Scholarly article on topic 'GC-MS analysis of bio-active compounds in methanolic extract of Lactuca runcinata DC'

GC-MS analysis of bio-active compounds in methanolic extract of Lactuca runcinata DC Academic research paper on "Chemical sciences"

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Academic research paper on topic "GC-MS analysis of bio-active compounds in methanolic extract of Lactuca runcinata DC"

PHCOG RES

ORIGINAL ARTICLE

GC-MS analysis of bio-active compounds in methanolic extract of Lactuca runcinata DC

Lakshmi Kanta Kanthal, Akalanka Dey1, K. Satyavathi2, P. Bhojaraju3

Departments of Pharmacology, 2Pharmaceutical Chemistry, and Biotechnology, Koringa College of Pharmacy, Tallarevu Mandal, Andhra Pradesh, Department of Pharmacy, Annamalai University, Chidambaram, Tamil Nadu, India

Submitted: 04-04-2013 Revised: 25-04-2013 Published: 12-12-2013

ABSTRACT

Background: The presence of phytochemical constitutes has been reported from species of the Compositae (Asteraceae). Hitherto no reports exist on the phytochemical components and biological activity of Lactuca runcinata DC. Objective: The present study was designed to determine the bioactive compounds in the whole plant methanol extract of Lactuca runcinata. Materials and Methods: Phytochemical screening of the entire herb of Lactuca runcinata DC revealed the presence of some bio-active components. Gas chromatography-mass spectrometry (GC-MS) analysis of the whole plant methanol extract of Lactuca runcinata was performed on a GC-MS equipment (Thermo Scientific Co.) Thermo GC-TRACE ultra ver.: 5.0, Thermo MS DSQ II. Results: The phytochemical tests showed the presence of alkaloids, cardiac glycosides, flavonoids, phenols, phlobatannin, reducing sugars, saponins, steroids, tannins, terpenoids, volatile oils, carbohydrates, and protein/amino acids in methanolic extract of L. runcinata. The GC-MS analysis has shown the presence of different phytochemical compounds in the methanolic extract of Lactuca runcinata. A total of 21 compounds were identified representing 84.49% of total methanolic extract composition. Conclusion: From the results, it is evident that Lactuca runcinata contains various phytocomponents and is recommended as a plant of phytopharmaceutical importance.

Key words: GC-MS analysis, Lactuca runcinata, phytochemical screening, whole plant methanol extract

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www.phcogres.com DOI: 10.4103/0974-8490.122919 Quick Response Code:

INTRODUCTION

Use of plants as a source of medicine has been inherited and is an important component of the health care system. India is the largest producer of medicinal herbs and is appropriately called the botanical garden of the world.[1] Plants used for traditional medicine contain a wide range of substances that can be used to treat chronic as well as infectious diseases.[2] Lactuca runcinata DC [L. runcinata, Synonym-Lactuca heyneana DC.] commonly known as Undirachakam[3] or Atheli is an annual erect herb belonging to the family Compositae (Asteraceae). Traditionally this plant finds its wide applicability as diuretic and in chronic obstruction of liver and bowel.[3] Lactuca runcinata DC also has been reported to be a valuable source of essential nutrients, such as carbohydrate, protein, fat etc.,

and micro-nutrients like calcium, iron, phosphorous etc., Chewing Lactuca runcinata with betel leaf is useful to cure the blisters of mouth and tongue.

In the last few years, gas chromatography mass spectrometry (GC-MS) has become firmly established as a key technological platform for secondary metabolite profiling in both plant and non-plant species.[4-6] A detailed literature review on the plant in investigation has shown that so far there are no published reports worldwide, related to the possible chemical components of "Lactuca runcinata". So, the present study was aimed to investigate the possible chemical components by first preparing the methanolic extract and separation and identification of the compounds by subjecting it to GC-MS analysis.

MATERIALS AND METHODS

Collection of plant material

The entire herb of Lactuca runcinata DC plant (healthy and disease free plant samples) was collected from the

Address for correspondence:

Asso. Prof. Lakshmi Kanta Kanthal, Department of Pharmacology,Koringa College of Pharmacy, Korangi - 533 461,Tallarevu Mandal, Andhra Pradesh, India. E-mail: lkkhaldia@gmail.com

nearby area of Thoothukudi District fields (Tamil Nadu) in December 2011, identified and authenticated by Dr. V. Chelladurai (Retired Research Officer-Botany, Central Council for Research in Ayurveda and Sidda, Govt. of India), Tiruneveli, Tamil Nadu, India. Herberium of the plant, Lactuca runcinata, was prepared and preserved in the Department of Pharmacognosy, Koringa College of Pharmacy, Korangi, East Godavari District, Andhra Pradesh, India.

