Study on physical properties of Indian based ayurvedic medicine : Abhrakh bhasma as nanomaterials by employing modern scientific tools

Abhrakh bhasma is an Ayurvedic medicine widely used for treating several diseases such as; pernicious, sickle cell anemia, Bell’s palsy, hepatic dysfunction, leukemia, sex debility, cystic fibrosis, post encephalic dysfunction and cervical dysplasia etc. An Ayurveda bhasma is a medicine, which is used since ancient time and claimed to be very useful medicine at Nano metric scale. However there is a need of scientific evidence of such Ayurvedic products using modern scientific techniques of 21st century. The physical behavior was measured by X-ray diffractometer (XRD), fourier transform spectroscopy (FTIR), vibrating sample magnetometer (VSM), scanning electron microscopy (SEM), and photoluminescence (PL) spectrometer. Abhrakh bhasma is prepared by using mica as a base materials. X-ray diffraction and SEM analysis revealed that this bhasma is in nanocrystalline form and, can be considered as nanomedicine. Magnetic hysteresis (M~H) loop study by using the VSM reveals the super paramagnetic nature. Luminescence in bhasma was observed in visible region upon excitation of 200 nm laser source. This investigation reveals that, the Abhrakh bhasma may not only used as nanomedicine but also can be employed a materials for various other technological applications due to both magnetic as well as luminescence properties. Preliminary testing of Abhrakh bhasma nanoparticles was examined on Staphylococcus aureus and Escherichia coli bacteria.


Introduction
An Ayurveda bhasma is Indian traditional Ayurvedic medicine obtained naturally.It being used since ancient time in India and also some other parts of the world.This medicine is accepted well in India.However, these medicines were being used on practice by BAIDYA (Ayurvedic practioner) in ancient India.Recently there is a considerable interest lies on scientific community to understand the physical properties of these Ayurvedic bhasma.The recent study reveals that the size of those bhasma materials almost lies between 1-100 nm range, as confirmed by X-ray diffractometer and Scanning Electron Microscopy [1][2].Ayurvedic Abhrakh bhasma is the mineral of mica [K(Mg,Fe)3(AlSiO10)(OH)2 and widely used for treating several diseases such as; pernicious, sickle cell anemia, Bell's palsy, hepatic dysfunction, leukemia, sex debility, cystic fibrosis, post encephalic dysfunction and cervical dysplasia etc.Its primary use is as aphrodisiac, astringent and restorative (powerful cell regenerator).It is also called for its penetrative and spreading property in the whole body and various micro tissues.It is an alternative, aphrodisiac, anti-pyretic, carminative, hematinic and proven rejuvenator [3][4][5].According to some new findings, bhasma are claimed to be biologically produced nanoparticle using Ayurvedic methods [6][7][8].The details of Ayurvedic methods for preparation of Abhrakh bhasma are explained in detail Ayurvedic text [9][10].Modern scientific tools such as; X-ray diffractometer (TTRX-III-Rigaku, Japan), Fourier Transform Infrared spectroscopy (Frontier, PerkinElmer, UK), Vibrating sample magnetometer (7410, LakeShore, USA), Scanning Electron Microscopy (EVO 18 Research, Zeiss UK), and Photoluminescence (LS-55, PerkinElmer, Germany) Spectrometer have been employed to characterize the prepared Abhrakh bhasma.The structural, size, chemical constituents, magnetization, colour in the bhasma are reported.The preliminary testing of antimicrobial activity was examined on Staphylococcus aureus and Escherichia coli microorganism which is reported here.Hence, it is interesting to note that the present article may help to Ayurvedic doctors to implement Abhrakh bhasma in a better way for the treatment.Also the other applications of the bhasma can be explored.

