In Ayurveda Sneha Kalpana is considered as an important dosage form where, lipid subsatnces like oils, ghee, animal fats are processed with herbs, minerals or any other drugs[1]. This particular dosage form is considered as an augmented processing of Panchavidha Kashaya kalpana[2] (the five basic dosage forms) which is expected to have specific pharmacological properties and more shelf life compared to the latter[3]. It is generally based on the principle of  embedding the phytoconstituents of herbs introduced either as kalka (bolus) and drava dravya into a complex lipid structure. Studies have hinted that sneha kalpana is the process of embedding hydrophilic bioactive phytochemicals inside lipid molecules[4], representing a liposomal drug delivery system[5] Ghrita, which is clarified butter from milk is considered the best among Sneha Dravya[6]. This is because Ghrita has property to assimilate the drug which is processed in it[7], thereby acting as a carrier to deliver those bioactive compounds into the tissues. Murcchana is advised as mandatory for all lipid compounds   before using in any formulation[8]. Murcchana process is a way to refine raw lipid resources before incorporating it into the preparation of any formulation. Murcchana includes usage of specific drugs like Haritaki, Vibhitaki, Amalaki, Musta, Haridra and Matulunga swarasa[9] used to prepare kalka (Bolus) processed to transfer the active phytoconstituents from kalka davya and fortified into ghrita. This process makes the ghee more stable and effective[10]. Sunthi Ghrita is a formulation described in Bhaishajya Ratnavali[11], it is indicated in all types of sotha and amaja grahani roga[12]. Even though it is widely used in clinical practice, a thorough analysis or phytochemical profiling is not available till present. The present study compares all 4 types of Ghrita; Go Ghrita, Murcchita Ghrita, Murcchita Sunthi Ghrita and Amurcchita Sunthi Ghrita, with an objective of proving the effect of Murcchana and phytochemical screening of Sunti Ghrita. 

The study compacts with the preparation of ‘Sunti Ghrita’ consuming plain ghee and pre-processed ghee (Murcchit Ghrita) followed by its comparative organoleptic, physicochemical, HPTLC -phytochemical evaluation. These four samples of ghee were analysed which includes. Sample:1- Go ghrita, Sample:2- Murcchit Ghrita, Sample:3- Murcchit Sunthi Ghrita and Sample:4- AmurcchitSunthi Ghrita. 

Sample 1: Go Ghrita

Plain cow ghee purchased from the local market was used for the preparation Sunti ghrita, the same ghee was of allotted as sample 1.

Sample 2: Preparation of Murcchit Ghrita[13, 14]

Cow ghee was melted in a clean steel vessel and allowed for self-cooling. To this ghee water was added and heated in mild fire. The kalka prepared was then added to the ghee mixture and heating was continued until Sneha Siddhi Laksana’s appeared (wick like formation of kalka, absence of cracking sound of kalka on igniting, lack of froth in ghee), indicating proper Murcchana. At the end the ghee was filtered and measured. 92 ml was filtered out which was stored in airtight container. 

Sample 3: Preparation of Murcchit Sunthi Ghrita[15]

Dashamoola kwatha was prepared by boiling 100 g of course Dashamoola in 1.6 L of water over mild fire and reducing to 400 ml.

The procedure was carried out, as per the guidelines of Sarangadhara Samhita. To the Murcchita Ghrita taken in a clean steel container, Dashamoola kwatha and Sunthi kalka was added and heated under mild fire. On attaining Sneha Siddhi Lakshana’s, the ghee was filtered and stored in a clean dry airtight container and labelled.

Sample 4: Preparation of Amurcchita Sunthi Ghrita The same procedure was followed using plain Go Ghrita to obtain Amurcchita Sunthi Ghrita.

The study samples were undergone analysis based on three benchmarks 

a) Organoleptic, b) Physicochemcial and c) HPTLC phytochemical 

On undergoing analysis, the results obtained were as follows: 

1. Analytical Study (HPTLC Profile Summary)

Sample preparation:  Methanolic extracts of four ghee samples were prepared.

Stationary phase: Merk, HPTLC Silica gel 60 F₂₅₄ plate.

Mobile phase: Toluene: Diethyl ether (8:2 v/v).

Visualization: UV visualization 

Murcchana of lipid bases such as Ghrita (Ghee), Taila (oil), Vasa (Fat) is considered to be essential before preparing them into any suitable dosage form. The Methanolic extracts of the four samples taken in this study namely Go ghrita (plain ghee), Murcchita Ghrita, Murcchita Sunthi Ghrita, and Amurcchita Sunthi Ghrita on HPTLC profiling reveals the phytochemical refinement as a result of the Murcchana process and Sunthi fortification[17].

At 254 nm, the chromatogram of Murcchita Ghrita revealed four distinct peaks with Rf values 0.196, 0.532, 0.682, and 0.921, which are likely to be gallic acid, quercetin, bisdemethoxycurcumin, and curcumin respectively (Ashok Mateti et al.)[18]. These compounds are primarily derived from Triphala, Musta, and Haridra used during the Murcchana process. Their presence indicates successful extraction of phenolic and flavonoid components into the ghrita thereby improving the antioxidant and anti-inflammatory potential of the formulation. The correlation of these Rf values with literature (Ashok Mateti et al.)[18], (Dinesh Kumar et al.)[19] supports the identity of these bio actives.

