The GC MS Study of one Ayurvedic Medicine, Vasakadyaristam

 

Muttevi  Hyagreva Kumar¹, Prabhu K², Mudiganti Ram Krishna Rao^3*, R. Lakshmi Sundaram⁴, Sampad Shil⁵, Arun Kumar S⁵

¹Research Scholar, Sree Balaji Medical College and Hospital, Bharath University, Chrompet, Chennai - 600092.

²Associate Professor, Department of Anatomy, Sree Balaji Medical College and Hospital, Bharath University, Chrompet, Chennai - 600092.

³Professor, Department of Industrial Biotechnology, Bharath Institute of Higher Education and Research, Chennai-600073.

⁴Scientific Officer, Central Research Facility, Sri Ramachandra Medical College and Research Institute, Purur, Chennai - 600116.

⁵Student, Department of Industrial Biotechnology, Bharath Institute of Higher Education and Research, Chennai-600073.

 

ABSTRACT:

The present study deals with the GC MS analysis of one Ayurvedic prepaation, Vasakadyarishtam, which is a mucolytic medicine used for the treatment of cough, cold, chest congestion asthma and related diseases like bronchitis, inflammation and hemoptysis. The most important constituent of this medicine is Vasaka or Adhatoda vesika plant, which is known for its anti-cough and cold usage in Ayurveda. This study was to understand the main biomolecules present in this medicine by GC MS analysis. The results indicted the presence of some important molecules such as Eucalyptol, Phenylethyl alcohol, Camphor, Ethyl hydrogen succinate, Succinic acid, cyclobutyl ethyl ester, 3-Cyclohexen-1-ol,4-methyl-(1-methylethyl)-(R)-,. alpha.-Terineol, 5-Hydroxymethyl furfural, p-Anisic acid,4-nitrophenyl ester, 2-Propen-1-0l,3-phenyl, Benzeneethanol, 4-hydroxy- Propiophenone, 2’-(trimethylsioxy)-, Hexadecanoic acid, 2-hydroxy-1-(hydroxymethyl)ethyl ester and Octadecanoic acid and 2,3 –dyhydroxypropyl ester. These molecules are known for their various medicinal properties which indicate their roles in this formulation.

 

 

 

INTRODUCTION:

Ayurvedic, Sidhha and other alternative medicinal systems are age old and time tested practices. The molecular mechanism of action of these medicines must be studied to understand their efficacy in the modern medical parameters. This study will help in the better management of these medicines at each level, like procurement, preparation, packing, dose specification and possible side effects and their elimination. The present study is one step in this direction.

 

Vasakadyarishtam is a popular as a mucolytic medicine used for the treatment of cough, cold, chest congestion asthma and related diseases like bronchitis, inflammation and hemoptysis. The main constituent of this medicine is Vasaka, which is known for its mucolytic properties. 12 to 25 ml of this liquid is taken once or twice daily under the supervision of an Ayurvedic practitioner. The patients are asked to avoid sour foods. This medicine is prepared as per the Ayurvedic treatise, Gada Nigraha Prayoga Khanda, Asava Adhikara 152 – 154

 

The medicine, Vasakadyaristam ingredients are:

Vasaka Plant – Adhatoda vasica Syn. Justica adhatoda – 9.6 kg

 

Dhataki - Woodfordia fruticosa - Flowers - 384 g

Cinnamon - Cinnamomum zeylanicum - bark -48 g

Cardamom - Elettaria cardamomum - Seeds48 g

Patra - Cinnamomum tamala – leaves – Stamens - 48 g

Nagakeshara – Mesua ferrea – Stamens - 48 g

Kankola – Piper cubeba – Fruits - 48 g

Pepper - Piper nigrum - Fruits -16 grams

Long pepper- Piper longum – fruits -16 g

Ginger – Zingiber officinaes – Dry Ginger rhizome - 16 g

Hribera – Pavonia odorata – 48 g

 

Vasaka plant – Adhatoda vasica is cut into coarse powder – 9.6 kg added to 24.576 liters of water, boiled and reduced to 6.144 liters and filtered to obtain Vasa Kashayam. To this 4.8 kg of Jaggery is added. Other ingredients are added in the following proportion and allowed to ferment in closed container for 15 days. After 15 days the liquid obtained is filtered and packed in bottles to be used as medicine.

 

It is appropriate to mention the medicinal roles of each of the ingredient to know their overall influence on the medicinal role of the medicine.

