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Introduction

The global interest in Ayurvedic herbomineral medications is growing steadily(1). Their rising demand is attributed to their therapeutic effectiveness, safety, and logical pharmacological outcomes. Parada (Mercury) and Gandhaka (Sulphur) are key components of Rasasindura however, their ratios can differ across various formulations, with a range from 1:1/6 to 1:6(1). The Kupipakwa Rasayana technique is frequently utilized by Ayurvedic practitioners to address a variety of conditions, including Prameha (Polyuria), Shoola (colicky pain), Bhagandara (fistula in ano), and Jwara (fever). This method serves functions such as Rasayana (promoting healthy tissues) and Vajeekarana (enhancing vitality), aiding in issues like digestive disorders, fever, urinary tract infections, respiratory illnesses, as well as promoting sexual health and rejuvenation(2). Rasasindura is an exceptional mercurial preparation crafted using the Kupi-pakwa Rasayana technique, where the material involved is placed in a narrow-mouthed, short-necked glass container covered with several layers of clay-coated muslin cloth. This mixture is then digested by gradually applying increasingly intense heat to achieve the desired quality of the final product. Nowadays, heavy metal toxicity is regarded as a significant concern. the sublimed mercury compounds commonly used in Ayurvedic treatments, such as Shadgunabalijarita Rasasindura(3), are considered safer than other options .Numerous researchers have confirmed findings from toxicity studies. When preparing Kupipakwa medicine, increasing the ratio of sulfur to mercury complicates the preparation process. Failures in the preparation of Kupipakwa medicines have often been observed when sulfur and mercury are mixed in equal proportions (Samagunabalijarita)(4). The likelihood of failure tends to rise with a higher amount of sulfur. In this current study, an effort has been made to establish a standardized method for preparing Shodhana). Consequently, this study aims to conduct pharmaceutical standardization and document the actual yield following Shadguna Gandhaka Jarana. Historically, the "Shadguna" (sixfold) sulfur variant is recognized for its enhanced therapeutic qualities, which are believed to stem from the Gandhaka Jarana process. While traditionalists favor a method involving sequential additions, there is also the option of a Single-Step approach method aims to standardize the heating cycle to ensure uniform sublimation and bond formation in a controlled environment.

Materials and Methods

The necessary raw materials of the necessary quality were gathered from the neighborhod marketplace. They were verified and put through the Kupipakwa process of Shodhana (purification) and Pachana (digestion).The preparation of Shodhana) Rasasindura shall follow the instructions in the classical source "Rasa Tarangini(5)."Utilizing physicochemical   metrics and organoleptic characteristics, the created formulation will be evaluated.The experi-mental investigation will be conducted in compliance with OECD criteria(6) .

1. Collection of Raw Material from certified seller
2. Drug Cleaned and Dried Shodhan of Parad and Gandhak
3. Preparation of Kajjali in 1:6 (Parad:Gandhak) Ratio
4. Bhavana of Kumari swarasa to Kajjali Mardan
5. Filing of Kupi with Kajjali
6. Put Kupi in EMF Preparation of Shadgunabalijarita Rasa Sindoor.

 Raw Material Purification (Shodhana)(7)

Before the pharmaceutical process, the raw materials underwent rigorous purification:

Shodhan(8) of Parad(9)

Materials

A. Parad  B. Lashuna Kalka C. Khalwayantra

1. To eliminate Parad Doshas, Parad Shodhan will be performed for three to seven days by taking Lashun Kalka (paste) using Khalva Yantra.

2. The Parad Samhita mentions Shodhan with Lashun as one of many Parad Shodhan techniques.

3. Lashun is natural remedy for Parad Bandh, contains sulfur & is an antidote for heavy metal toxicity.

4. Shodhan process reduces toxicity of 

Shodhan of Gandhak(10)

Purification of raw sulphur. In a steel wok coated with cow ghee, raw sulphur was melted entirely. The melted sulphur was then poured through a cotton cloth into a vessel filled with lukewarm cow milk, allowed to sink to the bottom, and then collected. It was then properly cleaned in hot water multiple times and allowed to dry in the shade.

Materials

1. Ashuddha Gandhak 2. Goghruta 3. Godugdha 4. Water as required 5. Vastra

Process

1. Godugdha will be taken in a stainless-steel vessel to cleanse Gandhak (sulphur). Goghrita, a type of ghee derived from cow's milk, will be spread over a four-layer muslin cloth and attached to the container's mouth.

2. Gandhak will reach a temperature of 115°C to 123°C, which is its melting point.

3. Gandhak will be melted and then put into a jar filled with Godugdha after being passed through muslin material that will have been smeared with Ghrita.

4. Gandhak will be removed from the container when it will be settled to the bottom.

5. After two more repetations of this procedure, the finished product will be removed.

6. The greasy portion of the Ghrita will be completely removed by repeatedly washing it in hot water. After that, it will be dried and ground into Shuddha Gandhak powder.

7. After being gathered, the product will be sealed in an airtight glass container to keep out moisture and light.

Kajjali Preparation(11)

Preparation of Shadguna Kajjali with Bhavana of Aloe vera swarasa(12).

