Pharmacognostical and Phytochemical Studies of Delonix regia

Shantha Sheela Nagarajan*, Muthusamy Periyannan, Radha Ramalingam

Department of Pharmacognosy, College of Pharmacy, Madras Medical College, Chennai, 600003.

*Corresponding Author E-mail: sheelacute08@gmail.com

ABSTRACT:

In the present study, an attempt was made to investigate pharmacognostical and phytochemical screening of Delonix regia seeds of family Leguminosae. The seeds were collected from its pods after authentication, shade dried and various pharmacognostical and phytochemical analysis were performed. The pharmacognostical parameters studied were macroscopy, microscopy, physicochemical analysis and chromatography. The coarsely powdered seeds were extracted with various extract and phytochemical screening were performed. The phytochemical screening revealed that the presence of secondary metabolites such as Flavonoids, sterols, triterpenoids, saponins and fixed oils in Delonix regia seeds. All the parameters were studied according to the WHO and Pharmacopoeial guidelines. HPTLC and TLC analysis were also performed. These pharmacognostical data are helpful to determine its identity, where as the Photochemical studies are useful to bring out the various pharmacological activity it may posses, based on the active constituent present in them.

KEYWORDS: Delonix regia, , Pharmacognostical, Phytochemical, Chromatography.

INTRODUCTION:

Herbal medicine also known as botanical medicine or phytomedicine- refers to using plants seeds, flowers, roots for medicinal purpose. Even today plant materials continue to play a major role in primary health care as therapeutic remedy in many developing countries. In the last few decades, medicinal plants have been subjected to pharmacognostical studies¹. This will help us to identify the correct plant for further study. Royal Poinciana is a deciduous, large tree with fern-like leaves². Delonix regia is also known as Gulmohar or flame tree or peacock tree³. It is mostly planted for their shade-giving properties and as an ornamental tree⁴. The major medicinal properties of Delonix regia include anti-diarrhoeal, anti-inflammatory, anti-diabetic, anti-oxidant, hepatoprotective, anti-microbial, anthelminitic, wound healing and gastro protective activity⁵.

It is used for the diabetes treatment in Bangladesh folk medicine⁶. Sterols, triterpenoids and saponin are reported to have anti- diabetic property.

MATERIALS AND METHODS:

Sample collection, processing, storage and method of extraction:

The dried seeds of Delonix regia were collected from Nugambakkam, Chennai, TamilNadu in June 2015 and authenticated by Dr. M. Palanisamy Scientist D-In-charge of Botanical Survey of India, Southern circle, TNAU Campus, Coimbatore (BSI/SRC/5/23/ 2015/Tech.1696). The seeds were collected from seed pods by applying mechanical strength and spreader on a neat sheet for shade drying and powdered using mechanical grinder. The powdered seeds of D. regia were stored in an air tight container. The coarsely powdered seeds were subjected to successive solvent extraction using petroleum ether, ethyl acetate, ethanol and water. The extracts were concentrated under reduced pressure and stored in a refrigerator for further use. Percentage yield of the various extracts were described in Table 1.

Table 1: Percentage yield of extract of D. regia seeds

S. No	Extract	Method of Extraction	Physical Nature	Colour	Yield (%W/W)
1	Hexane	Successive solvent extraction	Semi-solid	Green	0.6
2	Ethyl acetate		Sticky	Brownish green	1.3
3	Ethanol		Semi-solid	Green	4.6
4	Aqueous		Solid	Brown	5.7

PHARMACOGNOSTICAL EVALUATION:

Macroscopy:^{7 - 9}

Organoleptic evaluation of D. regia seeds were done to identity the nature of the plant. Macroscopical studies were carried out as per WHO guidelines. The parameters such as colour, odour, taste, size and shapes were measured and shown in the Fig 1 and Table 2 & Table 3.

Table 2: Organoleptic characters of seed and seed pod of Delonix regia seeds

S. No	Parameters	Observations
1	Nature	Soft and glossy
2	Colour	Brown
3	Odour	Odurless
4	Taste	Tasteless
5	Length	2.2 – 2.6cm
6	Width	0.6 – 0.8cm
7	Type	Most hard seed
8	Shape	Oblong
9	Arrangement	Transversely Mottled with a bony testa
Seed pod
1	Length	25 - 35cm
2	Width	4 – 5cm
3	Colour	Dark brown

Table 3: Organoleptic characters of crude powder

S. No	Parameters	Observation
1.	Nature	Coarse powder
2.	Colour	Greenish brown
3.	Odour	Pleasant
4.	Taste	Bitter

Fig 1: Macroscopy of seeds and seed pods.

Microscopy:^{10, 11}

The seed was cut and fixed in FAA solution (Formalin 5ml + Acetic acid 5ml + 90ml of 70% Ethanol). The specimen was dehydrated after 24 hours of fixing. The seeds were graded with series of tertiary butyl alcohol. The sections were stained as per the method published by O’Brein et al. The paraffin embedded specimens were sectioned with the help of Rotary Microtome. The thickness of the section was 10-20µ. The sections were stained using Toludine blue. Microscopical structures were shown in the Fig 2, Fig 3.

