ADALYA JOURNAL ISSN NO: 1301-2746

PRELIMINARY PHYTOCHEMICAL SCREENING AND FTIR ANALYSIS ON HUMILIS L. (MIXTURE).

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

The present investigation was focused on the preliminary phytochemical and Fourier Transform Infrared Spectral analysis of Rivina humilis L. The aqueous and organic solvent extract (petroleum ether, chloroform, ethyl acetate, methanol, water) were tested for availability of alkaloid, flavonoid, phenol, saponin, steroids, tannins. The FTIR spectrum showed the presence of primary/secondary amines, alkane, alkene, amino acid, nitro compounds etc. Isolation of medicinal components would however help to find new drugs. The results of this study offer a platform of using Rivina humilis as herbal alternative for various diseases.

Keywords: Phytochemical screening, FTIR, Rivina humilis, methanol, functional group

Introduction

Plants produce bioactive molecules in a diverse range making them a rich source of different types of medicine (Kala et al., 2011; Jeeva et al., 2012; Joselin et al., 2012; Florence et al., 2012; Florence et al., 2014). Phytochemicals are bioactive chemicals of origin. They are regarded as secondary metabolites and they are naturally synthesized in all parts of plant body (ie) any part of the plant body contain active compounds (Criagg and David; Tiwari, 2011). Medicinal have recently received the attention of the pharmaceutical and scientific communities and various publications have documented the therapeutic value of natural compounds in a bid to validate claims of their biological activity (Ncubeet al., 2008). The different phytoconstituents present in medicinal plants such as flavonoid, alkaloid, phenol and tannin, carboxylic acid, terpenes and amino acids and inorganic acids (Florence et al., 2014). These phytoconstituents present specific distinctiveness and properties and it has shown protective effects against diseases without reducing their therapeutic efficacy and

Volome 8, Issue 9, September 2019 208 http://adalyajournal.com/ ADALYA JOURNAL ISSN NO: 1301-2746

contains a wide range of bioactive compounds that can be used to treat various infectious diseases (De Britto et al., 2012).

Plants are the richest bio-resources of drugs of traditional system of medicine, modern medicines, neutraceuticals, food supplements, folk medicines, pharmaceutical intermediates and chemical entities for synthetic drugs (Hammer et al., 1999).

The Fourier Transform Infrared (FT-IR) spectroscopy allows the analysis of relevant amount of compositional and structural information in plants. Moreover, FT-IR spectroscopy is an established time saving method to characterize and functional groups (Grubeet al., 2008).

Rivina humilis (Blood or Rouge berry) is a species of in the family . It can be found in the the , and tropical . Rouge berry is a herbaceous . In these plant colourful bright shiny berries it is more attractive in than in . The red are gathered from the wild to be used for dyeing. The plant is widely grown as an ornamental in the tropics and subtropics. The aim of the present study is to identify the phytochemicals and functional groups present in the Rivina humilis L. (mixture) through screening of phytochemical and FTIR spectroscopy.

Methodology Collecting and processing of plant material

Rivina humilis plant was collected from Nagercoil, Kanyakumari district. The root, stem and were separately shade dried cut into small pieces, and fine powder was prepared by grinding, powdered was prepared by grinding, powdered material was stored in airtight container for further uses.

Preparation of extract

In shade dried root, stem and of Rivina humilis (at 25C) were powdered, in mechanical grinder. 20 gm of powdered sample of each extract was weighed. 150 ml of ethanol was added and kept for 3 days. The extract was filtered using Whatman No. 1 filter paper and the supernatant was collected. The residue was again extracted two times (with 3 days of interval for each extraction) and supernatant was collected. The supernatants were pooled and evaporated (at 1 room temperature 28 ± 1C) until the volume was reduced to 150 ml. (Geethu et al., 2014).

Volome 8, Issue 9, September 2019 209 http://adalyajournal.com/ ADALYA JOURNAL ISSN NO: 1301-2746

Qualitative phytochemical analysis

The extracts were tested for the presence of bioactive compounds by using following standard methods (Harborne, 1973).

