Pharmaceutico-Analytical Profile of Sardha Sama Guna and Dwi Guna Madanodaya Rasa - A Kupipakwa Rasayana

: Introduction: Madanodaya Rasa (MDR) is a Sagandha, Bahirdhuma Kupi Pakwa Rasayana (~ Mercury and Sulphur containing Herbo-mineral formulation) which is prepared out of Sardha Sama Guna Kajjali (S.S.G.K:- 1:1½ ratio of Mercury and Sulphur) as per Rasa Manjari and Dwi Guna Kajjali (D.G.K :-1:2 ratio of Mercury and Sulphur) as per Rasa Yoga Sagara. Both the preparations have the same bhavana dravya (~triturating media) as Rakta Kamala Pushpa Swarasa (~ Red lotus flower) and utility is vrishya(~aphrodisiac). Both are unavailable in the market and its comparative study and standardization is not carried out yet, hence the present study was carried out to standardize the temperature pattern and to do instrumental analysis. Materials and Methods: S.S.G and D.G.MDR were prepared as per classical reference and the samples were subjected to XRD, FTIR and SEM-EDS. Results:- The duration of trituration of S.S.G and D.G.MDR Kajjali was 236 hr and 260 hr respectively. The duration of heating was 22 hrs., and 19 hrs and total yield was 28 % and 14.5 % respectively. XRD showed Cubic crystalline structure with the presence of HgS in both the Kajjali and Hexagonal crystalline structure with HgS in both MDR. SEM shows the S.S.G.MDR has smallest particle size of 18.46 nm and D.G.MDR has 27.53nm. EDS shows Hg and S as the major elements. FTIR report confirms that both the batches of MDR K and MDR contain organic functional groups. Conclusion : XRD, FTIR and SEM-EDS showed not much difference in the crystalline structure.


INTRODUCTION
Madanodaya Rasa is one of the unique Herbo-mineral compounds mentioned in Rasa Manjari [1] and Rasa Yoga Sagara prepared by Kupi Pakwa Rasayana method having vrishya property.As per Rasa Manjari, it is a Sardha Sama Guna and as per Rasa Yoga Sagara [2] it is a Dwi Guna , Saagni Kandastha Bahirdhuma Kupi Pakwa Rasayana prepared under Kramagni tapa for a specific period.Madanodaya is prepared by Parada(~Mercury) and Gandhaka (~ Sulphur) as the main ingredient with a unique bhavana dravya Rakta • Email: editor@ijfmr.comIJFMR230610721 Volume 5, Issue 6, November-December 2023 2 Kamala Pushpa Swarasa.Sardha Sama Guna Kupi Pakwa Rasayana is a rare Kupi pakwa preparation and the specific bhavana dravya, Rakta kamala pushpa swarasa, mainly indicated in Male infertility .Kupi Pakwa Rasayana is a unique Herbo-mineral preparation , prepared in Kacha Kupi (~ glass bottle) by maintaining Kramagni Paka for a specific time in either Valuka Yantra or in Vertical Muffle Furnace [3] .Among Chaturvidha Kalpana's, Kupi Pakwa Rasayana is considered as most potent mercurial preparation than Kharaliya Rasayana and Parpati Kalpana which is quicker in action even in a smaller dosage and having strong chemical bond compared to another Kalpana's.Madanodaya Rasa is a variety of Rasa Sindura and similar preparation with different ratio of Parada and Gandhaka with same bhavana dravya are mentioned by different acharyas which is mainly described as a Vajikarana dravya(~Aphrodisiac drug).Rasa Sindura is a very popular drug which is available in the market , but Madanodaya Rasa is an uncommon drug, and no comparative research works of different ratios has been carried out yet regarding its pharmaceutical preparation and its aphrodisiac activity.
Hence the present study was carried out to standardize the temperature pattern and to provide scientific analytical evidence by conducting Physico-chemical and instrumental analysis regarding the preparation.

AIMS AND OBJECTIVES
This study mainly focuses on preparing Madanodaya Rasa as per Rasa Manjari and Rasa Yoga Sagara and to compare the difference in the temperature pattern and the analytical parameters. 1.To prepare Sardha Sama Guna Madanodaya Rasa and Dwi Guna Madanodaya Rasa. 2. To carry out Physico-chemical analysis and Instrumental analysis like X-ray Diffraction (XRD), Scanning electron microscope and energy-dispersive X-ray spectrometry (SEM-EDS) and Fourier Transform Infrared Spectrometry (FTIR) of the samples.

