Pharmaceutico-analytical study of Makardhwaj - A Classical Mercury based Ayurvedic rasayoga

 

A Singh1*, S Ota1, N Srikanth1, B Sreedhar2, Galib3, A Sheetal1, KS Dhiman1

1Central Council for Research in Ayurvedic Sciences, New Delhi.

2All India Institute of Ayurveda, New Delhi.

3Indian Institute of Chemical Technology, Hyderabad.

*Corresponding Author E-mail: arjunccras@gmail.com, Sarada_ota@yahoo.com, ccras_tec@nic.in, galib14@yahoo.co.in, sreedharbojja@gmail.com, draarti.sheetle@gmail.com, dg-ccras@nic.in

 

ABSTRACT:

Makaradhwaja is a mercury-based formulation, frequently being used by Ayurveda professionals in the management of various diseases. However, quality and safety concerns are being witnessed in the recent past on these traditional formulations, which are letting down reputation of the system. Thus manufacturing of quality formulations, establishing pharmaceutical and analytical profiles become an important task. Makaradhwaja is prepared by Kupipakwa method using processed Swarna (gold), Parada (mercury) and Gandhaka (sulphur) in a proportion of 1:8:16. Preliminary physico-chemical analysis, assay of elements, XRD and XPS were performed for evaluation of analytical profile and chemical characterization. Chemical analysis revealed that Makaradhwaja contains 83.60% of Mercury and 13.55% of Sulphur along with other minor elements. Though, large quantity of gold was used in the preparation, very little (33.35ppm) was found in the finished sample. Makaradhwaja is tri/hexagonal (alpha) form of mercury sulphide and XRD samples of three batches confirmed this form.

 

KEYWORDS: Makaradhwaja, physico-chemical analysis, Mercury, Sulphur and XRD.

 

 


INTRODUCTION:

Makaradhwaja is one of the most commonly used organo-mercurial formulations in Ayurveda, prepared by varying proportions of processed gold, mercury and sulphur, subjecting to gradual heating process in a glass bottle. Ayurveda physicians use this formulation along with suitable herbal ingredients and adjuvant in the management of respiratory disorders, pyrexia, diabetes, rheumatoid arthritis, neurological disorders1 etc. It is also used as Rasayana.2 Its immunomodulatory activity3-4 and usefulness as an aphrodisiac agent5 have been well known. Though this is an important formulation, its manufacturing method, preliminary analytical profiles comprising of physico-chemicaland instrumental parameters19-27 so are not reported. Thus, an attempt has been made here.

 

MATERIALS AND METHODS:

Preparation of Makaradhwaja:

Raw materials were procured from Maharishi Ayurveda Pharmacy, Faridabad and authenticated by Dravyaguna expert and Botanist. Metal / mineral components were certified by Rasa Shastra expert. The composition of Makaradhwaja is placed at Table 1.

 

Table 1: Composition of Makaradhwaja

 

Ingredient

Botanical Name/Scientific name

Proportion

Wt. As per proportion

1.                     

Shuddha Swarna

Processed gold

1

148 gms.

2.                     

Shuddha Parada

Processed mercury

8

1184 gms.

3.                     

Shuddha Gandhaka

Processed sulphur

24

3552 gms.

Bhavana Dravya

4.                     

Rakta Karpasa (Flowers)

Gossypium herbaceum Linn.

Q.S.

Q.S.

5.                     

Kumari (Leaf)

Aloe barbadensis Mill.

Q.S.

Q.S.

 

 

The process was done in 04 steps6

1.     Shodhana of Swarna patra, Gandhak and Hingulottha parada

2.     Preparation of Kajjali,

3.     Bhavana with Kumari, Rakta karpasa puspha swarasa

4.     Valuka yantra paka (Heating in Sand Furnace)

 

Shodhana of Swarna Patra, Gandhaka and Hingulottha Parada:

Shodhana of Swarna Patra:

Table 2: Ingredients in the Shodhana of Swarna Patra

S. No.

Materials used

Botanical Name/ Scientific name

Proportion

1.

Swarna Patra

Gold leaf

1

2.

Tila taila

Sesame oil

Q.S. for 3 times Quenching processes

3.

Takra

Buttermilk

-Do-

4.

