Bioactive Compound Profiling of Theriophonum minutam (Willd.) Baill. using GC-MS: Implications for Pharmaceutical and Therapeutic Application

Introduction

Medicinal plants are crucial in both the pharmaceutical and food sectors. Phytochemical substances derived from these plants have been converted into commercial medications and continue to be a source for discovering new drugs. [1] Numerous plant species contain a variety of chemical components traditionally utilised for managing illnesses. Many drug formulations are rooted in the use of these substances in customary medicine.[2]

The genus Theriophonum (Araceae), consisting of seasonally tuberous perennials, is commonly found in India and Sri Lanka. Theriophonum minutum is a wild edible plant exhibiting natural diversity and offering higher nutritional values than standard food sources. Several studies indicate that Theriophonum minutum has been analysed regarding its phytochemical composition and pharmacological effects, although it is primarily noted for its exceptional nutritional benefits.[3] Ethnomedical practices show that the tubers of Theriophonum minutum (Willd.) Baill. are prepared by boiling with tamarind leaves, rinsed, and then cooked as a vegetable by the Konda reddis and Koyas from the East Godavari district of Andhra Pradesh, as well as by the Koyas of the Lankapalli Forest Reserve in Khammam district, Telangana. Tuber powder mixed with honey is used to treat gastrointestinal issues, small intestinal injuries, and stomach pain.[4]

 In Maharashtra, the Indian vernacular names for this species are Doda and Tangya. In the Vidarbha region, it is called undirkani; the Koyas of Andhra Pradesh refer to it as “adavi champak.” The leaves of T. fischeri Sivad. They are blended with turmeric and applied externally to address skin conditions and wounds, as well as to alleviate rheumatic pain. [5] The boiled tubers are consumed for two weeks to treat piles and ease body aches.[6] Indigenous people of Gadchiroli District, Maharashtra, use the leaves of Theriophonum minutum to prepare chille (leaves chopped finely and made into chapati). This is often complemented by additional wild foods. Different communities consume these species based on local availability. Various preparations of plant species are created and marketed in tribal markets.[7] Modern pharmacology highlights the significance of natural products in the development of new medications. Numerous natural compounds have served as the basis for creating drugs and are still employed today to treat a range of illnesses. Nevertheless, using contemporary medications presents various challenges, such as significant side effects and resistance to antibiotics or even cancer treatments, necessitating the development of new drugs.[8] A thorough literature review indicates that Theriophonum minutum has not yet been examined for GC-MS analysis or the identification of bioactive compounds. This study represents the initial effort to identify different bioactive compounds using GC-MS and explore their pharmaceutical and therapeutic applications.

Material & Methods

Plant Collection and Extraction

The fresh plant specimen was collected from the area of Ghot, District Gadchiroli, Maharashtra (Fig. A & B) with latitude 19.81247⁰& Longitude 79.991825^0.A voucher specimen was deposited atthe Department of Botany, Bhawabhuti Mahavidyalaya, Amgaon. (voucher numberBMV 731). The plant specimen was verified by a field botanist of the same institute uponcollection from the wild. The mature leaves (500gm) were cleaned, air-dried for 5 days,homogenized using a heavy-duty blender, and subjected to Cold Maceration & Extracted with Methanol after extraction Sample was allowed to dry in room temperature for 5-6 days until semisolid crud drugs were from after that 1mg sample was dissolve in 10 ml of methanol & sample was send for GC-MS analysis.

GC-MS Analysis

The Crude drug of Theriophonum minutam was diluted in methanol and subsequently sent to the Sophisticated Analytical Instrument Facility at IIT Madras. The sample was injected (1 µL) into a GC-MS system consisting of a gas chromatography system (Agilent 8890) coupled with a mass spectrometry detector (Agilent 5977B). An HP-5MSultrainert (30 m × 250 µm ×0.25 µm) capillary column was used with 0.25 µm film thickness of coated material. The injector temperature was set to 350 ◦C, and the temperature program followed was as follows: it started at 60 ◦C and held for 0.5 minutes, then ramped from 60 ◦C to 350 ◦C at a rate of 0.5 ◦C/min, followed by a 0.5-minute hold. A post-run phase at 50 ◦C for 3 minutes was conducted to prepare for the subsequent injection.Gas chromatography was conducted in splitless mode, utilising helium gas as the carrier at a consistent flow rate of 1 mL/min. Compounds were identified by referencing the NIST database, and their compositions were calculated based on the peak abundances observed in the chromatogram. The entire analysis was completed in triplicate.

