Available online on 15.01.2022 at http://jddtonline.info

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 

Profile of bioactive compounds in Rosmarinus officinalis 

A. Jeevalatha¹, R.V. Kalaimathi², A.N. Basha², C. Kandeepan²*, S. Ramya³, T. Loganathan⁴, R. Jayakumararaj⁵

¹ Department of Zoology, GTN College, Dindigul, TamilNadu, India

² Department of Zoology, Arulmigu Palani Andavar College of Arts & Culture, Palani-624601, TN, IN

³ The Thavaram Trust, Thiruppalai - 625014, Madurai, TamilNadu, India

⁴ Department of Plant Biology & Biotechnology, LN Government College (A), Ponneri-601204, TN, IN

⁵ Department of Botany, Government Arts College, Melur – 625106, Madurai, TamilNadu, India

 

 

INTRODUCTION 

Rosemary (Rosmarinus officinalis L.), belongs to family Lamiaceae, is aromatic, evergreen, usually erect, bushy shrub up to 2m tall and wide. Stem indistinctly quadrangular finely grey pubescent. Leaves opposite, tufted on the branches, sessile to short, petiolate, blade linear, 1-5cm×1-2mm, base attenuate, margin entire but revolute, apex obtuse, leathery, dark glossy, sea green and sub glabrous above, white felted tomentose beneath, aromatically fragrant when crushed. Inflorescence Racemose, axillary 5-10 flowered, 0.5-2.5 cm long, terminating short lateral branches, pedicle 2-5mm long, calyx campanulate, 2-lipped, 5-6mm long, densely stellate tomentose, upper lip small and 3 dentate, lower lip 2–lobed, corolla tabular, 2 lipped, 10- 13 mm long, pale blue or white, upper lip or curved , 2-lobed, ovate, about 4mm long, lower lip 3-lobed above 7mm long, with large concave middle lobe; 2 anterior stamens perfect, 7-8 mm long, ascending under the base of the upper lip, two posterior stamens reduced to hardly visible staminodes, pistil with deeply 4-parite ovary style incurved, 1.5 cm long ending into 2 short, unequal branches with stigma. Fruit composed of 4sub-globose to obovoid nut-lets, above 2mm long, glabrous and smooth; Flowering – Nov - Jan^1,2. 

Since antiquity foliage is used as a common household culinary spice for flavouring. Rosemary extracts, derived from leaves, are used as flavouring and antioxidant agents in food processing and cosmetics. Rosemary has been used in traditional and complementary alternative medicine for its digestive, tonic, astringent, diuretic, and diaphoretic properties. It has been linked to a broad range of beneficial health benefits³. 

Main constituents of ROEO are camphor (5.0–21%), 1,8-cineole (15–55%), α-pinene (9.0–26%), borneol (1.5–5.0%), camphene (2.5–12%), β-pinene (2.0–9.0%) and limonene (1.5–5.0%) in proportions that vary according to the vegetative stage and bioclimatic conditions⁴. Rosemary has long been used in traditional medicine to cure a variety of ailments⁵. Phytochemicals in R. officinalis include rosmarinic acid, camphor, caffeic acid, ursolic acid, betulinic acid, carnosic acid and carnosol⁶. ROEO composed of phenolic compounds, di and triterpenes and essential oils. In traditional medicine ROEO is used to treat wounds, rashes, headache, dyspepsia, circulation problems, and as expectorant, diuretic and anti-spasmodic in treatment of renal colic⁷ which is attributed to the class of chemical compounds. Polyphenols are antioxidants primarily responsible for fruit colouring, and are classified as phenolic acids, flavonoids and non-flavonoids. Epidemiology evidence indicates that a diet rich in antioxidant fruits and vegetables significantly reduces the risk of many oxidative stress related diseases viz. cancers, diabetes and cardiovascular diseases (CVD). In addition to their antioxidant properties, ROEO play a very important role in plant defences against herbivores, pathogens and predators; therefore, used to control infectious agents in humans. Polyphenols in ROEO are apigenin, diosmin, luteolin, genkwanin and phenolic acids (>3%), especially rosmarinic acid, chlorogenic acid and caffeic acid. furthermore, biological activities exhibited by phenolic acids in There are numerous epidemiological and experimental evidences present describing the protective role of phenolic acids in degenerative diseases such as cardiovascular, cancer, diabetes, inflammation and many more⁹. The ability of plant secondary metabolites depends on the bioavailability which accounts for the proportion of their absorption, digestion, and metabolism after entering in the circulation system^10-28.