Gomutra

Cow urine

-Do-

5.

Kanji

Rice gruel

-Do-

6.

Kulatha Kashaya

Decoction of horse gram

-Do-

 

148gms of Swarna patra having a width of 1.5cm and thickness of 2mm was collected. Total 12 leaves were collected. Each leaf was rolled into the bolus; the boluses were taken into ladle and subjected to heat till they became red hot. These red-hot boluses were quenched immediately in 100ml sesame oil. After self cooling boluses were taken out from the vessel, heated and quenched. The process was repeated for two more times in sesame oil. The same process was repeated in other liquid medias viz. Takra, Gomutra, Kanjika, and in Kulatha kashaya in a succession.7 For every quenching process fresh and same amount of media was used. The pH of the quenching medias used during the procedure was noted before and after processes. (Table - 3) Weight of gold boluses neither decreased nor increased after the process.

 

Table3: Ph of quenching media before and after Shodhana

Sl.

No.

Quenching

Media

Ph

Before Shodhana

After three times of Shodhana

1.

Taila

NA

NA

2.

Takra

3.85

4.12

3.

Gomutra

7.15

7.60

4.

Kanji

3.69

3.65

5.

Kulattha Kashaya

4.88

4.80

 

Shodhana of Gandhaka:

Coarse powder of Ashudha Gandhaka (raw sulphur) was melted on low flame by addition of Goghrita and poured into milk through a cotton cloth smeared with Goghrita. This procedure was repeated for two more times and every time fresh Goghrita and Godhughda were used. At the end of the third process, it is washed with hot water to remove sticky fat content. This was further dried under sunrays and powdered.8

Ashuddha Hingula (Cinnabar, HgS) was reduced to fine powder in a Khalva yantra (mortar and pestle) and levigated with Nimbu swarasa (lime juice). Likewise, the levigation process was repeated for 06 times. Small chakrikas (pellets) of processed Hingula were prepared, dried and placed in Damaru yantra and the mouth of both pots was sealed with mud smeared cloth. Heat was given to the bottom of lower pot having Hingula, whereas the bottom of upper pot was covered with cloth drenched with water. Kramaagni was given for 06 hours. After the apparatus gets cool on its own, the inner surface of upper pot was scrapped carefully with a clean cloth to collect ashes and mercury globules.9 This was sieved carefully through a cloth to obtain Parada in globular form. The process was repeated in more batches.

 

Preparation of Kajjali:

148 gms of Shuddha swarna patra and 1184gms of Hingulothha Parada were taken in a Khalva yantra. The contents were levigated thoroughly in the end runner till the mixture got amalgamated. 3552gms of Shuddha gandhaka was then added to the amalgamated mixture, triturated till it attains black color, soft, lusterless fine powder like collyrium (Kajjali). Mardana was done for 24 hrs till the formation of fine, soft, Nishchandra kajjali. Prepared Kajjali was soft on touch and no shining of mercury was observed.10

 

Bhavana with juices of Kumari (Aloe barbadensis Mill.) and Rakta karpasa kusuma (flowers of Gossyppium herbaceum Linn.):

4.8kg of Ghrita kumari (Aloe barbadensis Mill.) was collected from the local garden, washed under tap water and pulp was separated. This pulp was added to Kajjali in the end runner and levigation (Bhavana) was carried out till the material dries completely. This was followed by levigation with Rakta karpasa kusuma swarasa. The flowers were collected from local garden, washed under tap water, processed to obtain juice. This was added to Kajjali in the end runner and levigation process was carried out till the material completely dried. The dried Kajjali was filled in the glass bottles to prepare Makaradhwaja.

 

Valuka yantra paka (Heating in sand furnace):

4872gms. of Kajjali was divided in to 15 batches (considering 324gms. in each batch) and filled in pre-treated glass bottles through a funnel. Pre-treatment of glass bottles include wrapping with seven uniform layers of mud smeared cloth and sun drying. These bottles with Kajjali in them were placed in the center of the Valuka yantra.11 Heating process was carried out in a Kramagni paka (Gradual increasing sequence of heating) over a period as per the schedule i.e. Mandagni, Madhyamagni, and Tivragni. At regular intervals, the temperature of the furnace was recorded with the help of digital pyrometer. (Graph 1)

 

Graph 1: Temperature pattern observed in the preparation of Makaradhwaja

 

Emission of mild white Sulphur fumes was observed that gradually increased and the colour changed to light yellowish to dark yellowish - brown later. These fumes were allowed to escape. At the end of Madhyamagni phase, dark yellowish-brown fumes get converted into bluish flames. During the course of heating in Tivragni phase, the neck of the bottle was frequently cleared to prevent blocking from subliming Sulphur with Tapta shalaka (hot iron rod).

