The GC MS Analysis of one Ayurvedic Medicine “Balarishtam”

 

Sharmila D1, L. Jeyanthi Rebecca2, Mudiganti Ram Krishna Rao3*

1Asst. Professor, Dept. of Industrial Biotechnology, Bharath Institute of Higher Education and Research, Chennai.

2Professor and Head, Dept. of Industrial Biotechnology, Bharath Institute of Higher Education and Research, Chennai.

3Professor, Dept. of Industrial Biotechnology, Bharath Institute of Higher Education and Research, Chennai.

*Corresponding Author E-mail: mrkrao1455@gmail.com

 

ABSTRACT:

Balarishtam is an Ayurvedic nerve tonic. The present study deals with the GC MS analysis of the medicine to understand the biomolecules present and their possible nerve related activity. Balarishtam was subjected to GC MS analysis. The presence of some important molecules such as Eucalyptol, 2-Isopropoxyethylamine, p-menth-1-en-8-ol, Eugenol, N-Acetyl-. alpha.-methyl-4-ethynyl-2-thiazolemethanamine etc. has given a clue towards the nerve tonic effect of Balarishtam. Thus this study could lead to the identification of marker biomolecules which could help in validation of this drug as a nerve tonic.

 

KEYWORDS: Balarishtam, Ayurvedic, Eugenol, Eucalyptol, Marker, Biomolecule.

 

 

INTRODUCTION:

Ayurveda and Sidha are two traditional medical practices of India with rich and classical literature. Sidha and Ayurvedic medicines have not been tested for their efficacy, short term and long term side effects, mechanism of action at different levels of pharmacology, standardization etc. as compared to modern medicines. Once these traditional medicines are proven and validated, this will lead to the availability of cheap, easily affordable and safe medicines.1-14

 

The present study is one step in this direction in which the GC MS analysis of one Ayurvedic medicine, Balarishtam is being reported. The knowledge of the types of bio molecules present as indicated by GC MS analysis could throw some light on the mechanism of action of this medicine. Balaristham is an Ayurvedic tonic for nerve related diseases like neuralgia, hemiplegia, paraplegia and also for arthritis, spondylosis. This tonic is prepared from nine herbal plants and jaggary according to the reference of Ayurvedic treatise named Bhaishaja Ratnavali Vatavydhi-569-572.

 

Balarishtam is prepared by the following ingredients.

Bala (Sida cordifolia) – root/whole plant – 4.8kg; Ashwagandha (Withania somnifera) – root – 4.8kg
Water for decoction – 49.152 liters, boiled and reduced to 12.288 liters. Jaggery – 14.400kg; Dhataki (Woodfordia fruticosa)– flower – 768g; Payasya (Ipomea digitata) – root/whole plant 96g; Eranda (Ricinus communis) – root – 96g; Rasna (Pluchea lanceolata) – root - 48g; Ela (Cardamom) (Elettaria cardamomum)– 48g; Prasarini (Paederia foetida) – root – 48g; Usheera (Vetiveria zizanioides) – 48g; Gokshura (Tribulus terrestris) – whole plant/fruit – 48g. Coarse powders of Bala and Ashwagandha are added to water, boiled and filtered. To this Kashayam, jaggery is added, filtered for impurities. Rest of the ingredients is added and kept closed in an air tight container for a month time. After fermentation, the liquid is filtered and stored in air tight container. The standard manufactures of Balarishtam are Dabur, Baidyanath, Arya vaidya Sala, AVP etc. Very scanty literature is available on the scientific validation of this medicine. (Rajalakshmy and Sindhu, 2011; Tiwari, 2014)15,16

 

MATERIAL AND METHODS:

The medicine which is available in liquid form was subjected to GC MS analysis after necessary procedure.

 

 

The metabolites in the samples were identified using a P2010 gas chromatography with thermal desorption system TD20 coupled with mass spectroscopy (Shimadzu). The identification of metabolites was accomplished by comparison of retention time and fragmentation pattern with mass spectra in the NIST spectral library stored in the computer software (version 1.10 beta, Shimadzu) of the GC-MS. The relative percentage of each extract constituent was expressed with peak area normalization.