Preparation of extracts

A portion of dried aerial parts (100 g) of Lactuca runcinata DC was placed in a soxhlet apparatus. Extraction was performed with 750 ml of methanol for 48 h at a temperature not exceeding the boiling point of the solvent. Extract was filtered through a 45 |m filter.[7] The resulting solution was concentrated in vacuum to dryness to give methanol extract (9 g). The extract was stored in a refrigerator at 4°C for further use.

Preliminary phytochemical screening

The methanol extract was tested for alkaloids, anthroquinones, flavonoids, phenols, steroids, tannins, terpenoids,[8] cardiac glycosides, saponins,[9] phlobatannin,[10] reducing sugars,[11] volatile oils,[12] carbohydrates and protein/amino acids.[13,14]

GC-MS (Gas Chromatography-Mass Spectrometry) analysis

The phytochemical investigation of methanolic extract was performed on a GC-MS equipment (Thermo Scientific Co.) Thermo GC-TRACE ultra ver.: 5.0, Thermo MS DSQ II. Experimental conditions of GC-MS system were as follows: TR 5-MS capillary standard non-polar column, dimension: 30Mts, ID: 0.25 mm, Film thickness: 0.25|m. Flow rate of mobile phase (carrier gas: He) was set at 1.0 ml/min. In the gas chromatography part, temperature programme (oven temperature) was 40°C raised to 250°C at 5°C/min and injection volume was 1 | l. Samples dissolved in chloroform were run fully at a range of 50-650 m/z and the results were compared by using Wiley Spectral library search programme.

RESULTS AND DISCUSSION

After the successful conventional hot soxhlet extraction of the whole part of the plant in investigation, the preliminary phytochemical study revealed that methanolic extract of Lactuca runcinata contains alkaloids, cardiac glycosides, flavonoids, phenols, phlobatannin, reducing sugars, saponins, steroids, tannins, terpenoids, volatile oils, carbohydrates, and protein/amino acids. Anthraquinones were absent in the L. runcinata methanolic extract, as summarized in [Table 1].

Table 1: Preliminary phytochemical evaluation of methanol extracts of Lactuca runcinata

Phytochemical Test/Reagents Result

constituents

Alkaloids

Anthraquinones Cardiac glycosides Flavonoids Phenols Phlobatannin Reducing sugars Saponins Steroids Tannins Terpenoids

Volatile oils Carbohydrates Protein/Amino acids

+ =Present; - =Absent

The results pertaining to GC-MS analysis of the methanolic extract of Lactuca runcinata DC lead to the identification of a number of compounds. These compounds were identified through mass spectrometry attached with GC. The various components present in the entire herb of Lactuca runcinata DC that were detected by the GC-MS are shown in [Table 2]. Carbetapentane, 2-Propenoic acid, 2-methylpropyl ester, Bornyl Acetate, Anti (9, 10)-tricyclo [4.2.1.1 (2, 5)] dec-3-en-9-endo-ol, Methanone (1-hydroxycyclohexyl) phenyl, 1 (3H)-Isobenzofuranone, 3-ethoxy, 4-Hydroxy-6-(4-methoxyphenyl)-2-pyranone, 1-Benzoyl-3-(4'-nitrophenyl) pyrrolo [2,1-a] phthalazine, Pyridine-3-carboxamide, 2,4,6-trichloro-N - (4-methoxyphenyl) - 5-nitro, Ethyl1-benzyl-4,6-dibromo-3-methyl-2-oxoindoline-3-carboxylate, 5à-Cholestan-19-oic acid, 2à- hydroxy-, acetate, (5â) Pregnane-3,20a-diol, 14à,18à-[4-methyl-3-oxo-(1-oxa-4-azabutane-1,4-diyl)]-, diacetate, O, O-Diethyl (1 formylpentyl) selenophosphate, 3-n-Pentadecyl-2,4-dinitrophenol, 1,3-Bis (4-nitrobenzyl)-5,6 dihydrobenzo [f] quinazoline, 1,2-Benzenedicarboxylic acid, bis (2-ethylhexyl) ester, Dibenzoxazabicycloundecane, 4-[p-Bis (2-hydroxyethyl) amino] phenyl-1-bromoisoquinoline, Ergost-25-ene-3,6-dione, 5,12-dihydroxy-, (5à,12â), Lup-20 (29)-en-3-ol, acetate, (3a)-, E-Ethyl (Z)-3-(4- Acetylphenylthio) cinnamate were present in the methanolic extracts of Lactuca runcinata. The composition determined for this methanolic extract corresponds to 86.63% of the entire GC-MS chromatogram.

