GC-MS Analysis and Quality Evaluation of Dashmoola Kashaya

INTRODUCTION

Dashmoola Kashaya is classical ayurvedic polyherbal decoction prepared from the roots of the ten medicinal plants collectively known as Dashmoola. Dashmoola Kashaya is traditionally indicated in a wide spectrum of disorders including Fever (Jwara), Respiratory disorders (Shwasa), Cough (Kasa), Inflammatory conditions (Shotha), Abdominal masses (Gulma), Facial palsy (Ardita), Hemiplegia (Pakshaghata), and various musculoskeletal disorders like non-specific low back ache and Sciatica (Gridhrasi). Internally it acts by pacifying aggravated Vata and Kapha by reducing systemic inflammation. Externally Dashmoola decoction is used in procedures such as Parisheka, Nadi sweda and Sthanik basti therapies where it employs local anti-inflammatory and analgesic action (AFI, 2003).

Gas Chromatography-Mass Spectrometry (GC-MS) is a robust and sensitive analytical platform widely employed for qualitative and semi-quantitative identification of volatile and semi-volatile phytochemicals in herbal formulations. The technique enables structural elucidation based on retention time, mass fragmentation patterns and spectral library matching, thereby facilitating quality control, bioactivity correlation and standardization of complex polyherbal preparations.

The medicinal efficacy of Ayurveda formulation is depended on the quality and consistency of the ingredients used (Nhawkar et al., 2014). There is a need for analysis of herbal formulation through modern pharmacological analytical technic to establish its therapeutic effect. The present study aims to analyse the physiochemical composition of freshly prepared Dashmoola Kashaya using GC-MS and to correlate the identified metabolites with their biological activities. Such analytical validation may contribute to evidence-based integration of classical formulations into contemporary clinical practice.

MATERIALS AND METHODS

Materials

Ingredients of Dashmoola Kashaya

The formulation comprises (Table 1) Bilwa (Aegle marmelos), Agnimantha (Clerodendrum phlomidis), Shyonaka (Oroxylum indicum), Patala (Stereospermum suaveolens), Gambhari (Gmelina arborea), Brihati (Solanum indicum), Kantakari (Solanum xanthocarpum), Shalaparni (Desmodium gangeticum), Prishniparni (Uraria picta) and Gokshura (Tribulus terrestris) (Taru et al., 2022).

Procurement of Dashmoola Kashaya Choorna (Coarse powder)

The Sadvaidyashala Private Limited, Nanjangud-571301, Mysore, Karnataka, India.

Methods

Preparation of Dashmoola Kashaya

As per the classical reference Dashmoola Kashaya was prepared (Savrikar, 2020) The roots of Dashmoola are individually taken and physical impurities were removed Coarse powder of each of the above mentioned 10 herbs are taken in equal quantities. The combination is boiled with 16 time the water in a big vessel. It is boiled until 4 times (parts) water remains i.e. reduced to ¼ of the content.

Place of Physicochemical analytical study

The Sadvaidyashala Private Limited, Analytical Laboratory, Nanjangud-571301, Mysore, Karnataka, India.

Ethical Statement

Not applicable.

Statistical Analysis

Not applicable.

Method of analysis

All of the tests listed were carried out in accordance with API standard guidelines (API, 2008).

Place of gas chromatography

Gas Chromatography and Mass Spectrometry (GC-MS): Amrith Labs (Nisargam Private Limited), Shimoga, Karnataka.

Instrument for Gas chromatography

GCMS-QP 2010SE Gas chromatography Shimadzu model.

Sample Preparation

By using above mentioned instrument, 0.1 mL of sample was diluted with 10 mL of petroleum ether and 1micro litre of extract was injected. After agitation of the sample for 10 seconds, the sample was taken for gas chromatography for analysis.