Material and methods
Abhrakh bhasma has been prepared by using mica as a base materials.The pieces of mica are heated till they become red and it dipped in the decoction of cow urine and cooled it.This process is repeated for seven times after that mica powder cover with paddy and it was kept in a blanket and tied firmly.Then it is kept in sour gruel for a day and thereafter, it was rubbed with hand so that mica entirely gets out into the sour gruel.This is taken out and washed and dried in the sun light.After dried, it is ready for incineration.Equal amount of jaggery is mixed with dhanyabhrakh and was processed with the leaves juice of Eranda and made into small discs which were dried in the sun light and, heated in the sealed saucers.The leaves of vata (Ficus bengalensis) were placed above and beneath the discs before heating.This process is repeated for three times, a lusterless Abhrakh bhasma powder was get ready.The details of preparation of such bhasma also can be found in the Ayurvedic [9][10].

Structural and microstructural studies
An X-ray Diffraction (PXRD) technique uses this principle to elucidate the crystalline nature of materials.The scattering of X-rays from atoms produce a diffraction pattern that contains information about the atomic arrangement in crystal.The sample kept on sample holder and take crystallography.The crystallographic phase analysis of prepared Abhrakh bhasma was carried out by employing the X-ray diffractometer, with CuKα radiation source (λ = 1.5418Å) and, are shown in figure 1 (a) and (b) for clear visibility of the peaks.It is divided into two parts (a) 10 0 to 35 0 and (b) 35 0 to 80 0 of 2θ values.There are several diffraction peaks are observed, which could be identified by using the ICDD PDF no.98-007-4571, 98-006-8756, 98-006-0684 and 98-008-0684 and these are correspond to different elements which are enlisted in table 1.The maximum intensity peak in XRD pattern of Abhrakh bhasma shown in figure 1(a-b) is indexed to Mg2O4Si1.Hence, it is assumed that the Abhrakh bhasma is mostly a byproduct of magnesium.Similar result has been reported by other research group [11].The XRD analysis reveals that the Abhrakh bhasma contains various other elements such as iron, magnesium, potassium, calcium, aluminum, silica and sodium with oxygen.XRD pattern has been separated for the better presentation and visibility only The average crystallite size of the Abhrakh bhasma is calculated by employing the Sherrer's formula [12] d = Kλ /β cos θ Where K is a constant which is equal to 0.9, β is the full width half minimum (FWHM), θ is the Braggs' angle and λ is the wavelength of Cu-Kα radiation.
The crystallite size calculation was carried out with considering all intense diffraction peaks.The above equation yields the average crystalline size ~24±4 nm.Thus the XRD study concludes the Abharakh bhasma is in nanocrystalline form.

FTIR analysis
FTIR measurement of Abrakh bhasma was observed by the KBr palate methods.In this method, obtained sample proper grind and mixed with potassium bromide (KBr) at approximately 1:20 mass ratio with the help of mortar and pestle.The mixture is then pressed to make the thin pallets by palatalizer and applied pressure up to 6.5 tons.Spectra were observed with Frontier, Perkin-Elmer Singapore at room temperature.The FTIR spectrum of nanocrystalline Abhrakh bhasma is shown in figure 2. The absorption bands in transmission mode are in the range of 400 to 1700 cm - 1 at room temperature.The functional groups available in Abhrakh bhasma with its corresponding wave number are enlisted in table 2. The functional groups identified in the FTIR spectra are well supported to the XRD analysis.The above two techniques reveals that the prepared Abhrakh bhasma is made of organic molecules from the herb which play a major role in different deceases as discussed in the previously reported work and also it is reported by several research groups [13][14][15][16][17].The present investigation may help to Ayurvedic doctors and practitioners to treat the several diseases with the action of Abhrakh bhasma in safe way.