 

After disappearance of the flame and observing the positive copper coin test and sheeta shalaka (cold iron rod) test; the mouth of the Kupi was corked. Copper plate / coin test is an important test to confirm the phase of the process. If, white precipitation over the plate/ coin on placing it on the mouth of the bottle appears, it indicates sublimation of Mercury. The temperature was increased and it was maintained for the next 6 hrs.12 The bottles were allowed to be in Valuka yantra and left for self-cooling.

 

After Swangasheeta (self-cooling), the glass bottles were taken out from the furnace; outer coverings (mud layered cloth) were removed. A thread, soaked in kerosene was tied an inch below from the deposited final product at the neck of the bottle and ignited. A few drops of water were sprinkled over the bottle, which leads in crackling of the Kupi. The bottle was opened carefully to collect Makaradhwaja from the neck of the Kupi avoiding glass pieces, and was preserved in airtight container.

 

Valuka yantra paka was started at 12 noon of the day. Mild white fumes started to arise from the neck of the bottle from 2.00 pm onwards. Light yellow fumes started to arise from 3.30 pm onwards. Dense yellow fumes started to arise from 4.15 pm onwards. Density of fumes increased and changed to dark brownish yellow colored from 6.15 pm onwards. Dense fumes converted into the yellow flame appeared at neck at 6.30 pm. Flame was continuous until 10.45 pm. Escaping of fine shiny particles from the neck of the bottle was observed at 10.45 pm.

 

Around 75 gms of Makaradhwaja was collected from the neck of the Kupi. This was triturated in Khalva yantra to collect fine red colored powder. Total 1100 gms. of the end product was collected from 15 bottles. The various steps involved in the preparation of Makaradhwaja are shown in figure 1.


 

Fig.1: Various steps involved in the preparation of Makaradhwaja


 

Chemical Analysis:

Reagents and standards:

All chemicals, reagents and solvents were used analytical grade and obtained from authentic suppliers.

 

Physico-chemical Analysis:

Physicochemical analysis, viz. description, estimation of Loss on drying, Ash content, Acid insoluble ash, Water/Alcohol soluble extractive, pH, qualitative/quantitative elemental testing etc. were carried out by following standard methods as per Ayurvedic Pharmacopoeia of India (API) guidelines13-16.The quantitative estimation of Metals viz. Hg, Zn, Mg, B, Ca and Au were carried out by Atomic Absorption Spectrometer (Perkin Elmer (USA) Analyst 400) and ICP-AES (THERMO ELECTRON Corporation’s model IRIS INTREPRID II XDL). However Sulphur were quantified by using CHNS analyzer as well as conventional methods19. X-ray diffraction, XPS survey scans, and CHNS analysis were performed to evaluate the elemental composition ratio17-18.

 

X-ray diffraction:

Powder X-ray diffraction (XRD) analysis of Makaradhwaja was carried out using Rigaku Ultima-IV X-ray diffractometer with CuKα radiation = 1.54A°) operating at 40 kV and 30mA. Pattern was recorded for angle (2θ) ranging from 10-100° at a scanning rate of 1°/second and scan step of 0.1°. Sample identification was done by matching d-spacing with the standard database.

 

X-ray photoelectron spectroscopy (XPS):

XPS measurements of Makaradhwaja were obtained on a KRATOS AXIS 165 instrument equipped with dual aluminum–magnesium anodes using Al Ka radiation. The X-ray power supply was run at 15 kV and 5 mA. The pressure of the analysis chamber during the scan was 10-9 Torr. The peak positions were based on calibration with respect to the C 1s peak at 284.6 eV. The obtained XPS spectra were fitted using a nonlinear square method with the convolution of Lorentzian and Gaussian functions after the polynomial background subtraction from the raw data.

