Formulation and Evaluation of Herbal Mosquito Repellent Spray
Shital D. Godse1, Sabeeha A. Kamal2, Aniket S. Nimbalkar3, Vinod Shinde4*
1Department of Pharmaceutical Quality Assurance, Vishwakarma University School of Pharmacy, Kondhwa, Pune.
2,3Department of Pharmaceutical Quality Assurance,
Smt. Kashibai Navale College of Pharmacy, Kondhwa (Bk), Pune 411048, Maharashtra, India.
4Department of Pharmacology, BKL Walawalkar Rural Medical College, Dervan, Chiplun, Maharashtra-415606.
*Corresponding Author E-mail: vshinde62@gmail.com
ABSTRACT:
Mosquitoes, particularly those of the Anopheles, Culex, and Aedes genera, pose a significant global health threat, transmitting diseases like malaria, dengue, and encephalitis. To combat these insects, humans have historically relied on plant-based repellents. This study explores the potential of herbal formulations to create safe and effective mosquito repellents. Due to its known mosquito-repellent properties, Citronella oil, extracted from lemongrass, has been a focus of attention. By combining citronella oil with other essential oils such as eucalyptus, clove bud, and sweet orange peel, this research aims to develop novel repellent formulations. These oils were extracted using methods like steam distillation and cold pressing. Previous studies have highlighted the efficacy of plant extracts and essential oils against malaria vectors. This research seeks to build upon this knowledge by investigating citronella oil's impact on Aedes aegypti mosquitoes. The goal is to develop environmentally friendly alternatives to synthetic repellents, especially for regions with prevalent mosquito-borne diseases.
KEYWORDS: Mosquito repellent, Traditional practice, Herbal mosquito repellent, Essential oils, Deadly disease.
INTRODUCTION:
Mosquito-borne diseases, including malaria, dengue, and Zika, constitute a formidable global health challenge, resulting in approximately 750,000 fatalities annually. The expanding geographical range of mosquito species, exacerbated by climate change, is anticipated to further escalate the incidence of these diseases.1,2 To effectively mitigate this growing threat, the development of safe and efficacious mosquito repellents is imperative. Mosquitoes are particularly attracted to humans due to carbon dioxide and lactic acid emissions, which are detected by specialized chemoreceptors located on the insect's antennae.
While commercially available repellents often employ masking agents to obscure the human scent, their potential adverse effects on human health cannot be overlooked.3,4 This study aims to address this issue by formulating a plant-based mosquito repellent that prioritizes human safety and environmental sustainability. Citronella oil, renowned for its distinctive aroma, owes its insect-repellent properties to a combination of key constituents, including citronellal, eugenol, geraniol, and limonene. By harnessing the power of these compounds, this research endeavors to create a citronella-based spray capable of effectively deterring Aedes aegypti mosquitoes, a primary vector for a multitude of diseases. The essential oils for the repellent formulation will be extracted through the distillation process. This study seeks to establish the efficacy of citronella oil as a potent insect repellent while also exploring its potential to reduce mortality rates caused by Aedes aegypti mosquitoes. By developing a sustainable and human-friendly repellent, this research contributes to the ongoing efforts to combat the global burden of mosquito-borne diseases.5,6 Studies have reported the encapsulation of botanical repellents in different controlled release systems, resulting in positive effects such as prolonged action and low permeation.7-10
MATERIALS AND METHODS:
Materials:
The selection of plants was based on their availability as raw materials, scientific evidence supporting their efficacy, and their traditional use as mosquito repellents.
