The GC MS Study of one Ayurvedic Medicine, Khadirarishtam

 

Hassan Mohammad1, Prabhu K2, Mudiganti Ram Krishna Rao3*, R. Lakshmi Sundaram4, Sampad Shil5, N. Vijayalakshmi5

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.

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

 

ABSTRACT:

The present study of GC MS analysis of one Ayurvedic formulation, Khadirarishtam, was undertaken to know the types of biomolecules present in it. This medicine is used to treat chronic skin ailments. The GC MS profile of Khadirarishtam indicated the presence of some important biomolecules such as  Eucalyptol, Phnylethyl alcohol, Ethyl hydrogen succinate, .alpha.-Terpineol, 5-Hydroxymethylfurfural, Acetic acid, chloro-,3-phenylpropyl ester, Eugenol, Benzeethanol,4-hydroxy-, Propanoic acid, anhydride,  Benzoic acid, 4-hydroxy-, Disulphide, methyl (methylthio)phenylmethyl, Hexadecnoic acid, 2-hydroxy-1-(hydroxymethyl) ethyl ester, Octadecanoic acid and  2,3-dihydropropyl ester etc. These biomolecules have been reported to have the medicinal properties which represent those of Khadirarishtam. Further study to establish the molecular mechanism of these compounds is warranted.

 

KEYWORDS: GC MS, Ayurvedic, Khadirarishtam, Eucalyptol, Phnylethyl alcohol, Ethyl hydrogen succinate, .alpha.-Terpineol, 5-Hydroxymethylfurfural.

 


 

 

 

 

INTRODUCTION:

Khadirarishta or Khadiraristam is an Ayurvedic medicine used mainly for the treatment of chronic skin diseases.  It is also used as a good heart tonic, helps in curing anemia and liver diseases, tumors, cysts and cough. The refrence of this medicine in found in the Ayurvedic treatise Bhaishajya Ratnavali 54/365-370, Sharangdhara Samhita, Madhyama Khanda 10/60-65. This medicine is prescribed at a dosage of 12– 24 ml. one or two times a day, usually after food or under the supervision of qualified Ayurvedic practitioner. If needed, it can be mixed with equal quantity of water. This medicine is made from the following ingredients.

 

 

 

 

Khadirarishta ingredients: 

Ist set

Khadira – Acacia catechu – heart wood – 2.4 kg

Devadaru – Cedrus deodara – heart wood – 2.4 kg

Bakuchi – Psoralea corylifolia – Seed – 576 grams

Darvi – Berberis aristata – 960 g

Haritaki – Terminalia chebula – 960 g

Vibhitaki – Terminalia bellirica – 960 g

Amalaki – Amla – Emblica officinalis – 960 g

 

2nd Set

Makshika – honey – 9.6 kg

Sharkara – Sugar candy – 4.8 kg

Fine powders of each of

Dhataki – Woodfordia fruticosa – Flower – 960 g

Kankola – Piper cubeba – Fruit – 48 grams

Nagakeshara – Mesua ferrea – Stamen – 48 g

Jatiphala – Myristica fragrans – Seed – 48 g

Lavanga – clove - Syzygium aromaticum 48 g

Ela – cardamom - Elettaria cardamomum - 48 g

Twak – Cinnamon – Cinnamomum zeylanicum- 48 g

Patra - Cinnamomum tamala - leaves – 48 g

Krishna - Pippali - Long pepper - Piper longum - 192 g

 

The first set of herbs in coarse powder form is taken, added with 98.304 lit of water, boiled and reduced to 12.288 liters. It is than filtered and to the filtrate, honey and sugar sandy are well and filtered again. Then rest of the ingredients are added and kept in an air tight container which is smeared from inside with ghee for fermentation. After a month, it is taken out and filtered which is filled in bottles to be used as medicine. The medicine is manufactured by many companies such as Zandu, Sandu, Dabur, Baidyanath, Arya Vaidya Sala, AVP etc.

 

The medicinal roles of each of the ingredients are reported by various workers. Some of the important properties of each plant are mentioned below:

 

1. Khadira – Acacia catechu

The medicinal values of this plant were reviewed by Stohs and Bagchi, 2015.1 This plant has medicinal roles such as antioxidant, anti-inflammatory and chemo protective properties.

