Introduction {#sec1-1}
============

*Makaradhwaja* is a popular *Kupipakwa Rasayana*, prepared with the *Swarna* (gold), *Parada* (mercury) and *Gandhaka* (sulfur) in a specified ratio.  It was first described by Rasendra Chintamani\[[@ref1]\] by the name of *Chandrodaya Rasa*, although the word *Makaradhwaja* was first coined by Rasaratnakara.  The term *Makaradhwaja* is composed of two words, i.e. *Makara* and *Dhwaja*.  The term *Makaradhwaja* is also a synonymous of *Kamadeva*,\[[@ref2]\] the God of beauty.  When it is used for therapeutic purposes, it produces *Rasayana* and *Vrishya* (in literary meaning, *Makara* indicates the aphrodisiac because *Makara Retasa* has been described as best *Shukrala*\[[@ref3]\] while *Dhwaja* \[straight rod or flagpole\] indicates its sexual potency.  It is directly correlates the *Dhwajabhanga*, i.e. non-erection of penis.)  property.  A total of 30 formulations\[[@ref4]\] are found described by the name of *Makaradhwaja* and *Chandraodaya*, and two types of instruments are used for its preparation, i.e. *Valuka Yantra* and *Khalwa Yantra*.  Nowadays, *Makaradhwaja* is generally prepared as per the reference of Rasendra Chintamani due to easy and convenient preparation method; here, *Swarna, Parada* and *Gandhaka* are used in 1:8:16 ratio, but in different *Rasa* classics it is found described that the amount of *Gandhaka* is directly proportional to the therapeutic activity.\[[@ref5]\] Therefore, in this study, the ratio was followed as per the *Bhaishajya Ratnawali*, i.e. 1:8:24. 

Aims and objective {#sec2-1}
------------------

The aim of the study was to develop the Standard Manufacturing Process (SMP) for *Makaradhwaja* prepared by *Swarna-Patra-Varkha* and *Bhasma* by Electrical Muffle Furnace (EMF). 

Materials and Methods {#sec1-2}
=====================

*Swarna* was collected from a local authentic hallmark-certified Jeweler from Jamnagar. * Hingula, Tila Taila, Gandhaka* and *Kulattha* were collected from the Pharmacy, Gujarat Ayurved University, Jamnagar. * Takra, Kanji, Kulattha Kwatha, Japakusuma Swarasa, Kumari Swaras, Nimbu Swarasa* etc.  were taken as per the classical reference and processed through prescribed methods.  The whole process has been divided into the following:

Shodhana of raw materials {#sec2-2}
-------------------------

*Kantakavedhi Swarna Patra* was prepared from 24 carat gold biscuits on the machine that had 2.5 cm breadth, 150 cm length and 0.5 mm diameter.  For *Samanya shodhana*, these *Kantakavedhi Swarna Patra* were heated up to red hot through a Gas blower and dipped into prescribed liquid media three times.\[[@ref6]\] *Gandhaka*\[[@ref7]\] was melted along with *Goghruta* and poured into the *Godugdha*.  Then, *Swedana* (heating under liquid bath) was done for 3 hrs.  After that, it was washed with hot water, dried and powdered \[[Table 1](#T1){ref-type="table"}\]. 

###### 

Observation of *Shodhana* of the raw materials

![]( Ayu-32-109-g001)

Hingulottha Parada Nirmana {#sec2-3}
--------------------------

*Shodhana* of *Hingula* was done by giving *Bhavana* of lemon juice three times.\[[@ref8]\] After that, it was washed, dried and stored.  The required amount of *Parada* was extracted from *Hingula* by *Nada Yantra* method.  Fine powder of *Shuddha Hingula* was wrapped in cotton cloth (equal weight of *Hingula*) and burnt under the pot.  Thus, due to heat, the sulfur part of *Hingula* burns and leaves the *Parada*, which gets evaporated as vapor and is collected on the inner side of the pot. * Parada* was collected by rubbing with cloth and then washing with hot water and filtering through four folders of cloth.  This *Parada* was used to prepare *Makaradhwaja* \[[Table 2](#T2){ref-type="table"}\]. 