GC-MS protocol

0.1 mL of the Kashaya sample was diluted with 10 mL of petroleum ether and a non-polar extract was prepared, filtered through a syringe filter (Nylon 13 mM 0.2 μM) and injected into GC-MS. The GC-MS column consists of DB-5MS (30 mm × 0.25 mm diameter × 0.25 micrometre thickness). Analysis was performed by injecting 1 μL of the sample with an injection temperature of 280.00ºC and a split ratio of 1:10. Helium gas (99.999%) was used as a carrier gas with a flow rate of 19.5 mL/min. The analysis was performed in the EI (electron impact) mode with 70 eV of ionisation energy. The injector temperature was maintained at 280ºC. The compounds are identified by GC-MS Library (NIST and WILEY). The ion source temperature was set to 200ºC and the interface temperature was pointed at 300ºC. The solvent cut time was 1.40 min, minimising solvent interference and the detector gain mode was set relative to the tuning result with a gain of 0.95 kV.

Oven temperature

Mass Spectrometry, Group 1- Event

These conditions are set and allowed for the precise separation and identification of analysts.

RESULTS

Results of Organoleptic and Physicochemical Analysis of Raw Drugs

The sample had a brownish colour, a distinct taste, and a distinctive odour according to conventional quality characteristics (Table 2). There was no evidence of extraneous material. It was discovered that the physicochemical characteristics fell within the limits of in-house specifications mentioned in Table : Analysis of Raw Drugs used in Dashmoola Kashaya Choorna. Low moisture is shown by the drying loss (4.99% W/W), which is significantly less than the maximum limit (NMT 10% W/W). The total ash measurement (2.90% W/W) indicates few inorganic contaminants and is within acceptable limits (NMT 5% W/W). The alcohol-soluble extractive value (12.05% W/W) satisfies the minimal criterion (NLT 12% W/W), whereas the acid-insoluble ash was 0% W/W (Table 2).

Result of GC-MS Analysis of Dashmoola Kashaya

The chemicals found were (Table 3 and Figure 1) 9,12-Octadecadienoyl chloride, (Z, Z)-, (+)-Sesamin, Cholest-5-en-3-ol (3β)-, tetradecanoate. 2H-Pyran, 2-(7-heptadecynyloxy) tetrahydro-. (+)-Sesamin, Cholest-5-en-3-ol (3β)-, tetradecanoate, (R)-6-Methoxy-2,8-dimethyl-2-((4R,8R)-4,8,12-trimethyltridecyl)chroman, Undec-10-ynoic acid, undecyl ester, Oleic acid, 15-Tetracosenoic acid, (Z)-, TMS derivative, 3-O-Methyl-d-glucose, 9,12-Octadecadienoic acid, methyl ester, (E,E)-, 15-Octadecenoic acid, methyl ester, (2-Cyclohexylpropyl)(methyl)amine, 3,7-Dimethyl-6-nonen-1-ol acetate, 9,12-Octadecadienoyl chloride and (Z, Z)-, 9-Octadecen-12-ynoic acid, methyl ester. These compounds are recognised for their anti-inflammatory, anti-oxidant, analgesic effect. The result of GC-MS analysis of Dashmoola Kashaya includes retention time, Possible compound, molecular weight and percentage peak area are mentioned in Table 3.

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DISCUSSION

Discussion on Physicochemical Analysis

Physicochemical (Table 2) analysis can be used for identification and assessing herbal mixture. The formulation has brownish color, distinctive flavor and odor which anticipates its quality. The coarse particle size was appropriate for the preparation of the formulations like Kashaya.

The loss on drying (4.99% w/w) was within the standard limit (10%w/w), suggesting that the sample had a low moisture content, this is important for preventing microbial development, enzymatic degeneration, and formulation deterioration during storage.

Standard acceptable total ash value (2.90% w/w) and acid-insoluble ash (0% w/w) indicates that the formulation has minimal impurities such as, siliceous elements or mineral matter, demonstrating an excellent and purity of the raw material utilised in the preparation.

The alcohol-soluble extractive value (12.05% w/w) which is less than normal (12% w/w) suggest presence of alcohol-soluble phytoconstituents such as flavonoids, alkaloids, phenolic compounds and other active components.

DISCUSSION

Discussion on GC-MS Analysis

9,12-Octadecadienoyl chloride, (Z, Z)-

9,12-octadecadienoyl chloride (Z, Z) (Table 3 and Figure 2) is a linoleic derivative with antimicrobial and anti-quorum sensing properties (Singh and Singh, 2019). Such chemicals inhibit bacterial communication (QS pathways), lowering biofilm formation and virulence expression. The fatty acids from this class exhibits membrane modulating action, which contribute to antibacterial activity (Begum et al., 2025).