In Murcchita Sunthi Ghrita, a more complex chromatographic profile was observed, with six peaks at 254 nm (Rf 0.212, 0.406, 0.546, 0.693, 0.792, and 0.917). These are supposed to be attributed to gallic acid, zingerone, quercetin, bisdemethoxycurcumin, eugenol, and curcumin respectively. The additional bands for zingerone and eugenol signify the incorporation of phytoconstituents from Sunthi (Zingiber officinale). Both compounds are known for their antioxidant, carminative, and anti-inflammatory properties, validating the therapeutic enrichment achieved through the Murcchana process followed by Sunthi fortification[19].

Amurcchita Sunthi Ghrita displayed similar spots (Rf 0.218, 0.408, 0.554, 0.685, 0.792, and 0.910), but the intensity of peaks was comparatively lower, suggesting the potential of Murcchana to enhance the extractive capability of ghrita and influence the stability of active components within the ghee base. The result also points towards the significance of the Murcchana process in improving lipid-phase solubilization of herbal phytochemicals.

At 366 nm, additional peaks were observed in both Murcchita and Amurcchita Sunthi Ghrita extracts at Rf 0.037 and 0.031 which is tentatively identified as rutin, a polar flavonoid reported by (Sunil P. Waghmare et al.)[20]. Other peaks (Rf 0.022, 0.214, 0.575, 0.690, and 0.975) represented compounds of varying polarity, indicating a broad spectrum of photoactive constituents. The occurrence of both polar and non-polar components demonstrates the compatibility of ghrita as a biphase extraction medium capable of retaining diverse phytochemicals[21].

Comparatively, Murcchita Sunthi Ghrita showed the most enriched phytochemical fingerprint, confirming synergistic enhancement between the Murcchana procedure and Sunthi processing. This combination yielded a well-balanced formulation containing phenolic acids (gallic acid)[17], flavonoids (quercetin, rutin), and curcuminoids (curcumin derivatives) with volatile compounds like zingerone and eugeno[19, 22]. These bio actives compounds have proven pharmacological effects like antioxidative and anti-inflammatory. Thus, these analytical results showed that Murcchana not only detoxifies and stabilizes ghrita but also potentiates its therapeutic efficacy. 

Sneha Kalpana is aimed to facilitate effective distribution of polar phytoconstituents beyond the lipid medium supporting a lipid envelop[6] which may improve stability, absorption, and therapeutic efficacy of dosage form. The observed change in colour from pale yellow to greenish-yellow and the development of strong aroma in Murcchita Sunthi Ghrita indicate efficient extraction of herbal constituents and removal of undesirable characteristics, which are classical features of Murcchana[23].

All samples exhibited slightly acidic pH (5–5.6), typical of lipid-based preparations[24], however, the reduced acid value in Murcchita samples suggested that the lower free fatty acid content and decreased oxidative degradation which indicates enhancement in stability. Slight increase in the specific gravity reflects the incorporation of phytoconstituents, while the minimal changes in refractive index confirm preservation of the lipid structure[23]. Additionally, lower moisture content and peroxide value in Murcchita Sunthi Ghrita signify better resistance to rancidity and oxidative deterioration[25].

Iodine value remained within the normal range which suggests that the fatty acid unsaturation is not adversely affected by processing, while the high saponification value suggests the presence of lower molecular weight of fatty acids, contributing to improved digestibility and bioavailability[26]. The high iodine value is due to its high content of unsaturated fatty acids[27]. Increased unsaponifiable matter further confirms the enrichment with bioactive compounds such as sterols and antioxidants[6].

Thus, based on the outcomes of physicochemical parameters, Murcchita Sunthi Ghrita, is expected to exhibit superior stability and quality among all the four formulations. This may also support the classical view that Murcchana might act as a refining process that enhances potency of Sneha Kalpana like Grita, Taila etc.[26]. Moreover, Sunthi ghrita prepared from Murcchita Ghrita may exhibit superior therapeutic properties compared to that prepared from plain ghrita, a difference attributable to its enriched profile of bioactive phytoconstituents, including gingerols, zingerone, and other phenolic compounds.

Among the two samples of Sunti ghrita, based on optimal analytical outcomes, Sunti ghrita prepared form Murcchita ghrita stands prime through the synergistic effect of drugs adopted for Murcchana. The physicochemical findings conclude that the parameters like Acid value, Iodine value, Peroxide value, Moisture content of Murcchita Sunthi Gritha are in optimum values suggesting its resistance to physicochemical variations like oxidation, lipid degradation etc. The phytochemical constituents present in the drugs of Murcchana like gallic acid, quercetin, bisdemethoxycurcumin, and curcumin from drugs Triphala, Musta, and Haridra are proved to be antioxidant, anti-inflammatory which enables activity like free radical scavenging, arresting lipid degradation, which may enhance the shelf life of ghrita.

Acknowledgements:

The authors would like to express their sincere gratitude to their institution for providing the necessary facilities and support to carry out this research work. We also thank all individuals who directly or indirectly contributed to the completion of this study.

Source of Support and Fundings:

The authors received no financial support for the research, authorship, and/or publication of this article. This study was conducted without any external financial support or funding from public, commercial, or not-for-profit organizations.

Conflict of Interest statement:

The authors declare that there are no conflicts of interest related to this study.

Author Contribution Statement:

Both authors contributed equally to the conception, design, execution, and writing of the manuscript, and have approved the final version.

Consent to Participate:

Participation was voluntary, and informed consent was obtained from all participants in accordance with ethical standards.