MATERIALS AND METHODS

Collection of Plant material: Rosmarinus officinalis L. (Rosemary)  leaves were collected from Palani Hills, Western Ghats (2000 m above the mean sea level), and identity of the plant was confirmed by Botanical Survey of India, Southern circle, Coimbatore, Tamil Nadu. The collected leaves samples were rinsed with tap water dried and powdered and then stored at 4 °C. Plant extracts preparation 5g of each sample of R. officinalis was extracted with 100 ml of methanol using Soxhlet apparatus. The extract was filtered and methanol was evaporated by rotary evaporator and then stored at 4°C for future use. 

Phytochemical Screening

The methanolic extracts were subjected to chemical tests for the detection of different phytoconstituents using standard procedures^21,27. 

Test for Phenols

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

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

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

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 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 test1 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 test1 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 test1 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 ferricchloride solution. This was underlayed with 1 ml of concentrated sulphuric acid. A browning of the interface indicates a deoxy sugar characteristic of cardenolides. 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

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

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 colouration of the inter face was formed which indicates the presence of terpenoids. 

Test for Steroids

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

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 each 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.

Preparation and extraction of sample 

Protocol for preparation of sample was according to the methods previously described²⁷, with modifications wrt temperature and duration of drying the sample. A 100 g leaf was weighed and dried in an oven at 60°C. Dried sample was ground into powder using Thomas-Willey milling machine and sieved on a wire mesh screen (3 × 3 mm²). Sample was stored at 4°C in air-tight container with screw caps. Sample was prepared according to the methods previously described²⁴. 25 g of sample was suspended in 250 mL of distilled water in stoppered flasks and allowed to stand for 24 h, filtered with Whatman No 24 filter paper, concentrated in a rotary evaporator for 12 h at 50°C and dried in vacuum desiccator and subjected to GC-MS analysis.

GC-MS Analysis 

Phyto-components were identified using GCMS detection system as previously with minor modification, whereby portion of the extract was analysed directly by headspace sampling. GCMS 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 follows: 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 libraries.

RESULTS AND DISCUSSION 

Chemical Properties and Identifier

Chemical kingdom	Organic compounds
Superclass	Lipids and lipid-like molecules
Class	Prenol lipids
Subclass	Monoterpenoids
PubChem Identifier	170833
Synonyms	ISOPULEGOL; ALPHA-TERPINEOL;
Canonical SMILES	C[C@@H]1CC[C@H]([C@@H](C1)O)C(=C)C
InChI Key	ZYTMANIQRDEHIO-KXUCPTDWSA-N

GCMS analysis of Rosmarinus officinalis (Rosemary) 

