COMPARATIVE PHARMACEUTICO-ANALYTICAL STUDY OF SWARNAMAKSHIKA BHASMAS

Dr. Sushant Kumar 1 , Dr. (Mrs) Prabha Kumari 2 , Dr Amarendra Kumar Singh 3 , Dr. Prabhat Kumar Dwivedi 4 and Dr. Ajay Kumar Singh 5 . 1. Asst. Prof, Dept of RS & BK, Sri Sai Ayurvedic P.G. Medical College & Hospital, Aligarh (UP). 2. Asso. Prof., Dept of Prasuti Tantra & Stri Roga, Govt. Ayurvedic College & Hospital, Chaukaghat, Varanasi (UP). 3. Reader, Dept of Roga Nidana & Vikriti Vigyana, Govt. Ayurvedic College & Hospital, Patna. 4. Reader, P.G. Dept of Rasa Shastra & Bhaishjya Kalpana, Govt. Ayurvedic College & Hospital, Patna. 5. Prof. & HOD, P. G. Dept of Rasa Shastra & Bhaishjya Kalpana, Govt. Ayurvedic College & Hospital, Patna. ...................................................................................................................... Manuscript Info Abstract ......................... ........................................................................ Manuscript History


ISSN: 2320-5407
Int. J. Adv. Res. 5 (9), 1741-1750 1742 various classics of Ayurveda vi . It contains extrinsic factors vii such as copper and iron required for the formation of haemoglobin. Copper is an essential trace element exists in the diet and is needed to absorb and utilize iron viii . The importance of copper as a supplement to iron for haemoglobin regeneration in anaemic rats has already been established ix . This discovery revealed the utility of small amount of copper in the treatment of Microcytic Hypochromic Anaemia. Being a Saumyakalpa x of iron it is easily digestible and hence widely used in infants, pregnant, lactating women and frail old people. Easy availability and cheaper price makes it a drug of choice for the treatment of Pandu Roga i.e. IDA.
In Rasa Shastra, Shodhana (Purification/ Detoxification/ Potentiation) is the process of removing the impurities and toxicities of metals and minerals xi by means of Swedana, Bharjana, Bhawana, Mardana, Prakshalana, Avapa, Nirvapa etc and potentiating the efficacy by adding the qualities of liquid media in which it is done. So in this study, two different methods of Shodhana (i.e. Swedana in Kadalikanda Swarasa and Bharjana in lemon juice) have been done. Marana or Bhasmikarana (Incineration/ Calcination) is the process of converting the already purified metals and minerals into ashes i.e. Bhasma. The whole process of Marana can be summarised in the following manner. The purified material is subjected to Bhawana (Wet levigation in the specified liquid media) → formation of Chakrika (pelletization) → Sharava Sampootikarana (Drying and closing in between two earthen lids)→ Putapaka (specialised conventional heating system) → Bhasma formation. Through the process of Marana, the purified material becomes detoxified, easily digestible, absorbable, assimilable and finally excreted if consumed in extra quantity which are quintessential for any drug xii . Incineration of metals with mercury results in uttama (top quality) bhasma, with herbs results in madhyama (middle quality) bhasma, with sulphur and other substances results in adhama (low quality) bhasma xiii . Incineration with arilohas and other substances results in durguna (causing harmful effect) bhasma. Keeping this in mind, three different ways of marana were adopted here to evaluate their comparative pharmaceutical and analytical study.

Pharmaceutical study:-Aims and Objectives:-
The prime objective of the present study was to develop a SOP (Standard Operative Procedure) for the preparation of Swarnamakshika Bhasma by adopting different methods of shodhana and marana. Further it was also aimed to produce a safe, effective and quality drug for the treatment of Pandu Roga i.e. Iron Deficiency Anaemia. The materials used and the methods adopted was decided on the basis of references in the classical literatures, easy availability of raw materials, cost feasibility of the methods, traditional experiences as well as expert opinions. Red Brownish Red Swarnamakshika Bhasma obtained Analytical study:-Analytical study of a product provides some standards to judge its quality as well as interprets the pharmacokinetics and pharmacodynamics of the same. Main aim of this study is to find out the physico-chemical changes and interpret the effect of different samskaras (shodhana, marana etc) during the pharmaceutical processing.

