Spectroscopic Analysis of Bioactive Components of Ichnocarpus frutescens R. Br.

 

C. Yogeshwari1*, P. Kumudha2

1Research Scholar, PG and Research Department of Botany, Vellalar College for Women (Autonomous), Thindal, Erode – 638012, Tamil Nadu, India

2Associate Professor, PG and Research Department of Botany, Vellalar College for Women (Autonomous), Thindal, Erode – 638012, Tamil Nadu, India.

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

 

ABSTRACT:

The present study was carried out to characterize the bioactive constituents present in the ethanolic extracts of Ichnocarpus frutescens using UV and FTIR spectroscopy. Qualitative phytochemical analysis revealed the presence of saponins, phenols, terpenoids, anthraquinones, quinones, alkaloids, flavonoids, carbohydrates, glycosides and absence of fixed oil, coumarins, tannins, proteins and amino acids. The UV-VIS profile of Ichnocarpus frutescens plant extract was taken at the wavelength ranging from 190 nm to 1100 nm which showed the peaks at 239.00 nm and 276.00 nm with the absorption values 3.961 and 1.111 respectively. The FTIR analysis confirmed the presence of the functional groups of nitriles, alkanes, ketones, phenols, alkyl halides and aliphatic amines.

 

KEYWORDS: Ichnocarpus frutescens, FTIR, UV, phytochemical, spectroscopy.

 

 


INTRODUCTION:

Medicinal plants are the richest bio resource of drugs. For many centuries, plants have been a rich source of therapeutic agents and provided basis for several synthetic drugs1. The different phytoconstituents present in medicinal plants such as flavonoid, alkaloid, phenol and tannins, carboxylic acids, terpenes, amino acids and inorganic acids present specific distinctiveness and properties to plants2. UV and Infrared (IR) spectrometry provides a useful method for herbal analysis as well as for quantitative analysis of drugs3 and also serves as a cost-effective4 and timesaving method to characterize phytocomponents5.

 

Ichnocarpus frutescens is a large much branched twining shrub belonging to the family Apocynaceae. The plant possesses medicinal properties such as anti-inflammatory, antioxidant, antidiabetic, antitumour, analgesic6 and hepatoprotective activity7. Traditionally, the leaves are boiled in oil and applied for headaches and fevers. They are also applied to treat wounds8. The aim of the present work is to characterize the bioactive constituents in Ichnocarpus frutescens plant extract.

 

MATERIALS AND METHODS:

Collection of Plant Material:

Aerial part of the fresh and healthy plant materials of Ichnocarpus frutescens were collected from the Aanaikatti hills, Coimbatore district (a part of the Western Ghats of Western Tamil Nadu). The collected study plant was identified with the help of the Flora9.

Extraction of Plant Material:

Freshly collected plants were cleaned, shade dried and were mechanically ground to coarse powder. Cold extraction of powdered plant materials was carried out using the solvents viz., petroleum ether, acetone, ethanol and water. About 10gm of powdered material was soaked in 100ml of each solvent for 24 hrs accompanied by continuous shaking. They were filtered using standard Whatmann filter paper No.1 and the filtrate was allowed to evaporate at low temperature (10°C). The extracts were stored in refrigerator and used for further analysis.

 

Qualitative Phytochemical Analysis:

Phytochemical screening of different successive solvent extracts was carried out following the standard procedures10-12. Carbohydrates, proteins and amino acids, alkaloids, flavonoids, tannins, phenolic compounds, terpenoids, coumarins, saponins, quinones, anthraquinones, glycosides and fixed oils were qualitatively analysed.

 

Spectroscopic analysis:

Spectroscopic analysis by UV-Vis spectroscopy:

UV Visible spectroscopy uses light in the visible ranges or its adjacent ranges. The ethanolic extract of the plant was scanned in the wavelength range of 190-900 nm using GBC UV/ VIS 918 model UV spectrophotometer and the characteristic peaks were detected and the values of the peaks were recorded.

 

Spectroscopic Analysis by FTIR spectroscopy:

FTIR has proved to be a valuable tool for the characterization and identification of compounds or functional groups (chemical bonds) present in an unknown mixture of plant extract13. In addition, FTIR spectra of pure compounds are usually so unique that they are like a molecular "fingerprint"14.

 

Dried powder of ethanolic extract was used for FTIR analysis. 10 mg of the dried extract powder was encapsulated in 100 mg of KBr pellet in order to prepare translucent sample discs. The powdered sample was then loaded in Perkin Elmer spectrophotometer and the characteristic peaks were detected.

