PHYTOCHEMICAL STUDIES, FTIR AND GC-MS ANALYSIS OF HARDWICKIA BINATA ROXB. (FABACEAE / CAESALPINIACEAE)

PHYTOCHEMICAL STUDIES, FTIR AND GC-MS ANALYSIS OF HARDWICKIA BINATA ROXB. (FABACEAE / CAESALPINIACEAE)

S. V. Deshmukh ^* and N. A. Ghanawat

Department of Botany, Yashavantrao Chavan Institute of Science, Satara - 415001, Maharashtra, India.

ABSTRACT: Hardwickia binata belongs to family Fabaceae / Caesalpiniaceae and commonly known as ‘Anjan’. The present study includes phytochemical screening, FTIR and GC-MS analysis. Shade dried powdered leaves, seed and husk of H. binata extracted in petroleum ether, chloroform, ethyl acetate, acetone, methanol, ethanol, distilled water using Soxhlet apparatus and used for phytochemical analysis. Crude powder of plant used for FTIR analysis and ethanolic extracts was analyzed using GC-MS. The phytochemical screening of H. binata revealed the presence of carbohydrates, proteins, amino acids, steroids, glycosides, flavonoids, lipids, tannins, phenolic compounds, quinones, mucilage, volatile oils, fats, and fixed oils whereas alkaloids, emodins and resins were not detected in any extracts of leaves, seed, and husk. The majorities of compounds are present in acetone, methanol, ethanol and distilled water and glycosides, lipids and mucilage are present in all extracts. FTIR analysis showed the presence of alcohol, phenols, amines, amides, carboxylic acids, aromatics, alkenes, alkanes, aliphatic amines, esters, ethers, alkynes, alkyl halides in leaves, seed and husk; saturated aliphatic found in only seed and aldehydes only in husk and GC-MS analysis confirmed ten different chemical compounds 1,1-diethoxy-ethane, tetradecamethyl-cycloheptasiloxane, linoleic acid, ethyl ester, ethyl oleate in leaves, seed and husk; hexadecanoic acid and ethyl ester found in only leaves and seed; 1-methyl-4 -(1-methylethyl)-Benzene, 2-methyl-5-(1-methylethyl)-Phenol, 3, 7, 11, 15-tetramethyl-2-hexadecen-1-o1 are found in only leaves; dodecamethyl-cyclohexasiloxane only in seed; thymol only in husk shows various biological activities. The different extracts of H. binata revealed several phytochemical components as a rich source of pharmaceutically and biologically important bioactive compounds that may build up new medicines with more efficiency.

Hardwickia binata, Phytochemicals, FTIR, GC-MS analysis

INTRODUCTION: The plant kingdom is a significant part of potential drug an inexhaustible source of bioactive constituents. Preliminary phytochemical screening is a valuable step in the detection of the bioactive compounds present in plants.

Phytochemicals are known to provide protection against insect attacks and plant diseases. Hardwickia binata is a deciduous, moderate to large-sized tree. The bark of saplings almost silvery-white and smooth. Leaves are alternate, bifoliolate resembling Bauhinias. Leaflets are almost kidney-shaped and grayish-green in color, sessile, entire, obliquely ovate and coriaceous.

Flowers are small, yellowish-green in an axillary and terminal lax panicled raceme. The fruit is a flat, samaroid pod con­taining one seed near the apex. Seed is flat; straight to slightly bended pointed at one end and rounded at the other, with a fairly hard testa ^{1, 2}. Previous researchers worked out tannins from the bark are used to produce medicines for the treatment of diarrhea, worms, indigestion and leprosy and also produce an appetizer ³. The leaves, pods and bark contain tannins ⁴.

Bark used for gums and resins ⁵. The leaves are used for fodder and manure ⁶. The leaves extract showed activity against both gram-positive and gram-negative bacteria and fungi. Bioactive substances showed antimicrobial agents for the treatment of various bacterial and fungal infections including gonorrhea, pneumonia, eye infections and mycotic infections and tannins have been traditionally used for protection of catarrh, wounds, hemorrhoids, diarrhea and antidote in heavy metal poisoning. Flavonoids possess anti-inflammatory, anti-allergic, antithrombotic, anti-microbial and vasoprotective effects ⁷. The leaves are used for headache and treatment of constipation ^{1, 7}. The leaves had crude protein ^{8, 9}. The bark and leaf used as a source of medicine for rheumatism ¹⁰. In H. binata an investigation was made of the antioxidants and their protective mechanism in chloroplasts against SO₂ pollution ¹¹. The phenolic compounds' role in giving protection to the plants against deleterious effects of UV rays and certain phytopathogenic microorganisms. The protein, phenols and flavonoids can be used as chemical markers in taxonomic studies ¹². Phytochemical analysis studies the presence of active compounds. Hence, the present investigation was undertaken to work out preliminary phytochemical studies, FTIR and GC-MS analysis with the aim to report the functional components of leaves, seed and husk of Hardwickia binata Roxb.

