1. INTRODUCTION:

Luffa acutangula is commonly known as Ridge gourd. It is one of the regular vegetable took by many villagers in Tamilnadu in their food. It can be averagely grown in around 40 days with quite a good supply of water. Ridge gourd is popular as a vegetable in various regional cuisines in Asia, but the fruit is only edible before it ripens¹. Profiling of bioactive principles from edible and medicinal plants provides useful insights into their chemical diversities, medicinal potentials and toxicity². The natural constituents that are present in the plant are derived from any parts of plant like leaves, roots, bark, flowers, fruits and seeds i.e., active components can be present in any parts of the plant³. Gas chromatography (GC) is used to seperates mixtures into individual components using a temperature-controlled capillary column.

Mass spectrometry (MS) is used to identify the various components from their mass spectra. Gas Chromatography (GC) and Mass Spectroscopy (MS) associated with particular detection techniques has become a sophisticated means for analysis of various compounds⁴. Several phytochemical studies have been carried out throughout the world. Plant based drugs offering great emphasis on preventive action of diseases⁵. To the best of our knowledge, the phytochemical analysis of Luffa acutangula is very limited. The present study aims the qualitative analysis and GC – MS profiling of aqueous extract of Luffa acutangula peel

2 MATERIALS AND METHODS:

All the chemicals used in the phytochemical analysis were procured from Hi media, India, of analytical grade with high purity.

2.1 Collection and Standardization of Plant sample:

The Luffa acutangula was collected from the fields of Tirupattur, Vellore District, Tamilnadu, India and used for the study.

2.2 Preparation of plant sample:

Luffa acutangula obtained were washed thoroughly with double distilled water to devoid of any impurities and minerals. The skin of cleaned Luffa acutangula was peeled off carefully with a skin peeler. The peeled skin was shade dried for dehydration for about a week. The completely dried skin was made into a fine powder using electric mixing grinder. The ground powder was sieved, stored in an air tight container and used whenever it was needed.

2.3 Preparation of Peel extract:

Ten grams of Luffa acutangula peel powder was weighed and added to 100ml of double distilled water in the conical flask. This was kept in boiling water bath at 100°C for 60 minutes. The conical flask was removed after that period and allowed to cool to room temperature. Then the contents were filtered through whatman No:1 filter paper. The filterate can be used for further studies by storing in refrigerator at 4°C not more than a week.

2.4 Preliminary phytochemical analysis:

The Luffa acutangula peel aqueous extract was screened for phytochemicals by following Harbone and Kokate method^6,7 with slight modifications.

2.5 GCMS analysis:

Gas chromatograph coupled with mass spectrometer and auto sampler was used. The Gas chromatograph (GC) was equipped with an Agilent HP-5MS (5% Phenyl methyl siloxane) capillary column of thickness 30m× 0.25 mm ID×0.25µm was used. Helium with high purity 99.999% was used as carrier gas with constant flow rate of 1 mL/min. The temperatures of injector and mass selective detector transfer line were 250 ̊C and 280 ̊C respectively. The column temperature was programmed as follows: initial column temperature was 50 ̊C with hold on time of 2 min. Then with multiple ramp rates of 25 ̊C /min the temperature was raised to 200 ̊C with hold on time of 2 min and then raised to 280 ̊C at the rate of 10 ̊C/min with hold on time of 7 min. The mass spectrometer operating parameters were used in electron ionization (EI) mode with an ion source temperature of 230 ̊C and quadruple temperature of 150 ̊C. The MS scan range was set at full mass range of 30 – 500 mass to charge ratio (m/z). The electron ionization voltage was 70eV.

The prepared peel extract of Luffa acutangula was dried to a powder. The sample for running GCMS was prepared by 1% dried extract in methanol. The methanolic sample volume of 1.0µL was injected in split less mode. The GC running time was totally 35 min.

The spectrum of Luffa acutangula peel extract obtained by running GCMS was compared with the spectrum stored in the National Institute of Standards and Technology (NIST) mass spectral library.