 

RESULTS AND DISCUSSION:

The GC MS profile is represented in Figure 1. The identification of metabolites was accomplished by comparison of retention time and fragmentation pattern with mass spectra in the NIST spectral library stored in the computer software (version 1.10 beta, Shimadzu) of the GC-MS along with the possible pharmaceutical roles of each bio molecule as per Dr. Duke’s Phytochemical and ethnobotanical data base (National Agriculture Library, USA) and others as shown in Table 1.17-26

 

Figure 1: Showing the GC MS profile of Balarishtam.

 

Table 1: Indicates the various details of the GC MS profile along with medicinal role of each molecule.

Sl. No

Retention Time

Compound

% Peak Value

Medicinal role

1

4.062

Butane, 1-(1-methylethoxy)-

1.18

Not Known

2

4.062

1-Propanol, 2-ethoxy-

1.18

Not Known

3

4.062

12-[Trifluoromethyl]-3,6,9-trioxat ridecan-1-ol

1.18

Oligosaccharide provider

4

4.191

Glycerin

12.95

Glycerin is a common chemical used for cosmetic application for smoothness of skin

 

4.368

Diglycerol

0.65

Not known

 

4.368

N-Methoxy-N-methylacetamide

0.65

Anaphylactic, antitumor, Arylamine-N-Acetyltransferase-Inhibitor,  Decrease Norepinephrine Production,  Down regulation of nuclear and cytosol androgen re uptake, GABA-ergic, Increase Natural Killer cell activity, inhibit production of Tumor Necrosis factor, myo-neuro stimulant, NADH oxidase inhibitor, NADH-Ubiquinone-Oxidoreductase-Inhibitor

5

4.790      

Propane, 2-cyclopropyl-

1.79

Not known

6

4.790

4-Methyloxazolidine

1.79

Not known

7

4.790

Butanoic acid, 2-oxo-

1.78

Acidifier, inhibitor for Arachidonic acid, decreases the production of uric acid, acidifies urine and increases aromatic amino acid decarboxylase activity.

8

4.873

Eucalyptol

5.51

Used in mouth washes and cough suppressants and helps contractile activity of smooth muscles.

9

5.025

2-Isopropoxyethylamine

0.92

Acidifier, inhibitor for Arachidonic acid, decreases the production of uric acid, acidifies urine and increases aromatic amino acid decarboxylase activity, adrenocortical stimulant, analgesic, ANS- stimulant, anticancer and anti-diabetic.

10

5.025

Acetoacetic acid, 1-thio-, S-allyl ester

0.92

Acidifier. Acidulant, Arachidonic acid-Inhibitor, Increase Aromatic Amino Acid Decarboxylase Activity, Inhibit Production of Uric Acid,  'Smart-Drug', Adrenocortical-Stimulant, anti-diabetic

11

5.025

Xylitol

0.92

Used for otitis media, dental caries, intravenous nutrition, and osteoporosis.

12

5.190

Butanoic acid, 3-hydroxy-, methyl ester

1.14

Acidifier, inhibitor for Arachidonic acid, decreases the production of uric acid, acidifies urine and increases aromatic amino acid decarboxylase activity, as inducer for enzyme testosterone hydroxylase, as inhibitor for enzymes like 17-beta-hydroxysteroid dehydrogenase, aryl-hydrocarbon hydroxylase, catechol-o-methyl hydroxylase etc.

13

5.190

Butanoic acid, 2-oxo-

1.14

Acidifier, inhibitor for Arachidonic acid, decreases the production of uric acid, acidifies urine and increases aromatic amino acid decarboxylase activity.

14

5.190

Propanol, methoxy-, acetate

1.14

Not Known

 

15

6.517

Phenylethyl Alcohol

5.24

Alcohol dehydrogenase inhibitor, alcohol detoxificant and antimicrobial.