###### 

Observation of extraction of *Parada* from *Hingula*

![]( Ayu-32-109-g002)

Preparation of *Swarna Varkha* {#sec2-4}
------------------------------

*Shodhita Swarna Patra* was cut into small pieces (2 cm × 4 cm length).  Then, these *Patra* were continuously hammered with the help of an iron hammer in the leather bags up to the *Varkha* formation.  This procedure was performed in Ahmedabad under supervision.  The thickness of the *Varkha* was not measured because it was too thin.  This *Varkha* was then used in the preparation of *Swarna Pishti*. 

Preparation of *Swarna Bhasma* {#sec2-5}
------------------------------

*Shuddha Swarna Patra* was cut into fine pieces and triturated with the *Hingulottha Parada* up to *Pishti* (amalgam) formation and then this amalgam was kept between *Shuddha Gandhaka* in *Sharava*, i.e. *Swarna Pishti* was covered completely by *Shuddha Gandhaka*. * Sharava Samputa* was prepared and after complete drying, it was subjected to heat.  After each *Puta, Parada* was reduced by 1/16^th^ from the quantity of the initial *Parada* and the *Gandhaka* was in equal quantity of *Swarna* up to the last *Puta*.  But, here, a total of 30 *Puta* were required for the preparation of *Swarna Bhasma* up to *Kumkum* colored without shining of particles.\[[@ref9]\]

Preparation of *Swarna Pishti* {#sec2-6}
------------------------------

*Shudha Swarna patra* were cut into small pieces. * Hingulottha Parada* was taken in *Simaka Khalvayantra* and, after that, *Shuddha Swarna Patra* pieces were added one by one to it with proper trituration.  The mixture was triturated up to a homogeneous form.  The *Nimbu Swarasa* was added for proper preparation of amalgamation.  Thus, semisolid, soft *Swarna Pishti* (amalgam of gold) was formed.\[[@ref10]\]

The second batch of *Swarna Pishti* was prepared from *Swarna Varkha* and *Hingulottha Parada*; Nimbu Swarasa was not needed because it was prepared within few minutes by simple trituration. 

The third batch of *Swarna Pishti* was prepared from *Swarna Bhasma* and *Hingulottha Parada*, with the addition of *Nimbu Swarasa*.  But, for this, a large quantity of *Nimbu Swarasa* and a longer period of trituration was required even it was not properly prepared so that an equal amount of *Saindhava Lavana*\[[@ref11]\] was added to it and again triturated till the formation of *Pishti* \[[Table 3](#T3){ref-type="table"}\]. 

###### 

Observations of *Swarna Pishti* from *Swarna Patra, Varkha* and *Bhasma*

![]( Ayu-32-109-g003)

Preparation of *Kajjali* of *Makaradhwaja* {#sec2-7}
------------------------------------------

The *Swarna Pishti* (amalgam of gold) was taken in a *Simaka Khalvayantra* and *Shuddha Gandhaka* was added to it in the prescribed quantity. * Mardana* was done for 24 hrs till fine, soft, *Nishchandra Kajjali* was formed \[[Table 4](#T4){ref-type="table"}\].  This was subjected to *Bhavana Dravya* of *Japakusuma Swarasa*, which was added to it in an adequate amount to wet the *Kajjali (Rasapankavat)* semisolid and *Mardana* was carried out for 3 hrs, till the homogeneous, soft mass was formed and then it was dried in sunlight\[[@ref8]\] \[[Table 5](#T5){ref-type="table"}\].  The same procedure was repeated by adding the *Bhavana Dravya*, i.e. *Kumari Patra Swarasa*, and it was taken 1/4^th^ of the total amount of *Kajjali* as it was found to be sufficient to wet the total material.\[[@ref10]\] The observations of *Kupibharana* of *Makaradhwaja* are shown in [Table 6](#T6){ref-type="table"}. 