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(+)-Sesamin

Sesamin, (Table 3 and Figure 3) a main compound found in Sesamum indicum, has potent antioxidant, anti-inflammatory, hypolipidemic and cardioprotective properties. It regulates NF-κB signalling, improves hepatic fatty acid oxidation, and lowers oxidative stress markers. Some clinical and experimental studies demonstrate better lipid profiles and Cytokine inhibition and endothelial protection property. Sesamin also shows hepatoprotective and neuroprotective effects through ROS scavenging and cytokine modulation (Dalibalta et al., 2020).

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Cholest-5-en-3-ol (3β)-, tetra decanoate

Sterol metabolism (Table 3) is related to this cholesterol ester derivative. Cholesterol metabolites affect inflammatory signalling pathway and glucose homeostasis. Sterol derivative studies have shown that they can modulate lipid metabolism and may have anti-inflammatory properties (Maganga et al., 2024).

2H-Pyran, 2-(7-heptadecynyloxy) tetrahydro-

Pyran derivatives, (Table 3) identified in phytochemical investigations of Erythrina variegata which have antibacterial and antioxidant activity (Amutha et al., 2014). These heterocyclic compounds have membrane-protective and radical scavenging characteristics. Anti-inflammatory and cytoprotective effects implied by experimental models (Muthukrishnan et al., 2016). Their biological function is often attributed to oxygen-containing ring structures.

R-6-Methoxy-2,8-dimethyl-2-((4R,8R)-4,8,12-trimethyltridecyl) chroman

This is a chroman derivative (Table 3), shares structural similarities with tocopherol analogues, which are substances that resemble vitamin E. Research emphasizes the potential of antioxidants and enzyme inhibitors, such as scavenging free radicals and preventing lipid peroxidation. It exhibits anti-aging property, cryoprotection, and potential anti-inflammatory effects (Velmani et al., 2025).

Undec-10-ynoic acid, undecyl ester

This molecule, derived from 10-undecynoic acid (Table 3), has potent antibacterial and antibiofilm properties, particularly against oral Streptococcus species. It acts by inhibiting microbial adherence and biofilm development (Goc et al., 2019). Antifungal and topical antimicrobial treatments contain comparable chemicals.

Oleic acid

It is monounsaturated fatty acid (Table 3) in plant. It improves the gut-liver axis and controls lipid metabolism through modulating SCD1. Some studies established its insulin-sensitizing, cardioprotective and anti-inflammatory activities. It promotes endothelial health and lowers LDL oxidation (Sener et al., 1999).

15-Tetracosenoic acid, (Z)-, TMS derivative

It is a long chain unsaturated fatty acid (Table 3), which can reduce inflammation by modulating the NF-κB pathway and suppressing cytokine production. Fatty acid derivatives can lower inflammatory mediators and oxidative damage (Li et al., 2025). They may also help maintain membrane structural integrity.

3-O-Methyl-d-glucose

A glucose analogue (Table 3) was investigated for transport and metabolic pathway studies. It is useful for assessing glucose absorption pathways involving GLUT transporters (Jay et al., 2018). While not directly therapeutic, it provides information about carbohydrate metabolism and cellular glucose regulation.

9,12-Octadecadienoic acid, methyl ester, (E, E)-

A methyl ester of linoleic acid (Table 3) with antioxidant and anti-inflammatory properties. Studies show that free radicals are scavenged and lipid peroxidation is inhibited (Muzahid et al., 2022). It may help to protect the cardiovascular system.

15-Octadecenoic acid, methyl ester

It is said to be an anti-inflammatory chemical found in plant extracts that controls inflammatory mediators and lowers cytokine expression (Table 3) (Dong et al., 2000). Fatty acid methyl esters frequently exhibit antioxidant and membrane-stabilizing effects.

(2-Cyclohexylpropyl)(methyl)amine

Plant extracts have been reported to have an anti-inflammatory chemical that controls inflammatory mediators and lowers cytokine expression (Table 3) (National Centre for Biotechnology Information, 2026) Methyl esters of fatty acids frequently exhibit membrane-stabilizing and antioxidant characteristics.