 

RESULTS AND DISCUSSION:

In the present study, the preliminary qualitative phytochemical screening of different solvent extracts of Ichnocarpus frutescens showed the presence of saponins, phenols, terpenoids, anthraquinones, quinones, alkaloids, flavonoids, carbohydrates, glycosides and absence of fixed oil, coumarins, tannins, proteins and amino acids. The results are tabulated in Table-1.

 


 

Table – 1: Phytochemical analysis of Ichnocarpus frutescens

S.No.

Phytoconstituents

Inference

Petroleum ether

Acetone

Ethanol

Water

1.

Saponins

+

-

+

+

2.

Oil & Fat

-

-

-

-

3.

Anthraquinones

-

+

+

-

4.

Coumarins

-

-

-

-

5.

Quinones

+

+

-

-

6.

Gum & Mucilage

-

-

+

-

7.

Alkaloids

i)Wagner’s test

+

+

+

+

8.

Flavonoids

-

+

+

+

9.

Terpenoids

-

-

+

-

10.

Phenols

i)Lead acetate

+

-

+

+

11.

Tannins

i)Ferric chloride

-

-

-

-

12.

Carbohydrates

i)Barfoed’s test

-

+

+

+

13.

Proteins &Amino acids

i)Biuret test

-

-

-

-

 

ii)Ninhydrin test

-

-

-

-

(+): Present, (-): Absent

 


Table - 2: UV-Vis peak values of ethanolic extracts of Ichnocarpus frutescens

S. No.

Wavelength (nm)

Absorption

1.

1050.00

0.104

2.

933.00

0.094

3.

817.00

0.089

4.

770.00

0.091

5.

667.00

0.094

6.

537.00

0.096

7.

468.00

0.110

8.

276.00

1.111

9.

239.00

3.961

10.

1063.00

0.099

11.

966.00

0.092

12.

840.00

0.086

13.

797.00

0.087

14.

691.00

0.083

15.

647.00

0.084

16.

604.00

0.086

17.

458.00

0.108

18.

259.00

0.847

 

Figure - 1: UV-Vis analysis of ethanolic extract of Icnhnocarpus frutescens

 

Table - 3: FTIR peak values of ethanolic extracts of Ichnocarpus frutescens

S. No.

Peak values

(cm-1)

Types of stretching

Functional group

1.

510.18

C-Br group

Alkyl halides

2.

668.82

C-Cl group

Alkyl halides

3.

1031.92

C-O group

Aliphatic amines

4.

1457.23

C=O group

Ketones

5.

1515.1

C-H stretching

Alkanes

6.

1647.7

C=O group

Ketones

7.

1687.73

C=O group

Ketones

8.

1743.66

C=O group

Ketones

9.

2358.96

C=N group

Nitriles

10.

2858.05

C-H Stretching

Alkanes

11.

2924.59

C-H Stretching

Alkanes

12.

3434.28

-OH group

Phenols

13.

3740.49

Unknown

Unknown

 

 

Figure - 2: FTIR analysis of ethanolic extract of Icnhnocarpus frutescens

 

The UV-VIS profile of Ichnocarpus frutescens plant extract was taken at the 190 nm to 1100 nm wavelength due to the sharpness of the peaks and proper baseline. The profile showed the peaks at 239.00 nm and 276.00 nm with the absorption value of 3.961 and 1.111 respectively (Table-2, Fig.-1).

 

The FTIR spectrum was used to identify the functional groups of the active components present in the Ichnocarpus frutescens plant extract based on the peak values in the region of IR radiation. When the extract was passed into the FT-IR, the functional groups of the components were separated based on its peaks ratio15. IR spectrum is most frequently used in phytochemical studies as a “Fingerprinting” device16. Results of FTIR spectroscopic analysis are summarised in Table-3 & Fig.-2. FTIR analysis of ethanolic extract of Ichnocarpus frutescens showed major peaks at 3434.28, 2924.59, 2858.05, 2358.96, 1743.66, 1687.73, 1647.7, 1515.1, 1457.23, 1031.92, 668.82 and 510.18 cm-1. The broad band at 3434.28 cm-1 was attributed to -OH stretching in phenol group. The peaks around 2924.59 cm-1, 2858.05 cm-1 and 1515.1 cm-1 corresponds to C-H stretching in alkanes group and 2358.96 cm-1 for C=N stretching in nitrile group. The bands around 1743.66 cm-1, 1687.73 cm-1, 1647.7 cm-1 and 1457 cm-1 was attributed to C=O stretching in ketones group. The peak at 1031.92 cm-1 corresponds to C-O stretching in aliphatic amine group. The bands around 668.82 cm-1 and 510.18 cm-1 were attributed to C-Cl and C-Br stretching respectively in alkyl halide group. Thangarajan et al. (2012) identified the presence of trace elements like calcium, magnesium, silicon, chloride, potassium, carbon and the functional groups like amino acids, amides, amines, carboxylic acid, carbonyl compounds and organic hydrocarbons in Ichnocarpus frutescens2. Similar results were obtained in the present study and confirmed the presence of the functional groups of nitriles, alkanes, ketones, phenols, alkyl halides and aliphatic amines. In addition, UV-VIS and FTIR spectroscopy is proved to be a reliable and sensitive method for detection of biomolecular composition17.

