BIOLOGICALLY ACTIVE COMPOUNDS OF LALLEMANTIA ROYLEANA (BENTH.) BENTH. AND THEIR SIGNIFICANCE IN MODERN MEDICINE

ISSN: 2582-4686 SJIF 2021-3.261,SJIF 2022-2.889, 2024-6.875 ResearchBib IF: 9.948 / 2024 VOLUME-5, ISSUE-10 416 BIOLOGICALLY ACTIVE COMPOUNDS OF LALLEMANTIA ROYLEANA (BENTH.) BENTH. AND THEIR SIGNIFICANCE IN MODERN MEDICINE Pulatova Aziza Ramazon kizi Makhkamov Trobjon Khusanboyevich Allaberdiyev Rustamjon Khamrayevich 1Department of Botany and Genetics, National University of Uzbekistan, Tashkent, Uzbekistan 2Department of Forestry and Landscape Design, Tashkent State Agrarian University, Kibray district, Tashkent region, Uzbekistan Abstract: This study presents the results of a comprehensive investigation into the chemical composition of the essential oil extracted from the aerial parts of Lallemantia royleana (Benth.) Benth. The qualitative and quantitative analysis of the volatile constituents was performed using gas chromatography coupled with mass spectrometry (GC–MS). The plant material was collected during the active vegetative growth phase in May 2024 from the experimental plot of the Botanical Garden of the National University of Uzbekistan named after Mirzo Ulugbek (Tashkent city). A total of 70 volatile compounds were identified, forming the chemical profile of the essential oil. The major constituents of the oil were β-ocimene (15.67%), citral (12.59%), nerol (12.72%), and βcitral (9.15%). These compounds exhibit significant biological activities, including antimicrobial, antioxidant, and anti-inflammatory effects, which support the potential of L. royleana as a valuable source of natural bioactive substances. The high content of monoterpenes and aldehydes in the essential oil highlights its aromatic and therapeutic properties, opening up broad prospects for its use in the pharmaceutical, food, cosmetic, and perfume industries. The findings confirm the significant potential of L. royleana as medicinal and technical raw material and emphasize the need for further research focused on standardization, biological evaluation, and industrial application of its essential oil. Keywords: Uzbekistan, Lallemantia royleana, essential oil, GC-MS, citral, ocimene, hydrocarbons, phytotherapy, aromatic compounds. Introduction In recent years, the study of biologically active compounds found in medicinal plants and their application in modern medicine has become one of the most pressing topics on a global scale. The growing demand for natural remedies, the adverse side effects associated with synthetic drugs, and the increasing emphasis on environmental safety have significantly boosted interest in plant-based pharmaceuticals (World Health Organization). The rising demand for natural agents in modern medicine and the pharmaceutical industry has enhanced the significance of many plant species, including Lallemantia royleana (Benth.) Benth. (Mahmood S. et al., 2013; Ghannadi A. et al., 2015; Bozorgi M., Vazirian M., 2016; Al-Snafi A.E., 2019; Sardarodiyan M. et al., 2019; Saleem A. et al., 2022). The seeds of L. royleana have been identified as promising for medical use due to their healing and antioxidant properties, high total phenolic content, and their effectiveness in reducing cholesterol and triglyceride levels in blood serum (Mahmood S. et al., 2013; Bozorgi M., Vazirian M., 2016; Sardarodiyan M. et al., 2019; Saleem A. et al., 2022; Ghannadi A. et al., 2015). ISSN: 2582-4686 SJIF 2021-3.261,SJIF 2022-2.889, 2024-6.875 ResearchBib IF: 9.948 / 2024 VOLUME-5, ISSUE-10 417 The seeds of L. royleana contain 25.60% protein, 18.27% fat, 1.29% fiber, as well as alkaloids, anthraquinones, flavonoids, glycosides, phlobatannins, volatile oils, mixed fatty acids, and terpenoids. These compounds have demonstrated a wide range of pharmacological effects, including antimicrobial, antioxidant, antidepressant, anxiolytic, sedative, antiemetic, hypolipidemic, and protective activities. Some studies suggest that these compounds could be useful in the treatment of skin diseases (Al-Snafi A.E., 2019; Massey S. et al., 2022; Saleem A. et al., 2022). Traditionally, L. royleana seeds have been used