Preparation And Evaluation of Herbal Ointment Using ACACIA NILOTICA Bark Extract

A plant, is traditionally known for its antimicrobial, anti-inflammatory, and wound healing activities.

AIM

•           Preparation and evaluation a herbal ointment containing Acacia nilotica bark extract for wound healing activity.

OBJECTIVES

•      To collect and authenticate the bark of Acacia nilotica.

•      To prepare the bark extract using a suitable extraction method (maceration with ethanol). To carry out preliminary phytochemical screening of Acacia nilotica bark extract.

•      To formulate a herbal ointment by incorporating the bark extract into a suitable ointment base.

•      To evaluate the prepared herbal ointment for physical parameters such as color, odor, and consistency.

•      To determine the physicochemical parameters, including pH, spreadability, viscosity, and Temperature.

•      To assess the stability of the formulated herbal ointment under different storage conditions.

•      To study the suitability of Acacia nilotica bark extract as a natural wound healing agent in topical for.mulations.

PLANT PROFILE

Scientific Classification Taxonomy

Kingdom: Plantae

Division: Magnoliophyta Class: Magnoliophyta Order:  Fabales

Family: Fabaceae Leguminosae Genus:  Acacia

Species: Acacia nilotica

Fig No:3

METHODS

The acacia nilotica bark powder is extracted with extraction method.

EXTRACTION:

•      The extraction process was carried out in maceration method with powder of the bark acacia nilotica.

•      Using 70% ethanol and 30% distilled water. Ratio of the solvents Ethanol:

•      Distilled water (70:30) separate for 24hrs and filter

•      The extract was stored in air free sterile container and perform various chemical analysis.

Fig No:4

Maceration:

This is an extraction procedure in which coarsely powdered drug material, either leaves or stem bark or root bark, is placed inside a container: the menstruum is poured on top until completely covered the drug material. The container is then closed and kept for at least three days. The content is stirred periodically, and if placed inside bottle it should be shaken time to time to ensure complete extraction. At the end of extraction, the micelle is separated from mare by filtration or decantation. Subsequently, the micelle is then separated from the menstruum by evaporation in an oven or on top of water bath. This method is convenient and very suitable for thermolabile plant material.

Methods:

Step 1: Preparation of Acacia nilotica Extract

PREPARATION OF BARK POWDER

The bark are collected and shade dried. It was powdered in a mixer. The coarse powder was sieved and stored in close container.

Physical Evaluation                                pH Measurement

Fig No:5                                          Fig No:6

Procedure:

CHEMICAL TEST RESULT:

The ethanolic extract of Acacia nilotica bark was subjected to preliminary phytochemical tests to detect the presence of:

•           Tannins

•           Flavonoids

•           Alkaloids

•           Saponins

•           Phenolic compounds Standard qualitative chemical tests were performed.

HPLC apparatus and conditions

A Waters HPLC system, consisting in two module pumps, model 1525, a manual injector (Breeze 7725i, Rheodyne) and UV-Vis detector (Waters 2487) at 277 nm, was used. The analyses were carried out on an Eclipse Plus Zorbax C18 Agilent (150 mm×4.6 mm i.d., 5 μm particle size) column as a stationary phase. The mobile phase was a mixture of 5% acetic acid aqueous solution and methanol (80:20, v/v) used in mode isocratic elution. Twenty microliters of sample was injected into the HPLC system. The overall run time was 7.0 min and the flow rate was 1.0 ml/min. All the analyses were carried out at room temperature. Results were acquired and processed by internal software (Empower, Waters, Milford, MA, USA).

Preparation of solutions:

Standard preparation:

The standard solution (12.5 mg) was accurately weighted and transferred into a 25 ml amber volumetric flask and diluted with a solvent mixture of 5% acetic acid aqueous solution and methanol (80:20, v/v) (mobile phase) by 30 min sonication. This standard solution had a concentration of 500 μg/ml. Aliquots of the solution prepared above were transferred to 10 ml amber volumetric flasks to generate solutions to the final concentration required and were then supplemented with purified water. Solutions were filtered through a 0.45 μm membrane filter (Sartorius, Texas, USA) prior to injection.

The standard solutions are Andrographolide, 14-Deoxy-11,12-didehydroandrographolide, Neoandrographolide, 1,3 Triethoxy-Propane, Nobiletin and Tangeritin.

Sample preparation:

For the analysis of extraction Andrographis paniculata, was accurately weighed into a 25 ml amber volumetric flask. The volume was completed with a solvent mixture of 5% acetic acid aqueous solution and methanol (80:20, v/v) (mobile phase). The resulting solution was sonicated during 30 min to enable complete dissolution of sample and filtered using Whatman filter paper n°1. This sample solution had a concentration of 500 μg/ml. Aliquots of this solution were accordingly diluted with purified water in 10 ml amber volumetric flask in order to obtain solutions with final concentration required. These solutions were filtered through a 0.45 μm nylon filter before injections.

CONFIRMATORY TEST

Fig No:7