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RESEARCH ARTICLE

 

 

 

Phytochemical Analysis of   Terminalia  catappa  Stem using GC-MS/MS

 

M. Krishnaveni1*, G. Krishna Kumari2, C. Ragina Banu2, M. Kalaivani2

1Assistant Professor, Department of Biochemistry, School of Bio-Sciences, Periyar University, Salem

2M.Phil  students, Department of Biochemistry, School of Bio-Sciences, Periyar University, Salem

*Corresponding Author E-mail: krishnavenim2011@gmail.com

ABSTRACT:

Stem stores nutrients. It also supports leaf, flower, fruit growth, asexual reproduction.  Hence, for the present study, Terminalia catappa stem was used for the analysis of phytochemicals, antimicrobial activity. The results of phytochemicals assessed in ethanol extract of Terminalia catappa stem through GC-MS/MS showed 13 compounds.  Highest peak was observed with Propane, 1,1-diethoxy- (48.69) and t-Butyl hydrogen phthalate (33.26). All the other compounds showed moderate peaks in the range of 2.85 to 0.37. The antimicrobial nature of Terminalia catappa stem was possible with most of the phytochemicals  identified.  The antimicrobial activity was higher with fungi Aspergillus niger, Candida albicans on comparison with bacteria. The microbial activity was confirmed with zone of inhibition. The zone of inhibition observed was higher for fungi showing its antifungal potency.

 

KEYWORDS: Anti fungal, Anti bacterial, Analytical Technique, Plant, Stem, Terminalia catappa.

 


INTRODUCTION1-10

Plants that have healing properties are termed as medicinal plants or herbs. It is a tree native to India, Indo china, Thailand present in Andaman and Nicobar islands also. Wood of  Terminalia catappa trees are used  in the manufacture of furniture and also as  fuel. Good growth of this tree requires 1000mm rain, sandy soil for its fast growth. The maximum height of the tree is 800m and in India it grows up to 300m. Medicinal  plants forms a base for the development of novel drugs which are cheaper, safe than synthetic medicine. Phytotherapy uses plant extract for medicinal purposes leading to healthy life. Hence, the present study was planned to identify the compounds present in Terminalia catappa stem using analytical technique- GCMS/MS and its antimicrobial activity.

 

 

 

 

 

 

 

Received on 19.07.2015          Modified on 18.08.2015

Accepted on 28.08.2015        © RJPT All right reserved

Research J. Pharm. and Tech. 8(9): Sept, 2015; Page 1281-1283

DOI: 10.5958/0974-360X.2015.00232.2

 

MATERIALS AND METHODS:

Sample collection:

Fresh Terminalia catappa stem samples were collected, shade dried, powdered. 25grams of powdered stem sample was used for ethanol extraction. The extracted sample was used for phytochemical analysis through GC-MS/MS, anti-microbial activity. The plant was authenticated by Dr. A. Balasubramanian. The authentication number was AUT/PUS/070 dated 17/12/2014.

 

Analytical Method:

GC-MS/MS was performed on a Scion 436-GC Bruker carrying Triple quadruple mass spectrophotometer with fused silica capillary column BR-5MS (5% Diphenyl95% Dimethyl poly siloxane), 30m x 0.25mm ID x 0.25m df. The column oven temperature program was as follows: 80°C hold for 2 min, Up to 160°C at the rate of 20°C/min-No hold, Up to 280°C at the rate of 5°C / min-No hold, Up to 300°C at the rate of 20°C/min-10 min hold, Injector temperature 280°C, Total GC running time was 41 min. The inlet temperature was set at 280°C, source temperature 250°C; ionization mode, ionization at 70-eV ionization energy; For single scan analysis, the scan range was set from m/z 40 to 600; Solvent Delay: 0-3.5 min; and the injection volume was 2μl. The GC-MS/MS was performed by Institute of crop processing technology, Tanjavur.  Here, GC-MS/MS was used as it identifies compounds in complex at trace levels i.e lower than the limits of GC-MS. 