 

RESULTS AND DISCUSSION:

The Organo-leptic observation shows that the prepared Makaradhwaja is in the form of Reddish brown fine powder having no characteristic odour and taste. The qualitative analysis shows the positive test for the presence of Mercury and Sulphur. The Chemical analysis revealed that it contains 83.60% of Mercury and 13.55% of Sulphur on average together with minor elements viz. Boron, Zinc, Magnesium, Calcium, and Gold. Moisture content 0.06% was found when determined loss on drying at 1050C. Total ash content (approx 0.20%) is left after burning of volatile matter. The observations show that water soluble (0.45%) and alcohol soluble (2.25%) matter are also present in this formulation. (Details are mentioned table 4). Particles size is about 10μm-650μm and is homogeneously distributed which showed the presence of micro fine particle.

 

Table 4: Observations of three batch analysis

S. No.

Parameter Tested

Observed results of three batch analysis

1.

ORGANOLEPTIC CHARACTERS

a.

Colour

Reddish brown

b.

Taste

Tasteless

c.

Odour

Odorless

d.

Appearance

Fine powder

2.

PHYSICO-CHEMICALS

 

Identification

Yields the reaction characteristics of sulphur and mercury

a.

Loss on drying(%)

0.01-0.10

b.

Total Ash (%)

0.10-0.30

c.

Acid Insoluble Ash (%)

0.01-0.10

d.

Water soluble extractive (%)

0.30-0.60

e.

Alcohols soluble extractive (%)

1.50-3.0

f.

pH

6.0-7.0

g.

Volatile matter

99.60±0.57

h.

Specific gravity

0.9988±0.0006

i.

Particle size distribution

10%,

50%,

90%

 

9.85-261.73 μm

20.62-413.21 μm

47.30-650.15 μm

3.

Assay (%)

a.

Mercury

83.0-84.2

b.

Sulphur

13.0-14.10

c.

Zinc

0.60-0.80

d.

Boron

0.01-0.20

e.

Magnesium

0.20-1.10

f.

Calcium

0.80-2.0

g.

Gold

30.30-36.40ppm

 

XRD: The three Makaradhwaja samples (of 3 batches) have near identical XRD patterns. The incorporated gold cannot be detected in the XRD. The small amounts of gold (in parts per million levels of concentration) incorporated into the crystal structure of cinnabar. Fairly large quantity of gold is used in the preparation of Makaradhwaja but very little i.e. 33.35ppm is found to be incorporated into the sample. Makaradhwaja, is tri/hexagonal (alpha) form of the mercury sulphide and the XRD of the samples of three batches confirmed to this phase. Usually prepared by the Kupipakva procedure, the free sulphur (as is in kajjali) is completely absent in these samples, due to high temperature volatilization of the sulphur; (even if present the amount is too low to be detected in XRD). The XRD pattern is shown in Fig.2 and Fig.3.


 

Fig. 2: XRD pattern of Makaradhwaja (batch –I, II, III)

 

 

Fig. 3: Overlay of XRD pattern of Makaradhwaja (batch –I, II, III)


ESCA (XPS): XPS survey scans do not show any remarkable changes for all the three batches. The atomic concentration quantification report for the elements Hg and S for batches I, II and III shows a ratio of 49:51(Hg:S), 49:51 and 52:48 respectively.

 

Hg to S ratio is thus standardized as 49-52: 48-51.

 

Fig. 4: Typical ESCA (XPS)

 

The observed binding energy peaks for Hg 4f are deconvoluted into two peaks at 100.59 eV and 104.62 eV characteristic of Hg 4f7/2 and Hg 4f5/2 that can be attributed due to Hg bonded to S in the form HgS.

 

The observed binding energy peaks for S 2p are curve fit into two peaks at 162.18 eV and 163.45 eV characteristic of S 2p3/2 and S2p1/2 respectively, due to S in HgS. The high-resolution narrow scan for Au 4f in Makaradhwaja is also shown in the Fig.5.

 

Fig. 5: Binding energy peaks of different orbitals

 

As can be seen the 4f peaks at 84 eV and 88 eV which are due to Au 4f7/2 and Au 4f5/2 are deconvoluted into two peaks each and the observed peaks are tabulated in the Table.5.

