Phytochemical Characterization of Hexane Extract of Hygrophila Auriculata Leaves Through GC–MS Analysis

For thousands of years, medicinal plants have played a central role in human healthcare, providing a major source of therapeutic molecules and constituting the basis of traditional medical systems across the world¹. Ancient cultures including Egyptian, Greek, and Chinese civilizations, as well as Indigenous groups in Africa, the Americas, and Asia, have long exploited the healing potential of plants to manage various diseases². Owing to their diverse bioactive constituents, these plants have been employed against common disorders, infectious diseases, and severe conditions, establishing themselves as a vital component of healthcare worldwide. Their relevance extends beyond history, as medicinal plants continue to be a dynamic and evolving area influencing contemporary traditional and modern medicine.³ While medicinal plants have been used in healthcare for centuries, scientific scrutiny of their properties is a relatively recent development. The progress of modern pharmacology has highlighted the importance of validating traditional knowledge and investigating the intricate phytochemical profiles of plants with long histories of medicinal use.⁴ Despite its promising therapeutic profile, Hygrophila auriculata remains an underexploited medicinal species requiring further systematic investigation. A member of the Acanthaceae family, it is widely distributed in South and Southeast Asia including Nepal, India, Myanmar, Malaysia, and Sri Lanka.⁵ The plant is a rich source of phytochemicals such as glycosides, polyphenols, phytosterols, fatty acids, alkaloids, proanthocyanidins, enzymes, amino acids, and terpenoids. It demonstrates a broad spectrum of pharmacological effects including cardioprotective, anti-stress, diuretic, anti-inflammatory, anticancer, hepatoprotective, nephroprotective, and neuroprotective activities. In addition, its traditional use in Bengali culinary preparations underscores its potential for translation into functional foods and nutraceuticals that integrate traditional wisdom with modern science.⁶

Vernacular names:⁷

Marathi  : Kolshinda Talimkhana

Sanskrit : Kokilaksa

Bengali  : Kuliyakhara

Gujrati   : Ekharo

Hindi     : Talmakhana

Kannada: Kolavali

Marathi  : Talikhana Kalsunda

Tamil     : Golmidi, Neermulli

Urdu       : Talmakhana.

Taxonomy⁸

• Kingdom      : Plantae
• Class             : Magnoliopsida
• Subkingdom  : Viridiplantae
• Superorder    : Asteranae
• Infrakingdom : Streptophyta
• Order            : Lamiales
• Superdivision: Embryophyta
• Family          : Acanthaceae
• Division        : Tracheophyta
• Genus            : Hygrophila R. Br.
• Subdivision     : Spermatophyta
• Species          : Hygrophila auriculata

Traditionally, various parts of the plant including roots, leaves, and seeds have been utilized in Indian medicine as diuretic agents and for managing conditions such as jaundice, dropsy, rheumatism, anasarca, and urinogenital tract diseases. Ayurvedic literature describes kokilaksha as an effective remedy for arthritis. The plant is also credited with improving strength and appetite and with therapeutic efficacy against edema, ascites, excessive thirst, bladder calculi, ophthalmic disorders, and dysentery. Its root decoction exhibits diuretic properties, and a formulation containing its leaves and roots with flowers of Stuea frondosa is employed for leucorrhoea. Additionally, it is traditionally used to address impotence, spermatorrhea, and seminal weakness.^8,9 GC–MS analysis of the hexane extract of the leaves of Hygrophila auriculata was performed to identify the phytochemical constituents present in the extract.

MATERIALS AND METHODS

Collection of Plant material

Hygrophila auriculata leaves were collected in the month of April 2025 from Tirunelveli District, Tamil Nadu. The whole plant was authenticated by Dr. M. Syed Ali Fathima, Assistant Professor and Head, Sadakathullah Appa Arts and Science College, Tirunelveli. (Specimen Number: SAC/BOT/2025).