in folk medicine for the treatment of various conditions, particularly digestive disorders, respiratory inflammations, and skin diseases (Saleem A. et al., 2022). In recent years, extracts and bioactive components derived from this plant have increasingly been incorporated into modern pharmaceutical formulations, further underscoring its medical relevance (Mahmood S. et al., 2013; Atabaki R., Hassanpour-Ezatti M., 2014). A chemical analysis of essential oil obtained from the aerial parts of Lallemantia royleana (Benth. in Wall.) Benth., cultivated in Isfahan province, Iran, using gas chromatography (GC) and gas chromatography–mass spectrometry (GC-MS), revealed 46 components. The major constituents were identified as verbenone (16.4%), trans-carveol (9.8%), and β-cubebene (8.9%) (Ghannadi A., Zolfaghari B., 2003). Furthermore, literature analysis indicates that the essential oil composition of the vegetative organs of L. royleana remains poorly studied. However, scientific interest in this plant has grown in recent years, especially due to the pharmacological potential of its volatile compounds (essential oils). This article presents an analysis of the chemical composition of the biologically active compounds in L. royleana, their pharmacological mechanisms of action, and directions for their application in modern medicine. The potential for practical implementation of these findings is also discussed based on current scientific literature. Materials and Methods Plant material collection and identification The object of this study is Lallemantia royleana (Benth.) Benth., a medicinal and essential oil-bearing plant belonging to the Lamiaceae family, widely cultivated in various regions of the world. It is commonly known as "Balangu" in Iran, "Tukh Malanga" in Pakistan, and "Mallachoy" in Uzbekistan. Plant samples were collected during the juvenile stage in May 2024 from the experimental plot of the Botanical Garden, Faculty of Biology and Ecology, National University of Uzbekistan named after Mirzo Ulugbek (Tashkent). The species was identified by Professor Zafar Muhammad, from Quaidi-Azam University. Voucher specimens were deposited in the Department of Botany and Genetics, National University of Uzbekistan. Essential oil extraction A total of 266 grams of fresh aerial parts of the plant were crushed and subjected to hydro-distillation for 5 hours using a Clevenger-type apparatus. The essential oil obtained was dried over anhydrous sodium sulfate and stored in sealed amber vials at +4 °C in a dark environment until further analysis. GC-MS analysis method Gas chromatography–mass spectrometry (GC-MS) analysis was performed using an Agilent 7890B GC system coupled with a 5977A mass selective detector. The system was equipped with a DB-5MS capillary column (30 m × 0.25 mm i.d., film thickness 0.25 µm). Injection volume was 1.0 µL in split mode (split ratio 1:50), with helium as the carrier gas at a constant flow rate of 1.0 mL/min. The oven temperature program was as follows: initial temperature of 50 °C (held for 2 minutes), increased at a rate of 5 °C/min to 250 °C, and held for 39 minutes. ISSN: 2582-4686 SJIF 2021-3.261,SJIF 2022-2.889, 2024-6.875 ResearchBib IF: 9.948 / 2024 VOLUME-5, ISSUE-10 418 Mass spectrometry was performed in electron ionization (EI) mode at 70 eV with a scan range of 40 to 550 m/z. Compounds were identified by comparing their mass spectra and retention indices with data from the NIST08.L, W9N11.L, and W8N05ST.L spectral libraries. The relative percentage of each component was calculated based on peak area normalization without applying correction factors. Results GC-MS analysis revealed a total of 70 compounds, primarily corresponding to terpenes and hydrocarbons. The major constituents identified in the essential oil included α-terpinene, limonene, β-phellandrene, ocimene, p-cymene, geraniol, citral, nerol, pinocarvone, germacrene-D, and farnesol. The most abundant components were β-ocimene (15.67%), nerol (12.72%), citral (12.59%), pinocarvone (9.08%), and germacrene-D (5.69%) (Table 1). Table 1. Chemical constituents of L. royleana essential oil identified by