 

7.256

4H-Pyran-4-one, 2,3-dihydro-3,5-dihydroxy-6-methyl-

0.70

Catechol-O-Methyl-Transferase-Inhibitor, Methyl-Donor, Methyl-Guanidine-Inhibitor, 11B-HSD-Inhibitor, 17-beta-hydroxysteroid dehydrogenase-Inhibitor,  5-HETE-Inhibitor, 5-HT-Inhibitor, 8-HETE-Inhibitor, Anti-5-HT, Anti-HIV-Integrase, Antidote, Aryl-Hydrocarbon-Hydroxylase-Inhibitor, HDL-genic,  Hemagglutinator

 

7.256

3-Hydroxy-N,N-dimethylpropanamide

0.70

Anaphylactic, antitumor, Arylamine-N-Acetyltransferase-Inhibitor,  Decrease Norepinephrine Production,  Down regulation of nuclear and cytosol androgen re uptake, GABA-ergic, Increase Natural Killer cell activity, inhibit production of Tumor Necrosis factor, myo-neuro stimulant, NADH oxidase inhibitor, NADH-Ubiquinone-Oxidoreductase-Inhibitor

 

7.256

Oxirane, 2,3-dimethyl-

0.70

Oxirane, 2,3-dimethyl-

 

7.644

3-Acetyl-2-oxo-1,3-oxazolidine

0.42

Acetyl-choline-antagonist, Acetyl-CoA-Carboxylase-Inhibitor, Arylamine-N-Acetyltransferase-Inhibitor.  Inhibit Acetyl Coenzyme A

 

7.644

Propane, 1-isothiocyanato-

0.42

Not  Known

 

7.644

1-[3-Hydroxypropyl]-aziridine

0.42

Not  Known

16

7.967

Ethyl hydrogen succinate

13.97

Hydrogen peroxide inhibitor and Succcinate dehydrogen inhibitor.

17

7.967

4-Ketopimelic

13.97

Not Known

19

8.102

2-Oxazolidinone, 3-methyl-

2.63

catechol-o-methyl hydroxylase and Methyl guanidine inhibitor and a methyl donor

20

8.278

Oxirane, [(1-methylethoxy) methyl]-

1.72

Not Known

21

8.278

p-menth-1-en-8-o

1.72

Oligosaccharide provider, decrease endothelial, adhesion, encephalopathic, endo anesthetic, tonic, enteroprotective, enterostimulant and energizer.

22

8.278

3-Cyclohexene-1-methanol,  alpha., .alpha.4-trimethyl-

1.72

Inhibitor of many enzymes like Alpha amylase, Alpha glucosidase, 5-alpha reductase, interlucin 1-alpha etc and increases the activities of Alpha Mannosidase.

23

12.053

Phenol, 2-methoxy-3-(2-propenyl)-

44.79

Not Known

24

12.053

3-Allyl-6-methoxyphenol

44.79

Anticancer activity.

25

12.053

Eugenol

44.79

Antifungal, fat soluble antioxidant, anticonvulsant, local anaesthetic, antistress, bacteriostatic, bactericidal, Anticarcinogenic and has CNS activity.

 

12.317

1H-Imidazole-4-carboxylic acid

0.25

Acidifier, inhibitor for Arachidonic acid, decreases the production of uric acid, acidifies urine and increases aromatic amino acid decarboxylase activity

 

12.317

3-Deoxy-d-mannoic lactone

0.25

Deoxypyridinoline-Inhibitor, smart drug, 17-beta-hydroxysteroid dehydrogenase-Inhibitor, Alcohol-Dehydrogenase-Inhibitoranticancer, antidote, Antileukotriene-D4, Circulatory-Depressant, coronary dialator, Cyclin-D1-Inhibitor, Decalcifier, Decarboxylase-Inhibitor, Decongestant, Decrease C-Teleopeptide Excretion, Decrease Endothilial Leukocyte Adhesion

 

12.317

2-Butenediamide, 2-methyl-, (Z)-

0.25

Not known

 

16.703

1,3,5-Triazin-2(1H)-one, 4-amino-6 -(ethylamino)-

0.26

Not known

 

16.703

cis,trans-2-Methyl-1-thiadecalin

0.26

Catechol-O-Methyl-Transferase-Inhibitor, Decrease Glutamate Oxaloacetate Transaminase, Glucosyl-Transferase-Inhibitor, Glucosyl-Transferase-Inhibitor, Glutathione-S-Transferase-Inhibitor,  Increase Glyoxalate Transamination, Reverse-Transcriptase-Inhibitor