Fig 2: Palisade epidermis and bearer cells in transverse section (EP-Epidermis, OSct - Outer seed coat, ISct – Inner seed coat)

Fig 3: a). Palisade epidermis layer, b). bearer cells in surface view- pigment layer, c).Epidermis of flat face and palisade cells of cotyledon in transverse section

Powder microscopy:^{10, 11}

Microscopic examination of the powdered seeds of Delonix regia was performed. The crude powder passed through sieve no 60 was used for powder microscopy. The powdered drug was separately treated with phloroglucinol and hydrochloric acid solution, glycerin and iodine solution to determine the presence of lignified cells, calcium oxalate crystals and starch grains. Both stained and unstained specimens were used to identify and confirm the microscopic structures. Structures of the powder of D. regia were shown in Fig 4.

Fluorescence analysis:¹²

The powdered seeds of D. regia when treated with different chemical reagent showed different colour reactions, in accordance to the nature of the constituents present in it. Many plants constituents show fluorescence in visible light and some of the metabolites shows

fluorescence only when they are exposed to ultraviolet light. Fluorescence characteristics of the powder and extracts were described in Table 4 and Table 5 respectively.

Table 4: Fluorescence analysis of the powders of D. regia seeds

Treatment	Day light	UV light
Treatment	Day light	254nm	365nm
Powder as such	Brownish green	Dark brown	Pale brown
Powder + water	Pale brown	Dark brown	Pale brown
Powder + NaOH	Brownish green	Dark green	Pale brown
Powder + Hcl	Dark green	Brownish green	Pale brown
Powder + Acetic acid	Pale brown	Dark brown	Pale brown
Powder + Alc.NaOH	Brownish green	Dark green	Pale brown
Powder + Picric acid	Brownish yellow	Brownish yellow	Yellowish brown
Powder + Sulphuric acid	Pale brown	Brownish green	Pale brown
Powder + Nitric acid	Brownish green	Dark brown	Pale brown
Powder + Iodine	Dark brown	Dark brown	Dark brown

Table 5: Fluorescence analysis of the extracts of D. regia seeds

Extracts	Day light	UV light
Extracts	Day light	254nm	365nm
Hexane extract	Dark greenish brown	Dark brown	Dark brownish
Ethyl acetate extract	Dark brown	Dark brownish green	Reddish brown
Ethanol extract	Dark brown	Dark brown	Reddish brown
Aqueous extract	Dark brown	Dark brown	Dark greenish brown

RESULTS AND DISCUSSION:

Fig 4: a). Palisade epidermis, b). Modified parenchyma and sclerenchyma, c). Aleurone layer within seed-coat

Fig 4: a). Sclerotic seed coat, b). Isolated bearer cells, c). Trichomes, d). Calcium oxalate crystals

Table 6: Physicochemical parameters of D. regia

		Results (%W/W)
Ash values	Total ash Sulphated ash Water insoluble ash Acid insoluble ash	2.53 15.0 1.1 0.7
Loss on drying		6.5
Extractive values	Water soluble extractive Alcohol soluble extractive Ether soluble extractive Non – volatile ether soluble extractive	11.0 5.0 2.9 3.6
Other parameters	Foaming index Swelling index	<100 NIL

Table 7: Preliminary phytochemical screening of D. regia

Phytoconstituents	Powder	Hexane extract	Ethylacetate extract	Ethanol extract	Aqueous extract
Alkaloids	-	-	-	-	-
Saponins	+	+	-	+	+
Glycoside	-	-	-	-	-
Carbohydrates	-	-	+	+	-
Tannins and phenolic compounds	-	-	-	-	-
Flavonoids	+	+	+	+	-
Phytosterols	+	+	+	+	-
Proteins and aminoacids	-	-	-	-	-
Triterpenoids	+	+	+	+	-
Fixed oils and fats	+	+	-	+	-
Gums and mucilage	-	-	-	-	-

Fig 5: Qualitative estimation of phytoconstituents of D. regia. a). TLC of ethanoic extract D. regia, b). HPTLC of D. regia and its chromatogram.

S. No	Rf	Height	Area	Lamda Max
1	0.11	29.1	861.9	668
2	0.20	2.3	23.7	507
3	0.25	3.7	100.2	507
4	0.32	1.8	27.0	507
5	0.41	1.8	55.2	507
6	0.48	1.5	32.1	507
7	0.65	4.1	53.7	507
8	0.67	7.6	111.2	507
9	0.68	8.0	144.0	507
10	0.74	37.7	701.2	615

CONCLUSION:

The present study showed that the presence of macroscopical and microscopical structures. Microscopical structures such as epidermis, pigment layers, palisade cell of cotyledons, modified parenchyma and sclerenchyma, aluerone grain, sclerotic seed coat, isolated bearer cells, trichomes, and calcium oxalate crystals were identified. In phytochemical studies, the presence of various phytoconstituents such as saponins, sterols and triterpenoids in the Delonix regia seeds of various extracts. From the overall study we conclude that the plant has various phyto constituents and it might be useful plant for treating various diseases such as diabetes, inflammations, diarrhoea, gastric ulcers and microbial infections. Further studies are needed for knowing the exact mechanism of the plant on various diseases.

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Received on 17.02.2016 Modified on 01.03.2016

Res. J. Pharmacognosy and Phytochem. 2016; 8(2): 70-74

DOI: 10.5958/0975-4385.2016.00013.3