Int J Pharm Pharm Sci, Vol 6, Issue 9, 83-85Original Article

PHYTOCHEMICAL SCREENING, FUNCTIONAL GROUPS AND ELEMENTAL ANALYSIS OF LEAF EXTRACT OF IPOMOEA OBSCURA (L) KER-GAWL

SARAVANA PRABHA P1, GOPALAKRISHNAN V. K 1,2*

1Department of Bioinformatics, Karpagam University, Coimbatore, Tamilnadu, India 641021, 2Department of Biochemistry, Karpagam University, Coimbatore, Tamilnadu, India 641021
Email: vkgopalakrishnan@gmail.com

Received: 14 May 2014 Revised and Accepted: 01 Sep 2014

ABSTRACT

Objective: The aim of the study was to find out the phytochemical constituents, and to evaluate the functional group and elemental analysis of ethanol leaf extract of Ipomoea obscura (L.).

Methods: The secondary metabolites was analyzed for various extracts by using standard techniques. The metabolic activites can be measured by using FTIR and SEM-EDX.

Results: Phytochemical screening revealed the presence of alkaloids, glycosides, steroids, terpenoids, flavonoids, saponins and fatty acid and lipids in the extract. FTIR was used to identify various functional groups. Scanning Electron Microscope (SEM) with an energy dispersive X- ray spectrometer (EDX) showed the presence of calcium, magnesium, silicon, chloride, oxygen and potassium in ethanol leaf extract.

Conclusion: The results suggested that the ethanol leaf extracts of Ipomoea obscura (L.) showed the presence of most of the secondary metabolites, functional groups and trace elements.

Keywords: Phytochemical screening, FTIR, EDAX, Ipomoea obscura.

INTRODUCTION

Plant based drug has been used worldwide in traditional medicines for treatment of various disease. The study of medicinal plants with a history of traditional use as a potential source of substance with significant pharmacological and biological activities such as antioxidant, anticancerous and hepatoprotective [1]. Phytochemical studies have attracted the attention of plant scientists due to the development of new and sophisticated techniques. These techniques played a significant role in the search for additional resources of raw material for pharmaceutical industry [2]. Ipomoea obscura (L.) Commonly known as ‘Laksmana’ in Ayurveda belongs to the family Convolvulaceae. It is small climbing vine, with small cordate leaves and acuminate apex. Corolla composed of five fully fused petals. Plant grows on fences or low ground cover as substrate in distributed areas [3]. Ayurveda has identified many medicinal properties of this plant and it is effectively used against dysentery, is applied on open sores and pustules. A paste of leaves is applied on ulcers, hemorrhoids and swellings [4]. It has also ornamental value as climber with attractive flowers. This plant also included as plants affecting central nervous system [5], and also actively used as an antioxidant [6]. The main objective of the study is to screen the ethanol leaf extract of Ipomoea obscura (L.) for its phytochemical constituents, and to evaluate the functional groups using FITR and elemental analysis through EDAX analysis.

MATERIALS AND METHODS

Plant collection

The whole plant of Ipomoea obscura (L.) Used for the investigation was obtained from Madurai district, Tamilnadu, India. The plant specimen was authenticated by Dr. G. V. S. Murthy, Botanical survey of India, TNAU Campus, Coimbatore. The voucher specimen was deposited in the laboratory for future reference (BSI/SRC/5/23/2010-11/Tech). Fresh plant material was washed under running tap water then air dried and powdered.

Extraction

The plant powder was extracted with successive solvent system, petroleum ether, chloroform, ethyl acetate, ethanol and water.

Totally 100 g of dried plant powder was extracted in 500 ml of corresponding solvents for 24 hr in occasional shaking at room temperature. The supernatant was collected and evaporated to make the final volume one-fifth of the original volume. It was stored at 4˚C in airtight bottles for further studies.

Phytochemical analysis

Phytochemical screening of plant extract was carried out according to the method adopted by Peach and Tracey [7].