MATERIALS AND METHODS:

Collection and Identification of Plant Material: The plant materials such as leaves and pods of Hardwickia binata were collected from different localities viz. Sangali (Tal-Atpadi, Swatantrapur vasahat), Dhule (tal-dhule, borvihir Area) and Jalgaon (Tal-Muktainagar, Muktainagar forest) Districts in April 2015. Taxonomic identification of the plant was done and herbarium was deposited to the Department of Botany, Shivaji University, Kolhapur.

Preparation of Extracts: The collected materials (leaves and pods) of Hardwickia binata were shade dried and coarsely powdered in a mixer grinder, sieved and used for extraction. The leaves, seed and husk powder were extracted separately with petroleum ether, chloroform, ethyl acetate, acetone, methanol, ethanol, and distilled water in the increasing order of their polarity by using Soxhlet apparatus. The extract was decanted, filtered through Whatman No.1 filter paper, concentrated and preserved in airtight bottles until further use.

Phytochemical Studies: The qualitative phytochemical tests for the different solvent extracts were used for the identification of active chemical constituents like carbohydrates, proteins, amino acids, steroids, glycosides, flavonoids, alkaloids, lipids, tannins, phenolic compounds, quinones, mucilage, volatile oils, fats and fixed oils. Preliminary phytochemical analysis tests for presence or absence of various phytoconstituents.

FTIR Analysis: The Fourier Transform Infrared (FTIR) spectroscopy analysis was performed to know the occurrence of chemical bonds or functional groups present in the crude powder of leaves, seed, and husk of Hardwickia binata. The peak values of FTIR were recorded and their corresponding functional groups were identified.

GC-MS Analysis: The chemical composition of ethanolic extracts of leaves, seed, and husk of Hardwickia binata was determined by gas chromatography-mass spectrometry (GC-MS) as it provides enhanced sample identification, higher sensitivity, an increased range of analyzable samples and faster results. This helps to constitute the chemical picture of plant extracts and by which the complex mixtures can be resolved into individual components.

RESULTS AND DISCUSSION:

Phytochemical Studies: The preliminary phytochemical screening of H. binata revealed the presence of carbohydrates, proteins, amino acids, steroids, glycosides, flavonoids, lipids, tannins, phenolic compounds, quinones, mucilage, volatile oils, fats and fixed oils whereas alkaloids, emodins and resins were not detected in any extracts of leaves, seed and husk Table 1. Previous results revealed that the presence of saponins, tannins, steroids, terpenoids, phenolic groups, coumarin, flavonoids and carbohydrates.

Alkaloids were found to be absent in petroleum ether, chloroform, and ethanolic leaves extracts of H. binata ¹. A similar type of analysis has been documented in ethanolic leaves extracts of H. binata but alkaloids and anthraquinones were not present ⁷. The phytochemical study of leaves and seeds of Bauhinia racemosa, Bauhinia purpurea and Hardwickia binata in ethanolic extracts are the presence of phenols, saponins, flavonoids, glycosides and tannins while alkaloids were not detected ¹².

The phytochemical screening of leaves extract of Bauhinia variegata with petroleum ether, chloroform, ethanol, and water showed the presence of carbohydrates, gums, proteins, amino acids, fats and oils, triterpenoids, steroids, glycosides, flavonoids, alkaloids, tannins ¹³. Similar type of analysis has been documented in Bauhinia racemosa leaves ^{14, 15}.

The various extracts of root bark exudates of H. binata show the presence of carbohydrates, glycosides, fixed oils and fats, proteins and amino acids, saponins, tannins, phytosterols, alkaloids, phenolic compounds, flavonoids while gums and mucilage are absent ¹⁶.

In present work confirmed the previous reports. The majorities of compounds are present in acetone, methanol, ethanol, and distilled water and glycosides, lipids and mucilage are present in all extracts.

