Phytochemical Testing of Azadirachtolide indica

 Corresponding author: Harsh Suryakant Chavan Copyright © 2025 Author(s) retain the copyright of this article. This article is published under the terms of the Creative Commons Attribution Liscense 4.0. Phytochemical Testing of Azadirachtolide indica Harsh Suryakant Chavan 1, *, Prachi Nandkumar Padwal 2, Chaitali Mohan Barhate 1 and Aditya Sampat Dere 1 1 Student of Samarth Institute of Pharmacy, Belhe, Pune, Maharashtra, India. 2 Assistant Professor, Department of Quality Assurance Technique, Samarth Institute of Pharmacy, Belhe, Pune, Maharashtra, India. World Journal of Biology Pharmacy and Health Sciences, 2025, 24(02), 210-214 Publication history: Received on 26 September 2025; revised on 03 November 2025; accepted on 06 November 2025 Article DOI: https://doi.org/10.30574/wjbphs.2025.24.2.0980 Abstract Neem Azadirachta indica is a useful traditional medicinal plant growing in Nigeria, India, and America. The phytochemicals and the biopesticide components present were ascertained. The results showed that saponins, steroids and terpenes were mostly present, while tannins and glycosides were moderately present, and alkaloids, flavonoids, phenols and oxalic acid were least present. The presence of this phytochemical could account for the therapeutic uses of neem. Keywords: Bio Pesticidal; Neem; Phytochemicals; Therapeutic 1. Introduction Neem plant is a native of India, where it is known as divine tree; “life giving tree”. It belongs to malice family. Away From India, it is commonly found in Africa and America. It occurs naturally in tropical region and sub-tropical zones. However, it can still be planted or cultivated. Neem tree is an incredible therapeutic plant that has been declared the Tree of the 21st century by the United Nations Numerous organic compounds found in the plant kingdom have been Figure 1 Picture of Azadirachtolide indica Crude Plant employed for medical purposes and may be the basis for the creation of new drugs with high efficacy in treating a range of pathological disorders in the years to come. The neem plant is regarded as one of the most abundant sources of pharmaceutical intermediates, chemical entities, nutraceuticals, dietary supplements, folk medicine, and medications World Journal of Biology Pharmacy and Health Sciences, 2025, 24(02), 210-214 211 for both modern and traditional medicine. Regarding synthetic drugs. Through extensive research, a few of the phytochemicals found in neem plants have been identified, separated, and measured. These bioactive chemicals have provided leads in the development of several life-Saving drugs, which are in use today. Ancient medical literature generally advocate extract from Azadi Acta indica, also known as Dogon Yaro in some regions of Nigeria. The leaves are used as a medication for fever, eczema, and diabetes. Determining the phytochemical components of the neem plant and connecting them to some of its traditional uses was the aim of this study. 1.1. Plant Profile Table 1 Plant Profile kingdom plantae order Rurales suborder Retinae family Melicia Sub family Meloidae Tribe Melee genus Azadirachtolide species Indica 1.2. Chemical constituents Azedarach indica L. (neem) shows therapeutics role in health management due to rich source of various types of ingredients. . Azadirachtin is the primary active ingredient, followed by sodium nominate, gaduin, solanine, quercetin, namonin, nimbi, nimbi din, and libido. Leaves contain ingredients such as nimbi, immanence, 6-desacetylnimbinene, Nimba diol, imboiled, ascorbic acid, n-hexaconazole and amino acid, 7-desacetyl-7-benzoylazadiradione, 7-desacetyl-7benzoylgedunin, 17-hydroxyazadiradione, and nimbies. Fresh neem leaves were used to purify the polyphenolic flavonoids quercetin and ß-sitosterol, which were shown to have antibacterial and antifungal qualities. The seeds of the plant also contain important components including gaduin and azadirachtin. 