 

Table. 5. Observed binding energy peaks for Au 4f

Au 4f7/2

Au 4f5/2

83.97 eV

87.99 eV

85.98 eV

90.09 eV

 

The peaks at Au 4f7/2 (83.97 eV) and Au 4f5/2 (87.99 eV) are attributed due to Au in Au 0 state, whereas the peaks at Au 4f7/2 (85.98 eV) and Au 4f5/2 (90.09 eV) are attributed due to Au observed for other Au (I) complexes. The area under the curves gives an approximate amount of Au present in Au (0) and Au (1) state which is observed to be in the ratio 1:4.

 

CONCLUSION:

Makaradhwaja prepared by Kupipakwa method is reddish brown fine powder having no characteristic odour and taste. A very little quantity of gold i.e. 33.35ppm is found to be incorporated into the sample together with Mercury and Sulphur. It is tri/hexagonal (alpha) form of mercury sulphide confirmed by XRD. The developed analytical profile shall be helpful for scientists and researchers to build comprehensive standards for further studies.

 

ACKNOWLEDGEMENT:

The authors express their heartfelt thanks and would like to acknowledge Dr S. K. Sharma, Former Advisor (Ayurveda), Ministry of AYUSH, Government of India; Dr M. M. Padhi, Former Dy. Director General, CCRAS; and Dr Ravinder Singh, Assistant Director (Chemistry) for valuable guidance; and Dr V. K. Singh, M/s. Maharishi Ayurveda Products Pvt Ltd, Noida, India, for technical inputs. The authors are thankful to Dr J. Arunachalam, Former Head, National Centre for Compositional Characterization of Materials (BARC), Hyderabad, India, for helping in data analysis and interpretation of the results.

 

Thanks also conveyed to Dr. Aarti Sheetal, Senior Research Fellow (Ayu.) of CCRAS for technical assistance.

 

CONFLICTS OF INTEREST:

There are no conflicts of interest.

 

ETHICAL APPROVAL:

Not applicable for this research study

 

REFERENCES:

1.      Acharya Sadanada Sharma, Rasa Tarangini, Translated by Shri Kashinatha Shastri, 11th ed Reprint. Motilal Banarsidas, New Delhi, 6/248; 2009: 149-151.

2.      Sinyorita S, Ghosh CK, Chakrabarti A, Auddy B, Ghosh R, Debnath PK. Effect of Ayurvedic mercury preparation Makaradhwaja in geriatric Canine- A preliminary study, Ind J Exp Biol, 49; 2011: 534-539.

3.      Patgiri BJ, Prajapati PK, Ravishankar B, A toxicity study of Makaradhwaja prepared by Ashtasamskarita Parada. Ayu, 27(3)4; 2006: 55-62.

4.      Dhundi S, Ashok BK, Ravishankar B, Patgiri BJ, Prajapati PK. Immunodulatory activity of Triguna Makaradhwaja - An Ayurvedic compound formulation, Indian journal of natural product and resources. 3(3); 2013: 320-327.

5.      Sen GD, Shastri AD, Shastri RD. Bhaishajyaratnavali 19th Ed. Varanasi (India): Chaukhamba Prakashan, 2008:1128.

6.      Government of India. The Ayurvedic Formulary of India—Part-I. 2nd ed. New Delhi, India: Ministry of Health and Family Welfare, Government of India, 2003:210.

7.      Sharangdhara virachita Sharangdhara Samhita Adhamala’s Dipika and Kashiram’s Gudhrthadipika. Varanasi: Chaukhamba Orientallia, Ma. Kha.11/2-4, 2005: 241.

8.      Acharya Sadanand Sharma, Rasa Tarangini 8/7-12, translated by Shri Kashinatha Shastri, 11th Ed. Reprint. Motilal Banarsidas, New Delhi, 2009: 176.

9.      Acharya Sadanand Sharma, Rasa Tarangini 2/31, translated by Shri Kashinatha Shastri, 11th Ed. Reprint. Motilal Banarsidas, New Delhi, 2009: 16, 202.

10.   Acharya Sadananda Sharma, Rasa Tarangini 2/27 translated by Shri Kashinatha Shastri, 11th Ed. Reprint. Motilal Banarsidas, New Delhi, 2009:16, 202.

