Qualitative and Quantitative Determination of Secondary Metabolites of Lagerstroemia Parviflora Roxb Leaves

Namita Bharadwaj, Anjna Chaturvedi*

Dr. C. V. Raman University, Kota, Bilaspur, (C.G.)

*Corresponding Author E-mail: manojkorba.aisect@gmail.com

ABSTRACT:

Herbal medicines as the major remedy in traditional system of medicine have been used in medical practices since antiquity. In addition to its ancient historical uses, Lagerstroemia parviflora Roxb (L. parviflora) is used in several systems of medicine for a variety of ailments. The bioactive compounds that are produced by plants are collectively called as Phytochemicals. The phytochemical ingredients are plant derived compounds which protect the plants from environmental stresses, including insects, bacteria and fungus and weather changes. Though phytochemicals are not considered essential nutrients, it has become apparent that they offer many health benefits to the plants. The aim of the present study is to examine L. parviflora leaves for phytochemical profile. Qualitative analysis of various phytochemical constituents and quantitative analysis of total flavonoids were determined by the well-known test protocol available in the literature. TLC analysis of the methanolic fraction of the leaves was carried out using the solvent system Toluene: Ethyl acetate: Formic acid (in 5:4:1 ratio). Phytochemical analysis revealed the presence of phenols, flavonoids, tannins, saponins, alkaloids, fixed oil and fats. It is expected that the important phytochemical properties recognized by our study in the indigenous medicinal plants will be very useful in the curing of various diseases when taken along with our food.

KEYWORDS: Lagerstroemia parviflora Roxb, Phytochemical profile, TLC analysis, Total flavonoids.

INTRODUCTION:

India is the largest producer of medicinal herbs and appropriately called the Botanical garden of the world¹. Since ancient times plants have been traditionally used in therapeutic practices for the treatment of different types of ailments^2-5. There are a number of crude drugs where the plant source has not yet been scientifically identified. A phytochemical is a natural bioactive compound found in plants foods that works with nutrients and dietary fibre to protect against diseases. Many researchers suggest that, phytochemical working together with nutrients found in fruits, vegetables and nuts. They can have complementary and overlapping mechanism of action in the body including antioxidant effect. L. parviflora (Lythraceae) commonly known as Landia in India and Seja in Bundelkhand; is a species widely distributed in almost all moist and dry deciduous tracts of India.

In view of its wide distribution, the tree can withstand great variation in climate. It is often found as a companion to natural sal and teak it occurs as a distinct species in forests of sub- Himalayan tracts, Assam, Madhya Pradesh, Orissa, Maharashtra, Gujarat, Andhra Pradesh, Karnataka and Tamil Nadu (except Nilgiris and arid regions). In Madhya Pradesh, it is common in all districts⁶. The tree is primarily use for timber the bark of L. parviflora contain tannin (7% - 10%) and is used locally for tanning and dyeing lather and for dyeing cotton thread. It also has some medicinal importance⁷. Mazumder et al. (2003) reported the antibacterial activities of the leaves of the plant⁸and Bhakuni et al. (1969) reported the antiasthmatic activity of the flowers of L. parviflora⁹. Mazumder et al. (2005) reported antipyretic potential of L. parviflora leaves in our laboratory¹⁰. The leaf juice of this plant is used in traditional medicine to treat fever in Jharkhand, India (Jain and Tarafdar, 1970)¹¹. The aim of this work was to determine the quality (types), quantity (amount) of bioactive compounds of leaf of L. parviflora.

MATERIAL AND METHOD:

Plant material:

The leaves of L. parviflora were collected from Bhimbetka Bhojpur, Bhopal (M.P.) in the month of Feb, 2019. Plant material (leaves) selected for the study were washed thoroughly under running tap water and then were rinsed in distilled water; they were allowed to dry for some time at room temperature. Then the plant material was shade dried without any contamination for about 3 to 4 weeks. Dried plant material was grinded using electronic grinder. Powdered plant material was observed for their colour, odour, taste and texture. Dried plant material was packed in air tight container and stored for phytochemical and biological studies.

Chemical reagents:

All the chemicals used in this study were obtained from Hi Media Laboratories Pvt. Ltd. (Mumbai, India), Sigma-Aldrich Chemical Co. (Milwaukee, WI, USA), SD Fine-Chem. Ltd. (Mumbai, India) and SRL Pvt. Ltd. (Mumbai, India).All the chemicals and solvent used in this study were of analytical grade.