Fourier Transform Infrared spectroscopic analysis (FT-IR)

A dry powder of root, stem and leaf (mixture) was encapsulated in KBr pellet, to prepare translucent sample discs. Small amount of plant part sample extract of ethanol solvent was respectively. The powdered sample of each extract was loaded in FTIR spectroscope with a scan range from 400 to 4000 cm-1. Results Preliminary phytochemical screening in the extract of Rivina humilis whole plant powder are shown in the table 1. The result shows steroids and phenols absent in all the extracts Alkaloids presented in all the extracts except petroleum ether. Flavonoids present in petroleum ether and chloroform extract. Saponins presented in petroleum ether and water extract, but it did not present in chloroform, ethyl acetate, methanol. Tannins presented in petroleum ether and chloroform extract but absent in ethyl acetate, methanol, water solvent. Table-1: Preliminary phytochemical screening in the extract of Rivina humilis whole plant samples. Phytochemicals screened in Rivina humilis

Whole plant

solvent extracts

used

Alkaloids Flavonoids Phenols Saponins Steroids Tannins 1. Petroleum ether - + + + + + - + + + + + 2. Chloroform + + + - + + + + + - + + 3. Ethyl acetate + - - - - - + - - - - - 4. Methanol + - - - - - + - - - - - 5. Water + - - + - - + - - + - -

Volome 8, Issue 9, September 2019 210 http://adalyajournal.com/ ADALYA JOURNAL ISSN NO: 1301-2746

The FTIR spectroscopy has been used to identify the functional groups of the active components present in the plant extracts based on peak values in the region of IR radiation. When the extract was passed through the FTIR, the functional groups of the components are separated based on its peak ratio (Arun et al., 2016).

The absorption spectra of crude powder of Rivina humilis whole plant powder are shown in the table 2. The results of plant powder confirmed the presence of 9 peak values showing different functional group. The 3288.04 cm1peak represent the primary / secondary amines. The peak at 2917.77, 2110.71 and 1622.80 confirms the presence of alkane, alkene, and amino acids. The peak at 1373.07, 1318.11 confirm the presence of alkane, nitro compounds respectively. The peak at 1240.97 indicated fluoride, alkyl halide. The band at 1019.19 shows aliphatic amines. The peak at 512.97 cm1 shows halogen, alkyl halide. TABLE 2: FTIR PEAK VALUE AND FUNCTIONAL GROUPS OF WHOLE PLANT SAMPLE OF RIVINA HUMILIS L. (MIXTURE) S.NO PEAK FUNCTIONAL GROUP VALUE

1 3288.04 PRIMARY /SECONDARY AMINES 2 2917.77 ALKANE 3 2110.71 ALKENE

4 1622.80 AMINO ACIDS

5 1373.07 ALKANE, NITRO COMPOUNDS 6 1318.11 ALKANE, NITRO COMPOUNDS

7 1240.97 FLUORIDE, ALKYL HALIDE. ALIPHATIC AMINES 8 512.97 HALOGENS, ALKYL HALIDE.

Volome 8, Issue 9, September 2019 211 http://adalyajournal.com/ ADALYA JOURNAL ISSN NO: 1301-2746

FT-IR Spectra of Rivina humilis L.

Discussion

Plants are important source of functional components for the development of new pharmaceutical agents. In the phytochemical investigation of ethanolic extract of Rivina humilis revealed the presence of various phytochemicals such as alkaloid, flavonoids, saponins, tannins. Previous studies described that plant phenols are highly effective free radical scavengers and act as antioxidants (Maisuthisakul et al., 2007). In Unani and Ayurvedic system of medicine, phytomedicine have been used for the treatment of diseases. The phytochemicals screening serves as the initial step in predicting the types of potential active compounds (Suhumaran et al., 2011). The present studies revealed in the ethanolic extract of Rivina humilis have higher contents of the phytochemicals.

FTIR is one of the most widely used methods to identify the chemical constituents and elucidate the compounds (Gopalakrishnan et al., 2012). FTIR allows infrared spectrum simultaneously providing speed and accuracy in measurements of whole range of biological specimens (Griffith et al., 1986). FTIR spectral analysis of Rivina humilis spectrum reveals the presence of peaks, which functional group. Many researchers applied for FT-IR spectrum as a tool for distinguishing the medicinal plants based on their chemical constituents (Asha et

Volome 8, Issue 9, September 2019 212 http://adalyajournal.com/ ADALYA JOURNAL ISSN NO: 1301-2746

al., 2014; Florence and Jeeva, 2015; Florence et al., 2015; Lincy et al., 2015; Joselin and Jeeva, 2016).

Conclusion

In conclusion Rivina humilis are discriminated in the presence or absence of some of the phytoconstituents by primary qualitative screening and a combined FTIR-PCA. Both the studies clearly reveal that the maximum class of phytoconstituents from the Rivina humilis sample mixture. The identification of phytochemical constituents of this plants having highest therapeutic efficacy and could be develop more novel drugs for various ailments.

References

Arun, K., Pingal, K &Somasundaram, S.T. (2016). Phenolic composition, antioxidant activity and FT-IR spectroscopic analysis of halophyte Sesuviumportulacastrum L. extract. International Journal of Biological Science, 5, 1-13.

Asha V, Jeeva S and Paulraj K, 2014. Phytochemical and FT-IR spectral anaylsis of CarallumageniculataGrev. Et Myur. An endemic medicinal plant, Journal of Chemical and Pharmaceutical Research, 6(7): 2083-2088.