MATERIALS AND METHODS Collection of raw drugs:
Main raw drugs used in the preparation of Madanodaya Rasa were purchased from an authenticated source ensuring Grahya Lakshanas mentioned in ancient texts and SEM-EDS and XRD were done before the study.Raw drug identification of Herbal drugs like Rakta Kamala Pushpa was done in Pg & PhD studies of Department of Dravya Guna Vijnana, G. A. M.C, Bangalore.1.5 Kg Hingula was procured from All India Kirana Stores, Mumbai.Parada was extracted from purified Hingula in the Dept. of Pg and PhD studies of Rasa Shastra and Bhaishajya Kalpana, G.A.M.C, Bangalore.1.5Kg Gandhaka was procured from Kajarekar Ayurvedic raw drug depot , Belgaum and purified.Rakta Kamala Pushpa was collected from K. R. Market ,Bangalore.Associated drugs for purification of raw drugs like Ksheera, ghee, nimbu and haridra churna were collected from local market, Bangalore.PURVA KARMA ( ~ PRE-PREPARATORY PHASE) a) Hingula Shodhana and Hingulotha Parada Nirmana 780 gm Hingula(~Cinnabar) was purified by levigating with Nimbu Swarasa(~lemon juice) for 7 times and allowed to dry [4] .Hingulotha Parada was prepared by Urdhwa Patana method(~sublimation process) in 3 batches [5] .303gm Parada was extracted from 475 gm Shudha Hingula.Parada was purified by doing mardana with Haridra Churna (~Curcuma longa.Linn) for 24hrs and then washed thoroughly with hot water, filtered through cloth, dried, and stored in an airtight glass container [6] .Images of Hingula Shodhana and Hingulotha Parada nirmana are illustrated in figures 1 and 2.

Figure . 1 Hingula
Temp. graph of Hingulotha Parada Nirmana is shown in Graph No 1. b) Gandhaka Shodhana 650gm Gandhaka was purified by Galana method with cow's milk 7 times and 525 gm pure Gandhaka was obtained [7] .Gandhaka was finely powdered and melted with equal quantity of ghee and filtered through a cloth tied around the mud pot filled with milk.After each Galana process Gandhaka was washed thoroughly with hot water to remove the content of milk and ghee, dried and powdered.After purification it was powdered and stored in an inert glass container.This purified Hingulotha Parada and Shudha Gandhaka were used for the preparations of two batches of Madanodaya Rasa.

PRADHAN KARMA (~MAIN PHARMACEUTICAL PREPARATION) Placement of Kupi in VMF
One insulation brick (Komi bricks) with a length and width of 11 cm and a height of 6 cm were placed inside the bottom of heating element of the VMF, to adjust the height of the Kupi, so that the neck of the Kupi was to be at the same level to that of the upper part of the VMF.The depth of the heating element was 30 cm, and the diameter is 15 cm.The Kacha Kupi, which was earlier filled with Madanodaya Rasa Kajjali, was properly placed in the heating element of the VMF .The Kajjali-filled Kupi had been kept at the centre of the heating element chamber at ½ an inch from the sides and small pieces of bricks were holding the Kupi firmly without any movement.A hole was made in the centre of the ceramic wool and placed over the neck of the Kupi so that to allow the passage of the Kupi and used to cover the furnace opening to avoid escaping heat from the furnace.K-type Thermocouple and Tapta Shalaka were placed inside the furnace nearer to the wall of the furnace by making two holes in the wool without disturbing the Kupi.Sardha Sama Guna MDR was prepared with 200 gm Kajjali by Vertical Muffle Furnace method.Kramagni paka was maintained throughout the procedure.Total time taken was 22 hours and yield were 56gm.Dwi Guna MDR was prepared with 200 gm Kajjali and time taken was 19 hours and yield were 26 gm by VMF method.In S.S.G.MDR corking was done after 18 hours and in D.G.MDR it was done after 12½ hours.After corking 4 hours heat was given to S.S.G MDR and 6 hours heat was given for D.G.MDR.After complete self-cooling Kupi was removed from VMF. Observations during the preparation of S.S.G.MDR , and D.G.MDR have been tabulated in Table 1 and Table 2 respectively         When finely powdered MDR was carefully sprinkled into a glass vessel containing water, MDR floats on the surface of the water.Nischandratva There were no shiny particles in the finely powdered MDR even when it was rubbed between wet thumb and index finger observed in the bright sunlight.

Nirdhoomatva
The MDR didn't emit any smoke when sprinkled over red-hot coal.SEM ANALYSIS [9] SEM reports show that the smallest particle size of S.S.G.MDR.K and S.S.G.MDR are 36.57nmand 18.46 nm respectively.The smallest particle size of D.G.MDR K and D.G.MDR are 36.91nmand 27.53 nm respectively.Thus, this report confirms that particle size is reduced, and it helps in easy absorption and increased bioavailability.[ Table 6] FTIR reports shows the presence of organic compounds which may added during purificatory processes, and this will reduce the toxicity and increases the bioavailability of the product.SEM reports shows the reduced particle size.EDS shows that the Mercury percentage of MDR is like Hingula and at the end there is no change in the chemical composition.