The chemical composition of ROEO depends on plant genetics, growth conditions, development stage at harvest, and processes of extracting active compounds. Different parts of the plant (bark, leaf, fruit and seed) have been extensively investigated for their bioactive phytochemical constituents in various plants (Ramya et al., 2012). GC-MS analysis revealed that the extract of Rosmarinus officinalis contained different volatile oils. Tricyclo[3.2.1.0(2,4)] octane,8-methylene (1.α.,2. α.,4.α., 5.α. )- (C₉H₁₂), 3.237, 2 hits; Benzene, 1-ethyl-2,3-dimethyl- (C₁₀H₁₄), 4.318 min, 10 hits; Cyclohexanemethanol, 4-hydroxy-.α.,α.,4-trimethyl- (C₁₀H₂₀O₂), 4.436 min, 10 hits; Cyclohexanol, 5-methyl-2-(1-methylethenyl)- (C₁₀H₁₈O), 0.508 min, 10 hits; Eucalyptol (C₁₀H₁₈O), 4.566, 6 hits; 1,8-Cineole; 470-82-6; 1,8-Cineol; (C₁₀H₁₈O), 4.655 min, 10 hits; Geranyl tiglate (C₁₅H₂₄O₂), 4.811, 2 hits; 3-Oxatricyclo[4.1.1.0(2,4)]octane, 2,7,7-trimethyl- (C₁₀H₁₆O), 4.885 min, 10 hits; -Naphthalenol, decahydro- (C₁₀H₁₈O), 4.959 min, 10 hits; 4-Cyclooctene-1-methanol (C₉H₁₆O), 5.02 min, 10 hits; 1,2,4,5-Tetrazine (C₂H₂N₄), 5.243 min, 10 hits; 1-Cyclopentene-1-methanol, .α.,α.,4,5- tetramethyl-, trans- (C₁₀H₁₈O), 6.045 min, 10 hits; Tricyclo[4.2.2.0(1,5)]dec-7-ene (C₁₀H₁₄), 6.159 min, 10 hits; (1S-(1Α,2α,β))-1-isopropenyl-4- methyl-1,2-cyclohexanediol (C₁₀H₁₈O₂), 6.208 min, 10 hits; Bicyclo[3.1.1]hept-3-en-2-one, 4,6,6-trimethyl-,(1S)- (C₁₀H₁₄O), 6.244 min, 10 hits; Linalyl isobutyrate (C₁₄H₂₄O₂), 6.454 min, 10 hits; Bicyclo[2.2.2]oct-2-ene, 1-methylamino- (C₉H₁₅N), 7.084 min, 10 hits; Benzenemethanol, 4-ethyl- (C₉H₁₂O), 7.092 min, 10 hits; Dicyclopentadiene diepoxide (C₁₀H₁₂O₂), 7.344 min, 10 hits; 1,8-Nonadiyne (C₉H₁₂), 7.5 min, 10 hits; 2,6,11,15-Tetramethyl-hexadeca-2,6,8,10,14- pentaene (C₂₀H₃₂), 20.122, 2 hits; Phthalic acid, di(6-methylhept-2-yl) ester (C₂₄H₃₈O₄), 35.075 min, 10 hits respectively (Table 1; Fig. 1). 

Secondary metabolites of ROEO have been reported for its antitumor, antioxidant, anti-infectious, anti-inflammatory, and analgesic activities and effects on the central nervous system, endocrine system, disorders such ascardiac remodeling after myocardial infarction, body weight changes, dyslipidemia, cerebral ischemia, hepato-nephrotoxicity, stress, and anxiety^3,6. Structure and molecular biological properties of selected compounds in R. officinalis has been the determinant of the biological activity as depicted in Fig. 2a-f. Similarly, Sienkiewicz et al.²⁶ reported that ROEO contains 1,8-cineole (46.4%), camphor (11.4%) and α-pinene (11.0%) and precursors of other significant bioactive molecules used for therapeutic, cosmetics, and food industries. ROEO used by Jiang et al¹⁸ had 1,8-cineole (26.54%) and α-pinene (20.14%) present significant antimicrobial activity. Bendeddouche et al.¹³ reported that camphor (37.6%), 1,8-cineole (10.0%), p-cymene-7-ol (7.8%) and borneol (5.4%) were the main bioactive metabolites. Anti-inflammatory activity of rosemary has been attributed to the synergistic activity of carnosol and carnosic, rosmarinic, ursolic, oleanolic, and micromeric acids⁷. Specifically, anti-inflammatory activity has been attributed to synergic effects of ursolic and micromeric acids present in ROEO. These natural drugs can be proposed for preclinical and clinical studies in different diseases and pathological conditions^29-31.