FTIR spectrum analysis

The ethanol extract of Ipomoea obscura (L.) was mixed with KBr salt, using a mortar and pestle, and compressed into a thin pellet. Infrared spectra were recorded on a Shimadzu FTIR Spectrometer 8000 series, between 4,000-400 cm-1.

Energy dispersive X-ray spectroscopy (EDAX Analysis)

The ethanol extract derived from plant sample of Ipomoea obscura (L.) was subjected to the elemental analysis using Scanning Electron Microscope (SEM) with an energy dispersive x-ray spectrometer (EDAX).

RESULTS

Phytochemical screening of plant materrials

Table 1 shows the phytochemical screening of Ipomoea obscura (L.). Preliminary phytochemical screening of ten different secondary metabolites (Alkaloids, Cardioglycosides, Saponin, Oils and fats, Tannin and phenolic compound, Terpenoids, Flavonoids, Aminoacids and proteins, Steroids, Carbohydrates) were tested in five different extracts. Among the five different extracts, ethanol extract show the presence of the maximum number of secondary metabolites than other solvents.

Fourier transform infrared (ftir) fingerprinting analysis

Figure 1 shows the results of FTIR spectroscopic studies have revealed the presence of various functional groups in ethanol leaf extract of Ipomoea obscura (L.).

The peak at 3989 and 3385.07 cm-1 is corresponding to hydrogen - bonded O-H stretching frequency respectively. The peak at 3385.07 cm-1 that peak covers the entire region with a very broad peak. The peak at 2929.87 cm-1 to assign H-C-H stretch. The peak at 2223.92 cm-1 to assign C≡N stretch. The absorptions can be seen as several distinct peak in this region. The bands between 3000 and 2800 cm-1 represent C-H stretching vibrations that are mainly generated by lipids [8,9]. The more intense bands occurring at 2200.78, 2181.49, 2152.56, 2133.27 corresponding to C≡C stretching indicate the presence of alkynes. The peak at 1710.86 cm-1 to assign C=O stretching indicate the carboxylic acid. The peak at 1627.97 cm-1 to assign C-C=C stretching indicate the alkenes.

The peak at 1263.37, 1055.06 cm-1 assigned to the C-O stretching vibration and indicate the esters. The weak absorption band of 796.60, 700.16, 617.22 cm-1 indicate the presence of chloride, bromide in our plant extract [10]. This O-H stretching indicate the phenolic compound, that have excellent antioxidant properties[11].

Table 1: Phytochemical screening of leaf extract of Ipomoea obscura (L)

Extracts AL SA TP FL ST CG OF TN AP CH
Petroleum ether + + + + - + - - - -
Chloroform + - - + + - - + - +
Ethyl acetate + - + + - + + + - +
Ethanol + + - - + + + + + +
Water + + - + - + - - + +

 ‘+’ Present

‘_’ Absent

AL

Alkaloids

CG

Cardioglycosides

SA

Saponin

OF

Oils and fats

TP

Tannin and phenolic compounds

TN

Terpenoids

FL

Flavonoids

AP

Aminoacids and proteins

ST

Steroids

CH

Carbohydrates

Table 2: The percentage of trace elements present in ethanol leaf extract of Ipomoea obscura (L)

Element App Conc. Intensity Corrn. Weight% Atomic%
O 17.36 0.884 76.03 87.55
Mg 0.48 0.632 2.96 2.24
Si 0.25 0.83 1.17 0.77
Cl 0.61 0.833 2.85 1.48
K 3.65 1.059 13.32 6.28
Ca 0.88 0.932 3.67 1.69
Totals 100.00

Fig. 1: FTIR Spectrum analysis of Ethanol leaf extract of Ipomoea obscura (L.)

Energy dispersive X-ray spectroscopy

Table 2 shows the results of the elemental composition of Ipomoea obscura (L.) Using SEM and EDX technique. The SEM - EDX spectra of the ethanol extract of the Ipomoea obscura (L.) Showed in figure 2.