 

CONCLUSION:

Our findings revealed that crude aqueous and organic solvent extract of the plant Ichnocarpus frutescens contains medicinally important bioactive compounds and it justifies its use in the traditional medicines for the treatment of different diseases.

 

ACKNOWLEDGEMENT:

The authors are thankful to Central Research Laboratory, Vellalar College for Women, Erode for providing necessary facilities to carry out this experiment.

 

 

REFERENCES:

1.       Faboro EO et al. Characterization of dichloromethane and methanol extracts from the leaves of a medicinal plant: Globimetula oreophila. Industrial Crops and Products. 2016: 391-399.

2.       Starlin T et al. Element and functional group analysis of Ichnocarpus frutescens R.Br. (Apocynaceae). International Journal of Pharmacy and Pharmaceutical Sciences. 4 (Suppl 5); 2012: 4-6.

3.       Bunaciu AA et al. FTIR spectrophotometric methods used for antioxidant activity assay in medicinal plants. Applied Spectroscopy Reviews. 47(4); 2012: 245-255.

4.       Anand T and Gokulakrishnan K. Phytochemical analysis of Hybanthus enneaspermus using UV , FTIR and GC- MS. IOSR Journal of Pharmacy. 2(3); 2012: 520-524.

5.       Renuka B, Sanjeev B and Ranganathan D. Evaluation of phytoconstituents of Caralluma nilagiriana by FTIR and UV-VIS spectroscopic analysis. Journal of Pharmacognosy and Phytochemistry. 5(2); 2016: 105-108.

6.       Pandurangan A, Khosa RL and Hemalatha S. Evaluation of anti-inflammatory and antioxidant activity of Ichnocarpus frutescens root. DARU. 17(1); 2009: 1-5.

7.       Dash DK et al. Evaluation of hepatoprotective and antioxidant activity of Ichnocarpus frutescens (Linn.) R. Br. on paracetamol-induced hepatotoxicity in rats. Tropical Journal of Pharmaceutical Research. 6(3); 2007: 755-765.

8.       Kumarappan CT and Mandal SC. Antitumor activity of polyphenolic extract of Ichnocarpus frutescens. Experimental Oncolology. 29(2); 2007: 94-101.

9.       Gamble JS. Flora of the Presidency of Madras. Neeraj Publishing House, Delhi. 2014.

10.    Kokate CK, Purohit AP and Gokhale SB. Pharmacognosy. Nirali Prakashan, Pune. 2014.

11.    Harborne JB. Phytochemical Methods - A Guide to Modern Techniques of Plant Analysis. Springer (India) Private Limited, New Delhi. 2013.

12.    Evans WC. Trease and Evans Pharmacognosy. Elsevier, New Delhi. 2007.

13.    Hazra KM et al. Isolation of antibacterial pentahydroxy flavones from the seeds of Mimusops elengi Linn. African Journal of Biotechnology. 6(12); 2007: 1446-1449.

14.    Sasidharan S et al. Extraction, Isolation and Characterization of bioactive compounds from Plants’ extracts. African Journal of Traditional and Complementary Medicines. 8(1); 2011: 1-10.

15.    Nithyadevi J and Sivakumar R. Phytochemical screening and GC-MS , FT-IR analysis of methanolic extract leaves of Solanum torvum Sw . International Journal of Research Studies in Biosciences. 3(9); 2015: 61-66.

16.    Dhanasekaran M et al. Evaluation of Phytochemical constituents of Indian medicinal plant Hydnocarpus alpina WIGHT . 1(3); 2013: 23-28.

17.    Sahaya Sathish S, Janakiraman N and Johnson M. Phytochemical Analysis of Vitex altissima L . using UV-VIS , FTIR and GC-MS. International Journal of Pharmaceutical Sciences and Drug Research. 4(1); 2012: 56-62.

 

 

Received on 14.12.2017       Modified on 28.02.2018

Accepted on 12.04.2018       ©A&V Publications All right reserved

Res.  J. Pharmacognosy and Phytochem. 2018; 10(2): 171-174.

DOI: 10.5958/0975-4385.2018.00026.2