GC MS study on siddha formulation Kizhanelli Thylam

 Corresponding author: N VENKADESAN Copyright © 2025 Author(s) retain the copyright of this article. This article is published under the terms of the Creative Commons Attribution Liscense 4.0. GC MS study on siddha formulation Kizhanelli Thylam N VENKADESAN 1, *, R ANTONY DURAICHI 2, G ESSAKKY PANDIAN 3 and S SRIPRIYA 4 1 PG Scholar, Post Graduate Dept of Gunapadam, Govt Siddha Medical College Palayamkottai, Tamilnadu, India. 2 Assistant Professor, Post Graduate Department of Gunapadam, Govt Siddha Medical College Palayamkottai, Tamilnadu, India. 3 Professor,HOD, Post Graduate Department of Gunapadam, Govt Siddha Medical College Palayamkottai, Tamilnadu, India. 4 PG Scholar Post Graduate Department of Gunapadam, Govt Siddha Medical College Palayamkottai, Tamilnadu, India. World Journal of Advanced Research and Reviews, 2025, 27(01), 2018-2028 Publication history: Received on 14 June 2025; revised on 21 July 2025; accepted on 23 July 2025 Article DOI: https://doi.org/10.30574/wjarr.2025.27.1.2733 Abstract The present study deals with GC MS analysis of one siddha medicine formulation KIZHANELLI THYLAM which is prescribed for pitha related diseases such as kirukiruppu(giddiness),vayil neerural(excessive salivation),kai kal erichal(peripheral neuritis),kannerichal(burning eyes) and pitha paithiyam(psychiatric diseases).It is used as external bath oil. KIZHANELLI THYLAM prepared as per siddha literature Patharthaguna villakkam (Meteria Medica).KIZHANELLI THYLAM underwent standard procedure of GC MS analysis.GCMS analysis revealed the presence of 14 preliminary chemical components present in kizhanelli thylam.These components are D-Limonene, Ocimene, 3Carene, Cyclohexonal,5-methyl-2-(1-methylethenyl), Cyclohexane,1-ethenyl-1-methyl-2,4-bis(1-methylethenyl)-, Caryophyllene, Azulene,1,2,3,4,5,6,7,8-octahydro-1,4-dimethyl-7-(1-methylethylidene)-(1s-cis) and Caryophyllene oxide.The relationship between the claimed medicinal roles and the presence of biomolecules, as indicated by GC MS seems encouraging. Keywords: KIZHANELLI THYLAM; Bath oil; GC MS Analysis; Pitha Diseases 1. Introduction Siddha system is the most ancient and indigenous system of indian medicine.Thylam is important siddha formulation used as medicine in both internal and external application.The medicated oils are used as bath oil.Oil bath is one of the imperative practice.Regular practice of oil bath relaxes and rejuvenates us both physically and mentally.It is believed by the people of ancient time taking oil bath will improve and keep their good health. According to siddha literature Padhartha Guna Vilakkam ” Meteria Medica( vegetable kingdom)” KIZHANELLI THYLAM is one of the important thylam used as bath oil to treat pitha related diseases such as kirukiruppu(giddiness),vayil neerural(excessive salivation),kai kal erichal(peripheral neuritis),kannerichal(burning eyes) and pitha paithiyam(psychiatric diseases).However,the characteristics of traditional siddha medicine are systematize,multitarget and multichannel due to their complex chemical constituents and there is no study about the significant evidence of biomolecules present in it.This needs to be studied and scientifically understood. So,I took this study to find out the chemical components of KIZHANELLI THYLAM. 2. Material and methods Ingredients of KIZHANELLI THYLAM: World Journal of Advanced Research and Reviews, 2025, 27(01), 2018-2028 2019 • Kizhanelli (Phyllanthus amarus) • kadukkai (Terminalia chebula) • Neem resin (Azadirachta indica) • Gingelly oil (Sesamum indicum) • Cow’s Milk • water • Country sugar(nattu sarkarai) 2.1. Phyllanthus amarus (kizhanelli) 2.1.1. Therapeutic actions Kizhanelli plant has many pharmacological activities like antioxidant,antiviral, Hepatoprotective and Nephroprotective. 