Oxygen, Magnesium, Silicon, Chloride, Pottasium and Calcium are present in Ipomoea obscura (L.) CaCo3, Sio2, MgO, KCL, MAD, Ca wollastonites are used as the standards. In all these elements,Oxygen presented as high concentration while K and Ca presented as moderate amount. But Mg, Si, Cl presented only a trace quantities. Trace elements are estimated by determining the presentage abundance (%) of elements Mg, Si, Cl, K and Ca in the sample.

Fig. 2: The SEM EDX spectra of ethanol leaf extract of Ipomoea obscura (L.)

DISCUSSION

Phytochemical screening is of paramount importance in identifying new source of therapeutically and industrially valuable compound having medicinal significance, to make the best and judicious use of available natural wealth[12,13]. Phytochemical investigation of the ethanolic extracts of Ipomoea obscura (L.) revealed the presence of various phytochemicals such as steroids, flavonoids, tannins, saponins. The presence of cardiac glycosides and alkaloids in Ipomoea obscura (L.) may be associated with their use by traditional medicines for the treatment of different diseases. Phenolic have attracted a great attention in relation to their potential for beneficial effects on health [14]. phytochemical analysis of plants for the presence of saponins are widely well known to have expectorant and antitissue activity[15,16].

FTIR spectroscopy data analysis helps in understanding the chemical functionality of the compound in the plant sample and when run under IR region in the range of 400-4000 cm-1there was a variation in the peak in both the plant samples[17,18]. FTIR allows detecting whole range of infrared spectram simultaneously providing speed and accuracy in measurements of biological specimens[19]. The presence of characteristic functional groups phenols, carboxylic acid, alkanes, alkenes, esters, alkynes, fatty acid and lipids are responsible for various medicinal properties of Ipomoea obscura (L.) Based on the functional group analysis, Ipomoea obscura (L.) doesn’t contain any toxic compounds. In the present study, results of EDAX showed the presence of trace elements such as Mg, Si, Cl, K and Ca in Ipomoea obscura (L.). Deficiency of these trace elements in human subjects can occur under the most practical dietary conditions and in much diseased status[21]. Trace elements play both curative and preventive role in combating diseases. Calcium is needed in the development of bone and teeth and it regulate heart rhythm, help in normal blood clotting, maintain proper nerve and muscle functions and lower blood pressure[22]. Magnesium is important cofactor for the conversion of blood glucose into energy[23]. Potassium is essential for the transport of nutrients inside the cell. Without potassium, nutrients could not able enter into the cell that lead cell death. Silicon is also another important element to prevent the hardening of veins and arteries. Chloride works with sodium and potassium carry an electrical charge when dissolved body fluids and to regulate the pH in the body. Chloride is also important for digest the food properly and absorb many elements. The presence of these trace elements in Ipomoea obscura (L.) marks its use in therapeautic purpose.

CONCLUSION

The results obtained from the present study revealed that, the ethanol extracts of Ipomoea obscura (L.) showed the presence of most of the secondary metabolites in the plant leaves. The presence of functional groups and trace elements presents in the leaves of Ipomoea obscura (L.). The intensive study on the out - coming active constituents of Ipomoea obscura (L.) will lead to the discovery of a novel botanical - drug.

CONFLICT OF INTERESTS

Declared None.

ACKNOWLEDGEMENT

We, the authors are thankful to our Chancellor, Chief Executive officer, Vice-Chancellor and Registrar of Karpagam University for providing facilities and encouragement.

