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Journal of Drug Delivery and Therapeutics

Open Access to Pharmaceutical and Medical Research

Copyright   © 2022 The  Author(s): This is an open-access article distributed under the terms of the CC BY-NC 4.0 which permits unrestricted use, distribution, and reproduction in any medium for non-commercial use provided the original author and source are credited

Open Access  Full Text Article                                                                                                                                               Research Article 

Phytochemical Screening, GCMS Profile, and In-silico properties of Bioactive Compounds in Methanolic Leaf Extracts of Moringa oleifera

Kandeepan C.¹, Sabitha M.¹, Parvathi K.², Senthilkumar N.³, Ramya S.⁴, Boopathi NM.⁵, Jayakumararaj R.⁶*

¹ PG and Research Department of Zoology, Arulmigu Palaniandavar College of Arts and Culture, Palani – 624601, Dindigul District, TamilNadu, India

² Department of Zoology, Erode Arts and Science College (Autonomous), Erode – 638009, TN, India.

³ Institute of Forest Genetics and Tree Breeding (IFGTB), Indian Council of Forestry Research and Education (ICFRE), Coimbatore – 641002, TamilNadu, India 

⁴ PG Department of Zoology, Yadava College (Men), Thiruppalai - 625014, Madurai, TamilNadu, India

⁵ Centre for Plant Molecular Biology and Biotechnology, Department of Plant Biotechnology, Tamil Nadu Agricultural University, Coimbatore - 641 003, TN, India

⁶ Department of Botany, Government Arts College, Melur – 625106, Madurai District, TN, India

 

INTRODUCTION

Moringa oleifera Lam. (Family: Moringaceae) is a medicinal plant native to India, geographically distributed in tropical and sub-tropical climatic regions. However, has now been cultivated in other regions of the world¹. M. oleifera is considered to be ware-house of plant based natural products (PBNPs) further it has been recognized as economically and nutritionally important crop owing to its health benefits. The tree is endowed with splendid diversity and incredible richness of bioactive compounds that serve as main source of nutraceuticals in maintaining health and wellbeing and overcome the malnutrition problem. The use of this tree is encouraged as a nutritional supplement for infants and children². 

It has a wide range of culinary applications besides bioremediation, nutritional and medicinal properties³. Edible parts of this plant contain nutrients viz., proteins, essential and non-essential amino acids, vitamins, minerals, antioxidants and phenolic compounds. M. oleifera allele-chemicals [Amino Acids (Threonine, Methionine, Phenylalanine), Fatty Acids (Palmitic acid, Oleic acid, Linoleic acid), Phenols (Gallic acid, p-Coumaric acid, Ferulic acid) Flavonoids (Catechin, Quercetin, Kaempferol, Niazimicin) and other Bioactive Compounds, Vitamins (B, A, C, D and K)], Zeatin and Essential Macro (Potassium, Magnesium, Phosphorus) and Microelements (Iron, Zinc). Leaves, roots, seed, bark, fruit, flowers and immature pods of Moringa has been endowed with antioxidant, antidiabetic, antibacterial, antifungal, anti-tumor, anti-inflammatory, antiulcer, antispasmodic, diuretic, antihypertensive, hepatoprotective, antipyretic, antiepileptic, cardioprotective and cholesterol-lowering activities⁴. 

The plant is perennial tree; the fruit is known as DRUM-STICK and hence the plant is known as ‘drumstick tree’⁴. Though there are 13 species in the genus Moringa, M. oleifera is best known and probably the most widely distributed, and popular species due to its manifold uses⁵. In the meantime, it must be pointed out that the remaining 12 species in the genus have not yet been fully explored for their potential and medicinal properties⁵. This boils down to the fact that M. oleifera is the most extensively cultivated and undoubtedly exploited plant in the genus so far^6,7. 

