 1-2 drops of the freshly prepared reagent solution, mixed gently by tilting the slide and waited for 1-2 minutes. Then the slide was viewed in day light and (365 nm) ultraviolet radiations. The colors observed by application of different reagents in different radiations were recorded 23, 24. The plant material was subjected to fluorescence analysis in visible/ daylight and UV light (254 nm & 365 nm).
Preliminary Phytochemical Screening: Preliminary phytochemical screening was carried out on various extracts of Delonix regia stem bark powder like petroleum ether, chloroform, ethylacetate, acetone, methanol and aqueous extracts. The dried extracts were subjected to qualitative tests for identification of various plant constituents such as carbohydrates, alkaloids, glycosides, saponins, sterol and triterpenoids, flavonoids, tannins and phenolic compounds and proteins and amino acids 13, 25, 26, 27.
RESULTS AND DISCUSSION:
Macroscopical Studies: The colour of fresh bark at inner side was cream and from outer side grayish brown. The colour of dried bark from inner side was light brown and at outer side brown.
FIG. 1: BARK OF DELONIX REGIA RAFIN.
The shape of fresh bark was flat and of dried bark curved. The wrinkles were present and furrows were absent. The fracture was granular shown in Fig. 1. Bark was tasteless and odourless.
Microscopical Characters: The transverse section of stem bark consist of cork cells and numerous layers of thin walled flat polygonal cells. Few cells were filled with reddish brown masses. The parenchyma were observed in the cortex region. Occcasional cells were filled with microcrystal of calcium oxalate and starch. Secondary cortex was composed of wide zone of compact lignified strands of stone cells interspread in this region. Stone cells were rectangular to somewhat oval, highly thickened and lignified. Stone cells bearing numerous simple pits were also noticed. Secondary phloem of sieve tubes, companion cells, phloem parenchyma and stone cells were observed. Medullary rays 2 - 4 serriate were present shown in Fig. 2.
FIG. 2: TRANSVERSE SECTION OF STEM BARK OF DELONIX REGIA RAFIN
C: Cork; CC: Cork cambium; SC: Stone cells; SCT: Secondary cortex; PF: Pericyclic fiber; NLF: Non lignified fibers; MR: Medullary rays; PhF: Phloem fiber
Powder study of stem bark under microscope showed stone cells, cork cells and parquetry cells. It also showed lignified and non-lignified cells shown in Fig. 3.
FIG. 3: POWDER STUDY OF STEM BARK OF DELONIX REGIA RAFIN.
Histochemical Colour Reaction Tests: Transverse sections of Delonix regia stem bark when treated with various chemicals reagents for the tests of cell components showed change in colour as shown in Table 1.
TABLE 1: HISTOCHEMICAL COLOUR REACTIONS ON TRANSVERSE SECTION (STEM)
|5% Ferric chloride||Dark green||Tannins present|
|Picric acid||Green||Alkaloid present|
|Phloroglucinol+Hydrochloric acid||Xylem vessesls become Pink||Lignin present|
|Millon’s regaent||Violet||Proteins present|
Physico-chemical parameters: The various parameters such as total ash, acid insoluble ash, water soluble ash, sulphated ash, loss on drying were established and shown in Table 2. The extractive values with colour by successive extraction method are summarized in Table 3.
TABLE 2: ASH VALUES AND LOSS ON DRYING
|Acid insoluble ash||2.005|
|Water soluble ash||3.0|
|Loss on drying||45|
TABLE 3: SUCCESSIVE EXTRACTIVE VALUE AND COLOUR OF EXTRACT UNDER VISIBLE LIGHT
(60 - 80 0C)
Fluorescence Analysis: The plant material may be identified from their adulterants on the basis of fluorescence nature. The powder of stem bark was treated with different chemical reagents and results are reported in Table 4.
TABLE 4: FLUORESCENCE ANALYSIS OF POWDERED STEM OF DELONIX REGIA RAFIN.
|Visible/Day light||UV 254 nm
|UV 365 nm
|Powder as such||Yellowish white||Yellow||Pale yellow|
|Powder + 5% NaOH||orange brown||Light green||Yellow|
|Powder + acetic acid||Yellow||Yellow||Yellow|
|Powder + distilled water||Yellow||Yellow||Yellow|
|Powder + 5%
|Powder + 5% FeCl3||Yellow||Yellow||Black|
|Powder +dilute H2SO4||Reddish brown||Cream||Fluorescent Yellow|
|Powder + dilute HCl||Yellow||Yellow||Fluorescent Yellow|
|Powder+ conc. HCl||Brown||Fluorescent green||Black
|Powder + 5% KOH||Orange brown||Reddish brown||Fluorescent Yellow|
|Powder + ammonia||Reddish brown||Reddish brown||Yellow|
|Powder +ethyl acetate||Yellow||Yellow||Fluorescent Yellow|
|Powder +chloroform||Yellow||Yellow||Off white|
|Powder + ethanol||Yellow||Yellow||Yellow|
|Powder + potassium
Quantitative studies: The other quantitative studies for foaming index, swelling index, crude fiber content, hemolytic activity, foreign organic matter and bitterness value were performed. The results are tabulated in Table 5.
TABLE 5: QUANTITATIVE STUDIES OF DELONIX REGIA RAFIN.
|Foaming index||> 100|
|Crude fiber content||42%|
|Hemolytic activity||Non hemolytic|
|Foreign organic matter||0.60%w/w|
|Bitterness value||Non bitter|
Preliminary Phytochemical Screening: The various extracts of powdered stem bark like petroleum ether, chloroform, ethylacetate, acetone, methanol and aqueous extracts were subjected for investigation of various phytoconstituents. It revealed the presence of different phytoconstituents, like carbohydrate, protein, glycosides, flavonoid, sterol and triterpenoid, phenolic and tannins compounds in different extracts as in Table 6.
TABLE 6: PHYTOCHEMICAL SCREENING OF EXTRACTS OF DELONIX REGIA RAFIN.
|Phenolic compound and
|Protein and amino acid||-||-||-||-||+||+|
|Sterol and triterpenoid||+||-||-||-||+||+|
Here, P-petroleum ether extract; C-chloroform extract; E-ethylacetate extract; A-acetone extract; M-methanolic extract; W- water extract. Here, (+) is present and (-) is absent
CONCLUSION: The scientists from past few decades are keen and sincere to evaluate many ethno medicinally used plants, due to their specific healing properties, desirable action, easy availability and less toxicity. The stem bark of Delonix regia Rafin. is still used in treatment of various disorders by many populations. The pharmacognostical standardization on this plant gives the idea about identification, standardization and monograph of the plant. It is also important to evaluate the medicinal and therapeutic action of this plant.
ACKNOWLEDGEMENT: We are thankful to Guru Jambheswar University of Science and Technology, Hisar and AICTE, New Delhi for grant.
CONFLICT OF INTEREST: The authors have no conflict of interest.
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How to cite this article:
Singh S and Sonia: Pharmacognostical evaluation and phytochemical analysis of Delonix regia Rafin. Stem bark. Int J Pharm Sci Res 2018; 9(5): 1908-12.doi: 10.13040/IJPSR.0975-8232. 9(5).1908-12.
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S. Singh * and Sonia
Department of Pharmaceutical Sciences, Guru Jambheshwer University of Science and Technology, Hisar, Haryana, India.
18 July, 2017
15 December, 2017
23 April, 2018
01 May, 2018