 

1. Vasaka (Malabar Nut) (Adhatoda vasica):

One of the oldest medicinal plants with various medicinal functions is used in all streams of medical practice mostly used for its anti-cold and cough activities. (Hossain and Haq, 2016; Dhankhar et al, 2011)^{1, 2}

 

2. Dhataki – Woodfordia fruticose:

This plant is reported to have many important secondary metabolites like alkaloids, flavonoids, glycosides, phenols, saponins, sterols etc. which contribute to this plants medicinal role as antibacterial and antioxidant. (Dubey et al, 2014; Grover and Patni, 2014)^{3, 4}

 

3. Ela (Cardamom) (Elettaria cardamomum):

Cardamom is another important culinary ingredient used for its characteristic aroma. Apart from the aroma it has medicinal values. Verma et al, 2013, have reported blood pressure lowering, fibrinolysis enhancing and antioxidant activities of Cardamom.⁵   Khan have shown the pharmacological basis of cardamom as medicine for asthma Khan et al, 2011)⁶

 

4. Twak – Cinnamon - (Cinnamomum zeylanicum):

Almost every part of the cinnamon tree has some medicinal or culinary use. (Ranasinghe et al, 2013; Jayaprakasha, et al, 2011)^{7, 8}

 

5. Patra (Cinnamomum tamala):

This plant show medicinal activities such as anti-ulcer, anti-microbial, antioxidant, anthelminthic, antifungal, anti-hyperlipidemic, analgesic & antipyretic, anti-diarrheal, anti-aflatoxigenic and lipid lowering.^9-12

 

6. Nagakesara (Mesua ferrea L.):

This medicinal role of this plant was reviewed by Chahar et al, 2013).¹³ It has medicinal activities like antioxidant and hepato protective, analgesic, antispasmodic, anti-venom, cancer chemotherapy, Immuno-modulatory, anti-neoplastic, anti-convulsant, anti-inflammatory, anti-ulcer and anti-microbial.

 

7. Kankola (Piper Cubeba):

This plant has been reported to have many medicinal roles such as, antioxidant, anti-inflammatory, bactericidal, antibacterial and nephron-protective. (Alsaid et al, 2015; Nahak and Sahu, 2011; Ahmed et al, 2012)^14-16

 

8. Pepper (Piper nigrum):

In Ayurveda it is known as Kapha virodhini (works against Phlegm). The decoction of Pepper is used for curing cough, diarrhea, lack of appetite and dyspepsia. There are reports of its pharmacological activities such as antioxidant, anticonvulsant, sedative, insecticidal, pesticidal, muscle relaxant, antipyretic, anti-inflammatory, antifungal, hepato-protective, antimicrobial, antiulcer, antibacterial, lipolytic etc. (Shamkuwar et al, 2012; Meghwal and Goswami, 2012)^{17, 18}

 

9. Pippali (Long Pepper) (Piper longum):

Kumar et al, 2011, have reviewed the various health benefits of Piper longum. Piper longum has many important medicinal values such as anticancer, antioxidant, hepato-protective, anti-inflammatory, immune-modulatory, antimicrobial, anti-hyper-lipidemic, analgesic, antidepressant, antiamoebic, vasodialtory, bioavailability enhancer due the presence of piperine in it, anti-obesity activity, radio-protective, cardio-protective and antifungal.¹⁹

 

10. Sunthi (Ginger) Zingiber officinale:

Ginger is common household medicine for common cold, cough and indigestion.  Its medicinal values are well documented. (Zadeh and Ko, 2014)²⁰

 

11. Hribera (Pavonia odorata):

This plant has many medicinal values such as anticancer and antioxidant activities.^21-22

 

MATERIALS METHODS:

The medicine Vasakadyarishtam was procured from standard Ayurvedic vendor at Chennai, India. 50 ml of Vasakadyarishtam was taken and extracted with ethyl acetate solvent in separating funnel. The extracted material was filtered and concentrated in water bath at low temperature and was charged to have GC MS patterns by standard procedures.

 

Instrument:

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

 

GC-MS Principle:

In gas chromatography Helium gas is used as stationary phase. The principle of separation in GLC is partition. For GLC the compound should be volatile. They have to be heated to higher temperature and converted in to vapors in injector portion and mixed with gaseous mobile phase then the components are separated according to their partition co-efficient. In GLC the Mass spectrometer is used as detector. Mass Spectrometer is used for the determination of molecular weight of the compound and also for their structure elucidation. The compounds are identified by GC-MS Library (NIST & WILEY).

 

Sample Preparation:

100 micro litres sample Dissolved in 1 ml 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:

Column DB5 MS (30mm×0.25mm ID ×0.25 μm , 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; Auxilary 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.

 

RESULTS AND DISCUSSION:

The GC MS profile of Vasakadyarishtam is represented in Figure 1. The presence of various biomolecules present in the GC MS is represented in Table 1.

 

Figure1. Indicates the GC MS profile of Vasaka Dyarishtam

 

 

 

 

Table1. Indicates the retentions Time, the type of possible compound, their molecular formulae, molecular mass, peak areas and their medicinal roles of each compound as shown in the GC MS profile of Vasakadyarishtam.