Factually, M. oleifera is native to India, commonly found in the southern region of the India-Subcontinent and the South East Asian Region. It is cultivated in Sub-Himalayan North-Eastern Pakistan to West Bengal in India; Southern part of Deccan Plains, North-Western Gangetic Plains; Central India to Dry Regions of Peninsular India. It is also cultivated in other regions of the world including Asia, Africa, and Europe⁴. It is a deciduous tree with brittle stem, whitish-gray corky bark with branches; leaves pale green, bipinnate/ tri-pinnate with opposite, ovate leaflets⁸. This crop can be cultivated on marginal lands under water-scarcity and elevated temperatures⁹. Optimum growth is observed in soil with alkaline pH; temperature range of 25-35°C however, it can withstand elevated temperatures, and frost⁸. 

M. oleifera recognized as "The Miracle Tree" due to its versatile nutraceutical uses¹⁰. National Institute of Health (NIH) has declared this plant as “Botanical of the Year – 2007”³. The tree is a rich source of essential nutrients such as proteins, vitamins, minerals, carbohydrates, with a potential to overcome malnutrition. Apart from the nutritional aspect M. oleifera is a rich source of other phytochemicals. Nearly, all parts of the plant including leaves, roots, pods, seeds, and flowers have been explored for their nutraceutical properties and biomedical applications^2,8. 

Of all parts, the leaves are inexpensive and abundantly available but largely underutilized, ignored and often discarded. Different pharmaceutical products from this plant have been manufactured and marketed in both the Indian and worldwide markets due to these medicinal advantages. Recently, attempts have been made to evaluate the effect of processing methods on leaves to formulate Ready-to-Eat functional value-added foods. Furthermore, leaves are the most explored part of the tree due to the presence of nutrients, essential amino acids and minerals like Iron, Calcium, and Potassium¹¹. In addition, low calorific value of leaves makes them a suitable candidate for obese diet¹². Seeds have high lipid content (42%); however superior to soybean in composition⁸. 

M. oleifera has been traditionally utilized in folk remedies to cure conjunctivitis, and given to lactating mothers for enhancing milk production. The juice obtained from leaves is used to normalize blood pressure and blood glucose levels⁵. The accepted and safe dose of M. oleifera powder is 14 g daily¹³. Pharmacological studies have indicated that the extracts obtained from the plant have antioxidants¹⁴, anti-diabetic¹², anti-bacterial, anti-fungal¹⁵, and anti-carcinogenic¹⁶properties. 

No adverse effects have been reported on rabbits¹⁷ rats¹⁸ during toxicity study with aqueous leaf extract Moringa. Though, significant variation in composition of different species exists¹⁹ versatile nature of phytochemicals remains the key aspect of nutrition for people suffering from malnutrition and food security³. Due to overwhelming nutritive and medicinal use of the plant, it is indicated that Moringa can be widely exploited for its nutritionally important phytoconstituents in the development of functional foods, nutraceutical products and therapeutic agent on a commercial by fortification to eradicate malnutrition^20,21. Moringa possesses immense potential for betterment of nutrition, enhanced food security and promotes sustainable rural development. Prospecting BASM using in-silico ADMET predictions to chart a new road map for drug discovery is the basic aspect sustainable exploitation of bioactive natural products^22-27. The aim of this study is to identify phytochemicals by GC-MS analysis followed by. Compounds with maximum peak area in GCMS profile have been prospected for the molecular and biological properties so as to exploit them for the development of novel leads considering its nutritional, pharmaceutical and ethnopharmacological applications.

MATERIALS AND METHODS

Plant material: 

Fresh leaves of M. oleifera were collected from farmlands near Alagarkovil Reserve Forest (longitude/ latitude geographical coordinates 10.0748° N, 78.2131° E, Eastern Ghats) Dindigul District, Tamilnadu during Jun-Jul 2021, taken to laboratory, cleaned and preserved as Herbarium, part of the collected sample was shade dried, powdered and subjected to extraction. Botanical identity of the plant was established using flora and confirmed by Department of Botany, Government Arts College, Melur, Madurai, India. 