SEM Measurement
The scanning electron microscope images were taken at 14KV EHT with mm working distance and 35.57kx & 70.96kx magnification.The powders were coated with Gold and Palladium mixture in spin coater and then the images were taken.The SEM micrograph of Abhrakh bhasma is shown the figure 3 (a) and (b) with different magnifications.It is observed that the average particle sizes are below 200 nm.But earlier in XRD analysis, crystalline size was found to be 24nm.Both the experimental techniques analysis indicates that agglomeration of nanocrystallites in the samples.However, it is observed that the microstructure is uniform which must in contest is as concern for the disease treatment i.e. the action of medicine should be uniform for better action.The present study reveals that the present method of bhasma preparation can be used as a standard method to produce uniform particle size of Abhrakh bhasma for better use in disease treatment.It reveals that the Abhrakh bhasma is magnetic in nature.However the coercivity is very low.It could be due to nanocrystalline nature of the Abhrakh bhasma, which is also observed from XRD analysis.The maximum saturation magnetization is found to be ~1.3 emu/g, which is comparable to very good magnetic materials.The 'S' shape curve of M-H hysteresis loop reveals the super paramagnetic nature of the bhasma.However it needs further magnetic investigations for confirmation such as field cooled or zero field cooled magnetization measurement, ac-susceptibility etc. which is our future research problem which are beyond the scope of the present study.The coercivity is found to be around 1400 Oe, which is very good for using this as a natural magnet for magnetic memory device applications.So, the present study reveals that the present bhasma not only can be used for diseases treatment but also can be explored the green technological and engineering applications as, it has been prepared from the natural products.

Figure 4. The M~H hysteresis curve of Abhrakh bhasma as nanomaterials
Similar study on magnetic measurement has been reported by the other research groups and their magnetic parameters values are suitable for electronics materials, high frequency applications, memory devices etc. [18][19][20].More important is that, the present study opens a window to motivate this natural bhasma for high contrast MRI (Magnetic resonance imaging) techniques.It needs further investigations.Hence, the present study opens a new window for natural Ayurvedic medicines.

Luminescence study
The powder samples kept on sample holder and measure the emission spectra.Figure 5 shows the room temperature PL spectrum and it is interesting to note that there are 5 peaks with different intensities which are in visible spectrum.
The peaks are also in UV and NIR as shown in the figure also.Each element has optical spectrum.XRD analysis confirms the presence of Fe, K, Al etc. So, emission peaks may correspond to these elements or their oxides.Luminescence due to metal oxide compounds are also reported by other research groups [21][22].Each bhasma have some color and such luminescence may be due to presence of different elements present in the compounds.

Conclusions
Abhrakh bhasma Indian based An Ayurvedic medicine can be prepared by the natural method.X ray diffraction and Scanning Electron microscopy results confirm the nanocrystalline size of the prepared Abhrakh bhasma.The XRD and FTIR techniques reveal that the prepared Abhrakh bhasma is made of organic molecules from the herbs which play a major role in different deceases.SEM results observed that the microstructure is uniform which must in contest is as concern for the disease treatment i.e. the action of medicine should be uniform for better treatment.The present study reveals that the method used in the present article can be used as a standard method to produce uniform particle size of Abhrakh bhasma.Magnetic hysteresis measurement shows that the coercivity is very low and the maximum saturation magnetization is found to be about 1.3 emu/g, which is comparable with the good magnetic materials.The 'S' shape curve of M-H hysteresis loop reveals the super paramagnetic nature of the Abhrakh bhasma.The coercivity is found to be around 1400 Oe, which is very good for using this natural product for magnetic memory devices applications.Therefore, the present study reveals that the Abhrakh bhasma not only can be used for diseases treatment but also, it can be explored for the technological and engineering applications without natural hazards.More important is that, the present study opens a window to motivate use of this natural bhasma for high contrast MRI (Magnetic resonance imaging) techniques.Hence, the present study opens a new window in natural Ayurvedic medicines.

Figure 1 .
Figure 1.XRD patterns of Abhrakh bhasma (a) from 2θ= 10 degree to 35 degree and (b) from 35 degree -80 degree.XRD pattern has been separated for the better presentation and visibility only

Table 1 .
The ICDD database card numbers with chemical formula for Abhrakh bhasma