###### 

Observations during preparation of *Kajjali* of *Makaradhwaja*

![]( Ayu-32-109-g004)

###### 

Observations during Bhavana processing of *Kajjali* of *Makaradhwaja*

![]( Ayu-32-109-g005)

###### 

*Kupikabharana* of *Makaradhwaja*

![]( Ayu-32-109-g006)

Preparation of *Makaradhwaja* Apparatus {#sec2-8}
---------------------------------------

Mortar and pestles, *Kanchakupi, Multani mitti*, cloth, *Loha shalakas*-2, kerosene oil, matchbox, thread, enamel tray, glass container, cork, copper coin, torch, knife; Electric muffle furnace: Outer length: 40 cm, breadth: 40 cm, height: 50 cm, inner hearth length: 15 cm, breadth: 15 cm, depth: 30 cm, *Kanchakupi*: Amber-colored beer bottle, capacity: 625 ml, total height: 28 cm, cylindrical part: Height: 14 cm; circumference: 24.5 cm; diameter of the bottom: 6.5 cm, conical part: Height: 14 cm; diameter of the mouth: 2cm, weight: Before *Kapadamitti*: 450 g; after *Kapadamitti*: 590 g.

### Procedure {#sec3-1}

The *Bhavita Kajjali* of *Makaradhwaja* was taken, triturated well in *Khalvayantra* and filled in *Kanchakupi* \[[Table 6](#T6){ref-type="table"}\].  The *Kupi* was placed exactly at the center of the electric muffle furnace and fixed in proper position with the help of firebrick blocks.  The heating process was carried out in a *Kramagni* pattern, i.e. increasing order but intermediate heating.  Heat was gradually increased over a period as per the schedule, i.e. 3 hrs *Mandagni* (120-250°C), 6 hrs *Madhyamagni* (250-450°C), 3 hrs *Tivragni* (450-600°C).  The temperature of the furnace was recorded after intervals of 30 min.  During the course of heating, the hot *Shalaka* was repeatedly inserted into the mouth of the *Kupi* to burn the accumulated sulfur at the neck of the bottle to prevent blocking.  After achieving the confirmative test, the mouth of the *Kupi* was corked and the temperature was increased up to 600°C and it was maintained for the next 2 hrs.  \[[Table 7](#T7){ref-type="table"}\].  Then, the Muffle furnace was switched off and left for self-cooling.  After *Swangsheeta, Kanchakupi* was taken out from the muffle furnace and the outer covering was removed; a thread (which was soaked in kerosene) was tied below 1 inch from the final product and ignited.  Then, a few drops of water were sprinkled, which leads to break the *Kupi*, and finally, *Makaradhwaja* was collected from the neck of the *Kupi.  Swarna* powder was collected from the bottom of the *Kupi*.  The *Makaradhwaja* was triturated well in *Khalva yantra* up to a fine red color powder \[[Table 8](#T8){ref-type="table"}\]. * Makaradhwaja* samples were analyzed by employing various possible organoleptic, physical and chemical parameters \[Tables [9](#T9){ref-type="table"} and [10](#T10){ref-type="table"}\].  The same procedures were followed for all three samples of the *Makaradhwaja*, i.e. for *Swarna Patra, Swarna Varkha* and *Swarna Bhasma*.  A total of 17 batches were prepared to determine the SMP \[Tables [8](#T8){ref-type="table"} and [11](#T11){ref-type="table"}\].\[[@ref10]\]

###### 

Observations during *Kupipaka* of *Makaradhwaja* (MP, MV and MB avg)

![]( Ayu-32-109-g007)

###### 

Results of preparation of *Makaradhwaja* of the different batches

![]( Ayu-32-109-g008)

