GC MS analysis of one Ayurvedic formulation “Nasika churnam”

 

Sharmila D¹, Poovarasan A², Pradeep E², Mudiganti Ram Krishna Rao^3*, Prabhu K⁴

¹Assit. Professor, Department of Industrial Biotechnology,

Bharath Institute of Higher Education and Research, Chennai.

² Student, Department of Industrial Biotechnology,

Bharath Institute of Higher Education and Research, Chennai.

³ Professor, Department of Industrial Biotechnology,

Bharath Institute of Higher Education and Research, Chennai.

⁴Associate Professor, Department of Anatomy, Sree Balaji Medical College and Hospital, Chrompet, Chennai

 

ABSTRACT:

The present study deals with the GC MS analysis of one Ayurvedic medicine, Nasika Churna which is used for the treatment of respiratory diseases. Nasika churnam was procured from a standard Ayurvedic vendor at Chennai and subjected to GC MS analysis following standard protocol. Some important molecules such as  Cyclobutane-1,1-dicarboxamide, N,N'-di-benzoyloxy-, Ar-tumerone, Ethyl p-methoxycinnamate, Tetradecanoic acid, cis-13-Octadecenoic acid, cis-Vaccenic acid, Dodecanoic acid, 2-phenylethyl ester, Stigmasterol, .gamma.‑Sitosterol, Succinic acid, di(3,7-dimethyloct-6-en-1-yl) ester etc. indicate the medicinal roles which are similar to the medicinal roles attributed to Nasika churna in Ayurveda. Further probing into the activities of each molecule is warranted.

 

 

 

INTRODUCTION:

There is need to bring contemporary and alternative medicinal forms to the fore of medical system due to their inherent advantages like easy affordability, low cost and less side effects. This need has become all the more imperative due to the high costs, less affordability and major side effects of the molecular medicines. Since last decade or so there is awareness towards this problem and some work in pouring in although much more need be done.^1-25 The present study is to find the types of molecules present in one Ayurvedic medicine, Nasika Churnam by subjecting it to GC MS analysis. Such knowledge could help in understanding the molecular mechanism of this medicine. Nitin et al, 2014 and Panda et al, 2017, have reviewed the standardization and efficacy of churnams in general.^26,27

 

This medicine contains many herbal ingredients and is used for the treatment of treating chronic rhinitis, allergic respiratory conditions, bad breath, upper respiratory tract infections, migraine, sinusitis, head tremors etc. The manufacturers of this medicine are Arya Vaidya Nilayam, Ashoka Pharmaceuticals, Ary Vaidya Sala Kottakkal, Sitaram Ayurveda Pharmacy etc. among others.

 

MATERIALS AND METHODS:

Nasika Churnam was obtained from standard Ayurvedic vendor at Chennai and was subjected to GC MS analysis by standard procedure.

 

Instrument:

Gas chromatography (Agilent: GC: (G3440A) 7890A. MS MS: 7000 Triple Quad GCMS,) was equipped with Mass spectrometry detector.

 

Sample Preparation

100 micro lit sample Dissolved in 1ml of suitable solvents. The solution stirred vigorously using vortex stirrer for 10 seconds. The clear extract was determined using gas-chromatography for analysis.

GC-MS protocol:

The GC MS Column consisted of DB5 MS (30mm × 0.25mm ID × 0.25μm, composed of 5% phenyl 95% methyl poly siloxane), Electron impact mode at 70 eV; Helium (99.999%) was used as carrier gas at a Constant flow of 1ml/min Injector temperature 280°C; Auxiliary Temperature: 290ᵒC Ion-source temperature 280°C.

 

The oven Temperature was programmed from 50°C (isothermal for 1.0 min), with an increase of 40°C/min, to 170°C C (isothermal for 4.0 min), then 10°C/min to 310°C (isothermal for 10min) fragments from 45 to 450 Da. Total GC running time is 32.02 min. The compounds are identified by GC-MS Library (NIST & Wiley).

 

RESULTS AND DISCUSSION:

Table 1 indicates the GC MS profile details of Nasika Churnam along with the possible medicinal role of each molecule. Figure 1 represents the GC MS profile of Nasika Churnam.