Preparations of Essential Oil from Plant Extracts:
A. Citronella Oil:
Citronella essential oil is commonly extracted from the leaves and stems of the citronella plant (Cymbopogon nardus) through a detailed procedure. First, the citronella plants are harvested when they reach maturity, typically after four to five months of growth. The optimal time for harvesting is in the morning when the concentration of oil in the leaves is highest. After harvesting, the leaves and stems are separated from the plant, and any debris or unwanted material is removed. The plant material is thoroughly washed to eliminate dirt or contaminants. Following this, the material is dried by spreading the leaves and stems in a well-ventilated area away from direct sunlight or by using a dehydrator set to a low temperature to speed up the drying process. Once dried, the plant material is chopped into small pieces or ground into a coarse powder to increase surface area, facilitating the extraction of essential oils. Several extraction methods are available, such as steam distillation, solvent extraction, and cold pressing, with steam distillation being the most commonly used for citronella essential oil. In steam distillation, water is heated to produce steam, which is passed through the chopped or ground citronella plant material. The heat from the steam causes the essential oil to vaporize. The steam and vaporized oil are then cooled in a condenser, where they condense into a liquid. The essential oil, being lighter than water, floats on top of the condensed water and is collected. The resulting liquid is the citronella essential oil, which can be further purified if needed. This method ensures the volatile essential oils are extracted while preserving their natural properties. The citronella essential oil is then stored in dark glass bottles to protect it from light and oxidation, which could degrade its quality. It is stored in a cool, dry place away from direct sunlight to maintain its potency and effectiveness over time. Before use, the oil undergoes rigorous quality control testing to verify its purity and potency and to ensure the absence of contaminants. After passing these checks, the essential oil is carefully packaged and labelled.11-15
B. Sweet Orange Oil:
Sweet orange oil is commonly extracted through the cold-pressed method, which preserves its distinct fruity aroma. The process begins with the preparation of fresh, sweet orange peels, along with a large natural sponge and a hard surface for pressing. The orange peels are placed beneath the sponge on the hard surface, and pressure is applied by bending the peels into the sponge, targeting the essential oil glands. As pressing continues, the oil is gradually released from the glands into the sponge. The sponge is then squeezed to transfer the oil emulsion into a separate container. Afterward, the oil emulsion is allowed to settle, during which the pure sweet orange peel oil separates and can be collected. The extracted oil is then stored in a suitable container to preserve its fruity aroma. This cold-pressed method is known for maintaining the natural scent of sweet orange peel oil, setting it apart from oils produced using alternative extraction methods.
C. Eucalyptus Oil:
Eucalyptus oil is commonly extracted through steam distillation using a Clevenger apparatus. The process begins with the preparation of fresh eucalyptus leaves, which are washed thoroughly to remove dirt and debris. The leaves are then roughly chopped or crushed to increase the surface area for better oil extraction. Next, the Clevenger apparatus is set up according to the manufacturer’s instructions, with the round-bottom flask filled with distilled water, ensuring it’s not overfilled. The chopped eucalyptus leaves are placed into the distillation flask, with enough distilled water added to cover the leaves. The distillation process is started by gently heating the flask using a hot plate on a low heat setting to avoid rapid boiling. As the water heats, steam rises and carries the essential oil vapors with it.
The vapor, along with the water vapor, travels through the condenser of the apparatus. As the vapor cools in the condenser, it condenses into liquid form, which is then collected in the separator funnel of the Clevenger apparatus. The essential oil, being lighter than water, floats on the top layer of the collected liquid. Once enough oil has been collected, it is carefully drained into a separate container. A glass dropper or pipette is used to extract the oil from the top layer. The collected essential oil is filtered using cheesecloth or a coffee filter to remove any impurities or plant particles. Finally, the filtered eucalyptus oil is transferred to a clean, dark-colored glass bottle for storage to protect it from light exposure and degradation. If a larger quantity of oil is needed, the distillation process can be repeated with fresh eucalyptus leaves and clean water.16-19
Table 1. Solvents and extraction techniques used for different constituents
|
Material |
Solvent used |
Process |
|
Citronella leaves |
Hexane |
Steam distillation |
|
Eucalyptus leaves |
Hexane |
Clevenger apparatus |
|
Clove bud |
Ethanol |
Hydro-distillation |
|
Sweet orange peel |
Hexane |
Cold pressing |
Table 2. Quantity of constituents of Herbal Repellent Spray
|
Sr. No. |
Constituents |
F1 (ml) |
F2 (ml) |
F3 (ml) |
Role of ingredient |
|
1 |
Citronella essential oil (Cymbopogon nardus) |
5.00 |
6.00 |
6.00 |
Strong scent and mosquito repellent |
|
2 |
Eucalyptus essential oil (Eucalyptus globulus) |
5.00 |
5.00 |
6.00 |
Masking the scent that attracts mosquito |
|
3 |
Clove bud essential oil (Eugenia caryophyllus) |
6.00 |
5.00 |
4.00 |
Strong scent and mosquito repellent |
|
4 |
Sweet orange peel essential oil (Citrus sinensis) |
5.00 |
5.00 |
5.00 |
The fragrance of the spray |
|
5 |
Neem extract (Azadirachta indica) |
5.00 |
5.00 |
5.00 |
Mosquito repellent |
|
6 |
Hexane |
2.00 |
2.00 |
2.00 |
Insect repellents |
|
7 |
Ethanol |
2.00 |
2.00 |
2.00 |
Solvent or carrier for the active ingredient |
|
8 |
Tween 80 |
20.00 |
20.00 |
20.00 |
Emulsifier |
|
9 |
Lavender oil |
10.00 |
10.00 |
10.00 |
Fragrance |
|
10 |
Distilled water |
40.00 |
40.00 |
40.00 |
Solvent |
Figure 1. F1, F2 and F3 spray formulations.