 

Antimicrobial assay:

The antimicrobial activity was assessed by means of Kirby- Bauer technique.1

 


Results and Discussion:

Table.1. Depicts the results of compounds identified in Terminalia catappa Stem.

S.No

RT

Name of  the compound

Molecular Formulae

MW

Peak area %

1.

4.73

Propane, 1,1-diethoxy-

C7H16O2

132

48.69

2.

5.84

2-Dodecyloxyethanol acetate (ester)

C16H32O3

272

2.33

3.

7.52

1,2,3-Benzenetriol

C6H6O3

126

1.45

4.

8.80

Benzenamine, N,N-dimethyl-4-[[(1-methylethyl)imino]methyl]-

C12H18N2

190

0.79

5.

9.87

β-l-Arabinopyranoside, methyl

C6H12O5

164

1.60

6.

12.92

Nonanoic acid, ethyl ester

C11H22O2

186

0.65

7.

15.67

Phthalic acid, pentyl tridec-2-yn-1-yl ester

C26H38O4

414

2.85

8.

16.34

Dodecanoic acid, ethyl ester

C14H28O2

228

2.36

9.

19.32

8-Methyloctahydrocoumarin

C10H16O2

168

1.80

10.

25.21

t-Butyl hydrogen phthalate

C12H14O4

222

33.26

11.

29.32

Cyclopropanecarboxylic acid, oct-3-en-2-yl ester

C12H20O2

196

0.37

12.

32.34

Stigmastan-3,5-diene

C29H48

396

2.26

13.

34.29

Stigmasterol

C29H48O

412

1.58

 

 

 


13 compounds were identified totally. Among the thirteen compounds identified, the highest peak was observed with propane, 1,1-diethoxy having molecular weight of 132, molecular formulae C7H16O2, showing peak area percent of  48.69. The other compound that was showing highest peak was t-Butyl hydrogen phthalate carrying molecular formulae C12H14O4, molecular weight 222, showing peak area percent 33.26. The following compounds showed moderate peak area percent Phthalic acid, pentyl tridec-2-yn-1-yl ester, Dodecanoic acid, ethyl ester, 2-Dodecyloxyethanol acetate (ester), Stigmastan-3,5-diene, 8-Methyloctahydrocoumarin, β-l-Arabinopyranoside methyl, Stigmasterol, 1,2,3-Benzenetriol, Their respective peak values are as follows: 2.85, 2.36, 2.33, 2.26, 1.80, 1.60, 1.58, 1.45. Their molecular formulae and molecular weight are: C26H38O4/414, C14H28O2/228, C16H32O3/272, C29H48/396, C10H16O2/168, C6H12O5/164, C29H48O/412, C6H6O3/126. The compounds showing least peak area percent are: Benzenamine, N,N-dimethyl-4-[[(1-methylethyl)imino]methyl]-(Molecular formulae: C12H18N2, Molecular weight: 190, Peak: 0.79%), Nonanoic acid, ethyl ester (Molecular formulae: C11H22O2 Molecular weight: 186, Peak: 0.65 %), Cyclopropanecarboxylic acid, oct-3-en-2-yl ester (Molecular formulae: C12H20O2, Molecular weight: 196, Peak:0.37 %). Terminalia catappa fruit, flesh, nut, shell was studied by Krishnaveni et.al for its phytonutrient, antioxidant, qualitative assay as well as GC-MS/MS analysis, air pollution tolerance index of Terminalia catappa  leaves.2-4,5&6-8 Likewise, air pollution tolerance index, antioxidant activity of Terminalia tomentosa was studied by Krishnaveni et.al.9,10.  


 

 

 

 

Fig.1 Shows the chromatogram of  Terminalia catappa  stem.

 


 

 

 

 

 

 

 

 

Antimicrobial activity:

Table.2 Antimicrobial activity of ethanol extract of  Terminalia catappa stem

Plant used

Microbes tested

Zone of inhibition (mm)

Terminalia catappa stem

Escherichia coli

Staphylococcus aureus

Aspergillus niger

Candida albicans

 8

 6

15

14

 

The results of antimicrobial activity was shown in Table.2. The antimicrobial activity was measured through zone of inhibition. From the results we could observe higher zone of inhibition for fungi such as Aspergillus niger and  Candida albicans. The bacteria like Escherichia coli, Staphylococcus aureus was showing lesser zone of inhibition. This confirms the tendency of ethanol extract of Terminalia catappa stem and its effect was found to be antifungal. 