GC-MS № Retention Time (min) Compound Name CAS Number Relative Abundance (%) Identification Quality 1 2.462 α-Terpinene 99-86-5 0.15 97 2 2.688 Limonene 138-86-3 0.38 98 3 2.785 β-Phellandrene 555-10-2 0.32 95 4 3.108 β-Ocimene 6874-10-8 15.67 97 5 3.218 γ-Terpinene 99-85-4 0.49 96 6 3.309 β-Ocimene Y 3779-61-1 2.95 96 7 3.509 p-Cymene 99-87-6 0.21 95 8 3.736 α-Terpinene 99-86-5 0.06 95 9 3.768 Isoamyl-2-methyl butyrate 27625-35-0 0.10 58 10 4.609 6-Methyl-5-hepten-2-one 110-93-0 0.78 95 11 5.017 Camphene 79-92-5 0.08 78 12 5.541 Neo-allo-ocimene 673-84-7 0.27 98 13 5.851 Cyclooctene 61233-78-1 0.09 72 14 5.993 Nonanal 124-19-6 0.06 90 15 7.035 1-Octen-3-ol 3391-86-4 0.07 53 16 7.462 γ-Terpinene 99-85-4 0.25 95 17 7.604 Isopentyl hexanoate 2198-61-0 0.04 64 18 8.503 Copaene 947-59-1 0.07 97 19 8.943 1-Hexene, 4,5-dimethyl 16106-59-5 0.19 53 20 9.034 Pinanone 15358-88-0 1.83 95 21 9.318 Cyclohexaneacetaldehyde 3991-38-6 0.08 50 22 9.473 Pinocarvone 30460-92-5 9.08 91 23 9.862 Bicyclohept-2-en-4-ol acetate 998166-346 5.35 89 24 10.767 γ-Terpinene 99-85-4 1.73 83 25 11.071 Myrtenal 564-94-3 1.35 98 26 11.705 trans-Pinocarveol 547-61-5 1.05 70 ISSN: 2582-4686 SJIF 2021-3.261,SJIF 2022-2.889, 2024-6.875 ResearchBib IF: 9.948 / 2024 VOLUME-5, ISSUE-10 419 27 12.359 cis-p-Mentha-2,8-dien-1ol 3886-78-0 0.07 70 28 12.501 p-Mentha-1,3,8-triene 18368-95-1 0.18 74 29 12.701 Tetracyclodecane 998041-929 0.32 68 30 12.857 Citral 106-26-3 9.15 96 31 13.277 3-Carene 13466-78-9 0.22 83 32 13.665 Germacrene-D 23986-74-5 5.69 98 33 13.995 Camphene 79-92-5 0.87 90 34 14.215 Citral 5392-40-5 12.59 97 35 14.836 Nerol 106-25-2 12.72 91 36 15.043 β-Citronellol 106-22-9 0.30 96 37 15.159 Bicycloheptane 79-92-5 0.05 46 38 15.237 Myrtenol 515-00-4 0.64 81 39 15.703 Nerol 106-25-2 3.42 89 40 15.890 (E)-1-Iodo-6-(4isopropenyl-1cyclohexenyl)-3-methyl-3hexene 998573-460 0.07 53 41 16.000 trans-1,2Dihydroperillaldehyde 998072-246 0.09 58 42 16.123 2,6-Dimethyl-1,3,5,7octatetraene, E,E460-01-5 0.06 86 43 16.460 Artemisia triene 18383-70-5 0.15 64 44 16.964 Nerol 106-25-2 7.00 91 45 17.449 Tetracyclodecane 100018558-7 0.06 59 46 18.607 α-Pinene 80-56-8 0.13 70 47 19.118 Geranyl acetate 105-87-3 0.04 74 48 20.321 Nonadecane 629-92-5 0.20 96 49 20.503 1,3,8-P-Menthatriene 18368-95-1 0.13 46 50 20.574 1(7),3,8-O-Menthatriene 000000-000 0.05 46 51 21.654 p-Menthatriene 18368-95-1 0.05 49 52 21.803 Bicyclo[4.2.0]oct-1-ene 998041-921 0.33 72 53 22.948 Fumaric acid, myrtenyl pentyl ester 998518-222 0.15 59 54 23.071 Camphene 79-92-5 0.07 52 55 23.181 3-Hexen-1-ol, benzoate 25152-85-6 0.21 90 56 24.423 Nerol 106-25-2 0.36 87 57 24.901 1-Octadecanesulphonyl chloride 998590-842 0.32 70 ISSN: 2582-4686 SJIF 2021-3.261,SJIF 2022-2.889, 2024-6.875 ResearchBib IF: 9.948 / 2024 VOLUME-5, ISSUE-10 420 58 25.063 γ-Cadinene 39029-41-9 0.08 50 59 25.716 δ-Cadinene 483-76-1 0.19 86 60 27.760 Farnesol 4602-84-0 0.06 72 61 29.145 Tricosane 638-67-5 0.19 93 62 30.070 Artemisia triene 18383-70-5 0.08 46 63 31.868 2-Thiophen-2-yl-1Hpyrrole 998064-456 0.03 72 64 33.110 Cyclotetradecane 295-17-0 0.31 89 65 33.505 Ethanol, 2-(3,3dimethylcyclohexylidene)- , (Z) 26532-23-0 0.09 81 66 33.996 (2E)-3,7,11,15Tetramethyl-2-hexadecene 14237-73-1 0.18 50 67 34.294 3Oxatetracyclo[5.3.1.0(2,4). 0(6,8)]undec-9-ene 63896-19-5 0.06 45 68 35.005 Pentafluoropropionic acid, octadecyl ester 999040-903 0.04 42 69 36.816 Octatetracontane, 1-iodo 40710-70-1 0.21 90 70 38.776 9-Octadecenoic acid (Z) (Oleic acid) 112-80-1 0.09 64 The main components of the essential oil consist of monoterpenes, sesquiterpenes, and aldehydes (Table 2). Table 2. Main chemical groups in the essential oil of L. royleana Chemical Group Content (%) Major Compounds Monoterpenes 40–45% Limonene, γ-Terpinene, α-Terpinene, β-Phellandrene Sesquiterpenes 10–15% Germacrene-D, Copaene, Farnesol Aldehydes 20–25% Citral, Nonanal Alcohols 15–20% Nerol, Myrtenol, trans-Pinocarveol Discussion A total of 70 chemical components were identified in the essential oil of Lallemantia royleana. Among these, the dominant compounds in the mixture are as follows: Citral (combined cis and trans isomers): 21.74% – Exhibits