Botanical Description of the Plant

Habit: Trees to 12 m tall; Stem: Cylindrical; Bark: pale smooth to rugose but not fissured; Leaves: petiolate, 3-pinnate, 25-60 cm, stalked, glands often exuding clear or amber liquid at base of petiole and leaflets; Leaflets: 4-6 pairs, ovate, elliptic, or oblong, 1-2 × 0.5-1.2 cm, puberulous - young but glabrous - maturity, base rounded to cuneate, apex round to emarginate; petiolules slender, 1-2 mm; Inflorescence: widely spreading panicle; Bracteate: 10-30 cm; Bracts: linear, ca. 1 mm; Flowers: white to cream, fragrant, resembling an inverted Fabaceae flower with 2 dorsal sepals and 1 dorsal petal usually remaining un-reflexed and forming a projecting "keel" while perianth reflexes down to form a "banner" at right angles to the "keel", each flower borne on a false pedicel 7-15 mm; Pedicel: 1-2 mm; Sepals: lanceolate to linear-lanceolate, 0.7-1.4 mm, usually puberulent; Petals: spatulate, 1-2 cm, glabrous or puberulent at base; Stamens: hairy at base; Ovary: hairy. Fruits: Pod, Capsule 3-valved, 20-50 × 1-3 cm, dehiscent; Seeds: sub-globose, 3-angled, 8-15 mm in diam. excluding wings; wings 0.5-1 cm wide, rarely absent; Fl. - year round, Fr. - Jun-Dec²⁸  (Fig. 1).

Sample preparation

Using direct method of extraction, approximately 10 g of powder was extracted with 100 ml of methanol. The extract was transferred in to glass vials. The process was repeated 3 times with fresh solvent. The solvent was removed by Rota-vapor. The extracted residue was re-dissolved in the solvent to yield a final volume of 10mg/ml and the content was stored in cold (at 4°C) until further use.

Phytochemical Screening 

The methanolic extracts were subjected to chemical tests for the detection of phytoconstituents using standard procedures^29-33.  

Test for Phenols (FeCl₃ Test)

To 1 ml of the extract, 3 ml of distilled water followed by few drops of 10% aqueous Ferric chloride solution was added. Formation of blue or green colour indicates the presence of phenols. 

 

Test for Flavonoids (Shinoda Test)

To 2 ml of the extract, 1 ml of 1% ammonia solution was added. Appearance of yellow colour indicates the presence of flavonoids. 

Test for Tannins (FeCl₃ Test)

To 1 ml of the extract, 1 ml of 0.008 M Potassium ferricyanide was added and then add 1ml of 0.02 M Ferric chloride containing 0.1 N HCl. Appearance of blue-black colour indicates the presence of Tannins. 

Test for Alkaloids (Wagner’s Reagent Test)

Approximately, 1 ml of crude extract was mixed with 2 ml of Wagner’s reagent. Reddish brown colour precipitate indicates the presence of alkaloids. 

Test for Carbohydrates (Fehling’s test, Benedict’s test)

Fehling’s test 

Equal volume of Fehling A and Fehling B reagents were mixed together and then add 2ml of crude extract in it and gently boiled. A brick red precipitate appeared at the bottom of the test-tube indicates the presence of reducing sugars. 

Benedict’s test

1 ml of crude extract was mixed with 2ml of Benedict’s reagent and boiled. A reddish brown precipitate was formed which indicates the presence of the carbohydrates. 

Test for Proteins (Millon’s Test, Ninhydrin Test)

Millon’s test

1 ml of crude extract was mixed with 2ml of Millon’s reagent; white precipitate appeared which turned red upon gentle heating that confirmed the presence of protein. 

Ninhydrin test

1 ml of crude extract was mixed with 2ml of 0.2% solution of Ninhydrin and boiled. A violet colour precipitate was appeared suggesting the presence of amino acids and proteins. 