Graph No 10
shows XRD Reports of Dwi Guna MDR Kajjali Graph No 11 shows XRD reports of S.S.G.MDR Graph No 12 shows XRD analysis of DWI GUNA MDR FTIR ANALYSIS [9] FTIR report confirms that both the batches of MDR K and MDR contain organic functional groups like Fluro compound, alkane, amine, alcohol, aldehyde ether, amine salt, carboxylic acid etc which are added during bhavana.[Graph no.13-16] Graph No.13 shows FTIR reports of S.S.G.MDR Kajjali Graph No 14 shows FT-IR reports of Dwi Guna MDR Kajjali Graph No 15 shows FTIR reports of Sardha Sama Guna MDR Graph No.16 shows FTIR reports of D.G.MDR.

Table No .1 shows observations during S. S. G .MDR Time duration Time Set Temp. Display Temp. Thermo- Couple Temp
GraphNo 2 shows temperature pattern of S.S.G.MDR.Table No .2shows observations during D.G.MDR.245.6 ̊ C Dense yellow fumes found.Bottom was not visible with torch light.

MDRK DG MDRK DGMDR Water ̊soluble ̊Åsh Graph No. 8 shows Loss on drying. Table No. 5 shows Physico-chemical analysis XRD ANALYSIS [8]
Samples S.S.G.MDR Kajjali, D.G.MDR Kajjali, S.S.G.MDR, D.G.MDR were subjected to X-Ray diffraction study.[Figures 24-27] XRD values of Hingula were compared with Standard Joint Committee Powder Diffraction File (JCPDS) and confirmed that the drug contains HgS in hexagonal crystalline structure.XRD values of S.S.G.MDR K and D.G.MDR K were compared with standard value and confirmed that both contains Mercury sulphide (HgS) with Cubic crystalline structure, whereas S.S.G.MDR and D.G.MDR are having Hexagonal crystalline structure and it contains HgS.[ Graph no 9-12] Graph

Table No 6 shows the SEM result .
The EDS report shows that the WT.% of Mercury and Sulphur in Hingula is 85.35% and 14.65% ; in S.S.G.MDR K is 63.29% and 36.71% ; in D.G.MDR K is 71.73% and 28.27% ; in S.G.G.MDR is 84.64% and 15.36% ; in D.G.MDR is 85.27% and 14.73% respectively.Wt.% of Mercury was reduced, and wt.% of Sulphur was increased in both the Kajjali whereas Mercury wt.percentage increased in both the product due to the evaporation of extra Sulphur during Kramagni paka.[Table7-8]

Table no 8 shows the EDX result of D.G. MDR Kajjali and D.G. MDR
DISCUSSIONKupipakwa Rasayana is a unique Herbo-mineral formulation prepared in specifically prepared glass bottle by gradual heating in Valuka yantra or EMF.In general, Kupi Pakwa Rasayanas have disease curing property, Rasayana and Vajikarana properties, are mainly prepared by Sagandha Murchana or Nir Gandha Murchana.Madanodaya Rasa is a Sagandha Bahirdhuma, Kandastha Kupi Pakwa Rasayana, described by Rasa Manjari and Rasa Yoga Sagara as a Vrishya yoga.Similar preparations are mentioned by Rasa Ratna Samucchaya, Bhaishajya Ratnavali, Rasa Ratnakara etc.The name itself suggests that it is suitable for increasing desire of love and affection, and it is suitable to treat male infertility.Mercury has great affinity towards Sulphur molecules and during the preparation of Kajjali it binds easily with Sulphur molecules and molecules of Mercury easily disintegrate.It is believed that Mercury is Siva Virya and Sulphur is menstrual blood of Goddess Parvathy which represents the easy assimilation and strong bondage between the two drugs.Sulphur has the tendency to reduce toxicity of Mercury and therefor if the quantity of Sulphur increases provides a good therapeutic effect.During the preparation of Madanodaya Rasa by two different ratios, it was noted that the time taken was reduced in D.G.MDR compared to S.S.G.MDR.It was found to be difficult to assess all the siddha lakshanas in both the preparations and after getting Copper coin test positive along with grey colour discoloration in the foil associated with mercury particles corking was done to avoid the escape of Mercury.In XRD analysis of S.S.G.MDRK a total of 36 peaks were identified at different angles from 15.361 to 86.292.10 strong peaks were chosen as strong with their relative intensity and compared to Standard X-Ray Powder diffraction file.(XPDF) and confirmed that it contains Mercury sulphide with Cubic crystalline structure.Total 25 peaks were identified in Madanodaya Rasa (22 Hours preparation) at different angles (2Ɵ) from 23.75 to 88.32 and 10 strong peaks were compared to Standard X-ray powder diffraction file.(XPDF) and it has Hexagonal crystalline structure which contains HgS .Total 41 peaks were identified in Dwi Guna Madanodaya Rasa Kajjali at different angles(2Ɵ) from 26.35 to 86.28 and identified as Mercury sulphide with Cubic crystalline structure.Totally 24 peaks were identified in Dwi Guna Madanodaya Rasa (19 Hours) preparation) at different angles (2Ɵ) from 23.829 to 88.41 and identified as Cinnabar with Hexagonal crystalline structure.Thus, the black sulphide of Mercury converted into red Sulphide of Mercury by gradual increase of heat.