Material and
In Ayurveda, use of metallic and herbomineral preparations for therapeutic purposes were mainly based on clinical observations. Ancient scholars had scientific vision regarding safe use of these preparations. So different parameters for examining the purity of bhasma have been mentioned in Textbooks of Rasa Shastra e.g.Varitara, Rekhapurnatwa, Apunarbhava, Uttama, Niruttha, Mrtaloha etc xviii .
Mere efficacy without safety is of no use. In this scientific era, people have become quite concerned regarding the safety of the drugs consumed by them. Hence it is necessary to know the nature of the drug, which we prescribe to our patients. For knowing whether it contains any harmful substance or not, analytical study of the metallic or herbomineral drugs is mandatory. Detection of the presence of any free metal in Bhasma is quite essential as free metals are considered highly toxic for the vital organs of the body.

X-Ray diffraction study (XRDS)
:-X-ray diffraction is a tool for investigation of the fine structures of the crystals. When a focussed X-ray beam interacts with planes of atoms, some part of the beam is transmitted, some is absorbed, some is refracted and some part is diffracted. X-rays are diffracted by each mineral differently, depending upon the atoms that make the crystal lattice and how these atoms are arranged.
This resulting analysis is described graphically as a set of peaks with one percent intensity on the Y-axis and goniometer angle on X-axis.

Field Emission Scanning Electron Microscopy (FESEM) study:-
It provides topographical and elemental information at magnification of 10x to 3,00,000x, with virtually unlimited depth of field. Compared with Scanning Electron Microscopy (SEM), it produces a clearer less electrostatically distorted images with spatial resolution down to 1.5 nm. It is 3-6 times better than the conventional SEM. So it is used for ultra high magnification imaging. The high resolution reached by FESEM (~ 2nm) allows the study of very small structural details. There is various application of this instrument but as far as Rasa Shastra is concerned, it is mainly used in the morphological analysis i.e. in knowing the shape and size of particles and qualitative elemental analysis. Besides this it is used in the localization of boundaries between regions of different atomic number. After going through the SEM study of both the samples (as received by the IIC, IIT, Roorkee) it was observed that- Most of the particles in all the photographs are in the range of 100nm-5µ.  There is large amount of 2-5µ sized homogeneous particles.  There are also very fine particles observed which are in the range of nano particle size.  Most of the particles are in the nano range less than 80nm.  There is agglomeration of fine nano particles.

Result:-
The methodology employed for the preparation of Swarnamakshika Bhasma by different methods yields nano particles in general.

Inductively Coupled Plasma Mass Spectrometry (ICP-MS):-
It is considered as the most reliable and latest method for the quantitative elemental analysis available till date. The primary goal of ICP is to get elements to emit characteristic wavelength specific light which can then be measured. The intensity of wavelength specific light is compared to previous measured intensities of the known concentration of elements and the concentration is computed. This method is used to determine Arsenic, Cadmium, Lead, Mercury, Copper etc in any herbomineral or metallic drug.

Observation and Results:-
The concentration of different elements (in %age) in the two samples of Swarnamakshika Bhasmas are given here. This analytical study through ICP-MS was done at IIC, IIT, Roorkee. 0.292 0.192 It can be concluded from the above data that %age of heavy metals are within the permissible limit as per the GMP guidelines in both the samples of Swarnamakshika Bhasma.