Test for Cardiac glycosides (Keller-Kiliani test) 

5 ml of extract was treated with 2 ml of glacial acetic acid containing one drop of ferric chloride solution. This was underlayed with 1 ml of concentrated sulphuric acid. A browning of the interface indicates a deoxy-sugar characteristic of carotenoids. A violet ring may appear below the brown ring, while in the acetic acid layer, a greenish ring may form just gradually throughout thin layer.

Test for Saponins (Foam Test)

2 ml of crude extract was mixed with 5 ml of distilled water in a test tube and it was shaken vigorously. Add some drops of olive oil. The formation of stable foam was taken as an indication for the presence of saponins.  

Test for Coumarin (Sodium hydroxide Test)

10 % Sodium hydroxide was added to the extract and chloroform was added. Formation of yellow color shows the presence of Coumarin.  

Test for Terpenoids (Salkowski test) 

5 ml of extract was mixed with 2 ml of chloroform and 3 ml of concentrated sulphuric acid was carefully added to form a layer. A reddish brown coloration of the inter face was formed which indicates the presence of terpenoids.  

Test for Steroids (Salkowski Test)

2 ml of acetic anhydride was added to 0.5 ml of crude extract containing 2 ml of sulphuric acid. The colour changed from violet to blue or green in samples indicates the presence of steroids. 

Test for Quinones (Sodium hydroxide Test)

Diluted sodium hydroxide was added to the 1 ml of crude extract. Blue green or red coloration indicates the presence of quinones. 

Test for Anthraquinones (Borntragers test) 

0.5 g of extract was boiled with 10% hydrochloric acid for few minutes in water bath. It was filtered and allowed to cool. Equal volume of CHCl₃ was added to the filtrate. Few drops of 10% ammonia was added to the mixture and heated. Formation of rose – pink color indicates of n-hexane, chloroform, ethyl acetate and methanol of the presence of the anthroquinones.

GC-MS Analysis 

Leaf samples of were collected from farmlands near Alagarkovil Reserve Forest (longitude/ latitude geographical coordinates 10.0748° N, 78.2131° E, Eastern Ghats) Dindigul District, Tamil Nadu, India. Phyto-components were identified using GC–MS detection system as described previously²⁶, however with modification, whereby portion of the extract was analysed directly by headspace sampling. GC–MS analysis was accomplished using an Agilent 7890A GC system set up with 5975C VL MSD (Agilent Technologies, CA, and USA). Capillary column used was DB-5MS (30 m × 0.25 mm, film thickness of 0.25 μm; J&W Scientific, CA, USA). Temperature program was set as: initial temperature 50°C held for 1 min, 5°C per min to 100°C, 9°C per min to 200°C held for 7.89 min, and the total run time was 30 min. The flow rate of helium as a carrier gas was 0.811851 mL/ min. MS system was performed in electron ionization (EI) mode with Selected Ion Monitoring (SIM). The ion source temperature and quadruple temperature were set at 230°C and 150°C, respectively. Identification of phyto-components was performed by comparison of their retention times and mass with those of authentic standards spectra using computer searches in NIST 08.L and Wiley 7n.l libraries²⁷. 