Defatting of plant material:

Powdered leaves of L. parviflora were shade dried at room temperature. The shade dried plant material was coarsely powdered and subjected to extraction with petroleum ether using maceration method. The extraction was continued till the defatting of the material had taken place.

Extraction by maceration process:

89gm of dried plant material were successively extracted with different solvent using maceration method for 48 hrs. The extracts were evaporated above their boiling points and stored in an air tight container free from any contamination until it was used. Finally the percentage yields were calculated of the dried extracts.

Phytochemical screening of the extract:

The extract of L. parviflora was subjected to qualitative analysis for the various phytoconstituents like alkaloids, carbohydrates, glycosides, phytosterols, saponins, tannins, proteins, amino acids and flavonoids^12,13.

Thin layer chromatography:

Thin layer chromatography is based on the adsorption phenomenon. In this type of chromatography mobile phase containing the dissolved solutes passes over the surface of stationary phase.

Distance traveled by solute

Rf= --------------------------------------------

Distance traveled by solvent

Total flavonoids determination:

The total flavonoid content was determined using the method of Olufunmiso et al¹⁴. 1ml of 2% AlCl₃ solution was added to 3ml of extract or standard and allowed to stand for 15 min at room temperature; the absorbance of the reaction mixture was measured at 420nm using UV/visible spectrophotometer. The content of flavonoids was calculated using standard graph of quercetin and the results were expressed as quercetin equivalent (mg/100mg).

RESULTS AND DISCUSSIONS:

The percentage yields of different extract obtained from L. parviflora are depicted in the Table 1. Preliminary phytochemical studies of the extract were done according to the published standard methods. These tests were broad in scope and used to determine the presence of flavonoids, saponins and diterpins but alkaloids was absents in the extract Table 2. The thin layer chromatographic analysis of the methanolic fractions of the leaf extract was carried out as explained. The chromatogram revealed various bands under normal light, long U.V and short U.V Table 3 and Figure 1. The content of total flavonoid compounds (TFC) was expressed as mg/100mg of quercetin equivalent of dry extract sample using the equation obtained from the calibration curve: Y = 0.06X+0.019, R²= 0.999, where X is the quercetin equivalent (QE) and Y is the absorbance Table 4 and Figure 2.

Table 1 % Yield of leaves of L. parviflora

S. No.	Solvents	Percentage Yield (%)
1.	Chloroform	2.53
2.	Ethyl acetate	2.47
3.	Methanol	3.67
4.	Aqueous	3.21

Table 2 Phytochemical screening of extract of L. parviflora

S. No.

Constituents

Chloroform extract

Ethyl acetate extract

Methanol extract

Aqueous extract

Alkaloids

A) Wagner’s Test:

B) Hager’s Test:

_-Ve

Glycosides

A) Legal’s Test:

_-Ve

_+Ve

_-Ve

Flavonoids

A) Lead acetate Test:

B) Alkaline Reagent Test:

_-Ve

_+Ve

_-Ve

Saponins

A) Froth Test:

_-Ve

_+Ve

Phenolics

A) Ferric Chloride Test:

_-Ve

Proteins and Amino Acids

A) Xanthoproteic Test:

_-Ve

Carbohydrate

A) Fehling’s Test:

_-Ve

Diterpenes

A) Copper acetate Test:

_-Ve

_+Ve

_-Ve

Table 3 Calculation of R_f.Value

S. No.

Compound

Extract

Rf Value

Quercetin

Toluene: Ethyl acetate: Formic acid

(5:4:1)

0.40

Figure 1: Photograph of T.L.C (Quercetin)

Table 4 Total flavonoid content of L. parviflora extract

S. No.	Extract	Total flavonoid (mg/100mg)
1.	Methanol extract	6.69

Figure 2: Graph of calibration curve of quercetin

CONCLUSION:

L. parviflora leaves have potential to act as a functional food and a source of useful drugs because of the presence of various phytochemical components. Methanolic extracts shows good results regarding presence of phytoconstituents hence these plants may directly use in medicine preparation or for the development of novel agents for various pathological disorders. Further research on the health benefits of phytochemicals in this plant may be warranted.

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Received on 04.12.2019 Modified on 31.01.2020

Research J. Pharm. and Tech. 2020; 13(10):4740-4742.

DOI: 10.5958/0974-360X.2020.00835.5