 

2. Devadaru - Cedrus deodara (ROXB.) LOUD

In Ayurveda this tree has very important medicinal roles. Pharmacological studies have revealed its role as antibacterial, mast cell stabilizing activity and immune-modulating, anti-inflammatory, anti-arthritic, anticancer, analgesic, anti-diabetic, anti-hyper-lipidemic, antioxidant, anxiolytic, anticonvulsant and antiulcer. (Saifulla et al, 2014; Chandur et al, 2011; Saxena et al, 2010; Zeng et al, 2012; Kumar et al, 2011)2-6

 

3. Bakuchi -Psoralea corylifolia

The plant possesses antibacterial, anti-depressent, antitumor, antioxidant, anti-inflammatory, antifungal and immunomodulatory activity (Chisty, 2016; Zang et al, 2016)7-8

 

4. Daruharidra - Berberis aistata

Berberis aristata is ethno botanically important herb that is used from time immemorial by mankind for the treatment of various ailments. Sharma et al, 2011, has reviewed this plant’s therapeutic role such as hepato-protective, hypoglycemic, anticancer, antimicrobial, anti-inflammatory, antioxidant etc. among many other medicinal values.9

 

5. Haritaki – Chebulic Myrobalan fruit rind – Terminalia chebula

One of the constituent of the common Triphala choornam, T. chebula bark, rind, galls etc. have been found to have activities like antioxidant, antimicrobial, anti- diabetic, hepato protective, anti-inflammatory, anti-arthritic, anti-mutagenic, anti-proliferative, radio-protective, cardio protective, hypo lipidemic, antispasmodic, Immuno-modulatory and antiviral, Bag et al, 2013.10

 

6. Belerica - Terminalia bellerica

Terminalia bellerica contain several phyto-constituents, which are found in seeds, leaves, peel of fruits and in whole plants; these phyto- chemicals includes flavones, steroids, tannin, ellagic acids, gallete, gallic acids, glycocides, terpenoid, saponin. Terminalia bellerica shows several pharmacological properties and functions these are anti-diabetic, anti- fertility, anti - fungal, analgesic, anti –cancer activity, anti-diarrheal activity, anti- hypertensive effect, anti-mutagenic effect, anti-oxidant, anti-secretory, anti- spasmodic, antithrombotic, anti-ulcer activity, immune modulatory activity, gastro intestinal, spleen, dysentery, cough and also wound healing (Saraswathi et al, 2012).11

 

7. Dhatri- Amla- Embelica officinalis

Amla has multifarious medicinal properties such as antipyretic, analgesic, as skin care lotion, antioxidant and also used to treat Gonorrhea, nausea, vomiting, indigestion, nose bleeding etc.12, 13

 

8. Dhataki – Woodfordia fruticosa

This plant is reported to have many important secondary metabolites like alkalaoids, 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)14, 15

 

 

 

9. 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)16-18

 

10. Nagakesara (Mesua ferrea L.)

This medicinal role of this plant was reviewed by Chahar et al, 2013). 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.19

 

11.  Nutmeg – Jathikkai - Myristica fragrans Houtt.

This is yet another spice used widely in Indian culinary practice for its characteristic fragrance. It has medicinal properties such as, antibacterial, antiviral, anti-diabetic, anti-leukemic etc. (Iyer et al, 2009; Chirathaworn et al, 2007).20-21

 

12. Lavanga – Clove – Syzigium aromaticum

Cloves are rich source of phenolic compounds like eugenol and gallic acid, which have medicinal properties such as antioxidant, antimicrobial, antiviral and cytotoxic.22

 

13. 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, 2009, have reported blood pressure lowering, fibrinolysis enhancing and antioxidant activities of Cardamom. Khan et al, 2011, have shown the pharmacological basis of cardamom as medicine for asthma.23, 24

 

14. 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). 25, 26

 

15. Patra - Cinnamomum tamala

This plant show medicinal activities such as anti-ulcer, anti-microbial, antioxidant, anthelminthic, antifungal, anti-hyper-lipidemic, analgesic and antipyretic, anti-diarrheal, anti-aflatoxigenic and lipid lowering.27-30

 

16. 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-hyperlipidemic, analgesic, antidepressant, anti-amoebic, vaso-dialtory, bioavailability enhancer due the presence of piperine in it, anti-obesity activity, radio-protective, cardio-protective and antifungal.31

MATERIALS AND METHODS:

The medicine Khadirarishtam was procured from standard Ayurvedic vendor at Chennai, India. 50 ml of Khadirarishtam 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 and WILEY).

 

Sample Preparation:

100 micro lit 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 Khadirarishtam is shown in Figure 1. The results obtained were tabulated and the medicinal values of the important compounds present were collected from various sources such as Dr. Duke’s Phytochemical and Ethnobotanical data base, NIST Chemical Library and available literature which are mentioned shown in Table 1.


 

Figure 1. Figure 1. Indicates the GC MS profile of Khadirarishtam.

 

Table 1. Indicates the retentions values, the type 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 Khadirarishtam.

Sl.

No

Retention Time

Compound Name

Mol. Formula

Mol Weight

Peak Area

Medicinal Role

1.

3.71

Eucalyptol

C10H18O

154.1

17955846

It helps contractile activity of smooth muscles.

2.

4.36

Phnylethyl alcohol

C8H10O

122.1

66393430

Alcohol dehydrogenase inhibitor, alcohol, detoxificant and antimicrobial.

3.

4.77

Ethyl hydrogen succinate

C6H10O4

146.1

82087967

Hydrogen peroxide inhibitor

4.