RESULTS 

Phytochemical analysis of MLEMO revealed the presence of alkaloids, carbohydrates, coumarins, flavonoids, glycosides, phenol, proteins, quinones, saponins, steroids, tannins and terpenoids. However, anthraquinones were not detected in the samples analyzed (Table 1). GCMS analysis revealed the presence of the following 41 phyto-compounds (listed in the decreasing order of abundancy) 1,3-Propanediol, 2-ethyl-2-(hydroxymethyl)- (C₆H₁₄O₃) 21.190; Propanoic acid, 2-methyl-, octyl ester (C₁₂H₂₄O) 15.027; 9,12,15-Octadecatrienoic acid, (Z,Z,Z)- (C₁₉H₃₀O₂) 10.006; Ethanamine, N-ethyl-N-nitroso- (C₄H₁₀N₂O) 5.216; 4H-Pyran-4-one, 2,3-dihydro-3,5-dihydroxy-6-methyl- (C₆H₈O₄) 4.180; Benzeneacetonitrile, 4-hydroxy- (C₈H₇NO) 3.476; 3-Deoxy-d-mannoic lactone (C₆H₁₀O₅) 3.294; n-Hexadecanoic acid (Palmitic acid) (C₁₆H₃₂O₂) 2.570; Monomethyl malonate (C₄H₆O₄) 2.568; Dihydroxyacetone (C₃H₆O₃) 2.465; Inositol (C₆H₁₂O₆) 2.054; 1,7-Diaminoheptane (C₇H₁₈N₂) 1.799; 9-Octadecenamide, (Z)- (C₁₈H₃₅NO) 1.469; 1,2,3-Propanetriol, 1-acetate (C₅H₁₀O₄) 1.437; Octadecanoic acid (C₁₈H₃₆O₂) 1.205; 9-Octadecenoic acid (Z)-, methyl ester (C₁₉H₃₆O₂) 1.030; (1S)-Propanol, (2S)-[(tert.butyloxycarbonyl)amino]-1-phenyl- (C₁₇H₂₆N₂O) 0.967; Phytol (C₂₀H₄₀O) 0.966; Oxazolidine, 2-ethyl-2-methyl- (C₆H₁₃NO) 0.900; Hexadecanoic acid, Methyl palmitate (C₁₇H₃₄O₂) 0.851; 4,5-Diamino-6-hydroxypyrimidine (C₄H₆N₄O) 0.643; Benzyl .β.-d-glucoside (C₁₃H₁₈O₆) 0.605; 3-Piperidinol (C₅H₁₁NO) 0.597; Formamide, N,N-dimethyl- (C₃H₇NO) 0.584; d-Talonic acid lactone (C₆H₁₀O₆) 0.565; 2-Oxoglutaric acid (C₅H₆O₅) 0.546; 1,3-Benzenediol, 2-methyl- (C₇H₈O) 0.545; N-Methoxy-1-ribofuranosyl-4-imidazolecarboxylic amide (C₁₀H₁₅N₃O₆) 0.534; 1-Nitro-.β.-d-arabinofuranose, tetraacetate (C₁₃H₁₇NO₁₁) 0.511; Sorbitol (C₆H₁₄O₆) 0.482; Cyclohexanemethanol, alpha-methyl-4-(1-methylethyl)- (C₁₁H₂₂O) 0.482; N,N-Dimethylacetamide (C₄H₉NO) 0.454; 2(4H)-Benzofuranone, 5,6,7,7a-tetrahydro-4,4,7a-trimethyl- (C₁₁H₁₆O₃) 0.449; 6-methoxy-3-Pyridinecarboximidamide (C₇H₉N₃O) 0.429; 4-Allyl-3-(dimethylhydrazono)-2-methylhexane-2,5-diol (C₁₂H₂₄N₂O₂) 0.425; Benzenebutanal, .gamma.,4-dimethyl- (C₁₂H₁₆O) 0.395; D-erythro-Pentose, 2-deoxy- (C₅H₁₀O) 0.331; Furan, 2,3-dihydro-4-methyl- (C₅H₈O) 0.210; 3,4-Furandiol, tetrahydro-, trans- (C₄H₈O₃) 0.174; 4,6-dimethyl-2-propyl-1,3,5-dithiazinane (C₈H₁₇NS₂) 0.169; 1,8-Diamino-3,6-dioxaoctane (C₆H₁₆N₂O₂) 0.117 (Table 2). Calculated values pertaining to the molecular properties and predicted bioactivity scores relating functions of dihydroxyacetone phosphate; methylmalonic acid;  1,3-Propanediol, 2-ethyl-2-(hydroxymethyl)-; Benzeneacetonitrile, 4-hydroxy-; Ethanamine, N-ethyl-N-nitroso-; Propanoic acid, 2-methyl-, octyl ester; 3-Deoxy-d-mannoic lactone; Inositol; n-Hexadecanoic acid (Palmitic acid); 9,12,15-Octadecatrienoic acid, (Z,Z,Z)- is provided in Fig. 2 A-K with their 3D structure. Data indicate that 9,12,15-Octadecatrienoic acid in the MLEMO holds significant GPCR ligand (0.33); Ion channel modulator (0.23); Kinase inhibitor (0.19); Nuclear receptor ligand (0.35); Protease inhibitor (0.13); Enzyme inhibitor (0.42) activity respectively (Table 3K). 