TABLE 1: QUALITATIVE PHYTOCHEMICAL INVESTIGATION OF H. BINATA

(+) Positive test, (-) negative test; L- leaves, S- seed, H- husk; P. E- petroleum ether, chlo-chloroform, E. A- ethyl acetate, D.W- distilled water

FTIR Analysis: The results revealed in the crude powder of leaves, seed and husk of H. binata gave the characteristic absorption peaks as represented in Fig. 1 and Table 2. The crude powder of H. binata FTIR confirmed the presence of alcohol, phenols, amines, amides, carboxylic acids, aromatics, alkenes, alkanes, aliphatic amines, esters, ethers, alkynes, alkyl halides in leaves, seed and husk; saturated aliphatic found in only seed and aldehydes only in husk. FTIR spectra of untreated and alkali-treated Hardwickia binata fibers were determined the chemical composition of cellulose, hemicellulose and lignin ¹⁷. The methanolic extracts of Myristica dyctyloides fruit have FTIR spectra showed the presence of functional group in all the extracts which have medicinal properties ¹⁸.

FIG. 1: FTIR SPECTRUM ANALYSIS OF CRUDE POWDER OF HARDWICKIA BINATA (A) LEAVES, (B) SEED, (C) HUSK

TABLE 2: FTIR ANALYSIS OF CRUDE POWDER OF H. BINATA

FTIR- Fourier transform infrared spectroscopy; P.V. - peak values

GC-MS Analysis: GC-MS analysis confirmed the results as shown in Table 3 and Fig. 2. It showed the presence of 10 major groups. 1, 1-diethoxy-ethane, tetradecamethyl- cycloheptasiloxane, linoleic acid, ethyl ester, ethyl oleate in leaves, seed and husk; hexadecanoic acid and ethyl ester found in only leaves and seed; 1-methyl-4-(1-methylethyl)-Benzene, 2-methyl-5-(1-methylethyl)-Phenol, 3, 7, 11, 15-Tetramethyl-2-hexadecen-1-o1 are found in only leaves; dodecamethyl-cyclohexasiloxane only in seed; thymol only in husk is the pharmaceutically important. The different biochemical compounds identified by GC-MS show various biological activities shown in Table 4.

TABLE 3: GC-MS ANALYSIS OF ETHANOLIC EXTRACTS OF LEAVES, SEED AND HUSK OF H. BINATA

GC-MS- Gas Chromatography - Mass Spectrometry, R.T. - Retention time, MF- Molecular Formula, MW- Molecular Weight

The GC-MS analysis of leaves of H. binata was carried out on the petroleum ether, chloroform and ethanolic extracts and among all the extracts, ethanolic extracts showed the presence of maximum chemical constituents ¹. The GC-MS analysis of methanolic extracts in root bark exudates of H. binata identified 22 compounds ¹⁶.

FIG. 2: GC-MS CHROMATOGRAM OF ETHANOLIC EXTRACT OF HARDWICKIA BINATA (A) LEAVES, (B) SEED, (C) HUSK

TABLE 4: ACTIVITY OF BIOACTIVE COMPOUNDS IDENTIFIED BY GC-MS STUDY OF LEAVES, SEED AND HUSK OF H. BINATA

CONCLUSION: The present investigation of H. binata revealed the presence of many phytochemical components as a rich source of pharmaceutically and biologically important bioactive compounds that may build up new medicines. FTIR spectra showed the various functional groups may serve as a new potential source of medicines with more efficiency. 10 compounds that have been screened by a GC-MS it will help to therapeutic activity. Various studies confirmed that phytochemicals contribute medicinal properties that are of great importance as a good source of new useful drugs. The quantitative analysis of these phytocomponents will be an interesting area for further study.

ACKNOWLEDGEMENT: Authors are thankful to the Principal, Yashavantrao Chavan Institute of Science, Satara for providing the facilities for research work. The authors also thank the Head, Department of Botany for their support and cooperation during the research work.

CONFLICTS OF INTEREST: Nil

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    How to cite this article:

    Deshmukh SV and Ghanawat NA: Phytochemical studies, FTIR and GC-MS analysis of Hardwickia binata Roxb. (fabaceae/ caesalpiniaceae). Int J Pharm Sci & Res 2020; 11(1): 233-40. doi: 10.13040/IJPSR.0975-8232.11(1).233-40.

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