Results& Discussion

GC-MS Results of Theriophonum Minutum vegetative leaf Methanol extract. Identified 57 unique compounds, as shown in Graph1 & table 1. The chromatograms, peak, Real time, Area, Area Percentage, Name of compound, Probability of compound, CAS number,Molecular formula & Molecular weight are represented in table No.1. Major phytocompounds identified include 7,9-Di-tert-butyl-1- oxaspiro(4,5)deca-6,9- diene-2,8-dione(92.9%),2-Pentadecanone, 6,10,14trimethyl(86%),6Hydroxy4,4,7atrimethyl5,6,7,7tetrahydrobenzofuran2(82.45),Tetradecanoicacid(63.15%),1,3Diazacyclooctane2thione(40.55%),Pyrimidine2,4,6trione,l(+)Ascorbicacid2,6dihexadecanoate(37.54%),1cyclohexyl5[(2piperazin1yl(36.02%),1Propanamine,2methylN(2methylpropylidene)26.89%,cyclohexanespiro-5-(2,4,4-trimethyl-2-oxazoline (23.62%)  These are the major compounds having a probability rate of more than 20%. Other 49 compounds have been found in Theriophonum minutum with a probability rate between 1 and 20%. A variety of compounds have been found in different classes.A few important classes of bioactive compounds, such as fatty acids, branched alkenes, saturated hydrocarbons, terpenoids, esters, aldehydes, flavonoids, and vitamin C, were detected in the extract of TM. Out of the 57 compounds, 9 fatty acids, 4 heterocycliccompounds, 4 Organic nitrogen compounds, 6 esters, 4 amines, 4 ketones, 3 terpenoids, 2 saturated hydrocarbons, 2 aldehydes, 2branched alkenes, 2 steroids,1 flavonoid,and 1 pyrimidinewere detected as major contributors to thePharmaceutical&Therapeutic application of TM. Several Compound has different pharmaceutical & biological role reported by other researches &  these compound also found in Theriophonum minutamsuch as 2-Cyclohexylpiperidine has used full for synthesizing novel medicine, particularly those used as analgesic, anaesthetics, antiparasitic, anti-inflammatory& antibacterial[9], 3,3,3-Trifluro-1-piperidin-1-yl-2-Trifluromethyl-propane-1 used for novel drug discovery, 1,3-Dicyclohexylurea shows  Potent inhibitory action against soluble epoxide hydroxylase and also antihypertensive effect, vasodilation & anti-inflammatory activity studied in the study on Pharmacokinetic evaluation of a 1,3-dicyclohexylurea[10].Role in creatine biosynthesis, reduces blood glucose, hormonal regulation,& Neuromodulation activity shows by Guanidineacetic acid.[11] Deoxyspergualine compound isolated from TM shows Immunosuppressive agent & used in Organ transplantation, autoimmune diseases& cancer treatment. [12]β-Cyclocostunolide compound shows antioxidative, anti-inflammatory, and antiallergic bone remodelling, Neuroprotective activity.[13] Most of the compounds reported to have Antibacterial, Antioxidant, Anti-inflammatory, Anticancerous, Neuroprotective, and hepatoprotective activity are listed & summarized in Table No.2

Conclusion: – In the current research focusing on the GC-MS analysis of the methanolic extract of Theriophonum Minutum(Willd.)Baill leaves, we explore a commonly used ethnobotanical herb found in peninsular India and Sri Lanka during the monsoon season. The review of existing literature indicated a lack of comprehensive studies regarding its phytochemical components, especially in terms of GC-MS analysis. This marks the first GC-MS report on Theriophonum Minutum, where we identified 57 bioactive compounds, highlighting the presence of various bioactive constituents in the plant we examined. TM contains a wide range of secondary metabolites, which are extensively utilised in traditional medicine to address numerous health issues, as well as in contemporary medical practices. The identified secondary metabolites, including fatty acids, hydrocarbons, terpenoids, steroids, esters, phenols, tannins, and flavonoids, play a role in alleviating various biological challenges, supporting the use of these plants by traditional healers. Leaves of Theriophonum Minutum are prepared in a specific manner to eliminate their irritating substances. This method of food preparation is a novel contribution to both science and society. Therefore, it is recommended for pharmaceutical and therapeutic uses. Additional research is necessary to determine its bioactivity.

Acknowledgement:Authors are thankful to SAIF Panjab University Chandigarh, India for providing GC-MS facilities. The authors are thanking to Shri Sureshbabu Asati, President, and Shri K. L. Mankar, Secretary, & Dr P. K. Rahangdale, Principal Bhawabhuti Mahavidyalaya Amgaon, for encouraging and supporting. The authors also Dr. Ladke, Principal, Nilkantrao Shinde Science & Arts College, Bhadrawati, for providing Laboratory facilities for this work.

Funding: – No Funding received

Ethics clearance and participant consent: – Not relevant.

Conflict of Interest: -We do not have any conflicts of interest to report.

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