DISCUSSION

Medicinal and biological activities of plant extract have been upheld by in-vitro assays^26,33-42. In the leaf extracts of M. oleifera contains significantly high phenolic compounds primarily responsible for antioxidant effects^14,33,36. Most of the compounds identified in methanolic leaf extract of M. oleifera ­are endowed with medicinal properties and some of them are commonly present in many other medicinal plants used by local tribal people^33-42. Dihydroxyacetone (DHA), rarely been detected in plant extracts have various applications; DHA is primarily used as an ingredient in sunless tanning products, synthesis of polymeric biomaterials⁴³. 4H-Pyran-4-one, 2,3-dihydro-3,5-dihydroxy-6-methyl- (DDMP) is a strong anti-oxidant in glucose–histidine Maillard reaction products⁴⁴. Li et al.⁴⁵ indicated that the control of DDMP formation in the Maillard reaction is important to improve the thermally treated food quality as a result of its intense bitterness and potential toxicity. 

M. oleifera leaves were reported to have anti-cancerous activity against HeLa cells by activating the apoptotic pathway⁴⁶. Further, it has been reported that Moringa leaves extract induce apoptosis by up-regulating BAX and down-regulating BCL-2 expression, enhancing caspase-3-activity. Palmitic acid present in leaves inhibit cancer cell growth⁴⁷. In G1 phase of cell cycle, D-allose present in MLEMO induces specific thioredoxin interacting protein (TXNIP) and stabilizes p27kip1 protein that inhibits cancer cells growth without affecting the normal cells in the system. As of now, there are evidences in the literature that support the fact that chronic inflammation may lead to malignancies of different organs including stomach, colon, breast, skin, prostate, pancreas⁴⁸. Inhibition of NF-κB (nuclear factor kappa-light-chain-enhancer of activated B cells) signaling has potential therapeutic application in cancer therapy and management of metabolic inflammations. 

Likewise, 4H-Pyran-4-one, 2,3-dihydro-3,5-dihydroxy-6-methyl has been reported to suppresses the NF-κB target antiapoptotic genes Bcl-2 (B-cell lymphoma 2) while it induces expression of apoptotic genes Bax (Bcl-2-associated X protein), cleaved caspase-3 and cleaved PARP (Poly ADP-ribose polymerase). Methyl palmitate possesses a strong anti-fibrotic effect⁴⁸. It inhibits NF-κB and consequent pro-inflammatory and oxidative stress response. Conjugated linoleic acids in methanolic leaf extracts of Moringa significantly decrease prostate cancer cell proliferation by down-regulating phorbol ester induced NFκB activation and subsequent COX-2(Cyclooxygenase2) expression^48,49. Likewise, 9-Octadecenamide, an amide of oleic acid suppresses lipopolysaccharide induced expression of iNOS (inducible NO synthase) and COX-2 through inhibition of NF-kappa-B activation. A rationale for design and evaluation of novel antioxidant drug for atherosclerosis⁵⁰ wherein it has been pointed out that Hexadecanoic acid, Methyl palmitate displays antioxidant properties that reduce atherosclerosis significantly. 

CONCLUSION

Therapeutic mechanism of a plant can be better understood with a proper investigation of its bioactive secondary metabolites. M. oleifera leaves remain an ideal sources of micro-nutrients and phytochemicals that can be used for the development of nutraceuticals and functional foods. Moringa leaves contain phytochemicals, which makes this plant ideal source of BASM. Therefore, food products based on the leaves contain more protein, dietary fibers, other nutrients, and antioxidants. The compounds identified by the GC-MS analysis of methanolic leaf extracts of M. oleifera in the present study relate their applications in folklore medicine. BASMs in Moringa prompt inspiration for further investigation to identify novel lead molecules in the drug discovery and design of novel herbal drugs of GRAS standard.