11.   Acharya Sadananda Sharma, Rasa Tarangini, 4/29-31 translated by Shri Kashinatha Shastri, 11th Ed. Reprint. Motilal Banarsidas, New Delhi, 2009: 52-53

12.   Prajapati, P.K., Joshi, D., Dube, G.P., Kumar, M., Prakash, B. (M.D. (Ayu.) dissertion). Pharmaceutical and experimental study on Makaradhwaja, Varanasi: BHU, 1994.

13.   Anonymous, Pharmacopoeial standards for Ayurvedic formulations, (CCRAS, Ministry of Health and Family Welfare, Govt. of India, New Delhi). Revised Edition. Delhi: The Controller of Publications, 1987.

14.   Protocol for testing of ASU medicine (PLIM, Deptt of AYUSH, Ministry of Health and Family Welfare, Govt. of India, New Delhi), 2007.

15.   Quality Control Manual for ASU Medicine (PLIM, Deptt of AYUSH, Ministry of Health and Family Welfare, Govt. of India, New Delhi), 2008.

16.   Laboratory Guide for Analysis of Ayurveda and Siddha formulations (CCRAS, Deptt of AYUSH, Ministry of Health and Family Welfare, Govt. of India, New Delhi), 2010.

17.   Kumar A, Nair AGC, Reddy AVR, Garg AN. Availability of essential elements in bhasmas: Analysis of Ayurvedic metallic preparations by INAA. J Radio Anal Nucl Chem, 270; 2006: 173–180.

18.   Krishnamurthy LV, Sane RT. Study on Ayurvedic Bahamas on the basis of modern analytical instrumentation techniques. Indian Res J Chem Environ, 5; 2001: 65-67.

19.   P. Selvakumar, Devi Kaniakumari, V. Loganathan. Physicochemical Analysis of Alocasia sanderiana W. Bull. Asian J. Pharm. Ana. 6(1): January- March, 2016; Page 31-34.

20.   K. Parameswari , T. Ananthi. Physico-Chemical and Phytochemical Analysis of Mukia maderaspatana L. Research J. Science and Tech 5(2): April- June, 2013 page 272-274.

21.   Saranya A, Nithya S.Assessment of Heavy Metal Induced Organ Toxicity in marketed Ayurvedhic Formulation and Report its LD50 value with Brine Shrimp Lethality Assay. Research J. Pharm. and Tech. 2017; 10(1): 263-268.

22.   Deepak Dwivedi. Cadmium and Mercury Pollution and its Preventive Measures. Research J. Engineering and Tech. 3(1): Jan.-Mar. 2012 page 06-08.

23.   Pratima Rani Dwivedi, M. R. Augur. Physico Chemical Analysis of Ground Water in Chirimiri Area of Korea District, Chhattisgarh. Research J. Engineering and Tech. 5(1): Jan.-Mar. 2014 page 38-41.

24.   Ashwani Awasthi, Shashi Kant Tripathi, Ashok Kumar Tiwari. Physico-chemical analysis of Ken river water in Panna District Madhya Pradesh, India. Research J. Science and Tech. 2018; 10(2):131-136.

25.   Pooja Singh Baghel, Ashok Kumar Tiwari. Comparative Physico-Chemical Analysis of Mandakini River Water at Chitrakoot, District Satna (MP), India. Research J. Science and Tech. 2016; 8(2):71-76.

26.   Vishal D Joshi, Prajwal R Shetty, Tekeshwar Verma, Vasant D Khasia, Narahari N. Palei. Physico-Chemical Analysis of Ground water of selected area of Rajkot City. Asian J. Research Chem. 2(3): July-Sept., 2009, page 275-277.

27.   Archana Chapolikar, Jagdish Bharad, Balaji Madje, Milind Ubale. Microbiological and Physico-Chemical analysis of Ground Water from Thane-Belapur Industrial Area, Mumbai, India. Asian J. Research Chem. 3(1): Jan.-Mar. 2010; Page 102-105.

 

 

 

 

Received on 25.07.2019           Modified on 17.12.2019

Accepted on 15.02.2020         © RJPT All right reserved

Research J. Pharm. and Tech. 2020; 13(12):5685-5691.

DOI: 10.5958/0974-360X.2020.00990.7