 

16.703

Acetamide, N-(2-dimethylamino-3-ox o-1-butenyl)-

0.26

Not known

26

16.750

Decahydroquinolin-10-ol

0.52

Oligosaccharide provider

26

16.750

Octahydropyrano[3,2-b]pyridin-6-on

0.52

Not known

26

17.085

N-Acetyl-.alpha.-methyl-4-ethynyl-, 2-

1.08

Inhibitor of Alpha amylase, Alpha glucosidase, 5- alpha reductase, testosterone 5 alpha reductase, interlucin 1-alpha, actyl-coenzyme A  increases the activities of Alpha Mannosidase, actylcholinergic,  Actyle cholin antagonist.

27

17.085

2 ,4'-Dihydroxy-3'-methoxyacetophenone

1.08

Not known

28

17.085

Imidazo(1,2-)pyrimidine, 6-ethyl  5-oxo-1,2,3,5-tetrahydro

1.08

Inhibitor of catechol-o-methyl transferese, ACE, and 1,2,3,5-tetrahydro-5-alpha reductase enzymes, methyl donor, and Methyl guanidine inhibitor and a methyl donor., absorbent, acidifier, 11B-HSD-Inhibitor, 17-beta-hydroxysteroid dehydrogenase-Inhibitor,  5-HETE-Inhibitor, 5-HT-Inhibitor, 8-HETE-Inhibitor, Anti-5-HT, Anti-HIV-Integrase, Antidote, Aryl-Hydrocarbon-Hydroxylase-Inhibitor,

29

20.120

1-Heptadec-1-ynyl-cyclopentanol

0.20

Not known

29

20.120

1,2-15,16-Diepoxyhexadecane

0.20

Not known

30

20.120

Bicyclo[2.2.2]octane, 2-methyl-

0.20

Not known

29

20.367

Phenazine, 1-methoxy-

4.09

Not known

30

20.367

Dodecanoic acid, 1,1'-biphenyl-4-ylcarbonylmethyl ester

4.09

Acidifier, inhibitor for Arachidonic acid, decreases the production of uric acid, acidifies urine and increases aromatic amino acid decarboxylase activity.

31

20.367

2-Fluorenamine

4.09

The substituted products of Fluorenamine inhibit the activity of human telomerase enzyme.

 

The medicinal roles of molecules present in Balarishtam  as shown in Table 1 indicates their roles which are in tune with the claimed medicinal role of Balarishtam as a nerve tonic. GABAergic, myo-neuro-stimulant, decrease of norepinephrine production, inhibition of arachidonic acid activities of the molecules present in Balarishtam could contribute to its anti-allergic, antioxidant and anti-inflammatory functions. 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).27,28 This role of the molecules present in Balarishtam must be arresting the conversion of xenobiotics present in the body, thus saving the system form allergies and maintaining the homeostasis of the system. 11 beta hydroxysteroid dehydrogenase and 17 beta hydroxysteroid dehydrogenase enzymes help in the regulation of steroid metabolism thus increasing the levels of steroids and their precursors (Anagnostis et al, 2013; Day et al, 2010).29,30 This role could be in helping the control of stress related diseases pertaining to nervous system. 

 

The presence of 5α-Reductase inhibitor activity helps in controlling the metabolically active androgen hormones thus keeping a low profile of these hormones to control androgen associated disorders. Such roles as GABAergic, Increased NK cell Activity, Myoneuron stimulant etc. of molecules present in Balarishtam can help in maintenance of brain functions.

 

CONCLUSION:

The GC MS analysis of Balarishtam indicated the presence of some very important biomolecuels which have direct or indirect influence on the CNS activity. Further work is in progress to establish the scientific validation of this medicine.

 

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Received on 25.02.2020           Modified on 10.05.2020

Accepted on 29.06.2020         © RJPT All right reserved

Research J. Pharm. and Tech. 2021; 14(8):4226-4230.

DOI: 10.52711/0974-360X.2021.00733