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23. Kalaimathi RV, Jeevalatha A, Basha AN, Kandeepan C, Ramya S, Loganathan T, Jayakumararaj R. In-silico Absorption, Distribution, Metabolism, Elimination and Toxicity profile of Isopulegol from Rosmarinus officinalis. Journal of Drug Delivery and Therapeutics. 2022; 12(1):102-8. https://doi.org/10.22270/jddt.v12i1.5188

24. Jeevalatha A, Kalaimathi RV, Basha AN, Kandeepan C, Ramya S, Loganathan T, Jayakumararaj R. Profile of bioactive compounds in Rosmarinus officinalis. Journal of Drug Delivery and Therapeutics. 2022; 12(1):114-22. https://doi.org/10.22270/jddt.v12i1.5189

25. Ramya S, Murugan M, Krishnaveni K, Sabitha M, Kandeepan C, Jayakumararaj R. In-silico ADMET profile of Ellagic Acid from Syzygium cumini: A Natural Biaryl Polyphenol with Therapeutic Potential to Overcome Diabetic Associated Vascular Complications. Journal of Drug Delivery and Therapeutics. 2022; 12(1):91-101. https://doi.org/10.22270/jddt.v12i1.5179

26. Soorya C, Balamurugan S, Ramya S, Neethirajan K, Kandeepan C, Jayakumararaj R. Physicochemical, ADMET and Druggable properties of Myricetin: A Key Flavonoid in Syzygium cumini that regulates metabolic inflammations. Journal of Drug Delivery and Therapeutics. 2021; 11(4):66-73 https://doi.org/10.22270/jddt.v11i4.4890

27. Sabitha M, Krishnaveni K, Murugan M, Basha AN, Pallan GA, Kandeepan C, Ramya S, Jayakumararaj R. In-silico ADMET predicated Pharmacoinformatics of Quercetin-3-Galactoside, polyphenolic compound from Azadirachta indica, a sacred tree from Hill Temple in Alagarkovil Reserve Forest, Eastern Ghats, INDIA. Journal of Drug Delivery and Therapeutics. 2021; 11(5-S):77-84. https://doi.org/10.22270/jddt.v11i5-S.5026

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29. Loganathan T, Barathinivas A, Soorya C, Balamurugan S, Nagajothi TG, Jayakumararaj R. GCMS Profile of Bioactive Secondary Metabolites with Therapeutic Potential in the Ethanolic Leaf Extracts of Azadirachta indica: A Sacred Traditional Medicinal Plant of INDIA. Journal of Drug Delivery and Therapeutics. 2021; 11(4-S):119-26. https://doi.org/10.22270/jddt.v11i4-S.4967

30. Krishnaveni K, Sabitha M, Murugan M, Kandeepan C, Ramya S, Loganathan T, Jayakumararaj R. vNN model cross validation towards Accuracy, Sensitivity, Specificity and kappa performance measures of β-caryophyllene using a restricted-unrestricted applicability domain on Artificial Intelligence & Machine Learning approach based in-silico prediction. Journal of Drug Delivery and Therapeutics. 2022; 12(1-S):123-31.

31. Ramya S, Jepachanderamohan PJ, Kalayanasundaram M, Jayakumararaj R. In vitro antibacterial prospective of crude leaf extracts of Melia azedarach Linn. against selected bacterial strains. Ethnobotanical Leaflets. 2009; 2009(1):32.