1.3. Medicinal uses • Treats Acne: -Neem has an anti-inflammatory property which helps reduces acne. Azedarach Indica also helps reduce skin blemishes. • Nourishes Skin: -Neem is a rich source of Vitamin E which help repair damaged skin cells. • Treats Fungal Infections: -Neem has scientifically proven antifungal property which helps treat fungal infections. • Useful in Detoxification: -Neem can prove useful in detoxification both internally and externally. Consumption of neem leaves or powder stimulates kidneys and liver increasing the metabolism and eliminating the toxins out of the body. Externally, neem scrubs or paste can be used to remove germs, bacteria, dirt, etc from your skin preventing rashes and skin diseases. • Increase immunity: -Neem's antibacterial and antimicrobial properties are well-known. These qualities are very important for increasing immunity. • Insect & Mosquito Repellent: - To keep the insects away, you can burn some neem leaves. Additionally, this works well against certain mosquito species. Neem is the most effective home remedy for treating the early stages of malaria among all of them. • Guards Against Digestive Disorders: -Because of neem's anti-inflammatory qualities, a number of gastrointestinal disorders, including flatulence, stomach ulcers, and constipation, are less likely to occur. • Handles Injuries: - Neem leaves are used to treat wounds because of their antibacterial qualities. • Reduces Dandruff: - Shampoos and conditioners frequently include neem. The antifungal and antibacterial qualities of Azedarach Indica assist to strengthen your hair and get rid of dandruff. • Lessens Joint Pain: - Application of neem oil or extract on the afflicted region might help alleviate pain and discomfort. As a result, it is frequently used to treat arthritis. • Skin exfoliation: - Neem works wonders as an exfoliator. It aids in clearing the skin's surface of dead cells, which inhibits the development of imperfections. World Journal of Biology Pharmacy and Health Sciences, 2025, 24(02), 210-214 212 2. Method and material • Material: -fresh neem leaves, grinder, distilled water, conical flask, filter paper, fan, bowl, Soxhlet apparatus, methanol, freezer. • Method: --The collected Azedarach indica leaves were washed under running tap water and rinsed with sterile distilled water to remove a dust and debris. The leaves were separated out from each after wash, segregated and shed dried. The shed dried leaves were powdered using a dry mechanical grinder. The dried powder was passed through the mesh sieve to obtain the fine powder. The fifty grams of leaves powder was added with 250 ml of distilled water. After being securely sealed, the conical flask was allowed to macerate for 48 hours at room temperature. During this time, the contents were shaking every two hours. A muslin cloth was used to filter the contents of the conical flask. Using the small amount of filtrate that was left in the flask, the contents were rinsed twice before being filtered using 1 filter paper one more. The filtrate was moved to a sterile evaporating bowl and left there with a fan to allow the solvent to evaporate. The extract that remained in the bowl after the solvent had completely evaporated This following the solvent's full evaporation. • Methanolic ExtractSoxhlet apparatus was used for extraction purpose with 80% methanol. Different concentration of plant material and solvent were taken. After filling the Soxhlet apparatus with plant material and solvent it was run at 60°C until it gets colorless and continuous flows of water to cool down the condenser. Finally, the extract was collected in airtight bottles and stored at 4°C. 2.1. Preliminary phytochemical screening Detect the occurrence of different natural substances in the plant. The plant material underwent extraction. Subsequently, these extracts were analyzed using specific phytochemical tests to identify the presence of compounds like sugars, starch, polyphenols, saponins, mucilage, steroids, alkaloids, and flavonoids. Table 2 Tests for phytochemical screening Test OBSERVATION INFERENCE (1) Test for alkaloids: a) Dragados’s reagent test: b) Wagner’s reagent test Orange to red colour Brown to flocculent precipitation Presence of alkaloids Presence of alkaloids (2) Test for glycosides: (a)Benedict’s reagent test appearance of brownish to red colour presence of glycoside. (3). TEST FOR DETECTION OF PROTEINS: (a)Xanthopterin test: (b)Biuret test: appearance of white or yellow Precipitate development of violet to pink colour. Presence of proteins presence of proteins 4. TEST FOR DETECTION OF TANNINS: (a)Lead acetate test: (b)Ferric chloride test formation of precipitation development of green colour presence of tannins. presence of tannins. (5) Test for Flavonoids: Pale yellow brown colour Presence of flavonoids (6) TEST FOR PHENOLS Violet colour Presence of phenols (7) TEST FOR SAPONIN Formation of froth Presence of saponin 2.2. Development of Thin Layer Chromatography TLC analytical plates covered with 0.2 mm thick silica gel-G were used to evaluate the methanolic extracts. A 4:1:1:1 v/v mixture of butanol, acetic acid, and water was employed as the solvent system. The capillary action causes this combination to migrate on the silica-coated plates. After the coated plate was fully produced, it was allowed to air dry World Journal of Biology Pharmacy and Health Sciences, 2025, 24(02), 210-214 213 before being heated for 20 to 25 minutes. To find the spots, a 0.2% freshly made ninhydrin solution was sprayed onto the plate. Its retention factor (Rf) represented the spots' mobility. 