Discussion:-
In the pharmaceutical study, a SOP (standard operative procedure) for the preparation of Swarnamakshika Bhasmas by adopting different methods of purification and incineration has been taken. Here two different methods of purification (Shodhana) and three different processes of incineration (Marana) have been undertaken for the final preparation of Bhasma. The hypothesis behind the selection of these different methods was made on the basis of easy availability of raw materials, minimum labour involvement, less total time in the whole procedure, cheap cost of processing. There was also a thought to increase the iron content in the manufactured drug. Finally its easy absorption through the gut was also kept in mind behind adopting these Shodhana and Marana procedures.
In the present study, two methods of purification were adopted (Table no-1). In the first method of Shodhana through Swedana (heating under liquid bath), total 1110 gms of shuddha Swarnamakshika was obtained from 1250 gms of ashuddha one. In this method 11.2% weight loss was observed. Total time taken was approximately 6 hrs. In the 2 nd method of Shodhana, 1090 gm shuddha swarnamakshika was obtained from 1250 gm ashuddha one. Here 12.8% weight loss was observed. The loss might have occurred due to the hammering of ashuddha Swarnamakshika to make it coarse powder before the initiation of the process of Shodhana. More loss was observed in the Bharjana process. The cause of it might be the total time taken in the procedure is more compared to the Swedana procedure. Further the acidic media of lemon juice in might have caused more loss in the Bharjana process. Some amount of the material might have been lost during the repetition of the process. In both the samples, trituration was done for two hours till it became thick paste to prepare pellets (Chakrikas). These Chakrikas were made having diameter 2cm and thickness 1cm so that every particle of it get equal and adequate amount of heat for incineration.
After drying of chakrikas, sharava sampootikarana was done and subjected to Puta. Conventional method of heating was done to prepare all the samples. 10-12 kgs of Vanyopal were taken for Puta. Bhasma collected after each puta was subjected to triturition with Nimbu Swarasa. Chakrikas prepared after triturition were subjected to puta. Total 6 putas were given in both the samples.
After 1 st puta, it was observed that Swarnamakshika chakrikas were brownish red in colour and soft. The brownish red colour of chakrikas was observed after 2 nd & 3 rd puta but the hardness comparatively decreased. After 4 th , 5 th & 6 th puta Swarnamakshika chakrikas became red like vermillion in colour. 170 gm in Sample I and 163.7 gm in Sample II Swarnamakshika Bhasma was obtained from 250 gm of shuddha Swarnamakshika. It meant average yield of Swarnamakshika Bhasma was 68% in Sample I and 65.48% in Sample II (Table no-2 & 3).
As per Table no-4, 750 gm of shuddha Swarnamakshika (kadalikand swarasa shodhita) was taken and 1/8 th part of it shuddha hingula was added in it. This mixture was triturated by adding sufficient quantity of lemon juice in into it. Same process was repeated after each puta for total 7 times and total 8 putas were given in this method. Ultimately, 505 gm bhasma obtained from 750 gm of shuddha swarnamakshika. It meant the yield was 67.33% through this process.

1749
In preparation of Sample no IV (Table no- 5), 750 gm of shuddha Swaranmakshika Bhasma (bharjita in lemon juice) was taken and triturated with adding 250 ml lemon juice in it. Pellets prepared after triturition, were dried, kept in shrava samputa and finally subjected to puta. Same process was repeated for 9 times and total 10 putas were given in similar fashion. After 10 th puta, Swarnamakshika bhasma thus obtained, passed all the classical parameters and colour of bhasma finally changed to red. 470 gm bhasma obtained from 750 gm shuddha Swarnamakshika that meant the average yield in this process was 62.66%. So as far as yield is concerned, methods adopted in Sample III is better than in Sample IV. This proves that marana through adding Rasa Bhasma (as done with adding hingula in sample III) is best method of marana.
As per Table no-6 and Table no-7, it can be said that Swarnamakshika Bhasma prepared by all the three methods were fulfilled the organoleptic and classical analytical parameters. pH of all sample was in slightly acidic (Table no- (Table no-10). Swarnamakshika Bhasma (Oxides of Iron and Copper) if taken in empty stomach (increased amount of gastric acids) with Amalaki Churna (richest source of Vitamin C) would convert ferric iron to soluble and easily absorbable ferrous form. This will lead to increased uptake by membrane trasnferrin receptors xix which will increase iron absorption and distribution in body. This increased Haemoglobinisation will ultimately cure IDA i.e. Microcytic Hypochromic Anaemia.

Conclusion:-
It can be concluded from the present study that Shodhana by Swedana in kadalikand Swarasa) and Marana by Rasa Bhasma i.e. higula & lemon juice is ideal way of making Swarnamakshika Bhasma. Bhasma obtained through above method (in sample III) will be quite effective in treating iron deficiency anaemia because it contains higher percentage of iron and negligible percentages of As, Pb, Mn, Cr like heavy metals. If used with amalaki churna, this could be the ideal drug of treating IDA because of easy availability and cheaper price. It is further suggested to evaluate haematological effect of these drugs by conducting Serum Ferritin and TIBC investigations.