CONCLUSION 

Rosemary contains a large variety of bioactive molecules with great therapeutic potential such as triterpenes (e.g., ursolic and oleanolic acid), tricyclic diterpenes (e.g., carnosic acid and carnosol), phenolic acids (e.g., caffeic acid and rosmarinic acid), and essential oils. These secondary metabolites have been formulated in topical dosages. ROEO has anti-inflammatory, antimicrobial, and antioxidant properties, which have been extensively reported in oral formulations. However, development of new formulations containing other less common ROEO extracts is warranted through ADMET to evaluate and establish the potentials of pharmacologically active phyto-compounds towards safety and efficacy, in treating various pathological conditions.

REFERENCES

1. Gamble JS, The flora of the Presidency of Madras, London: Adlard & Son Ltd. London. 1935

2. Matthew KM, The Flora of Tamil Nadu Carnatic (The Rapinat Herbarium, St. Joseph's College, Tiruchirapalli, India). 1983; Vol I-III

3. Alavi MS, Fanoudi S, Ghasemzadeh Rahbardar M, Mehri S, Hosseinzadeh H. An updated review of protective effects of rosemary and its active constituents against natural and chemical toxicities. Phytotherapy Research. 2020. https://doi.org/10.1002/ptr.6894

4. Satyal P, Jones TH, Lopez EM, McFeeters RL, Ali NA, Mansi I, Al-Kaf AG, Setzer WN. Chemotypic characterization and biological activity of Rosmarinus officinalis. Foods. 2017; 6(3):20. https://doi.org/10.3390/foods6030020

5. Heinrich M., Kufer J., Leonti M., Pardo-de-Santayana M. Ethnobotany and ethnopharmacology-Interdisciplinary links with the historical sciences. J. Ethnopharmacol. 2006; 107:157-160. https://doi.org/10.1016/j.jep.2006.05.035

6. Borges RS, Ortiz BL, Pereira AC, Keita H, Carvalho JC. Rosmarinus officinalis essential oil: A review of its phytochemistry, anti-inflammatory activity, and mechanisms of action involved. Journal of ethnopharmacology. 2019; 229:29-45. https://doi.org/10.1016/j.jep.2018.09.038

7. Andrade JM, Faustino C, Garcia C, Ladeiras D, Reis CP, Rijo P. Rosmarinus officinalis L.: an update review of its phytochemistry and biological activity. Future science OA. 2018; 4(4):FSO283. https://doi.org/10.4155/fsoa-2017-0124

8. Kumar N, Goel N. Phenolic acids: Natural versatile molecules with promising therapeutic applications. Biotechnology Reports. 2019 Dec 1; 24:e00370. https://doi.org/10.1016/j.btre.2019.e00370

9. Kumar N., Gupta S., Yadav T.C., Pruthi V., Varadwaj P.K., Goel N. Extrapolation of phenolic compounds as multi-target agents against cancer and inflammation. J. Biomol. Struct. Dyn. 2019; 37(9):2355-2369 https://doi.org/10.1080/07391102.2018.1481457

10. Al-Shalah LA, Hindi NK, Mohsen IH. Essential Oils. Essential Oils-Bioactive Compounds, New Perspectives and Applications. 2020.

11. Atanasov AG, Waltenberger B, Pferschy-Wenzig EM, Linder T, Wawrosch C, Uhrin P, Temml V, Wang L, Schwaiger S, Heiss EH, Rollinger JM, Schuster D, Breuss JM, Bochkov V, Mihovilovic MD, Kopp B, Bauer R, Dirsch VM, Stuppner H. Discovery and resupply of pharmacologically active plant-derived natural products: A review. Biotechnol Adv. 2015; 33(8):1582-1614. https://doi.org/10.1016/j.biotechadv.2015.08.001

