CIKITUSI JOURNAL FOR MULTIDISCIPLINARY RESEARCH ISSN NO: 0975-6876

GC-MS ANALYSIS OF ETHANOLIC EXTRACT OF L. (STEM).

1Kavitha A and 2Mary Kensa V

1. Reg. No: 17223152142006, P.G. Department of Botany and Research Centre, S.T. Hindu College, Nagercoil- 629002. M.S. University, Abishekapatti, Tirunelveli 627012, Tamil Nadu, India. Email.id: [email protected]. 2. PG and Research Department of Botany, S.T. Hindu College, Nagercoil – 629 002, Tamil Nadu

Abstract

Rivina humilis is a species of flowering in the family . It is a monotypic genus native to Mexico. Common name of the plant is Blood berry, pigeon berry. In the present study the ethanolic stem extract of Rivina humilis has been subjected to GC-MS analysis. The chemical composition of stem ethanol extracts of R. humilis were investigated using Perkin Elmer Gas chromatography mass spectroscopy. The result of this study revealed the existence of the 14 compounds. Such as Cinnamyl 3, 4- Dihydroxy-a-cyannocinnamate, L-proline, N-methoxycarbonyle, Isohexylester, L- Tetradecene, 1-Nonadecence, caffeine, n-hexadecanoic acid, oleic acid, octadecanoic acid etc. This plant sample contains various bioactive compounds and therefore has various medicinal properties which can be used for the treatment of various diseases.

Keywords: Rivina, phytochemical, GC-MS, ethanol and medicinal properties.

Introduction

From ancient times, medicinal have been used extensively for their tremendous healing properties and health benefits India has a treasure of medicinal plants due to the rich diversity in its agroclimatic condition (Maheswari, 2018). Awareness of medicinal plants usage is result of the many years of struggles against illness due to which man learned to pursue drugs in barks, seeds, fruit bodies, leaves and part o the plants (Biljanai, 2012). In India the medicinal systems using medicinal plants are Ayurveda, Siddha, Homeopathy etc. to treat various ailments (Puspangandan et al., 1984).

Rivina humilis L. is a herbaceous plants commonly found in wasteland of garden and plains. It is a monotypic to genus, native to mexico. It belongs to the family Phytolaccaceae.

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The plant is commonly known as blood berry or rouge plant. The investigation was carried out to determine the chemical components of Rivina humilis stem using Perkin Elmer Gas chromatography-Mass spectrometry. While the mass spectra of the compounds found in the extract were matched with the National Institute of Standards and Technology (NIST) library. GC-MS analysis of ethanol extract of Rivina humilis stem revealed the existence of plants are a rich source of secondary metabolites with interesting biological activities (Kensa and Neelamegam., 2016). Higher plants as source of bioactive compounds continue to play a dominant role in the maintenance of human health. Reports available on green plants represent a reservoir of effective chemotherapeutants these are non-phytotoxic, more systematic and easily biodegradable (Vyas., 1999; Kaushiket al., 2002; ChamanLal and Verma., 2006). If we can come back to our nature, culture and tradition of use of medicinal plants it can bring up a bright and health new generation (Kirtikar and Basu., 1918).

In GC-MS method the unknown organic compounds in a complex mixture can be determined by interpretation and also by matching the spectra with reference spectra (Ronald, 1997; Chauhan et al., 2014). GC-MS analysis of aerial part of this plant revealed the presence of many bioactive components. Sudha et al. 2013). This plant is also known to posses’ Antimicrobial activity, Antioxidant activity, anti-inflammatory activity, antibacterial activity, therapeutic agent, antimicrobial activity Nematicidal activity.

Materials and Methods Collection of plant sample

Stem of R. humilis L. was collected from Nagercoil, Kanyakumari district of Tamil Nadu, India and authenticated by Botanist Dr. R. Murugan, BSI, Southern circle, Kovai, India. A voucher specimen was deposited in the herbarium of the Botanical Survey of India Coimbatore; Herbarium code No. BSI/SRC/18/710-17/Tech. (Kavitha et al., 2019).

Plant sample extraction

Stems were cleaned, shade dried and pulverized to powder in a mechanical grinder. Required quantity of powder was weighted and transferred to stoppered flask and treated with ethanol until the powder is fully immersed. (Shanmugavel et al., 2015). Dark green residues were obtained after concentrating the extract under reduced pressure. The obtained extracts were stored in desiccators for further GC-MS.

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GC-MS Analysis

Gas chromotogrpahy (GC) analysis was carried out using Perkin Elmer Clarus. SQSC gas chromatography equipped with capillary PTV injector. The chromatograph was fitted with DB 5 MS capillary standard nonpolar column (30 m  0.25 mm i.d., film thickness

0.25 m). The injector temperature was set at 250C and the oven temperature was initially set at 70C then programme as follows.