 

Figure 1. Indicates the GC MS profile of Nasika Churnam.

 

Table1. Indicates the retentions values, types of possible compound, their molecular formulae, molecular mass, peak area and their medicinal roles of each compound as shown in the GC MS profile of Nasika Churnam.

 

The molecule Cyclobutane-1,1-dicarboxamide, N,N'-di-benzoyloxy- has been attributed to have properties such as anphylactic, anti tumor,  Arylamine-N-Acetyletransferase-Inhibitor, decrease nor epinephrine production, GABAergic,  increase NK cell activity, inhibit production of Tumor necrosis factor, myo-neuro-stimulant, NADH-Oxidase-Inhibitor, NADH-Ubiquinone-Oxido-reductase-Inhibitor etc. (Table 2).

 

Arylamine N-acetyltransferases (NAT) are phase II xenobiotic-metabolizing enzymes (XME) which catalyze the transfer of an acetyl group from acetyl-coenzyme A (Acetyl CoA) to the nitrogen or oxygen group of aromatic amine chemicals. These enzymes are important in detoxification and/or bio-activation of several aromatic amine drugs and carcinogens (Hein et al, 2000; Sim et al, 2012).^29,30 The role of this molecule present in Nasika churnam could be to protect the body from allergic agents inhaled through nose. GABAergic role of this molecule could be attributed towards better neurotransmission of allergic signals. Increasing NK cell activity could help clearing the respiratory path from cell debries. The NADH Oxidase inhibitor activity could protect the lungs or related tissues from oxidative bursts caused by this enzyme. TNF is involved in autoimmune and immune-mediated disorders such as rheumatoid arthritis, ankylosing spondylitis, inflammatory bowel disease, psoriasis, hidradenitis suppurativa and refractory asthma and TNF inhibitors are used in the treatment of these diseases. Many TNF inhibitors are available in the market but they are beset with many side effects (Scheinfeld, 2004).³¹ The TNF inhibitor activity of molecules present in Nasika churnam could help in treating inflammatory reactions related to respiratory tract.

 

NADH-Ubiquinone-Oxido-reductase-Inhibitory role be could be attributed to control the bacterial infections in the respiratory tract. Increase in aromatic Amino acid decarboxylase activity leads to decarboxylation of L-Dopa and 5-hydroxytryptophan thus increasing the production of catecholamines such as dopamine, norepinephrine, epinephrine and Serotonin. The presence of more catecholamines cause mood elevation, stress relief and increased blood circulation in the affected areas caused due to inflammation. The inhibition of Arachidonic acid render the inactivation of Cox 1 and cox 2 enzymes thus stopping the synthesis of prostaglandins. Thus these molecules present in Nasika churnam function as Non-steroidal anti-inflammatory drugs (NSAID), thus reducing pain.

 

PDEs are key enzymes in the adenosine and guanosine cyclic nucleotides (cAMP and cGMP) signaling cascade. Their inhibition increases cyclic nucleotide levels inside the cell. Thus, Nasika churnam could contribute to the inflammation of respiratory system (Martinez and Gil, 2014).³² gamma.‑Sitosterol, present in Nasika Churnam has PPAR‑gamma antagonist role.

 

Peroxisome proliferator-activated receptor gamma (PPARγ) protein is a regulator of adipocyte differentiation and is associated with the pathology of numerous diseases including obesity, diabetes, atherosclerosis and cancer. The PPARγ inhibitory role of. gamma.‑Sitosterol could be attributed to its anti-inflammatory activity in respiratory infections.  Similarly, Stigmasterol is also effective in controlling hormone regulated inflammatory conditions. Thus it is clear that so many molecules present in Nasika chirnam could contribute at various levels of physiology related to respiratory disorders for which this medicine is used by Ayurveda. The roles of some of the molecules shown in Table 2 are not reported yet and in will be worthwhile probing their contribution towards the medicinal role of Nasika chirnam.

 

CONCLUSION:

The various roles of molecules present in Nasika churnam is indicative of its overall efficacy in curing respiratory diseases.

 

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Accepted on 27.04.2020           © RJPT All right reserved

Research J. Pharm. and Tech 2021; 14(3):1400-1404.