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