###### 

Classical analytical tests of the *Makaradhwaja* samples

![]( Ayu-32-109-g009)

###### 

ICP analysis of the *Makaradhwaja* sample

![]( Ayu-32-109-g010)

###### 

Standard manufacturing procedure for the preparation of *Makaradhwaja*

![]( Ayu-32-109-g011)

Discussion {#sec1-3}
==========

For the preparation of *Kupipakwa rasayan*, preparation of *Kajjali* and heating pattern are the most important factors to obtain maximum quantity of yield and to increase efficacy of the product without any untoward effect.  As per classical texts, the *Kramagi*\[[@ref12]\] heating pattern should be provided during processing of any *Kupipakwa rasayana*.  It means temperature pattern should be an increasing order but intermediate heating process.  In can be divided into three stages, i.e. *Mridu, Madhya* and *Tivra agni*.  Here, *Mriduagni* indicates the melting stage of *Kajjali, Madhyam agni* indicates the boiling stage of *Kajjali* and *Tivra agni* means immense heating, which takes place a confirmative test of the final product.  One such effort had been done by Prajapati *et al*.,\[[@ref13]--[@ref15]\] who has given the temperature range for the particulars of *Agni*, such as-*Mridu Agni* 120-250°C (6 hrs), *Madhyama Agni* 250-450°C (6 hrs), *Tivra Agni* 450-630°C (6 hrs).  This standardization was done in an electric muffle furnace for the preparation of *Makaradhwaja* in the ratio of 1:8:16, and the same was also followed by Chinta Durga *et al*.\[[@ref16]\] and Patgiri *et al*.\[[@ref17][@ref18]\]

But, for the present study, to prepare the *Triguna Balijarita Makaradhwaja* in minimum heat duration, the heating pattern was changed.  As the proportion of *Gandhaka* increases with *Parada*, i.e. *balijarana*, it was observed that there is an increase in the heating duration for the *Jarana*.  Here, an attempt was made to prepare the same in minimum time with least consumption of energy while maintaining its therapeutic efficacy.  Therefore, the duration of *Mriduagni* and *tivraagni* period was decreased without disturbing the *madhyamagni* duration (i.e., *Mriduagni* for 3 hrs, *Madhyamagni* for 6 hrs and *Tivragni* for 3 hrs).  Because of a higher amount of *Kajjali* in *Makaradhwaja* prepared by the *Swarna Patra* sample, each batch was prepared with 424 g of *Kajjali* whereas in the *Varkha* and *Bhasma* samples, because of the lesser amount, it was prepared with 330 g of *Kajjali* in the same heat duration for the standardization purpose. 

The *Hingulottha Parada* was used because as per classical text, its properties are equal to those of *Astasamskarita Parada*.\[[@ref19]\] For the preparation of *Swarna Pishti* of *Varkha*, only 1 hr was required as compared with 12 hrs required for *Swarna Patra*, which may be because the particle size of *Swarna Varkha* is too fine as against that of *Swarna Patra*.  In the preparation of *Pishti* of *Swarna Bhasma* 32 hrs were required, which was too much in comparison with that required for *Swarna Varkha* and *Swarna Patra*; the reason behind this may be that the *Swarna Bhasma* is not in elemental form, which leads to the extra time taken for the amalgamation with *Parada* \[[Table 3](#T3){ref-type="table"}\].  The *Nischandratva* and *Rekhapurnatva* tests of *Kajjali* were passed in 16 hrs, but 24 hrs trituration was done for the fineness.  The weight of *Kajjali* was found to be increased (3.5% on an average) after *Bhavana*, which may be due to the added solid contents of *Japakusuma Swarasa* and *Kumari Swarasa* \[[Table 5](#T5){ref-type="table"}\]. 