• Purified Mercury (1part) and Purified Sulphur (6parts) were weighed & taken in the mortar and triturated till the total mass was converted into a fine black, smooth, lusterless powder.
• Obtained Kajjali was also taken into the mortar pestle, and Juice of the Aloevera(Kumari) was added till the mixture became muddy.
• Then this mixture was triturated for 24 hours [6 hours daily] until it became dry and fine.

Shadguna Kajjali Preparation(13):

Materials:

1.Shudha Parada:105 g. 2.Sudhha Gandhaka: 600g.

Method: Mardana

Equipments:

Khalva Yantra, Spatula, Steel plate, Steel Spoon etc.

Procedure:

1.Shodita Parada  and Shodita Gandhaka were taken in equal quantity and triturated in Khalva Yantra. Gradually the white color of Parada  and greenish yellow color of Gandhaka disappear and a black powder was formed.

2.Trituration was continued till the powder became jet black in color and very fine like Kajala and fulfilled all the criteria of Kajjali.

3.Average to and fro movements of Peshani were 20-22 times/ minute.

4.Mardana was done for 6hrs daily.

Observations:

1.After 12hours of trituration,the color of Gandhaka started transforming in to blackish yellow.

2.After 24hours of trituration, Parad particles almost disappeared and the mixture turned into dark black color. But when rubbed between the fingers a small particle wasseen.

3.After 48hours of trituration,it appeared that the Kajjali was prepared.But when it was examined under sunlight,luster of free Parad particles were observed.

4.After 72hours of trituration,the mixture completely turned into Kajjali and fulfilled all the criteria of Kajjali.

5.Thus, prepared Kajjali was fulfilling the test of Varitara and Rekhapurnatva too.

6.The entire powder became fine,black,smooth, lusterless.

Showing different phases of Shadguna Kajjali during preparation

Table No 1-Time and Phases of Kajjali Preparation (1:6 ratio)

Precautions:

1.To prepare Kajjali,Gandhaka should always be taken in fine powder form.

2.Trituration should be done slowly and cautiously to check the loss of Parada.

3.Khalava should be kept covered when the process is not in progress.

Results:

1.Weight of Shudha Parada :105g.

2.Weight of Shudha Gandhka: 562g.

3.Weight of Kajjali obtained: 685g.

Cause of weight Loss

1.Spilling of mixture during trituration.

2.Sometimes particles of Kajjali adhere to Khalva which is difficult to collect.

3.Some quantity of Kajjali was lost during performing the confirmatory test of the product.

Preparation of Shadgunabalijarita Rasasindura(14) :-

The whole procedure of Rasasindura preparation was categorized under 3 headings

1.Purva Karma

2.Pradhana Karma

3.Paschat Karma

Purva Krama

1.Preparation of Kacha Kupi

2.Filling of Kajjali into Kacha Kupi

3.Placing of Kacha Kupi in Valuka Yantra

Pradhana Krama

1.Heating Schedule (Kramagni tapa)

2.Observation and recording of temperature

3.Sealing with cork to Kacha Kupi and self-cooling of the apparatus

Pashchat Karma

1.Removal of Kacha Kupi from Valuka Yantra

2.Breaking of Kach Kupi

3.Collection of Final Product

Kupi Nirmana(15)

Equipment -Kacha Kupi, Clean Cloth according to size of Kacha Kupi, Multhanimitti Water.

Procedure:

1.A clean and dry Kacha Kupi with narrow mouth having capacity around 700ml was taken (green colour beer bottle).

2.A clean cloth was taken that was smeared with Multhanimitti.  This  mud  smeared  cloth  was wrapped around the Kupi from all the sides to cover it uniformly.

3.This was kept for complete drying, the whole procedure was repeated for 7 times,each wrapping was done after complete drying of previous layer.

4.It will take maximum 4hours for drying one layer of wrapped Multani mitti cloth.

Precaution:

1.Wrapping was performed without any fold in the layer of cloth and air gaps.

2.Stony particle in the Multani mitti was removed before applying to the cloth.

3.Tight packing was done especially over bottom.

Observation:

1.Initially for drying each layer time period was less compare to successive layer.

2.It will take maximum 4 hours for drying one layer of wrapped Multani mitti cloth.

3.Kupi went on becoming thicker and thicker.

KupiPoorna(16)

Materials: Prepared Kupi, funnel, weighing Machine, ShadgunaKajjali 200g.

Procedure:

1.A clean funnel was placed over the mouth of the Kacha Kupi, mixture was slowly added into the two Kacha Kupi through the funnel.

2.Mouth of the funnel was cleared using the spatula while filling the mixture.

3.Mouth of the Kupi was temporary covered by using cork.

4.Each Kupi is filled with 200gms.

Observation:

1.Usage of funnel facilitates easy filling of the mixture in to the Kupi without spillage.

2.Kajjali was again triturated for half an hour before filling the Kupi.

3.Inner aspect of the Kupi was cleaned and dried properly with a clean cloth tied over a stick.