Initial Temp C Rate C Hold Time (min) 70 - 3 150 10 2 220 5 1 250 10 5

Helium was used in carrier gas with the flow rate of 1 mL/min. One microliter of sample (diluted with 1: 4) injected in the split mode in the ratio of 1: 12.

The mass spectrometer was operated in the electron impact mode +ve. Ion source and transfer line temperature was kept at 220 & 250 C. The mass were obtained by centroid scan of the mass range from 40 to 650 amu. The extract was identified based on the comparison of Retention time (RT) and their obtained mass spectra to NIST library data of the GC-MS system and literature data.

Identification of bioactive components

Interpretation of mass spectrum GC-MS was made by using the database of National Institute of Standard and Technology (NIST) having more than 62000 patterns. (Selvamangai and Bhaskar, 2012). Spectrum of the unknown component was compared with the spectrum of the known components stored in the NIST library. Prediction of bioactivity of compound is done based on Dr. Duke’s phytochemical and Ethnobotanical database. The relative percentage amount of each phyto-component was calculated by comparing it average peak area to the total area. (Amaravani and Ramesh, 2017).

Results and Discussion

The GC-MS chromatogram of ethanolic extracts of stem of Rivina humilis revealed the presence of various compounds with corresponding peaks at different retention time

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(Govindaraj and Rajangam, 2017). GC-MS is one of the best techniques to identify the constituents in plants. The GC-MS analysis of R. humilis stem revealed the presence of 14 compounds.

Table 1, figure 1 showed the various bioactive compounds were characterized and identified).

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Table 1: phytocomponents detected in R. humilis stem

S. Compound Name RT Area% Activity No. Potent and selective inhibitor of platelet 1 Cinnamyl 3, 4-dihydrox – a –cyanocinnamate 3.539 0.492 Antimicrobial activity 2 1-Proline, N-methoxy carbonyl-, isohexyl ester 6.965 1.210 It helps cartilage and cushion joints 3 1-Tetradecene 9.776 0.770 Antioxidant It has a role as a plat metabolite and a bacterial 4 1-Nonadecene 13.588 0.600 metabolic Acts as a central nervous system stimulant, Manage 5 Caffeine 18.670 0.631 drowsiness headaches. 6 n-Hexadecanoic acid 21.135 5.382 Anti-inflammatory agents 7 Oleic acid 21.821 0.764 Antibacterial activity used as an emollient Antioxidant, cancer preventive Dr. Dukes 8 Octadecanoic acid 24.887 6.147 phytochemical and ethano botanical dataset (Anita et al., 2017) 9 7,8-Epoxylanostan-11-01,3-acetoxy- 30.969 0.614 Antimicrobial activity 10. 9-octadeconoic acid (z)-, 2,3-bis acetyloxy) propyl ester 31.214 4.106 Pharmaceutical solvent 11 Cyclohexane, 1,1-(2-tridecyl-1,3-propanediyl) bis- 32.500 0.531 Antibacterial activity 12 3-(octanoyloxy)propane -1, 2-diyl bis (decanoate 32.990 20.770 Nematicidal activity 13 Octadecane, 3-ethyl-5-(2-ethylbutyl)- 34.156 1.070 Bacterial metabolite and a plant metabolite 14 Cholest -22-ene-21, 3,5 dehydro-6-methoxy privalate 34.811 0..729 Antioxidant activity

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The identification of the phytochemical compounds was confirmed based on the peak area, retention time. The results revealed the Cinnamyl3,4-dihydroxy- a – Cyanocinnamate, 1-proline, N-methoxy carbonyl-, isohexyl ester, 1-Tetradecence, 1-Nonadecence, Caffeine, n- Hexadecanoic acid, Oleic acid, octadecanoic acid, 7,8 Epoxylanostan-11-01,3-acetoxy-9- octadeconic acid (z)-2,3-bis(acetyloxy)propyl ester, cyclohexane, 1,1-(2-tridecyl-1,3- propanediyl)bis-,3-(octanoyloxy)propane-1,2-diyl bis(decanoate),Octadecane,3-ethyl-5-(2- ethylbutyl)-,cholest-22-ene-21-01,3,5-dehydro-6-methoxy revealed the presence of various compounds with corresponding peaks at different retention time (Govindaraj and Rajangam, 2017). GC-MS is one of the best techniques to identify the constituents in plants. The GC-MS analysis of R.humilis stem revealed the presence of 14 compounds (phytochemical constituents).