REFERENCES

  1. Ahmedulla M, Nayar MP. Red data book of Indian plants, Calcutta: Botanical survey of India;1999. p. 4.
  2. Alston RE, Turner BL. Biochemical systematic: Prentice Hall New Jersey;1963.
  3. Eckart E. Solanaceae and Convolvulaceae – seconday metabolite 2008:1:637.
  4. Christophe W, Pharm D. Ethnopharmacology of medicinal plants: Asia and the Pactic: Humana press;2002:1:69.
  5. Shahina A. Handbook of Arabian medicinal plants: edd.-1 CRC publication; 1994. p. 90.
  6. Srinivasan R, Chandrasekar MJN, Nanjan MJ, Suresh B. Free radical scavenging activity of Ipomoea obscura (L.) Ker-Gawl. J Nat Rem 2008:7(2):184-8.
  7. Paech D, Tracey MV. Modern methods of plant analysis. 4thEd. Wieland: Springer Verlag;1955. p. 373-1.
  8. Wolkers WF, Hoekstra AF. Aging of dry desiccation-tolerant pollen does not affect protein secondary structure. Plant Physiol 1995;109:907-15.
  9. Wei ZL, Dong L, Tian ZH. Fourier transform infrared spectrometry study on early stage of cadmium stress in clover leaves. Pak J Bot 2009;41:1743-50.
  10. Muruganatham S, Anbalagan G, Ramamurthy N. FT-IR and sem-eds comparative analysis of medicinal plants, Eclipta alba HASSK and Eclipta prostrata linn. Rom J Biophys 2009;19;285-94.
  11. Shirwaikar A, Malini S, Kumari SC. Protective effect of Pongamia pinnata flowers against cisplatin and gentamicin iduced nephrotoxicity in rats. Indian J Exp Biol 2003;1:58-62.
  12. Ambasta SP, Ramachandran K, Kashyapa K, Chand R. Useful plants of India. Publication and information directorate. Council of Scientific and Industrial Research: New Delhi;1986. p. 443-7.
  13. Kokate CK, Purohit AP, Gokhale SB. Practical pharmacogonosy. 1st Ed. Vallabh prakashan: Delhi;1998.
  14. Narayana KR, Reddy MS, Chaluvadi MR, Krishna DR. Bioflavonoids classification, pharmacology, biochemical effects and therapeutic potential. Ind J Pharmacol 2001;33:2-16.
  15. Rao UP, Brahman M, Saxena HO. Phytochemical surrey of Marurbhanj, Ganjam and Puri Dist. (Orissa) for tannins, Saponins, Flavonoids. Ind Drug 1984;22:503-7.
  16. Sharma SD, Chishti AM, Koul MK. Phytochemical survey of plants from Kashmir-II. Indian Drugs 1984;22:187-95.
  17. Thenmozhi M, Bhavya PK, Rajeshwari S. Compound Identification Using HPLC and FTIR In Eclipt alba and Emilia sonchifolia. Int J Eng Sci Tech 2011;3:292-8.
  18. Kalaiselvi M, Gomathi D, Vidya B, Uma C. Evaluation of Antioxidant potential and fourier transform infrared spectroscopy analysis of Ananus comosus (L.) Merr peel. Int Res J Pharm 2012;3:237-42.
  19. Griffiths PR, De Haseth JA. Fourier transform infrared spectroscopy. John Wiley and Sons: New York;1986. p. 656.
  20. Lin HX, Sun SQ, Lv GH, Chan KK. Study on Angelica and its different extracts by Fourier transform infrared spectroscopy and two-dimensional correlation IR spectroscopy. Mol Biomol Spec 2006;64:321-6.
  21. Udayakumar R, Begum VH. Elemental analysis of medicinal plants used in controlling infectious diseases. Hamdard Med 2004;67:35-6.
  22. Bibi S, Dastagir G, Hussain F, Sanaullah P. Elemental composition of Viola odorata Linn. Pak J Pl Sci 2006;12:141-3.
  23. Bahadur A, Chaudhry Z, Jan G, Danish M, Rehman A, Ahmad R, et al. Nutritional and elemental analyses of some selected fodder species used in traditional medicine. Afr J Pharm Pharmacol 2011;5:1157-61.