12. Ayyappan P, Ganesan K, Jayakumararaj R Ethnobotanic information on uncommon anti-diabetic medicinal plants from Alagarkoil Forest Reserve: Evidence based strategic rationale in management of diabetics. Int J Pharm Res 2019; 16:515-26

13. Bendeddouche M.S., Benhassaini H., Hazem Z., Romane A. Essential oil analysis and antibacterial activity of Rosmarinus tournefortii from Algeria. Nat. Prod. Commun. 2011; 6:1511-1514. https://doi.org/10.1177/1934578X1100601026

14. Bozin B., Mimica-Dukic N., Samojlik I., Jovin E. Antimicrobial and Antioxidant properties of Rosemary and Sage (Rosmarinus officinalis L. and Salvia officinalis L., Laminaceae) essential oils. J. Agric. Food Chem. 2007; 55:7879-7885 https://doi.org/10.1021/jf0715323

15. Chávez-González ML, Rodríguez-Herrera R, Aguilar CN. Essential oils: A natural alternative to combat antibiotics resistance. Elsevier Academic Press: London Wall, London, UK. 2016:227-37. https://doi.org/10.1016/B978-0-12-803642-6.00011-3

16. De Oliveira JR, Camargo SE, De Oliveira LD. Rosmarinus officinalis L.(rosemary) as therapeutic and prophylactic agent. Journal of Biomedical Science. 2019; 26(1):5-23. https://doi.org/10.1186/s12929-019-0499-8

17. Habtemariam S. The therapeutic potential of rosemary (Rosmarinus officinalis) diterpenes for Alzheimer's disease. Evidence-Based Complementary and Alternative Medicine. 2016. https://doi.org/10.1155/2016/2680409

18. Jiang Y., Wu N., Fu Y.-J., Wang W., Luo M., Zhao C.J., Zu Y.G., Liu X.L. Chemical composition and antimicrobial activity of the essential oil of Rosemary. Environ. Toxicol. Pharmacol. 2011; 32:63-68 https://doi.org/10.1016/j.etap.2011.03.011

19. 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

20. Kayashima T, Matsubara K. Antiangiogenic effect of carnosic acid and carnosol, neuroprotective compounds in rosemary leaves. Bioscience, biotechnology, and biochemistry. 2012:1111292741 https://doi.org/10.1271/bbb.110584

21. 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

22. Loganathan T, Barathinivas A, Soorya C, Balamurugan S, Nagajothi TG, Ramya S, Jayakumararaj R. Physicochemical, Druggable, ADMET Pharmacoinformatics and Therapeutic Potentials of Azadirachtin-a Prenol Lipid (Triterpenoid) from Seed Oil Extracts of Azadirachta indica A. Juss. Journal of Drug Delivery and Therapeutics. 2021; 11(5):33-46. https://doi.org/10.22270/jddt.v11i5.4981

23. Ramya S, Neethirajan K, Jayakumararaj R. Profile of bioactive compounds in Syzygium cumini-a review. J. Pharm. Res 2012; 5(8):4548-4553

24. Rašković A, Milanović I, Pavlović N, Ćebović T, Vukmirović S, Mikov M. Antioxidant activity of rosemary (Rosmarinus officinalis L.) essential oil and its hepatoprotective potential. BMC complementary and alternative medicine. 2014; 14(1):225. https://doi.org/10.1186/1472-6882-14-225

25. Sabitha, M., Krishnaveni, K., Murugan, M., Basha, A.N., Pallan, G.A., Kandeepan, C., Ramya, S. and 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

26. Sienkiewicz M., Lysakowska M., Pastuszka M., Bienias W., Kowalczyk E. The potential of use Basil and Rosemary essential oils as effective antibacterial agents. Molecules. 2013; 18:9334-9351. https://doi.org/10.3390/molecules18089334