In the present study compounds have been identified from exthanol extract of the R. humilis stem. Plants synthesize an extensive array of secondary metabolites often highly compound structures. The chemical investigations of medicinal plants have largely been driven to find new drugs to treat human disease. The secondary metabolites have been of interest to humans as flavors, fragrance, dyes, pesticides and pharmaceuticals (Govindaraj and Rajangam, 2017).

The mass spectametry analysis was carried out to identify the compounds in stem Rivinahumilis eluted at different retention time. Similarly the bioactive compounds of stem bark of sennaalata (Ananthi and Subalakshmi, 2016). 18 compounds have been identified from ethanol extract of stem of Nothapodytes nimmoniana (Shanmugavel et al., 2015).

Conclusion

In this present study about 14 bioactive compounds are identified from ethanol extract of R. humilis by GC-MS method. The presence of various phytoactive compounds in this plant is responsible for the pharmaceutical properties. Therefore, it is recommended as a plant of phytopharamaceutical importance. Present study may be useful in the identification of

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novel drugs from stem of R. humilis. It is concluded that the ethanol can be used for extracting active compounds from plants and incorporating into medicinal food products. In addition further research is necessary to identify the active compounds responsible for therapeutic activity and animal study to evaluate the dosage of the identified chemical compounds. References

Amaravani P Kanjikar, Ramesh L Londonkar. 2017. GC-MS analysis of bioactive compounds in petroleum ether extract of Ficus krishnae. International research journal of pharmacy. 8 (10): 178-181.

Anita Sinha, H.P. Sharma, Binod Singh and Anitpatnaik. 2016. Phyto-chemical studies ofmethanol extracts of Tinosporacordifoliastem by GC-MS.. World Journal of pharmaceutical Research, 6(4): 1319-1326.

Biljana, B.P. (2012). Historical reviews of medicinal plants usage. Pharmacognosy Review. 6: 1-5.

ChamanLal, and Verma, L.R. 2006. Use of certain bio-products for insect-pest control. Indian Journal of Traditional Knowledge, 5(1): 79-82.

Chauhan, A. Goyal, M.K, Chauhan P. 2014. GC-MS technique and its analytical applications in science and technology. J. Anal .Bioanal. Tech; 5: 222.

GovindarajSabithira and Rajangam.Udayakumar, 2017. GC-MS analysis of methanolic extracts of Leaf and stem of Marsilea minuta (Linnn). Journal of complementary and Alternative medical Research, 3(1): 1-13.

Kaushik, J.C, Arya Sanjay, Tripathi, N.N. Arya, S. 2002. Antifungal properties of some plant extracts against the damping off fungi of forest nurseries. Indian Journal of forestry: 25: 359-361.

Kirtikar, K.R. Basu, B.D. 1918. Indian medicinal plants. Indian Press, p. 34-44.

Maheswari, J.2011. Patenting Indian Medicinal Plants and Products.Indian J. Sci. Technol., 4: 298-301.

Mary kensa V, Neelamegum R. 2016. GC-MS determination of bioactive constituents of Hydrilla verticillata (L.F.) Royle. collected from unpolluted and polluted water sources. Asian Journal of Biology, 1(1): 1-6.

Volume 6, Issue 9, September 2019 27 http://cikitusi.com/ CIKITUSI JOURNAL FOR MULTIDISCIPLINARY RESEARCH ISSN NO: 0975-6876

Pushpangandan P, Atal C.K. 0984. Ethno-neico-botanical investigation in Kerala I. Some herbal medicines. J. Ethnopharmacol, 1: 59-77.

Ronals Hites A. 1997. Gas chromatography-Mass spectroscopy: a handbook of instrumental techniques for analytical chemistry; P. 609-11.

Selvamangai G. Bhaskar, A. 2012. GC-MS analysis of phytocomponents in the methods extract of Eupatorium triplinerve. Asian Pacific Journal of Tropical Biomedicine, 5:1329-1332.

Shanmugavel Kavitha, Maria Louis PackiaLincy, Soosaimicheal Mary Jelastin, Kala, VeerababuRamaswamy Mohan and ArumugamMaruthupandian. 2015. GC-MS analysis of ethanol extract of stem of Nothapodytes nimmoniana. Sci.enves. 3(6): 1145-1150.

Sudha T, Chidambarapillai S and Mohan V.R. 2013. GC-MS analysis of bioactive components of aerial parts of Kirganelia reticulates poir (Euphorbiaceae). J. Curr. Chem .Pharm., 3(2):113-22.

Vyas, D.D.1999. Soil fertility deterioration in crop land due to pesticide. Journal of Indian Botanical Society; 78:177-178.

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