Criagg, G.M, David, J.N, 2001. Natural product drug discovery in the next millennium, J. PharmaBiol. 39: 8-17.

De Britto AJ, Sebastian SR and Sujin RM, 2012. Antibacterial activity of selected species of Lamiaceae against human pathogen, Indian Journal of Natural Products and Resources, 3(3): 334-342.

Florence A.R, Joselin J, Brintha T.S.S, Sukumaran S and Jeeva S, 2014. Preliminary phytochemical studies bioactive constituents, Bio science Discovery, 5(1): 85-96.

Florence AR, Joselin J, Jeeva S. 2012. Journal of Chemical and Pharmaceutical Research; 4(11): 4908-4914.

Florence, A.R, Sukumaran S, Joselin J, Brintha T.S.S and Jeeva S, 2015. Phytochemical Screening of Selected Medicinal Plants of the family Lythraceae, Bio science Discovery, 6(2): 73-82.

Florence, A.R. and Jeeva, S. 2015. FTIR and GC-MS spectral analysis of Gmelinaasiatica L. Leaves, Science Research Reporter, 5(2): 125-136.

Volome 8, Issue 9, September 2019 213 http://adalyajournal.com/ ADALYA JOURNAL ISSN NO: 1301-2746

Geethu, M.G., Suchithra P.S., Kavitha, C.H., Aswathy J.M. Dinesh Babu, K. Murugan 2014. Fourier transform Infrared spectroscopy Analysis of different solvent extracts of Water hyacinth (Eichhorniacrassipes Mart Solms) Allelopathic approach. World Journal of Pharmacy and Pharmaceutical Sciences. 3(6): 1256-1266.

GopalakrishnanVK.,Starlin T., Arul Raj, C., Ragavendhran, P., 2012. Phytochemical Screening functional groups and elemental analysis of Tylophorapauciflorawight and arn. International Research Journal of Pharmacy., 3 : 6.

Griffiths, P.R and De Haseth, J.A., 1986. Fourier transform infrared spectroscopy. John Wiley and Sons, New York, 656.

Grube M, Muter O, Strikauska S, Gavare M and Limane B, 2008. Application of FT-IR spectroscopy for control of the medium composition during the biodegradation of nitro aromatic compounds, Journal of Industrial Microbiology and Biotechnology, 35: 1545-1549.

Hammer KA, Carson CF and Riley TV, 1999. Antimicrobial activity of essential oils and other plant extracts, Journal of Applied Microbiology, 86(6): 985-990.

Harborne JB, 1973. Phytochemcial Methods, Chapman and Hall, London.

Jeeva S, Johnson M. Asian Pacific Journal of Traditional Biomedicine, 2012, S151-S154.

Joselin J, Brintha TSS, Florence AR, Jeeva S. 2012. Journal of Chemical and Pharmaceutical Research; 5(4): 106-111.

Joselin, J. and Jeeva, S. 2016. GC-MS and FT-IR analysis of a coastal medicinal plant.Hyptissuaveolens (L) poit, Journal of Coastal Life Medicine, 4(5): 380-385.

Kala S, Johnson M, Raj I, Bosco D, Jeeva S, Janakiraman N. 2011. Journal of Nature Conscientia; 2(5): 478-481.

Lincy, M. P, Mohan V.R and Jeeva S. 2015. Preliminary phytochemical screening, gas chromatography mass spectrum and Fourier transform infrared spectroscopy anlaysis of aerial part of Maeruaapetala Roth (Jacobs)., Chemical Science Review and Letters 4(16): 1275-1284.

Maisuthisakul, P., Pongsawatmanit, R. and Gordon, M.H., 2007. Characterization of the phytochemicals and antioxidant properties of extracts from teaw (CrotoxylumFormosum Dyer). Food Chemistry., 100: 1620-1629.

Volome 8, Issue 9, September 2019 214 http://adalyajournal.com/ ADALYA JOURNAL ISSN NO: 1301-2746

Neube, N.S, Afolayan, A.J and Okoh A.I, 2008. Assessment techniques of antimicrobial properties of natural compounds of plant origin: Current methods and future trends, African Journal of Biotechnology, 7(12): 1797-1806.

Sukumaran, S., Kiruba, S., Mahesh, M., Nisha, S.R., Miller, P.Z., Ben, C.P., 2011. Phytochemical constituents and antimicrobial efficacy of the of pterocarpum Asian Pacific Journal of tropical medicine, 735-738.

Tiwari, Kumar B, Kaur G, Kaur, H. 2011. Int. Pharm Science, 1: 98-106.

Volome 8, Issue 9, September 2019 215 http://adalyajournal.com/