Sl. No	Retention Time	Compound Name	Mol. Formula	Mol Weight	Peak Area	Medicinal Role
1.	3.71	Eucalyptol	C10H18O	154.1	20035667	It helps contractile activity of smooth muscles.
2.	4.36	Phnylethyl alcohol	C8H10O	122.1	36666635	Alcohol dehydrogenase inhibitor, alcohol, detoxificant and antimicrobial.
3.	4.64	Camphor	C10H16O	152.1	80770421	Analgesic, antiseptic, antispasmodic, anti- inflammatory, expectorant and as cold suppressant. It is used for asthma, bronchitis and rheumatism.23
4.	4.76	Ethyl hydrogen succinate	C6H10O4	146.1	46679771	Hydrogen Peroxide Inhibitor, Succinate dehydrogenase inhibitor
5.	4.63	Succinic acid, cyclobutyl ethyl ester	C10H16O4	200.1	12256033	Succinic-Dehydrogenase-Inhibitor, Arachidonic acid-Inhibitor, Acidulant, Inhibit Production of Uric Acid
6.	4.86	3-Cyclohexen-1-ol,4-methyl-(1-methylethyl)-(R)-	C10H18O	154.1	28493199	Antibacterial
7.	4.96	.alpha.-Terineol	C10H18O	154.1	8042125	5-Alpha-Reductase-Inhibitor, HIF-1-alpha inhibitor, Alpha Glucosidase inhibitor, IKappaB-Alpha Phosphorylation inhibitor, Increase alpha mannosidase activity, testosterone-5-Reductase inhibitor
8.	5.14	5-Hydroxymethyl furfural	C6H6O3	126	15928048	5-Hydroxymethylfurfural - It is reported to stop neuron apoptosis.24
9.	5.37	p-Anisic acid,4-nitrophenyl ester	C14H11NO5	273.1	9045250	17-beta hydroxysteroid dehydrogenase inhibitor, Testosterone Hydroxylase inhibitor, Arachidonic acid inhibitor, increase aromatic amino acid decarboxylase activity, inhibit uric acid production
10.	5.66	2-Propen-1-0l,3-phenyl	C9H10O	134.1	23068715	Oligosaccharide provider
11.	6.31	Benzeneethanol, 4-hydroxy-	C8H10O2	138.1	11506156	17-beta-hydroxysteroid dehydrogenase inhibitor, Aryl hydrocarbon hydroxylase inhibitor, Testosterone hydroxylase inhibitor
12.	6.92	Propiophenone, 2’-(trimethylsioxy)-	C12H18O2Si	222.1	18851497	Oligosaccharide provider
13.	7.08	(4-Methoxy—phenyl)-(2-nitrocyclohexyl)-methanol	C14H19NO4	265.1	17964022	Not known
14.	10.71	Hexadecanoic acid, 2-hydroxy-1-(hydroxymethyl)ethyl ester	C19H38O4	330.3	43014169	17-beta-hydroxysteroid dehydrogenase inhibitor, Aryl hydrocarbon hydroxylase inhibitor, Testosterone hydroxylase inhibitor
15.	11.35	Octadecanoic acid, 2,3 –dyhydroxypropyl ester	C12H42O4	358.3	24331229	Arachidonic acid inhibitor, increase aromatic amino acid decarboxylase activity, inhibit uric acid production

 

Vasakadyarishtam is a medicine used for cold related diseases. Most of the ingredients present in it like black pepper, long pepper, ginger, Vasaka, Patra, Nagakesara, cardamom, cinnamon bark are long been used for curing cold related symptoms. The GC MS analysis indicated the presence of some very important biomolecules such as Eucalyptol, Phenylethyl alcohol, Camphor, Ethyl hydrogen succinate, Succinic acid, cyclobutyl ethyl ester, 3-Cyclohexen-1-ol,4-methyl-(1-methylethyl)-(R)-,  .alpha.-Terineol, 5-Hydroxymethyl furfural, p-Anisic acid,4-nitrophenyl ester, 2-Propen-1-0l,3-phenyl, Benzeneethanol, 4-hydroxy-, Propiophenone, 2’-(trimethylsioxy)-, Hexadecanoic acid, 2-hydroxy-1-(hydroxymethyl)ethyl ester and Octadecanoic acid, 2,3- dyhydroxypropyl ester. The medicinal roles of each of them are tabulated in Table 1 which correlate with the activity of the medicine Vasakadyarishta. Further study to relate the medicinal activities of these molecules in relation to the cure of cold and other lung diseases is warranted.

 

 

CONCLUSION:

From the above discussion it is clear that the various molecules as indicated by GC MS analysis support the medicinal role of Vasakadyarishtam as claimed in Ayurveda.

 

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Accepted on 03.11.2018        © RJPT All right reserved

Research J. Pharm. and Tech 2019; 12(2)569-573.