3. Result According to the review, azadirachtolide indica has a wide range of pharmacological effects since it is a rich source of bioactive substances such terpenoids, flavonoids, glycosides, and tannins. The presence of these phytoconstituents in different plant portions was verified by extraction and chromatographic investigations. These substances have strong analgesic, hepatoprotective, antibacterial, antioxidant, and anti-inflammatory properties. The results indicate Azadirachtolide Indica’s potential for creating standardized herbal formulations and justify its traditional applications in Ayurvedic, Siddha, and Unani systems. 4. Conclusion With an emphasis on recent studies, this report summarizes the body of information regarding Azadirachtolide indica. Ayurveda and traditional medicine have made considerable use of the rhizome and its preparations for a variety of therapeutic uses. Azadirachtolide indica is a well-known Ayurvedic remedy with a variety of therapeutic uses that have been reported. Its conventional claims cannot be verified without scientific validation. More research is necessary to examine its therapeutic potential utilizing contemporary methods and parameters, given its significance in traditional Indian medicine. We can then fully utilize this herb’s advantages for humanity Compliance with ethical standards Acknowledgments I would like to express my profound gratitude to my mentors and colleagues for their invaluable guidance and insightful comments during the preparation of this review. Special thanks to Samarth Institute of Pharmacy (junior) for providing the necessary resources and support. I also appreciate the constructive feedback from the anonymous reviewers, which significantly improved the quality of this article. Any errors or omissions are my own responsibility. Disclosure of conflict of interest No conflict of interest to be disclosed. References [1] Evaluation And Determination of Azadirachtin and Fatty Acid Constants from The Leaves of Azadirachta Indica, Research Paper | Open Access Volume 02 | Issue 12 | Article Id IJPS/240211126 [2] Therapeutics Role of Azadirachta indica (Neem) and Their Active Constituents in Diseases Prevention and Treatment Hindawi Publishing Corporation Evidence-Based Complementary and Alternative Medicine Volume 2016, Article ID 7382506, 11 pages http://dx.doi.org/10.1155/2016/7382506 [3] Zong A., Cao H., Wang F. Anticancer polysaccharides from natural resources: a review of recent research. Carbohydrate Polymers. 2012;90(4):1395–1410. Doi: 10.1016/j.carbpol.2012.07.026. [DOI] [PubMed] [Google Scholar] [4] Efferth T., Koch E. Complex interactions between Phytochemicals. The Multi-Target Therapeutic concept of Phytotherapy. Current Drug Targets. 2011;12(1):122–132. Doi: 10.2174/138945011793591626. [DOI] [PubMed] [Google Scholar] [5] Al-Bukhari M. I., Al-Bukhari S. The Collection of Authentic Sayings of Prophet Mohammad (Peace Be upon Him), Division 71 on Medicine. 2nd. Ankara, Turkey: Hilal Yayinlari; 1976. [Google Scholar] [6] Brahmachari G. Neem—an omnipotent plant: a retrospection. ChemBioChem. 2004;5(4):408–421. Doi: 10.1002/cbic.200300749. [DOI] [PubMed] [Google Scholar] [7] 5.Ketkar A. Y., Ketkar C. M. Various uses of neem products. In: Schmutterer H., editor. The Neem Tree. Weinheim, Germany: John Wiley & Sons; 2004. Pp. 518–525. [DOI] [Google Scholar] World Journal of Biology Pharmacy and Health Sciences, 2025, 24(02), 210-214 214 [8] Govindachari T. R., Suresh G., Gopalakrishnan G., Banumathy B., Masilamani S. Identification of antifungal compounds from the seed oil of Azadirachta indica . Phytoparasitica. 1998;26(2):109–116. Doi: 10.1007/bf02980677. [DOI] [Google Scholar] [9] Phytochemicals of neem plant (Azadirachta indica) explains its use in traditional medicine and pest control Article DOI: https://doi.org/10.30574/gscbps.2021.14.2.0394