Preliminary Phytochemical Screening of Different Solvent Extracts of Leaves and Stem of Eriolaena Quinquelocularis (Wight and Arn) Wight

The beneficial efficacy of many indigenous plants for a variety of diseases has been depicted by traditional herbal medicinal practitioners since ancient time. Natural products are the source of synthetic and conservative herbal medicine. These medicines are highly secure as well as environment friendly. According to WHO, 80% of the population from developing and developed countries relies on conventional medicine for their chief health care1. They are bioactive chemicals of plant origin, which are considering as secondary metabolites. Naturally, these bioactive chemicals are manufactured in all parts of the plant body i.e., bark, leaves, stem, root, flower, fruits and seeds. The quantity and quality of bioactive chemicals present in plant parts may vary from one part to another. In fact, the biological activity of plants are highly depends on the distribution of bioactive chemicals (or active principles) which are more frequent in some parts of the plants. The successful determination of active principles isolated from plant material is predominantly dependent on the variety of solvent used in the extraction methods. Hence it emphasizes that numerous solvent attempt are required to screen the plant parts for phytochemicals. The Genus Eriolaena belongs to the family Sterculiaceae is characterized by actinomorphic, bisexual flowers lacking staminodes and ovary tipped with a long style and spreading stigmas developing into a woody dehiscent capsule with numerous winged seeds. Murali & Sukumar (1994) reported that E. quinquelocularis exhibits leaf flushing and flowering events successively within the dry season. Two species from the genus Eriolaena; E. hookeriana and E. lushingtonii are much studied for the pollination biology (K. Ramana & Hareesh Chandra 2013; Raju 2014). The oil from seeds of Eriolaena hookeriana passes malvic acid (25.8%) and sterculic (6.0%) acids in addition to normal fatty acids (M. Ahmad 1979). Gnananath K. (2011) screened E. hookeriana Wt. &Arn for the presence of phytoconstituents like alkaloids, flavonoids and phenolic compounds. They also reported the ethnobotanical uses of the species and conclude that the root of this species has potent anti-inflammatory, wound healing and moderate analgesic properties. Many plants from the family sterculiaceae are medicinally important. Eriolaena quinquelocularis are mentioned in folk medicine.  In the present study, various solvent extracts of leaves and stems of Eriolaena quinquelocularis were qualitatively screened for phytochemicals using standard tests.

MATERIAL AND METHODS:

Collection, Identification, and Authentication of plant material:

The individuals of E. quinquelocularis were collected from natural habitat at Waghjai temple area of Chinchawade (16.72366º N; 74.083742º E), Kolhapur district, Maharashtra, India. The plant specimen was botanically identified and authenticated by Dr. Dinesh L. shirodkar, Botanist Botanical Survey of India, WRC, Pune. A voucher specimen (DRPEQ-1) 14-11-2024 was deposited in Botanical Survey of India, WRC, Pune, Maharashtra, India for future reference.

Preparation of sample extract:

The leaf and stem of Eriolaena quinquelocularis were chopped into small pieces and then shade dried. The dried samples were ground into fine powder using a mortar and pestle. Further, all the powdered samples were subjected extraction with Ethanol, Methanol, Petroleum ether and Water. About 10 gm of leaf and stem powder was added in 100 ml of respective solvent and allowed for sonicating (20 min.25 kHz, 50?). All the extracts were then centrifuged at 5000 rpm for 15 min. to remove debris. The suspension was used for Preliminary phytochemical analysis.

Qualitative Analysis of Secondary Metabolites:

Several qualitative tests were conducted to identify the presence of various secondary metabolites in the plant extracts. Reagent such as Dragendroff’s reagent, Lead acetate, Ferric chloride, Bromine water, Iodine, Benedict’s reagent, conc. HCl, Ninhydrin, Copper sulphate, Acetic anhydride, Alpha naphthol were used to identify Carbohydrate’s, Proteins, Amino acids, Glycosides, Alkaloids, Flavonoids, Phenol, Tannins, Reducing sugar, Quinones, Steroids and Triterpenoids.