32. Kandeepan C, Kalaimathi RV, Jeevalatha A, Basha AN, Ramya S, Jayakumararaj R. In-silico ADMET Pharmacoinformatics of Geraniol (3, 7-dimethylocta-trans-2, 6-dien-1-ol)-acyclic monoterpene alcohol drug from Leaf Essential Oil of Cymbopogon martinii from Sirumalai Hills (Eastern Ghats), INDIA. Journal of Drug Delivery and Therapeutics. 2021; 11(4-S):109-18. https://doi.org/10.22270/jddt.v11i4-S.4965

33. Ramya S, Neethirajan K, Jayakumararaj R. Profile of bioactive compounds in Syzygium cumini-a review. Journal of Pharmacy research. 2012; 5(8):4548-53.

34. Ramya S, Krishnasamy G, Jayakumararaj R, Periathambi N, Devaraj A. Bioprospecting Solanum nigrum Linn.(Solanaceae) as a potential source of Anti-Microbial agents against selected Bacterial strains. Asian Journal of Biomedical and Pharmaceutical Sciences. 2012; 2(12):65.

35. Sundari A, Jayakumararaj R. Medicinal plants used to cure cuts and wounds in Athur region of Thoothukudi district in Tamil Nadu, India. Journal of Drug Delivery and Therapeutics. 2020; 10(6-s):26-30. https://doi.org/10.22270/jddt.v10i6-s.4429

36. Sundari A, Jayakumararaj R. Herbal remedies used to treat skin disorders in Arasankulam region of Thoothukudi District in Tamil Nadu, India. Journal of Drug Delivery and Therapeutics. 2020; 10(5):33-8. https://doi.org/10.22270/jddt.v10i5.4277

37. Rajasekaran C, Meignanam E, Vijayakumar V, Kalaivani T, Ramya S, Premkumar N, Siva R, Jayakumararaj R Investigations on antibacterial activity of leaf extracts of Azadirachta indica A. Juss (Meliaceae): a traditional medicinal plant of India. Ethnobotanical leaflets. 2008; 2008(1):161.

38. Ramya S. Jayakumararaj R In Vitro Evaluation of Antibacterial Activity Using Crude Extracts of Catharanthus roseus L.(G.) Don. Ethnobotanical Leaflets. 2008; 2008(1):140.

39. Shanmugam S, Sundari A, Muneeswaran S, Vasanth C, Jayakumararaj R, Rajendran K. Ethnobotanical Indices on medicinal plants used to treat poisonous bites in Thiruppuvanam region of Sivagangai district in Tamil Nadu, India. Journal of Drug Delivery and Therapeutics. 2020; 10(6-s):31-6. https://doi.org/10.22270/jddt.v10i6-s.4432

40. Ramya S, Alaguchamy N, Maruthappan VM, Sivaperumal R, Sivalingam M, Krishnan A, Govindaraji V, Kannan K, Jayakumararaj R. Wound healing ethnomedicinal plants popular among the Malayali tribes in Vattal Hills, Dharmapuri, TN, India. Ethnobotanical Leaflets. 2009; 2009(10):6.

41. Sivaperumal R, Ramya S, Ravi AV, Rajasekaran C, Jayakumararaj R. Herbal remedies practiced by Malayali to treat skin diseases. Environ We Int J Sci Tech. 2009; 4(1):35-44.

42. Kadhirvel K, Ramya S, Sudha TS, Ravi AV, Rajasekaran C, Selvi RV, Jayakumararaj R. Ethnomedicinal survey on plants used by tribals in Chitteri Hills. Environ We Int J Sci Tech. 2010; 5:35-46.

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45. Li H, Wu CJ, Tang XY, Yu SJ. Insights into the Regulation Effects of Certain Phenolic Acids on 2, 3-Dihydro-3, 5-dihydroxy-6-methyl-4 (H)-pyran-4-one Formation in a Microaqueous Glucose-Proline System. Journal of agricultural and food chemistry. 2019; 67(32):9050-9. https://doi.org/10.1021/acs.jafc.9b01182

46. Nair S., Varalakshmi K.N. Anticancer, Cytotoxic Potential of Moringa oleifera Extracts on HeLa Cell Line. J. Nat. Pharm. 2011; 2:138-142. https://doi.org/10.4103/2229-5119.86260

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