4.Care was taken to spread the Kajjali uniformly inside the Kupi.

Kupi Sthapana(17)

Materials: EMF(Electric muffle furnace), bottle filled with Shadguna balijariata Kajjali(18) .

Procedure(19)

1.Take an EMF of good quality.

2.The Kupi was then placed in it at given place.

Pradhana karma

The specific observations during Pradhana karma were as follows:

Heating Schedule:

The heating was provided via EMF using a graduated Agni (thermal) schedule:

1. Mridu Agni (Mild): 100°C – 250°C (11 hours)
2. Madhyama Agni (Moderate): 250°C – 450°C (28 hours)
3. Teekshna Agni (Intense): 450°C – 630°C (9 hours)

Table No 4- Stages of Shadgunabalijarita Rasasindura with Temp and Observation

Graph no 1. Temperature Variation Across Time

Precautions:

1.Kajjali was again triturated for ½hour before filling of into Kupi.

2.The Kupi was kept exactly in Space provided in machine so that homogenous temperature would be obtained(20).

3.Precautions were taken while Kupi mouth closed with cork.

4.Kupi was removed from furnace and broken carefully.

5.Eyes and nose were protected by wearing glass and mask throughout the practical.

Kupi Bhedana

Materials: Knife,Thread, Kerosene, Matchbox, Kupi containing Shadaguna Kajjali, clean container.

Procedure:

• The bottle was carefully removed from the EMF.
• The outer layer of the bottle was scrapped  carefully  with  the  help  of  a  knife  to remove the Multanimitti coating and then the Kupi was wiped with wet cloth.
• kerosene dipped thread was tied around the bottle and was set to fire. When the thread burnt, it was wrapped with wet  cloth  then  broke  the  bottle in two  halves.
• Shadgunabalijarita Rasasindura was obtained as a whole block just by tapping the bottle.

Observations:

1.After taking out the Kupi from EMF, the upper portion was black in colour.

2.After complete removal of layers, the bottle was cleared then shiny and dark colour sublimated product was observed.

3.There was a thick collection of Rasasindura in the neck region, where as the lower portion contained grey coloured residue.

4.Block of Shadgunabalijarita Rasasindura was shiny greyish red coloured.

Critical Observations:

• Fuming Stage: Profuse yellow fumes of sulphur were observed during the mid-stage of Madhyama Agni.
• Chakra Nanatva: The formation of a sulphur "ring" at the neck of the flask required frequent clearing with a hot iron rod (Shalaka).
• Copper Coin Test: Positive (no white deposit) after 16 hours, indicating the completion of the reaction.

Precautions:

1.The bottle was separated into two halves only after the breaking noise and no force  was applied to separate the bottle.

2.The upper part of bottle should tap carefully so that bottle should not crack.

3.The Shadgunabalijarita Rasasindura was weighed and powdered in porcelain mortar and procured in airtight container.

RESULTS:

1.Time taken for preparation:48 hrs.

2.Weightof Kajjali taken:200g.

3.Weight of Shadgunabalijarita Rasasindura obtained:28g.

4.Weight of residue obtained:02g.

Discussion

• The single-step method for Shadgunabalijarita Rasasindura preparation poses a unique challenge: the high volume of sulfur.
• Unlike the 1:1 ratio, the 1:6 ratio requires a significantly longer Madhyam Agni phase to prevent the "choking" of the flask neck by rapidly sublimating sulfur.
• Our evaluation shows that while the yield is comparable to traditional methods, the Single-Step approach results in a highly crystalline, cinnabar-red Rasasindura with a stable HgS bond.
• The excess sulfur acts as a catalyst for the "Jarana" process before being expelled as SO2 gas

Conclusion

1.Preparation of Samagunabalijarita Rasasindur was easier and yield was also more compared to Shadgunabalijarita Rasasindur.

2.From the pharmaceutical point of view,there was much difference between Samaguna and Shadgunabalijarita Rasasindura.

3.Difference was there in the ratio of ingredients, total duration of heat and quantity of yield. In case of Samagunabalijarita Rasasindura, duration of Pakawas less but the yield was more. In case of Shadgunabalijarita Rasasindura duration of heat was more but yield was less.

4. Shadgunabalijarita Rasasindura can be successfully prepared in a single-step Kupipakva cycle, provided the Madhyama Agni phase is strictly controlled to manage the high sulfur-to-mercury ratio.

5.This method reduces the labour intensity of the traditional Jarana process while maintaining the pharmaceutical integrity of the Rasayana.

6.According to Reference, time required to prepare Shadgunabalijarita Rasasindura is 7days in valuka yantra, but this study shows that in minimum hours with the help of traditional Electric Muffle Furnace it can be made.

Pharmaceutical Yield:

Table No 5-Product Output

Acknowledgements:

The authors are deeply grateful to their teachers and mentors for their constant guidance, encouragement, and support throughout this work. They also sincerely appreciate the institutional facilities and the academic environment that made the completion of this study possible.

Conflict Of Interest:

The authors declare no conflict of interest. This research did not receive any specific grant from funding agencies in the public, commercial, or not-for-profit sectors.