METHODS:
After evaluating the mosquito repellent properties of various plant extracts and essential oils, a mosquito repellent spray was formulated.20,21
RESULT AND DISCUSSION:
Out of the three formulations, the F3 was found to have better efficacy than the other two. Thus, the evaluation tests were performed on the F3 spray.
Evaluation of Formulated Herbal Mosquito Repellent Spray:
1. (pH) Test: The pH of the spray was found to be 6.4, which is slightly acidic ensuring that the spray is gentle on the skin.
2. Skin irritancy test:
The spray was applied to a patch, and the patch was placed on the forearm for 48 hours, and a reaction was observed. After 24 hours, no irritation was observed on the surface of the skin.
Figure. 3. Skin Irritancy test.
3. Field Test:
All three formulations of herbal spray were evaluated. The best one is determined by applying formulated spray in places where there are many mosquitoes or in fields.
The below table indicates the number of mosquitoes that bite volunteers' hands and legs without spray (indoors and outdoors) and with spray (indoors and outdoors) within 6 hours for 2 days.
Table 3. Observation Table of Field Test
|
Day 1 |
Day 2 |
|||||||
|
Time |
Indoor |
Outdoor |
Indoor |
Outdoor |
||||
|
Without spray |
With Spray |
Without spray |
With Spray |
Without spray |
With Spray |
Without spray |
With Spray |
|
|
5.00 - 6.00 |
2 |
0 |
3 |
0 |
4 |
0 |
4 |
0 |
|
6.00 - 7.00 |
1 |
0 |
2 |
0 |
3 |
0 |
3 |
0 |
|
7.00 – 8.00 |
3 |
0 |
3 |
0 |
1 |
0 |
5 |
1 |
|
8.00 - 9.00 |
2 |
0 |
1 |
0 |
3 |
0 |
1 |
0 |
|
9.00 -10.00 |
1 |
0 |
0 |
0 |
2 |
0 |
2 |
0 |
|
10.00 -11.00 |
0 |
0 |
1 |
0 |
1 |
0 |
0 |
0 |
Field trials conducted over two days, with six-hour sessions each day, indicated a promising 90% efficacy for the mosquito repellent spray in deterring mosquitoes. However, it's essential to acknowledge that many variables can influence such results. Factors such as the specific mosquito species present, the quality and composition of the plant extracts and essential oils within the spray, and individual characteristics of test subjects, including age, sex, and unique biochemical profiles, could all impact the repellent's performance. Moreover, the production of essential oils, a cornerstone of many repellents, is a complex process influenced by various factors, including plant species, cultivation conditions, harvesting practices, storage methods, and extraction techniques. These variables can significantly impact the final product's potency and efficacy. Consequently, while the initial findings are encouraging, further research is warranted to fully understand the repellent's capabilities and limitations under diverse conditions.
Figure 2. Evaluation of spray
4. Public Survey:
A public survey of the final product was carried out on 20 volunteers to study the safety and efficacy of the formulation. The F3 spray was distributed among all volunteers, and feedback was collected.
Table 4. Observation Table of Public Survey
|
Rating |
Quality and usefulness |
Easy handling of product |
|
No. of Volunteers |
No. of Volunteers |
|
|
Excellent |
12 |
15 |
|
Very good |
7 |
4 |
|
Good |
1 |
1 |
|
Average |
0 |
0 |
|
Poor |
0 |
0 |
CONCLUSION:
The study successfully formulated and evaluated a herbal mosquito repellent spray using essential oils and plant extracts, demonstrating 90% efficacy in repelling mosquitoes in both indoor and outdoor field tests. The key ingredients, including citronella, eucalyptus, clove bud, sweet orange peel, neem extract, and lavender oil, contributed to the spray's effectiveness and pleasant fragrance. Out of the three formulations tested, the F3 spray had shown the most efficient response against mosquitoes. The public survey indicated high satisfaction with the product's quality, usefulness, and ease of handling. Given the increasing threat of mosquito-borne diseases and the harmful effects of synthetic repellents, this herbal formulation offers a promising, environmentally friendly, and safer alternative for mosquito control.22-26
CONFLICT OF INTEREST:
The authors have no conflicts of interest regarding this investigation.
ACKNOWLEDGMENTS:
The authors would like to thank Smt. Kashibai Navale College of Pharmacy for granting access to the necessary facilities to conduct this research.
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Received on 31.07.2024 Revised on 25.12.2024 Accepted on 06.03.2025 Published on 01.07.2025 Available online from July 05, 2025 Research J. Pharmacy and Technology. 2025;18(7):3340-3344. DOI: 10.52711/0974-360X.2025.00483 © RJPT All right reserved
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