 

CONCLUSION:

Any agents whether it is a drug, compound, extract that have the ability to kill or inhibit microbes are called as antimicrobials. In the present study, the compounds in the ethanol extract are able to act against bacteria and fungi. But the activity was higher with fungi when compared to bacteria.  The pharmacological activity of stem was due to its compounds contained in it.

 

ACKNOWLEDGEMENT:

The author wishes her thanks to Honorable Vice-chancellor Dr. C. Swaminathan Avl, and Registrar Dr. M. Manivannan Avl, Periyar University, Salem for their administrative support and excellent infrastructure facilities provided and also coordinator, School of Bio-Sciences, Periyar University, Salem, Dr. A. Balasubramanian, ABS Botanical garden, Salem for his help in authenticating plants, Director Dr. K. Alagusundaram and Dr. S. Kumaravel, senior scientist,     Indian Institute of Crop Processing Technology, Thanjavur for analyzing the samples. The author would like to express her gratitude to her dedicated teachers.

 

REFERENCES:

1.       Bauer AW, Perry DM, Kirby WMM. Single disc antibiotic sensitivity testing Staphylococci. AMA Arch Intern Med 1959; 104(2): 208–216.

2.       Krishnaveni M, Dhanalakshmi R. Phytonutrient analysis in Terminalia catappa  fruit, flesh,  nut, shell. International Journal of Current Pharmaceutical Review Research 2015; 6(1): 28-35.

3.       Krishnaveni M. Invitro antioxidant activity of Terminalia catappa nuts. Asian Journal of Pharmaceutical and Clinical Research 2014; 7(4): 33-35.

4.       Krishnaveni M, Jasbin Shyni G, Dhanalakshmi R, Magesh P, Ponraj K,  Lavanya K, Kalimuthu R. A preliminary study on phytoanalysis, Antioxidant potential of Terminalia  catappa l. fruit flesh. International Journal of Pharmaceutical Sciences Review and Research 2014; 28(1): 83-87.

5.       Stanley Ejike U, Doris Uchenna E. Comparative study on phytochemical, phenolic and antioxidant profiles of leaf, root, stem barks of Terminalia glaucescens (Planch Ex. Beth). Indo American Journal of Pharmaceutical Research  2014; 4(12): 5801-5807.

6.       Krishnaveni M, Krishna kumari G, Ragina banu C, Kalaivani M. GC-MS/MS Analysis of phytochemicals in Terminalia catappa L, Antimicrobial assay. Indo American Journal of   Pharmaceutical Research  2015; 5(3): 1250-1254.

7.       Krishnaveni M, Magesh P. Air pollution tolerance index induced by biochemical components  in plants. International Journal of Pharmacy and Pharmaceutical Sciences 2014; 6 (5):  362-364.

8.       Krishnaveni M, Krishna kumari G, Kalaivani M. Ragina banu C, Gas chromatography-Mass spectrometry/Mass Spectrometry analysis of  Terminalia catappa  L. nut and antimicrobial  assay. Asian Journal of Pharmaceutical and Clinical Research, 2015, Vol. 8(4), 168-170.

9.       Krishnaveni M, Durairaj S, Madhiyan P, Amsavalli L, Chandrasekar R. Impact of Air Pollution in Plants near Thermal Power Plant, Mettur, Salem, Tamilnadu, India. International  Journal of Pharmaceutical Sciences Review and Research 2013; 20(2): 173-177.

10.     Krishnaveni M, Durairaj S, Madhiyan P, Amsavalli L, Chandrasekar  R. In vitro free radical  scavenging activity of aqueous leaf extract of plants near thermal power plant, Mettur, Salem. International  Journal of Pharmaceutical Sciences and Research 2013; 4(9): 3659- 3662.