2.2. Terminalia chebula (kadukkai) 2.2.1. Therapeutic actions The extracts of kadukkai plant exhibit Anti-inflammatory, Anthelmintic and antioxidant activities. 2.3. Neem Resin 2.3.1. Therapeutic action Neem resin have Antimicrobial Activity 2.4. Sesame oil (gingelly oil) 2.4.1. Therapeutic actions Sesame oil have various pharmacological activities such as Antiaging,Demulcent,Anticancer,antioxidant, Antimicrobial and Vasorelaxant. 2.4.2. Authentication of raw drugs Plant origin drug were identified and authenticated by the experts in PG Gunapadam Department,Government Siddha Medical College, Palayamkottai. 2.4.3. Procedure The whole plant of phyllanthus amarus was collected and dried. 875gm of plant was cut into small pieces and boiled with 5.2 liter of water in the mud pot till the decoction concentrated to 1.3 liter and filtered.Each 17.5gm of terminalia chebula and neem resin were grounded with sufficient quantity of milk and made into paste.The paste, 1.3 liter of gingelly oil and decoction mixed together.Then it was boiled well until the confection in the oil converted into the waxy stage(mezhugu padham).In that stage 17.5 gm of country sugar was sprinkled over the oil.Then this oil was filtered and cooled.Finally filtered oil was stored in closed container. GC MS Analysis of prepared sample of Kizhanelli Thyam was conducted at Sophisticated Analytical Instrument Facility,Indian Institute of Technology,Chennai. 2.4.4. GC MS ANALYSIS For the oil extracts, a small amount of concentrated sample was taken in a separating funnel and shaken by adding water and ethyl acetate in the ratio of 1:4. The upper layer was collected and concentrated in rotary evaporator to about 1.5 ml. Added 100µl N, O-Bis(trimethylsilyl)trifluoroacetamide and trimethylchlorosilane(BSTFA+TMCS) and 20µl pyridine and heated at 60°c for 30 minutes. For the layers which are separated from the crude extracts, a small amount of extract was taken and evaporated out totally. To this added acetonitrile and filtered into a conical flask. To the filtrate added 50µl BSTFA+TMCS and heated at 60°c in a water bath for 30 minutes. Filtered using 0.45µ membrane filter to a vial. World Journal of Advanced Research and Reviews, 2025, 27(01), 2018-2028 2020 2.4.5. GC-MS Procedure Gas chromatography (GC) analysis was carried out using Agilent 6890N gas chromatography equipped with photon multiplier tube as detector coupled to front injector type 1079. The chromatograph was fitted with HP 5 MS capillary column (30 m ×0.25 mm i.d., film thickness 0.25 _m). The injector temperature was set at 250°C, and the oven temperature was initially at 70 °C hold for 4 mins then programmed to 200°C at the rate of 10°C/min and finally held at 200 °C for 13 min. Helium was used as a carrier gas with the flow rate of 1.5 ml/min. 0.2 microlitre of the Drug MC (diluted with methanol 1:10) were injected in the splitless mode. GC–mass spectrometry (GC–MS) analysis of sample was performed using Agilent gas chromatography equipped with JEOL GC MATE-II HR Mass Spectrometer. GC conditions were the same as reported for GC analysis and the same column was used. The mass spectrometer was operated in the electron impact mode at 70 eV. Ion source and transfer line temperature was kept at 250°C. The mass spectra were obtained by centroid scan of the mass range from 50 to 600 amu. The compounds were identified based on the comparison of their retention indices (RI), retention time (RT), mass spectra of WILEY, NIST library data of the GC-MS system and literature data (Adams, 2009). 