DragendorfFs test +

Wagner test +

Borntrager's test -

Kellar-Kiliani test +

Shinoda test +

Phenol test +

1% HCl +

Fehling test +

Frothing test/Foam test +

Liebermann-Burchardt test +

Braemer's test +

Liebermann-Burchardt +

Salkowski test +

Strin test +

Molish's test +

Biuret test +

Ninhydrin test +

Table 2: Compounds identified in the methanolic extract of Lactuca runcinata In GC-MS

RT Name of the compound Molecular Molec ular Peak Compound

formula weight area % nature

3.13 Carbetapentane C20H31 NO3 333 0.45 Ester

8.10 2-Propenoic acid, 2-methylpropyl ester C7H12O2 128 0.83 Ester

14.34 Bornyl Acetate C^H^ 196 0.89 Ester

17.77 anti (9,10)-tricyclo [4.2.1.1 (2,5)] dec-3-en-9-endo-ol C10H14O 150 1.24 Alcohol

20.60 Methanone (l -hydro xycyclohexyl) phenyl C13H16O2 204 3.28 Ketone

23.00 1 (3H)-Isobenzofuranone, 3-ethoxy C10H10O3 178 7.79 Ketone

29.06 4-Hydroxy-6-(4-methoxyphenyl)-2-pyranone C12H1°O4 218 0.62 Ketone

33.40 l-Benzoyl-3-(4'-nitrophenyl) pyrrolo [2,1-a] phthalazine C24H15N3O3 393 0.64 Ketone

33.82 Pyridine-3-carboxamide, 2,4,6-trichloro-N-(4-methoxyphenyl)-5-nitro C13H8Cl3N3O4 375 0.55 Amide

34.56 Ethyl 1-benzyl-4,6-dibromo-3-methyl-2-oxoindoline-3-carboxylate Ci9H17Br2N03 465 0.68 Ester

34.89 5a-Cholestan-19-oic acid, 2a-hydroxy-, acetate C29H48O4 460 0.63 Steroidal ester

36.01 (5a) Pregnane-3,20a-diol, 14a, 18a- [4-methyl-3-oxo-( 1-oxa-4-azabutane-1,4-diyl)]-, diacetate C28H43N06 489 0.78 Steroidal ester

36.56 0,0-Diethyl (l-formylpentyl) selenophosphate Ci0H2iÜ4PSe 316 0.72 Phosphate ester

36.87 3-n-Pentadecyl-2,4-dinitrophenol C21 H34N2O5 394 0.93 Phenol

37.16 l, 3-Bis (4-nitrobenzyl)-5,6 dihydrobenzo [f] quinazoline 452 1.54 Aromatic

38.00 1,2-Benzenedicarboxylic acid, bis (2-ethylhexyl) ester C24H38O4 390 1.41 Ester

38.59 Dibenzoxazabicycloundecane 341 2.39 Hydrocarbon

41.66 4- [p-Bis (2-hydroxy ethyl) amino] phenyl-1 -bromoisoquinoline Ci9Hi9BrN202 386 0.54 Iso-quinoline

43.20 Ergost-25-ene-3,6-dione, 5,12-dihydroxy-, (5a, 12a) C28H44O4 444 10.46 Steroid

43.84 Lup-20 (29)-en-3-ol, acetate, (3a)- C32H52O2 468 15.11 Steroidal ester

45.13 E-Ethyl (Z)-3-(4-Acetylphenylthio) cinnamate C1!3H18O3S 326 33.01 Ester

RT=Retention time

The GC-MS spectrum confirmed the presence of various components with different retention times as illustrated in [Figure 1]. The mass spectrometer analyzes the compounds eluted at different times to identify the nature and structure of the compounds. The large compound fragments into small compounds giving rise to appearance of peaks at different m/z ratios. These mass spectra are fingerprint of that compound which can be identified from the data library.

The present study helps to predict the formula and structure of 21 biomolecules. Further investigation may lead to isolation of bio-active compounds and their structural elucidation and screening of pharmacological activity will be helpful for further drug development.

CONCLUSION

The presence of various bio-active compounds detected after GC-MS analysis using the methanolic extract of Lactuca runcinata DC justifies the use of whole plant for various elements by traditional practitioner.

However, isolation of individual phytochemical constituents and subjecting it to the biological activity will be definitely giving fruitful results and will open a new area of investigation of individual components and their pharmacological potency. From these results, it could be concluded that "Lactuca runcinata' contains various

Figure 1: GC-MS chromatogram of methanolic extract of Lactuca runcinata

bio-active compounds. Evaluation of pharmacological activity is under progress. Therefore, it is recommended as a plant of phytopharmaceutical importance.

ACKNOWLEDGEMENT

The authors are thankful to the Management of Koringa college of pharmacy, Korangi, Andhra Pradesh, India for availing all the facilities and are also thankful to Radiant Research Services Pvt. Ltd, Srinagar, Bangalore - 560 050, India for carrying out GC-MS analysis of the sample.

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Cite this article as: Kanthal LK, Dey A, Satyavathi K, Bhojaraju P. GC-MS analysis of bio-active compounds in methanolic extract of Lactuca runcinata DC. Phcog Res 2014;6:58-61.

Source of Support: Nil, Conflict of Interest: None declared.

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