Formulation & Evaluation of Polyherbal Anti-Microbial Ointment of Blumea Lacera and Moringa Oleifera

Herbal drug are formulated in the form of ointment and are used topically for several purposes, e.g. protectants, antiseptics, emollients, antipruritic, keratolytic and astringents, Ointment base are always anhydrous and generally contain one or more medicaments in suspension or solution or dispersion. Ointment bases may be hydrocarbon (oleaginous), absorption and water removable and water-soluble type. On the basis of their level of action, ointment is aimed to heal the incised and excised wounds. In an earlier study, medicinal plants have been reported to be very beneficial in wound care promoting the rate of wound healing with minimal pain, discomfort and scarring to the patient The use of natural products or natural product-based medicine is increasing all over the work especially in the developing countries, even though synthetic drugs are readily available and highly effective in caring various diseases, there are people who still prefer using traditional folk medicine because of their less harmful effects. Approximately 25% of the prescribed drugs in the world are basically plant origin. In the developing countries like India, approximately 80% people rely m traditional plant-based drugs for their primary health-care needs. Recent widespread interest in plant derived drugs reflects its recognition of the validity of many traditional claims regarding the value of natural products in health care. For quality control of traditional medicines, phytochemical investigations are mainly applied. Thus, it makes a great significance to investigate chemical constituents and study pharmacological activity on this plant for its medicinal uses, which will be very useful in the field of medicine as new emerging drug. According to the WHO, medicinal plant are the best sources to obtain a variety of new herbal drug. (1) (2)

The following two herbs are used in formulation of polyherbal antimicrobial ointment

Blumea lacera (Burm.f.) DC. (3) (4)

Blumea lacera is a fast-growing, aromatic herb widely distributed in tropical Asia. It belongs to the family Asteraceae and is traditionally used for its diverse medicinal properties including antimicrobial, antipyretic, analgesic, and wound-healing activities. The leaves contain bioactive compounds like flavonoids, alkaloids and terpenoids, which contribute to its pharmacological potential. Due to its ability to combat various pathogenic microbes, Blumea lacera serves as a potent candidate for antimicrobial formulation.  

Fig 1. Blumea lacera herb

Taxonomical classification of Blumea lacera:

• Kingdom: Plantae
• Subkingdom: Tracheobionta
• Division: Magnoliophyta
• Class: Magnoliopsida
• Order: Asterales
• Family: Asteraceae
• Genus: Blumea
• Species: Blumea lacera (Burm.f.) DC.

Moringa oleifera Lam. (5) (6) (7)

Moringa oleifera, commonly known as the drumstick tree or horseradish tree, is a versatile plant belonging to the family Moringaceae. It is widely used in traditional medicine for its antimicrobial, antioxidant, anti-inflammatory and wound-healing properties. Various parts of the plant, particularly the leaves, are rich in bioactive compounds such as phenolics, flavonoids, and glucosinolates, which support its inclusion in topical formulations for microbial infections.

Fig. 2. Moringo oleifera herb

Taxonomical classification of Moringa oleifera:

• Kingdom: Plantae
• Subkingdom: Tracheobionta
• Division: Magnoliophyta
• Class: Magnoliopsida
• Order: Brassicales
• Family: Moringaceae
• Genus: Moringa
• Species: Moringa oleifera Lam.

MATERIALS AND METHODS:

Plant material collection and extraction:

The fresh plant leaves were collected from Maharastra state by gently uprooting the plant. Authenticated by­ Vilas M. Suroshe Msc., Phd (Ethnobotanist). Leaves collected are allow for shade drying process later grinded into fine powder. Extracted by reflux extraction technique by using 70% ethanol as solvent.

Apparatus used:

Reflux condenser, Mortar-pestle, water bath

METHOD:

Preparation of Extracts:

The leaves of Blumea lacera and Moringo oleifera were collected from Thane district of Maharastra during the month of October 2024. Leaves of Blumea lacera and Moringo oleifera were shade dried and grinded to obtain fine powder which then stored in closed container in dark place. The 20 gm of powdered were subjected to reflux extraction in 200 mL of 70% ethanol for 2-4 hours. The obtained extract were filtered and filtrate were evaporated to viscous mass, % yield was calculated and used for further study. (8)

Preparation of herbal ointment:

Heat the simple ointment base at 60–70°C in a water bath. Gradually add Blumea lacera and Moringa oleifera extracts while continuously stirring. Mix thoroughly using a glass rod or mechanical stirrer to ensure uniform distribution. Allow the mixture to cool at room temperature while stirring to prevent phase separation. Transfer the prepared ointment into sterile containers and store in a cool, dry place. (9) (10)

Thin Layer Chromatography (TLC):

TLC studies were carried out for the better identification of different components of the ethanolic extract of Blumea lacera. The results of TLC shows the presence of spots in the solvent system Chloroform: Benzene: Formic acid (      )

TLC Parameter  

• Sample preparation : Ethanolic extract with water
• Sample application : Micropipette
• Solvent system : Chloroform : Benzene :Formic acid 
• TLC plate development: Silica Gel G.
• Rf value :  F1-0.7, F2-0.8, F3-0.76

Evaluation of Formulation: (11) (12)

Prepared blumea lacera + moringo oleifera ointment formulation were evaluated for the following parameter.

Organoleptic parameter:

Herbal ointment formulations were evaluated based on their texture, consistency and appearance. Texture was determine on the basis of smoothness. Texture was found to be smooth; it can be spreadable and washable easily.

pH: (13)

2.5gm ointment formulation sample of each batch was taken in 100 ml of beaker, and add 50 ml of water to it. Beaker was heated on water bath maintain at about 65? to 70? for 10 min, cooled to room temperature, and then centrifuged at 3000 rpm for 10 min. The pH measurement were done by using a digital type pH meter by dipping the glass electrode into the ointment formulation

Spreadability: (14)

The spreadability is expressed in term of time in second taken by two slide to slip off form ointment, placed in between two slides under the direction of certain load. Lesser the time taken for separation of two slides, better the spreadability of ointment.

Spreadability of ointment formulation was determined by using formulation

                     S=M x LT

Where S= spreadability, M=Weight tied to upper slide, Length of glass slides and T= Time taken to separate the slides.

Extrudability:

(Spreading Diameter Method)

Place a fixed amount of ointment (e.g 0.5g) on a glass plate. Place another plate of the the same size on top and apply a standard weight (e.g.,500g) After a set time (e.g., 5min) measure the diameter of the spread ointment.

A large diameter indicates better extrudability.

Loss on drying:

The loss in weight, in the sample so tested, principally is due to loss to loss of water and smell amount of other volatile material from it. Loss on drying was determined by placing the 1gm of ointment formulation of different batches in a petri dish on a water bath and dried until constant weight was obtained.

Washability: (15)

Herbal ointment formulation were applied on the skin and then washing with water was checked. Washability was checked by keeping applied skin area under the tap water for about 10 min.

Antimicrobial Activity: The compound works well against both types of bacteria—Gram-negative and Gram-positive—showing effects similar to the commonly used antibiotic Gentamicin. The size of the clear zones where bacteria were stopped (called zones of inhibition) ranged from 23 to 28 mm, which is close to Gentamicin’s range of 24 to 29 mm. This antibacterial effect is likely due to natural chemicals in the plant leaves, like alkaloids, flavonoids, phenols, and tannins. Using a semi-solid form, like an ointment, helps the medicine stay on the skin longer and work better. Ointments made with these plant extracts showed good antibacterial properties and could be useful for treating wounds and burns.

Result & discussion:

Table 1. Evaluation parameters of ointment

Table 2. Anti-microbial evaluation