27. Soorya C, Balamurugan S, Basha AN, Kandeepan C, Ramya S, Jayakumararaj R. Profile of Bioactive Phyto-compounds in Essential Oil of Cymbopogon martinii from Palani Hills, Western Ghats, INDIA. Journal of Drug Delivery and Therapeutics. 2021; 11(4):60-5. https://doi.org/10.22270/jddt.v11i4.4887

28. 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 Jul 15; 11(4):66-73 https://doi.org/10.22270/jddt.v11i4.4890

29. Stevanović ZD, Bošnjak-Neumüller J, Pajić-Lijaković I, Raj J, Vasiljević M. Essential Oils as Feed Additives-Future Perspectives. Molecules. 2018; 23(7):1717. https://doi.org/10.3390/molecules23071717

30. Tai J., Cheung S., Wu M., Hasman D. Antiproliferation effect of Rosemary (Rosmarinus officinalis) on human ovarian cancer cells in vitro. Phytomedicine. 2012; 19:436-443. https://doi.org/10.1016/j.phymed.2011.12.012

31. Zhou Y, Wu F, Li L, Shen X, Chen G, Wang X, Liang X, Tan M, Huang Z. Computational approaches in preclinical studies on drug discovery and development. Frontiers in Chemistry. 2020; 8:726 https://doi.org/10.3389/fchem.2020.00726

 

 

Table 1 Qualitative phytochemical analysis of methanolic extract of R. officinalis

PHYTOCONSTITUENTS	TEST	PRESENT/ ABSENT
Phenol	FeCl3 Test	+++
Flavonoids	Shinoda Test	++
Tannins	FeCl3 Test	++
Alkaloids	Wagner’s reagent Test	+
Carbohydrates	Fehling’s test, Benedict’s test	++
Proteins	Millon’s Test, Ninhydrin Test	++
Glycosides	Keller-Kiliani Test	+
Saponins	Foam Test	+
Coumarins	Coumarins Test	+
Terpenoids	Salkowski Test	++
Quinones	Quinone Test	+
Steroids	Salkowski Test	+
Anthraquinones	Borntragers Test	-

+++ = Copiously present; ++ = moderately present; + = slightly present; - = absent
 

Figure 1: GCMS analysis of Rosmarinus officinalis (Rosemary) essential oil

 

Table 2: GCMS profile of Rosmarinus officinalis (Rosemary) essential oil

S.No	Compound	Molecular Formula	Retention Time (min)	Percentage (%)
	α-Pinene	C10H16O	6.94	13.64
	Camphene	C10H16	7.38	2.42
	β-Myrcene	C10H16	8.88	1.19
	α-Terpinine	C10H16	9.70	0.41
	p-Cymene	C10H14	9.98	6.23
	trans-3-Caren-2-ol	C10H16O	10.10	0.20
	1,8-Cineole	C10H18O	10.38	41.75
	γ-Terpinene	C10H16	11.25	0.59
	α-Terpinolene	C10H16	12.30	0.35
	Linalool	C10H18O	12.78	1.19
	Isopulegol	C10H16O	14.44	13.66
	Eucalyptol	C10H18O	15.21	6.71
	Terpinen-4-ol	C10H18O	15.56	1.24
	2-Naphthalenol	C10H18O	16.14	6.35
	(-)-Myrtenol	C10H16O	16.27	0.16
	Verbenone	C10H14O	16.67	0.42
	Terpine	C12H20O2	19.42	2.80
	α-Copaene	C15H24	22.49	0.20
	β-Caryophyllene	C15H24	23.92	1.40
	γ-Cadinene	C15H24	27.16	0.34
	Caryophyllene oxide	C15H24O	28.90	0.32

 

 

a)	MF	C10H14
p-Cymene	Molecular Properties	Calculated Values
CCC1=CC=CC(=C1C)C	miLogP	3.65
	TPSA	0.00
	natoms	10
	MW	134.22
	nON	0
	nOHNH	0
	nviolations	0
	nrotb	1
	volume	150.53
	Biological Properties	Bioactivity Scores
	GPCR ligand	- 1.11
	Ion channel modulator	- 0.81
	Kinase inhibitor	- 1.41
	Nuclear receptor ligand	- 1.23
	Protease inhibitor	- 1.47
	Enzyme inhibitor	- 0.77