strong antibacterial and antifungal activity. Due to its citrus aroma, it is widely used in perfumery and cosmetic formulations. Additionally, it possesses anti-inflammatory and antiseptic properties [11]. Nerol: 19.72% – Characterized by a mild floral scent, this compound has calming and antidepressant effects. It is commonly used in cosmetics for its soothing and antiseptic action on the skin [12]. β-Ocimene: 15.67% – Demonstrates anti-inflammatory and insecticidal properties. In aromatherapy, it is utilized to relieve stress and tension. It also plays a significant role in the plant's natural defense system [13]. Secondary components present in the mixture include: ISSN: 2582-4686 SJIF 2021-3.261,SJIF 2022-2.889, 2024-6.875 ResearchBib IF: 9.948 / 2024 VOLUME-5, ISSUE-10 421 Pinocarvone: 9.08% – Exhibits antibacterial and antifungal properties. Widely used in the perfume industry and has potential application as a natural pesticide [14]. Germacrene-D: 5.69% – Possesses antimicrobial and insecticidal activities. It strengthens the natural defense mechanisms of plants and is also used in perfumery [15]. The GC-MS analysis of Lallemantia royleana essential oil revealed a complex mixture of volatile compounds, with a noticeable presence of terpenes such as cis-Ocimene, dl-Limonene, βPhellandrene, γ-Terpinene, and p-Cymene. High-quality identification scores for most of the detected compounds confirm the reliability of the results. This essential oil contains a wide array of aromatic compounds commonly found in plants and essential oils. Conclusion In conclusion, the high concentrations of Citral, Nerol, and Ocimene in the essential oil suggest its suitability for use in perfumery and aromatherapy products. Compounds such as Pinocarvone and Germacrene-D exhibit notable antimicrobial and anti-inflammatory properties, making them promising candidates for therapeutic applications. Additionally, Limonene, Ocimene, and other terpenoids are well known for their antibacterial and antioxidant effects. Citral, with its lemon-like aroma, is widely utilized as a flavoring agent in the food and beverage industries. The oil is predominantly composed of monoterpenes (e.g., Ocimene, Citral, Nerol), which contribute citrusy, floral, and sweet aromatic notes. Sesquiterpenes like Germacrene-D add woody and earthy tones to the oil’s scent profile. The primary component identified was cis-Ocimene, with the highest relative abundance of 15.67%. Other key compounds include dl-Limonene, β-Phellandrene, γ-Terpinene, and p-Cymene, which are recognized for their aromatic and potential therapeutic properties. Several peaks in the chromatogram correspond to unidentified compounds, indicating the need for further analysis or updates to spectral libraries for more accurate identification. REFERENCES 1. Bozorgi M., Vazirian M. Antioxidant activity of Lallemantia royleana (Benth.) seed extract //Traditional and integrative Medicine. – 2016. – S. 147-150. 2. Al-Snafi A. E. 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Preparation, characterization and biological evaluation of silver nanoparticles and drug loaded composites for wound dressings formed from Lallemantia royleana seeds’ mucilage //Journal of Biomaterials Science, Polymer Edition. – 2022. – T. 33. – №. 4. – S. 481-498. ISSN: 2582-4686 SJIF 2021-3.261,SJIF 2022-2.889, 2024-6.875 ResearchBib IF: 9.948 / 2024 VOLUME-5, ISSUE-10 422 8. Atabaki R., Hassanpour-Ezatti M. Improvement of lidocaine local anesthetic action using Lallemantia royleana seed mucilage as an excipient //Iranian Journal of Pharmaceutical Research: IJPR. – 2014. – T. 13. – №. 4. – S. 1431. 9. Ghannadi A., Zolfaghari B. Compositional analysis of the essential oil of Lallemantia royleana (Benth. in Wall.) Benth. from Iran //Flavour and fragrance journal. – 2003. – T. 18. – №. 3. – S. 237239. 10.https://www.who.int 11. https://pubchem.ncbi.nlm.nih.gov/compound/Citral 12. https://pubchem.ncbi.nlm.nih.gov/compound/Nerol 13. https://pubchem.ncbi.nlm.nih.gov/compound/beta-OCIMENE 14. https://pubchem.ncbi.nlm.nih.gov/compound/Pinocarvone 15. https://pubchem.ncbi.nlm.nih.gov/compound/Germacrene-D