3,7-Dimethyl-6-nonen-1-ol acetate

A terpenoid ester (Table 3) commonly identified in plant essential oils. Terpenoids show antimicrobial, anti-inflammatory, and antioxidant actions. They may also contribute to aroma-therapeutic and mild cytoprotective effects (Salisu et al., 2025).

9-Octadecen-12-ynoic acid, methyl ester

9-Octadecen-12-ynoic acid (Table 3 and Figure 4), methyl ester is showing its ability to cross the BBB and potentially reach the central nervous system. Based on the pharmacokinetic parameters 9-Octadecen-12-ynoic acid, methyl ester was selected as the best choices due to their BBB penetration ability, superior absorption profile and permeability (Ilyas et al., 2025).

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CONCLUSION

The present study is conducted to establish the quality and suitability of the ingredients of the formulation through. The organoleptic, physicochemical and GC-MS analysis. Acceptable moisture content, ash values, and extractive values indicate the stability of the formulation and the presence of bioactive constituents.

The formulation reveals the bioactive compounds through GC-MS analysis, serves anti-inflammatory, antioxidant, antimicrobial, and analgesic activities.

The compounds present in the Kashaya which were discovered through GC-MS analysis, they serve anti-inflammatory, antioxidant, antimicrobial, and analgesic activities, which support its traditional therapeutic effect in systemic inflammatory disorders, respiratory diseases, musculoskeletal and metabolic conditions.

The results provide scientific support for the traditional use of Dashmoola Kashaya and may validates its quality standards by emphasizing the importance of integrating modern analytical parameters like GC-MS analysis with Ayurvedic knowledge. Further pharmacological and clinical researches are recommended to validates its therapeutic efficacy.

ACKNOWLEDGEMENT

The author extends sincere gratitude to Sadvaidyashala Private Limited, Nanjangud-571301, Mysore, Karnataka, India and Amrith Labs (Nisargam Private Limited), Shimoga, Karnataka laboratories for providing the necessary facilities, guidance, and support to carry out this work successfully.

ABBREVIATIONS

GMP

Good Manufacturing Practice

NA

Not applicable

GC-MS

Gass Chromatography and Mass Spectrometry

ASU

Ayurveda, Siddha, and Unani

UV

Ultraviolet

API

Ayurvedic Pharmacopoeia of India

EI

Electron Impact

NIST

National Institute of Standards and Technology

NF-κB

Nuclear factor kappa-light-chain-enhancer of activated B cells

m/z

Mass-to-charge ratio

eV

Electron volt; BBB: Blood-brain barrier

QS

Quorum sensing

ROS

Reactive oxygen species

LDL

Low-density lipoprotein

GLUT

Glucose transporter

TMS

Trimethylsilyl

SCD1

Stearoyl-CoA desaturase-1

KAHER

KLE Academy of Higher Education and Research.

CONFLICT OF INTEREST

The authors declare that there is no conflict of interest.

AUTHOR CONTRIBUTION

Dr. Shreya Balasaheb Nikam: Collection of data, data analysis and interpretation, writing article, critical revision of the article. Dr. Rajashekhar V. Sanapeti: Research concept and design, collection of data, data analysis and interpretation, writing article, critical revision of the article, final approval of article. Dr. Anil Koralli: Data analysis and interpretation, writing article, critical revision of the article, final approval of article. Dr. Shubham Deepak Anuse: Data analysis and interpretation, writing article, critical revision of the article. Dr. Sushmita Sanjay Gengane: Data analysis and interpretation, writing article, critical revision of the article.

SUMMARY

In ayurveda traditional decoction of Dashmoola is recommended for Vata- Kaphaj diseases which is having analgesic, anti-inflammatory and anti-oxidant properties. Good quality and purity were confirmed by physicochemical examination of raw pharmaceuticals, which revealed all parameters within API norms. Suitable moisture, total ash and extractive values show the presence of active phytoconstituents. GC-MS analysis discovered the compounds like Fatty acid derivatives, sesamin, sterol esters, terpenoids and tocopherol analogues. These compounds are known their analgesic, antibacterial, antioxidant, and anti-inflammatory qualities. Overall, the results support the classical therapeutic claims and scientifically may validate the pharmacological relevance of the formulation.