3 RESULTS AND DISCUSSION:

3.1 Preliminary qualitative phytochemical analysis:

The phytochemical constituents in the aqueous extract of Luffa acutangula peel extract was studied by the qualitative phytochemical tests. According to the observation of changes in the respective carried out experiments, the presence or absence of the bioactive components are confirmed. The results of the preliminary phytochemical screening of the Luffa acutangula peel aqueous extract are shown in the Table – I. The tests for flavanoids, carbohydrates, aminoacids, proteins, phenols, quinones, terpenoids, coumarins, saponins, tannins and leucoanthcyanidines have shown positive results and thereby confirm the presence of those phytochemicals in the aqueous extract of Luffa acutangula peel extract. The tests for volatile oils, alkaloids and glycosides gave negative results and therefore they are absent in the aqueous extract of Luffa acutangula peel extract.

Many researchers done phytochemical studies in various fruit peel aqueous extract and Mamta Arora et al.,⁸ reported the presence of alkaloids, tannin and saponins in the aqueous orange peel extract. The phytochemical screening of aqueous Punica granatum peel extract was done by Jayaprakash et al. ⁹and they reported the presence of tannins, saponins, quinones, terpenoids, steroids, flavanoids, phenols, alkaloids and coumarins. Suyanthi et al¹⁰ showed the presence of alkaloids, terpenoids and saponins and absence of flavanoids, tannins and quinones in ethanolic extract of Luffa acutangula (L) Roxb’s fruit. Also aqueous leaf extract of Luffa aegyptiaca was analysed for phytochemicals by Mhya D H et al.¹¹and found the presence of flavanoids, tannins and saponins. In this study, it is found that most of the phytochemicals are present in the aqueous extract of Luffa acutangula peel extract.

3.2 GCMS of Luffa acutangula peel extract:

The phytochemicals present in the aqueous Luffa acutangula peel extract are found by preliminary phytochemical qualitative tests. The bioactive compounds present in the peel extract were analysed by Gas chromatography mass spectrometer. The GC – MS chromatogram of aqueous Luffa acutangula peel extract is depicted in Figure 1. The X axis of the GC- MS chromatogram represents the retention time in minutes that relates to the time for each compound to be eluted from the time of injection. The Y axis represents the % peak area that indicates the relative concentration of the compounds present in the aqueous extract of Luffa acutangula peel.

Table -1 Phytochemical screening of Luffa acutangula peel aqueous extract

S. No	Phytochemical	Test	Result
1	Flavonoids	Lead Acetate test	+
		Decolorization test	+
		Ammonia test	+
2.	Carbohydrates	Molisch test	+
		Fehling’s test	+
		Cellulose test	+
		Iodine test	-
3.	Amino Acids and Proteins	Millon test	+
3.	Amino Acids and Proteins	Ninhydrin test	+
4.	Phenols	Ferric Chloride test	+
4.	Phenols	Phospho Molybdic test	+
5.	Quinones	Chloroform – Ammonia test	+
6.	Terpenoids	Salkowski test	+
7.	Coumarins	Filter paper test	+
8.	Saponins	Froth test	+
9.	Tannin	Braemer’s test	+
10.	Leucoanthocyanidines	Leucoanthocyanidine test	+
11.	Sterols	Libermann-Buchard test	+
12.	Volatile oils	Precipitation test	-
13	Alkaloids	Mayer’s test	-
		Hager’s test	-
		Wagner’s test	-
14.	Glycosides	Legal test	-

Figure -1 GC-MS chromatogram of aqueous extract of Luffa acutangula peel

Table -2 Phytocomponents detected in aqueous extract of Luffa acutangula peel extract by GC-MS.

S. No	Retention time	Name of the compound	Molecular formula	Molecular weight	Peak Area %
1	5.488	Benzofuran, 2,3, dihydro	C₈H₈O	120.14	20.49
2	6.317	Ethanone, 1 (2 – hydroxyl – 5- methyl phenyl)	C₉H₁₀O₂	150.17	11.50
3	15.154	Hexadecanoic acid, methyl ester	C₁₇H₃₄O₂	270.45	24.73
4	17.851	Cis-13- Octadecenoic acid, methyl ester	C₁₉H₃₆O₂	296.48	15.02
5	29.155	Pectolinaringenin	C₁₇H₁₄O₆	314.29	22.22
6	29.191	4-methyl – 2- 4- bis (p-hydroxyphenyl) pent-1-ene, 2TMS derivative	C₂₄H₃₆O₂Si₂	412.72	16.66

GC-MS analysis of aqueous Luffa acutangula peel extract showed six major peaks revealing the presence of 6 major phytochemical compounds in the peel extract. The 6 phytochemical constituents present in the Luffa acutangula peel aqueous extract were identified by matching the peaks obtained with the NIST spectral library. The compounds identified are listed in the Table -2 with compound name, with its retention time, molecular formula, molecular weight and peak area %.