3. Results and discussion In the present study 14 chemical compounds have been identified.They were D-Limonene, Ocimene, 3-Carene, Cyclohexonal,5-methyl-2-(1-methylethenyl), imonene-1,2-epoxide(fr.1), Bicyclo(3.1.1)hept-3-en-2-ol,4,6,6-trimethyl, Limonene-1,2-epoxide(fr.1), Cyclohexane,1-ethenyl-1-methyl-2,4-bis(1-methylethenyl)-, Caryophyllene, Dimethyl phthalate, Azulene,1,2,3,4,5,6,7,8-octahydro-1,4-dimethyl-7-(1-methylethylidene)-(1s-cis), 1,2-benzenedicarboxylic acid ethyl methyl ester, Caryophyllene oxide and 1,4-benzenedicarboxylic acid diethyl ester.The active principles with their retention time and peak area % were tabulated in Table 1.Components and their medicinal roles were also tabulated in Table 2. Some of them namely D-Limonene shows Anti-inflammatory, anticancer activity. Ocimene shows Antiviral, anti-inflammatory, antifungal,anticonvulsant activities. 3-Carene shows Anti-inflammatory, antifungal,sedative,Antispasmodic activities and Cyclohexonal,5-methyl-2-(1-methylethenyl) shows Anti-epileptic, Anti-anxiety, Anti-inflammatory, Antiviral, anesthetic activity.GC MS graphs were given in Figure 1-15. Table 1 GC MS Profile Of KIZHANELLI THYLAM S.NO RETENTION TIME PEAK AREA PERCENTAGE Name of the Molecules Identified with NIST Library 1. 5.37 4.5 D-Limonene 2. 6.65 6.8 Ocimene 3. 7.2 3.0 3-Carene 4. 8.43 4.5 Cyclohexonal,5-methyl-2-(1-methylethenyl) 5. 8.94 4.5 Limonene-1,2-epoxide(fr.1) 6. 10.32 9.1 Bicyclo(3.1.1)hept-3-en-2-ol,4,6,6-trimethyl 7. 11.62 7.6 Limonene-1,2-epoxide(fr.1) 8. 12.5 5.3 Cyclohexane,1-ethenyl-1-methyl-2,4-bis(1-methylethenyl)- 9. 13 14.5 Caryophyllene 10. 13.92 6.8 Dimethyl phthalate 11. 14.57 8.4 Azulene,1,2,3,4,5,6,7,8-octahydro-1,4-dimethyl-7-(1-methylethylidene)-(1s-cis) 12. 14.98 8.8 1,2-benzenedicarboxylic acid ethyl methyl ester 13. 15.65 9.1 Caryophyllene oxide 14. 16 6.1 1,4-benzenedicarboxylic acid diethyl ester World Journal of Advanced Research and Reviews, 2025, 27(01), 2018-2028 2021 Table 2 Medicinal Role Of Components S.NO Name of the Components Promising Medicinal Roles 1. D-Limonene Anti-inflammatory, anticancer activity, Used in obesity. 2. Ocimene Antiviral, anti-inflammatory, antifungal,anticonvulsant activities. 3. 3-Carene Anti-inflammatory, antifungal,sedative,Antispasmodic activities. 4. Cyclohexonal,5-methyl-2-(1-methylethenyl) Anti-epileptic, Anti-anxiety, Anti-inflammatory, Antiviral, anesthetic activity. 5. Limonene-1,2-epoxide(fr.1) Not known 6. Bicyclo(3.1.1)hept-3-en-2-ol,4,6,6-trimethyl Not known 7. Cyclohexane,1-ethenyl-1-methyl-2,4-bis(1methylethenyl)- Antitumor activity 8. Caryophyllene Anti-inflammatory, Used in osteoarthritis, diabetes mellitus, anxiety, and liver fibrosis. 9. Dimethyl phthalate Not known 10. Azulene,1,2,3,4,5,6,7,8-octahydro-1,4-dimethyl-7- (1-methylethylidene)-(1s-cis) Anti-inflammatory, antibacterial, anti tumor, antiviral,antifungal activities. 11. 1,2-benzenedicarboxylic acid ethyl methyl ester Not known 12. Caryophyllene oxide Antifungal,Antiplatelet aggregation activity. 13. 1,4-benzenedicarboxylic acid diethyl ester Not known Figure 1 GC Profile of Kizhanelli Thylam World Journal of Advanced Research and Reviews, 2025, 27(01), 2018-2028 2022 Figure 2 D-Limonene Figure 3 Ocimene World Journal of Advanced Research and Reviews, 2025, 27(01), 2018-2028 2023 Figure 5 Cyclohexonal,5-methyl-2-(1-methylethenyl) Figure 6 Limonene-1,2-epoxide(fr.1) World Journal of Advanced Research and Reviews, 2025, 27(01), 2018-2028 2024 Figure 7 Bicyclo(3.1.1)hept-3-en-2-ol,4,6,6-trimethyl Figure 8 Cyclohexane,1-ethenyl-1-methyl-2,4-bis(1-methylethenyl)- World Journal of Advanced Research and Reviews, 2025, 27(01), 2018-2028 2025 Figure 9 Caryophyllene Figure 10 Dimethyl phthalate World Journal of Advanced Research and Reviews, 2025, 27(01), 2018-2028 2026 Figure 11 Azulene,1,2,3,4,5,6,7,8-octahydro-1,4-dimethyl-7-(1-methylethylidene)-(1s-cis) Figure 12 1,2-benzenedicarboxylic acid ethyl methyl ester