4.86

3-Cyclohexen-1-ol,4-methyl-1-(R)-

C10H18O

154.1

14993539

Not Known

5.

 

4.96

.alpha.-Terpineol

C10H18O

154.1

5819375

5, Alpha Reductase inhibitor, Alpha amylase inhibitor, Alpha Gucosidease inhibitor, Alpha reductase inhibitor, HIF 1alph inhibitor, Increae Alpha NMannosidse activity, Interluckin 1 alpha inhibitor, testosterone 5 alpha reducatase inhibitor TNF alpha inhibitor

6.

5.14

5-Hydroxymethylfurfural

C8H6O3

126

31444529

It is reported to stop

neuron apoptosis (Hai).32

7.

5.19

Acetic acid, chloro-,3-phenylpropyl ester

C11H13ClO2

212.1

5994532

Arachidonic acid inhibitor, increase aromatic amino acid decarboxylase activity, inhibit uric acid production

8.

5.92

Eugenol

C10H1202

164.1

 

It has medicinal roles such as antifungal, antioxidant, anticonvulsant and

local anaesthetic, antistress and bacteriostatic,

bactericidal, Anticarcinogenic, depresses activity

of central nervous depressant, anti- radiation,

antiviral, induces apoptosis in melanoma cells and

HL-60 leukemia cells.33-39

9.

5.97

1,2,3-Benzenetriol

C6H6O3

126

160563616

Not known

10.

6.30

Benzeethanol,4-hydroxy-

C8H10O2

138.1

29261617

17-beta-hydroxysteroid dehydrogenase inhibitor, Aryl hydrocarbon hydroxylase inhibitor, Testosterone hydroxylase inhibitor

11.

6.36

Propanoic acid, anhydride

C6H10)3

130.1

5518555

Arachidonic acid inhibitor, increase aromatic amino acid decarboxylase activity, inhibit uric acid production

12.

6.39

Thymine

C5H6N2O2

126

5731129

Not Known

13

6.70

Benzoic acid, 4-hydroxy-

C7H6O3

138

15072090

17-beta-hydroxysteroid dehydrogenase inhibitor, Aryl hydrocarbon hydroxylase inhibitor, Testosterone hydroxylase inhibitor

14.

7.08

Disulphide, methyl (methylthio)phenylmethyl

C9H12S3

216

5588619

Catechol O methyl transferase Inhibitor, Methyl donar, Methyl Guanidine inhibitor

15.

8.27

Isopsoralen

C11H6)3

181

27573873

Not Known

16

8.65

Ethyl gallate

C9H10O5

198.1

138570858

Not known

17

10.70

Hexadecnoic acid, 2-hydroxy-1-(hydroxymethyl) ethyl ester

C19H38O4

33.3

61077753

17-beta-hydroxysteroid dehydrogenase inhibitor, Aryl hydrocarbon hydroxylase inhibitor, Testosterone hydroxylase inhibitor

18

 

11.34

Octadecanoic acid, 2,3-dihydropropyl ester

C21H42O4

358.3

31298721

Arachidonic acid inhibitor, increase aromatic amino acid decarboxylase activity, inhibit uric acid production

19

11.80

Benzaldehyde, 2,4-dihydroxy-

C7H6O3

138

15855316

Not known

20

12.46

2(3H)-Furanone dihydro-3,4-bis[(4-hydroxy-3-methoxyphenyl)methyl]-,(3R-trans)-

C20H22O6

358.1

29761710

Not known

21

12.66

(-)-Nortrachelogenin

 

C20H22O7

374.1

99362845

 

Not known

22

12.82

1,4-Butanediol, 2,3-bis[(4-hydroxy-3-methoxyphenyl)methyl]-, [R-(R*,R*)]-

C20H26O6

 

362.2

30775686

Not known

 


From the GC MS profile of Khadirarishtam it is clear that there are a number of  biomolecules such as  Eucalyptol, Phnylethyl alcohol, Ethyl hydrogen succinate, .alpha.-Terpineol, 5-Hydroxymethylfurfural, Acetic acid, chloro-, 3-phenylpropyl ester, Eugenol, Benzeethanol,4-hydroxy-, Propanoic acid, anhydride,  Benzoic acid, 4-hydroxy-, Disulphide, methyl (methylthio)phenylmethyl, Hexadecnoic acid, 2-hydroxy-1-(hydroxymethyl) ethyl ester, Octadecanoic acid and  2,3-dihydropropyl ester, which indicate the  medicinal properties of Khadirarishtam. 

 

CONCLUSION:

From the above results it is concluded that Khadirarishtam contains some important biomolecules which indicate the medicinal properties of this medicine.

 

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Received on 16.09.2018          Modified on 14.10.2018

Accepted on 03.11.2018        © RJPT All right reserved

Research J. Pharm. and Tech 2019; 12(2):535-540.

DOI: 10.5958/0974-360X.2019.00094.5