The descending order of 6 major phytoconstituent’s retained from Luffa acutangula peel aqueous extract, was Hexadecanoic acid, Pectolinaringenin, Benzofuran, 4-methyl – 2- 4- bis (p-hydroxyphenyl)pent-1-ene, Cis-13- Octadecenoic acid and Ethanone, 1 (2 – hydroxyl – 5- methyl phenyl) with % of area of the peak 24.73, 22.22, 20.49, 16.66,15.02 and 11.50 respectively.

The presence of coumaran compound, Benzofuran 2, 3, dihydro in Pavetta crassicaulis Bremek. leaf extract was reported by Ashwathanarayana et al¹². Revathi et al ¹³ confirmed the presence of Benzofuran 2, 3 dihydro the ethanolic extract of mangrove plant, Bryguiers cylindrica.

Both Ethanone, 1 (2 – hydroxyl – 5- methyl phenyl) and Hexadecanoic acid, methyl ester were present in methanolic extract of Alysicarpus monilifer¹⁴ and alcoholic extract of Epiphyllum oxypetalum leaves¹⁵.

Maruthamuthu etal¹⁶ reported the presence of Ethanone, 1 (2 – hydroxyl – 5- methyl phenyl), Hexadecanoic acid, methyl ester and Cis-13- Octadecenoic acid, methyl ester in the methanolic crude extract used in the treatment of kidney stones, in G. Thumlappati, Battalagundu, Dindugal district Tamil Nadu, India. The presence of Cis-13- Octadecenoic acid, methyl ester was also reported by Mustapha et al¹⁷and Karthika et al¹⁸ in the chloroform extract of Albizia adianthifolia and methanolic extract of stem and tuber of Solena amplexicaulis respectively.

A type of flavanoid , Pectolinaringenin was found to be present in the hexane extract of white birch bud¹⁹ and chloroform extract of Clerodendrum phlomidis²⁰.

The presence of flavanoids ( Pectolinaringenin ) and phenolic compounds (Ethanone, 1 (2 – hydroxyl – 5- methyl phenyl) & 4-methyl – 2- 4- bis (p-hydroxyphenyl) pent-1-ene in the aqueous extract of Luffa acutangula peel extract was confirmed and they would help in the formation and stabilization of zinc oxide nanoparticles ²¹.

4 CONCLUSION:

The qualitative phytochemical analysis of aqueous extract of Luffa acutangula peel revealed the presence of Flavanoids, Carbohydrates, Aminoacids, Proteins, Phenols, Quinones, Terpenoids, Coumarins, Saponins, Tannins, and Leucoanthocyanidines. GC-MS analysis of aqueous extract of Luffa acutangula peel showed the presence of six major compounds, Hexadecanoic acid, Pectolinaringenin, Benzofuran, 4-methyl – 2- 4- bis (p-hydroxyphenyl) pent-1-ene, Cis-13- Octadecenoic acid and Ethanone, 1 (2 – hydroxyl – 5- methyl phenyl). Thus the study concludes that Luffa acutangula peel contains important bioactive compounds and therefore the simple aqueous extract can be further studied for their activity.

ACKNOWLEDGEMENT:

Funding:

This work was supported by Sacred Heart College, Vellore District, Tamilnadu, India, through Don Bosco Research Grant. We would like to show our gratitude to the Management of Sacred Heart College, Vellore District, Tamilnadu, India for supporting the research by granting Don Bosco Research Grant.

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Received on 30.05.2019 Modified on 24.06.2019

Research J. Pharm. and Tech. 2019; 12(12): 6071-6074.

DOI: 10.5958/0974-360X.2019.01054.0