It was observed during a trial and error study that the amount of *Gandhaka* is directly proportional to the burning period.  Therefore, as per the classical notes, a specific temperature pattern was mentioned for the *Samguna Kajjali Sindura*, i.e. *Mridu Agni, Madhyama Agni* and *Tivraagni* in an equal ratio.  In this, *Gandhaka* was just in the melting stage in the *Mriduagni* while in the *Madhyama Agni* stage, *Gandhaka* boils and burns and in the *Tivra Agni* period, *Parada* with *Gandhaka* sublimes.  Thus, the temperature required for the melting of *Gandhaka* and sublimation of the product is not dependent on the amount of *Gandhaka*.  Taking note of this, the period of Mridu and Tivra Agni was not changed but the period of Madhyama Agni was increased twice of the normal ratio due to which an excess amount of *Gandhaka* gets more period for the burning, which is the main aim.  Also, in the previous study, it was found that *Samaguna* and *Dwiguna Sindura* were prepared in 16-20 hrs.  Here, an attempt was made to prepare the *Triguna Balijarita Makaradhwaja* in a minimum heating period, which was 12 hrs \[Tables [6](#T6){ref-type="table"} and [7](#T7){ref-type="table"}\].  For this, many trial and error studies were carried out to finalize the temperature pattern.  During the *Kupipaka*, flame of sulfur occurred at the neck of the *Kupi* after 7.30 hrs on an average, which was continued for 1.45 hrs \[[Table 7](#T7){ref-type="table"}\]. 

An average of 94.22% of the *Makaradhwaja* was prepared by the *Swarna Patra* with residue (gold powder) of 4.6%, also 76.2% of *Makaradhwaja* and 6.1% of gold powder was prepared by *Swarna Varkha* with 87.8% of *Makaradhwaja* and 3.9% gold powder was prepared by *Swarna Bhasma* \[[Table 8](#T8){ref-type="table"}\]. 

The first sample of *Makaradhwaja* prepared by *Swarna Varkha* was not properly sublimed due to irregular electrical power so that the average percentage of the final product of *Makaradhwaja* was decreased and residue (gold powder) was increased compared with the other two samples. 

Analytically, there were no changes found in loss on drying, ash value, acid-insoluble ash and carbon disulfide extract. 

But, in the ICP analysis for gold content, it was found to be 268 ppm in *Makaradhwaja* prepared by *Swarna Varkha* whereas in a previous study (Patgiri *et al*.),\[[@ref20]\] *Makaradhwaja* prepared by *Swarna Patra* found only 7.5 ppm and also by author prepared *Makaradhwaja* prepared by *Swarna Patra* and *Bhasma* was found to be 131 ppm and 19 ppm.  This variation found in the gold content may be due to the particle size of the raw material (gold), because the particle size of *Swarna Varkha* is least compared with the *Swarna Patra* and, in the *Bhasma* form, gold is not as elemental as the form of gold.  Thus, here, it is observed that least particle size of elemental gold increases the gold content in the sublimed *Makaradhwaja*.  This increased concentration of gold content in sublimed *Makaradhwaja* enhances the therapeutic efficacy of *Makaradhwaja*, which has been also observed in comparative clinical trials of *Makaradhwaja* prepared by *Swarna Patra-Varkha* and *Bhasma* on *Madhumeha* (diabetes mellitus),\[[@ref21]\] and pharmacological studies also supports this. 

Conclusion {#sec1-4}
==========

*Makaradhwaja* has been standardized in terms of time and temperature for 330-424 g. *Kajjali* (average), i.e. *Mridu Agni*: 100-250°C (2.5 hrs), *Madhyamagni*: 250-450°C (5 hrs), *Tivragni*: 450-600°C (4.5 hrs). * Makaradhwaja* prepared by least particle size of elemental gold increases the concentration of gold content in the sublimed *Makaradhwaja* \[[Figure 1](#F1){ref-type="fig"}\]. 

![ The average temperature pattern for preparation of *Makaradhwaja* through the electrical muffle furnace](Ayu-32-109-g012){#F1}