 

 

 

 

 

 

 

 

b)	MF	C10H20O2
Terpin (Cyclohexanemethanol, 4-hydroxy-.alpha.,.alpha.,4-trimethyl-)	Molecular Properties	Calculated Values
CC1(CCC(CC1)C(C)(C)O)O	miLogP	1.61
	TPSA	40.46
	natoms	12
	MW	172.27
	nON	2
	nOHNH	2
	nviolations	0
	nrotb	1
	volume	184.55
	Biological Properties	Bioactivity Scores
	GPCR ligand	- 0.39
	Ion channel modulator	0.35
	Kinase inhibitor	- 1.12
	Nuclear receptor ligand	- 0.35
	Protease inhibitor	- 0.55
	Enzyme inhibitor	- 0.02

 

c)	MF		C10H18O
Isopulegol (Cyclohexanol, 5-methyl-2-(1-methylethenyl)-)	Molecular Properties		Calculated Values
CC1CCC(C(C1)O)C(=C)C	miLogP		2.65
	TPSA		20.23
	natoms		11
	MW		154.25
	nON		1
	nOHNH		1
	nviolations		0
	nrotb		1
	volume		171.55
	Biological Properties		Bioactivity Scores
	GPCR ligand		- 0.78
	Ion channel modulator		- 0.16
	Kinase inhibitor		- 1.59
	Nuclear receptor ligand		- 0.22
	Protease inhibitor		- 0.71
	Enzyme inhibitor		- 0.14
d)		MF		C10H18O
Eucalyptol (1,8-Cineole; 470-82-6; 1,8-Cineol)		Molecular Properties		Calculated Values
CC1(C2CCC(O1)(CC2)C)C		miLogP		2.72
		TPSA		9.23
		natoms		11
		MW		154.25
		nON		1
		nOHNH		0
		nviolations		0
		nrotb		0
		volume		166.66
		Biological Properties		Bioactivity Scores
		GPCR ligand		- 0.93
		Ion channel modulator		- 0.01
		Kinase inhibitor		- 1.60
		Nuclear receptor ligand		- 1.07
		Protease inhibitor		- 0.90
		Enzyme inhibitor		- 0.15

 

e)	MF	C10H16O
alpha-Pinene oxide (3-Oxatricyclo[4.1.1.0(2,4)]octane, 2,7,7-trimethyl-)	Molecular Properties	Calculated Values
CC1(C2CC1C3(C(C2)O3)C)C	miLogP	2.74
	TPSA	12.53
	natoms	11
	MW	152.24
	nON	1
	nOHNH	0
	nviolations	0
	nrotb	0
	volume	155.87
	Biological Properties	Bioactivity Scores
	GPCR ligand	- 0.40
	Ion channel modulator	- 0.41
	Kinase inhibitor	- 1.24
	Nuclear receptor ligand	- 0.17
	Protease inhibitor	0.15
	Enzyme inhibitor	0.34
f)	MF	C10H18O
2-Naphthalenol, decahydro- (-Naphthalenol, decahydro-)	Molecular Properties	Calculated Values
C1CCC2CC(CCC2C1)O	miLogP	2.50
	TPSA	20.23
	natoms	11
	MW	154.25
	nON	1
	nOHNH	1
	nviolations	0
	nrotb	0
	volume	167.06
	Biological Properties	Bioactivity Scores
	GPCR ligand	- 0.49
	Ion channel modulator	- 0.09
	Kinase inhibitor	- 0.98
	Nuclear receptor ligand	- 0.48
	Protease inhibitor	- 0.40
	Enzyme inhibitor	- 0.02

 

Figure 2a-f Structure (2D, 3D), molecular biological properties of compounds in R. officinalis