Pharmacognostic and Preliminary Phytochemical Screening of Ampelocissus indica including Antioxidant Activity

 

Bincy Raj1, Soosamma John2, Abdul Rasheed A. R.3

1Department of Pharmacognosy, Dayananda Sagar College of Pharmacy, Bangalore -560078

2East Point College of Pharmacy, Bangalore - 560049

3College of Pharmacy, Kannur Medical College, Kerala - 670612

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

 

ABSTRACT:

Ampelocissus indica is tuberous plant. It has many reported traditional uses like used for dyspepsia, gout, health tonic, ulcer, indigestion, hepatitis etc. Present study focuses on standardisation of Ampelocissus indica on the basis of morphological, microscopical, physical evaluation, phytochemical investigation and antioxidant activity. From preliminary phytochemical screening it is found that methanolic extract of Ampelocissus indica is having many important phytoconstituents like alkaloids, flavonoids, phytosterols, tannins, phenolic compounds and carbohydrates. The antioxidant activity was estimated by using 2, 2- diphenyl-picryl-hydrazyl (DPPH) free radical assay. And it is found that Ampelocissus indica is having strong antioxidant activity. Total phenolic content was measured by Folin-Ciocalteau (FC) by using Tannic acid as the calibration standard. In the DPPH radical scavenging assay, the IC50 value of the extract was found to be 20μg/ml. The total phenolic content was measured by Folin-Ciocalteau was found to be 2.5 for tannic acid. In the present study morphological evaluation, microscopical evaluation, physical evaluation and chemical evaluation is done along with antioxidant studies. These standardization parameters will be helpful for identification of this endangering plant.

 

KEYWORDS: Ampelocissus indica, DPPH, Phytochemical, Folin-Ciocalteau, Total phenolic content.

 

 

INTRODUCTION:

Plants are one of the most important sources of medicine (1). The medicinal plants are rich in secondary metabolites and essential oil of therapeutic importance. The biosynthesis of the secondary metabolites, although controlled genetically, is affected by environmental influence (2). Hence there is a need for standardisation of herbal crude drugs and their bio constituents in justifying their acceptability in modern system of medicine. Ampelocissus indica is tuberous plant.

 

It has many reported traditional uses like antidote, used for dyspepsia, gout, health tonic, ulcer, indigestion, hepatitis etc. Present study focuses on standardisation of Ampelocissus indica on the basis of morphological, microscopical, physical evaluation, phytochemical investigation and antioxidant activity.

 

MATERIAL AND METHOD:

Pharmacognostic studies:

Tubers of Ampelocissus indica was collected from kerala and authenticated from RRI Bangalore. Morphologically tubers are brown in colour and no characteristic taste. A thin hand section of tuber is taken and washed with chloral hydrate and stained with phloreglucinol. Ash value and extractive value is determined as per IP. The extracts were prepared with methanol. Extraction is done by soxhlet extraction at 500C. The % yield was calculated (3) (4) (5).

Preliminary phytochemical screening:

For preliminary phytochemical analysis, extract was prepared from 500mg of the dried powdered tubers of Ampelocissus indica and were subjected to hot successive continuous extraction with methanol. The extracts were filtered and concentrated. The extracts were dried over desiccator and the residue was weighed. The presence or absence of the primary and secondary phytoconstituents was detected (6) (7).

 

Antioxidant activity and total phenolic content of Ampelocissus indica:

Antioxidant activity by DPPH scavenging assay:

To a set of clean and dried test tubes 3ml of methanol and 150μl of 0.1% DPPH reagent was added and mixed thoroughly. Then allow the solution to stand for 30 minutes. The initial absorbance of each test tube was measured at 517nm. To these test tubes 1ml of aqueous solution of extracts were added in increasing concentration of 5-25μg/ml. The solution of ascorbic acid (5-25μg/ml) was taken as the standard. The solution were mixed and allowed to stand for half an hour at room temperature and final absorbance was measured at 517nm using a Spectrophotometer (UVvisible 1700 Pharma Spec) The experiment was performed in triplicate. The percentage reduction in absorbance was calculated from initial and final absorbance at each level. Concentration of the substance required for 50% reduction in absorbance (IC50) was calculated from the calibration curve (8) (9).

 

Determination of total phenolic compound:

Total phenolic content of methanolic extract of Ampelocissus indica was determined using Folin-Ciocalteau method. The concentrations of total phenolics were calculated using the standard curve of tannic acid (r=0.9965) and the result were expressed in milligrams of tannic acid equivalents (10) (11).

 

RESULT AND DISCUSSION:

Ampelocissus indica is a climber with tuberous root. Stem and branches are hollow and smooth. Leaves are circular or broadly heart shaped with acute apex. Odour is none and taste is bitter. T.S of Ampelocissus indica showed the presence of cork, cortex, vessel, xylem and phloem fibres. Powder microscopic characters showed in Fig 2 and T. S of Ampelocissus indica showed in Fig 1.

 

Ampelocissus indica was subjected to physical evaluation by evaluating moisture content, total ash value, water soluble ash value, acid insoluble ash value, water soluble extractive value and alcohol soluble extractive value the results are shown in Table No 1. Methanolic extract of Ampelocissus indica showed the presence of carbohydrates, flavonoids, tannins, alkaloids, saponins and phenolic compounds.

 

 

Fig 1. T.S of tubers of Ampelocissus indica

CK- Cork, COR –Cortex, SP-secondary phloem, MR-Medullary rays

 

The antioxidant activity was estimated by using 2, 2- diphenyl-picryl-hydrazyl (DPPH) free radical assay. And it is found that Ampelocissus indica is having strong antioxidant activity. Total phenolic content was measured by Folin-Ciocalteau (FC) by using Tannic acid as the calibration standard. The total phenolic content was measured by Folin-Ciocalteau was found to be 2.500 for tannic acid. Calibration curve is showed in Graph 1. In the DPPH radical scavenging assay, the IC50 value of the extract was found to be 20μg/ml. Results are given in Table No 2. In current study, we have made an effort to identify potential bioactive secondary metabolite and their antioxidant activities.  The revealed antioxidant property of extract may provide potential therapeutic intervention against oxidative threats and degenerative disorders.

 

Fig 2. Powder microscopic characters of Ampelocissus indica

 

Table 1. Physical parameters for Ampelocissus indica

Physical parameters

Sample Identity

Ampelocissus indica

Moisture Content %

3.0

Total Ash %

11.2

Acid insoluble ash %

10.2

Water soluble ash %

0.85

Alcohol soluble extractive (%)

4.1

Water-soluble extractive (%)

4.5

 

Graph 1. Standard calibration curve for Tannic acid

 

Graph 2. Antioxident activity of Ampelocissus indica

 

Table 2. Antioxident activity of Ampelocissus indica

Drug

Absorbance

Concentration (µg/ml)

IC50 µg/ml

Ascorbic acid

 

5

10

15

20

25

 

 

17

Initial

0.88

0.88

0.88

0.88

0.88

Final

0.58

0.45

0.34

0.28

0.21

%Reduction

30%

43%

54%

60%

67%

Ampelocissus indica

Final

0.61

0.48

0.42

0.32

0.27

20

%Reduction

25%

40%

46%

56%

61%

 

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Received on 07.03.2020         Modified on 11.04.2020

Accepted on 08.05.2020  ©AandV Publications All right reserved

Res. J. Pharmacognosy and Phytochem. 2020; 12(2): 80-82.

DOI: 10.5958/0975-4385.2020.00014.X