Figure 1: Leaves of Hygrophila auriculata

Figure 2: Chromatogram of Hexane extract of Hygrophila auriculata

Compound Name	Mol. Formula	Mol. weight	Structure
Phenol, 2-methyl-5-(1-methylethyl) or Carvacrol	C10H14O	150
(Z,Z) alpha Farnesene	C15H24	204
Trans- alpha-Bergamotene	C15H24	204
Phenol, 2-methoxy-4-(2-propenyl)-, acetate or Acetyleugenol	C12H14O3	206
Copaene or Tricyclo dec-3-ene	C15H24	204
Longifolene	C15H24	204
Caryophyllene or Bicyclo undec-4-ene	C15H24	204
Humulene or alpha Caryophyllene	C15H24	204
2(4H)-Benzofuranone, 5,6,7,7a-tetrahydro-4,4,7a-trimethyl	C11H16O2	180
Neophytadiene or 7,11,15-Trimethyl-3-methylenehexadec-1-ene	C20H38	278
beta-Sitosterol	C29H50O	414
beta-Amyrone	C30H48O	424
9,19-Cycloergost-24(28)-en-3-ol	C32H52O2	468
Lup-20(29)-en-3-one	C30H48O	424
Lupeol	C30H50O	426
Stigmasta-3,5-diene	C29H48	396
Alpha-Tocopherolquinone	C29H50O3	446
2.beta 4.beta 16.alpha Tribromoallopregn-16-ene-3	C21H27Br3O2	548
Xanthine, 8-[[4-[2-(4-aminophenyl)-1-carboxy] ethylamino] carbonyl] methoxyphenyl]-1,3-dipropyl	C28H32N6O6	548
Campesterol	C28H48O	400

CONCLUSION:

The present study successfully characterized the phytochemical composition of the hexane leaf extract of Hygrophila auriculata using Gas Chromatography–Mass Spectrometry (GC–MS). More than 60 phytochemical constituents were identified, demonstrating the chemical diversity of the extract. The detected compounds included phenolics, sesquiterpenes, diterpenes, triterpenoids, phytosterols, and quinone derivatives, with notable constituents such as carvacrol, (Z,Z)-α-farnesene, trans-α-bergamotene, acetyleugenol, caryophyllene, α-humulene, neophytadiene, β-sitosterol, β-amyrone, lupeol, campesterol, stigmasta-3,5-diene, and α-tocopherolquinone. These compounds are known to possess a broad spectrum of biological activities, including antioxidant, antimicrobial, anti-inflammatory, hepatoprotective, anticancer, and immunomodulatory properties. The predominance of terpenoids and phytosterols indicates that the hexane solvent efficiently extracted lipophilic bioactive metabolites, highlighting the therapeutic potential of the plant. The comprehensive GC–MS profile generated in this study provides valuable baseline information for the chemical standardization and quality assessment of H. auriculata leaf extracts. Furthermore, the findings scientifically support the traditional medicinal use of this species and emphasize its potential as a promising source of bioactive natural products for pharmaceutical, nutraceutical, and functional food applications. Future investigations should focus on the isolation and purification of the major phytochemicals, quantitative analysis, and in vitro and in vivo pharmacological studies to elucidate their mechanisms of action and validate their therapeutic potential.

REFERENCES

1. Ahmad, I., & Mehmood, Z. (2017). Bioactive compounds of medicinal plants and their uses in traditional medicine. Phytochemistry Reviews, 16(4), 899-912.
2. Ali, M. S., & Wang, S. (2020). Role of medicinal plants in modern pharmacology: Review. Pharmaceutical Biology, 58(1), 74-88.
3. Aslam, S., & Lee, S. Y. (2019). Advances in medicinal plants: New opportunities for therapeutic applications. Journal of Medicinal Plant Research, 13(2), 40-56.
4. Manisha, Ram Babu, Maajitha Begam, Kavita Shakya Chahal, Akshay Ashok Harale. Medicinal Plants and Traditional Uses and Modern Applications. Journal of Neonatal Surgery. 2025; 14 (3): 162-175.
5. Doss, A. Antimicrobial activity of Hygrophila auriculata (Schumach.) Heine and Pergularia daemia Linn. African Journal of Plant Science. 2013; 7: 137–142.
6. Megha Malpotra, Meenakshi Garg, Neha Singh, Susmita Dey Sadhu, Rajni Chopra et.al. An overview of bioactive components and phytopharmaceutical potentials of Hygrophila auriculata–A herbaceous medicinal plant. Phytomedicine Plus 5 (2025) 100737.
7. Dhanalakshmi, Harikrishnan, Srinivasan, Pandian, Tanisha, Tharun Kumar et.al. A Perspective Overview on Hygrophila auriculata. Pharmacognosy Journal. 2020; 12 (6): 1748-1752.
8. F Nishvanth, D Nagavalli and B Prem Kumar. Assessing the impact of solvent polarity on the phytochemical profile of Hygrophila auriculata leaf extracts. International Journal of Pharmacognosy and Clinical Research 2025; 7(2): 107-114.
9. Suresh Kumar Jebamalai, Nirmala Ramachandran, Venkatraman Arumugam. Phytochemical and antimicrobial studies on Hygrophila auriculata Schumach. International Journal of Current Research in Biosciences and Plant Biology. (2021) 8(12), 23-31.
10. Nishvanth F, Aswini B, Kalaiselvi G, Abinaya R, Sangeetha T. GC-MS-Based Profiling of Bioactive Phytoconstituents in the Methanolic Whole Plant Extract of Bryophyllum pinnatum (Lam.) Oken. Journal of International Research in Medical and Pharmaceutical Sciences. 2026; 21(3): 240–255.