Preliminary Qualitative Test Methods:

1. Carbohydrate: Molisch’s test: Two ml of extract, added two drops of alpha naphthol solution in alcohol, shake and added. one ml of conc. H?SO? from sides of the test tube to form violet ring at the junction of two liquids. This test revealed a violet ring indicating the presence of Carbohydrate.
2. Proteins: Biuret test: Two ml filtrate was taken to which one drop of 2 % Copper sulphate solution was added; one ml of 95 % Ethanol was added. Then it was followed by excess addition of KOH. The appearance of pink colour indicates the presence of protein.
3. Amino acids: Ninhydrin test: In two ml of sample added 2-3 drops of Ninhydrin reagent (0.1%) and heated it on boiling water bath. Bluish or purple colouration appeared to specify the presence of amino acids.
4. Steroids and Triterpenoids: Lieberman Buchard test: Two ml extract was mixed with few drops of acetic anhydride, boiled and cooled, conc. Sulphuric acid was then added from the sides of the test tube and observed for the formation of a brown ring at the junctions of two layers. This colour shows the presence of Steroids and Triterpenoids.
5. Glycosides: Bromine water test: Two ml of extract mixed with few ml of bromine water. This test resulted in formation of yellow precipitate indicating the presence of Glycosides.
6. Saponins: Foam test: Two ml test solution was mixed with water and shaken and observed for the   formation of froth indicating the presence of Saponins.
7. Alkaloids: Dragendroff’s test: Two ml of extract, added two ml of Dragendroff’s reagents. This test resulted in formation of Reddish-brown precipitate confirming the presence of Alkaloids.
8. Flavonoids: Lead acetate test: One ml of plant extract, added few drops of 10% lead acetate solution. This test resulted in formation of yellow precipitate indicating the presence of Flavonoids.
9. Phenol: Iodine test: One ml of extract, added few drops of diluted iodine solution. This test show transient red colour, confirming the presence of phenols.
10. Tannins: Braymer’s test: One ml filtrate, added three ml of distilled water and three drops of 10% Ferric chloride solution was added. This test resulted in Blue green colour, confirming the presence of Tannins.
11. Reducing Sugar: Benedict’s test: One ml filtrate, added one ml benedict reagent then boiled for two min. This test resulted in Green/Yellow/ Red colour, indicating the presence of Reducing sugar.
12. Lignin: Labat test: Two ml extract solution added with gallic acid. This test resulted in olive green colour, indicating the presence of lignin.
13. Quinone: Conc. HCL test: One ml extract mixed with few ml of conc. HCL formation of green colour resulting in presence of Quinones.
14. Gums and Mucilage’s: Alcohol test: One ml extract, added one ml water and mixed with two ml Ethanol (constant stirring) formation of white or cloudy precipitate indicating the presence of Gums ani Mucilage’s.
15. Oils: Spot test: Little quantity of plant extract is pressed in between to filter papers. Formation of oil spot on the filter paper is indicating presence of oils.

RESULT AND DISCUSSION:

Detection of primary and secondary metabolites by qualitative methods:

All qualitative test were performed in triplicate. qualitative tests on Eriolaena quinquelocularis revealed the presence of various primary and secondary metabolites, including carbohydrates, proteins, amino acids, steroids, triterpenoids, glycosides, saponins, alkaloids, flavonoids, phenols, tannins, reducing sugar, lignin, Quinones, oils, gums, and mucilage’s, in the examined plant.

Table1: Qualitative analysis Results of leaf of Eriolaena quinquelocularis

+++: Highly present, ++: Moderately present, +: Low, -: Absent.

Table 2: Qualitative analysis Results of Stem of Eriolaena quinquelocularis

+++: Highly present, ++: Moderately present, +: Low, - :Absent.

Fig. 1. Graphical representation of qualitative phytochemical analysis of leaf and stem of Eriolaena quinquelocularis in selected solvent system.