GC-MS Analysis of n-hexane Extract of Roots of Aerva lanata (Linn) Juss. ex. Schult.

 

Susikumar S¹, Nartunai G², Ilavarasan R³*

¹Research Scholar, Captain Srinivasa Murthy Regional Ayurveda Drug Development Institute,

Recognized by University of Madras, CCRAS, Ministry of AYUSH, Govt. of India, A. A. Hospital Campus, Arumbakkam, Chennai – 600106.

²Research Officer, Captain Srinivasa Murthy Regional Ayurveda Drug Development Institute,

Recognized by University of Madras, CCRAS, Ministry of AYUSH, Govt. of India, A. A. Hospital Campus, Arumbakkam, Chennai – 600106.

³Assistant Director (S-4) In-charge, Captain Srinivasa Murthy Regional Ayurveda Drug Development Institute, Recognized by University of Madras, CCRAS, Ministry of AYUSH, Govt. of India, A. A. Hospital Campus, Arumbakkam, Chennai – 600 106.

 

ABSTRACT:

Context: The roots of the plant Aerva lanata are extensively used in Indian System of Medicine to cure urinary stones, antidiabetic, diuretic and used as demulcent. Aim: To investigate the phytoconstituents from the n-hexane extract of the roots of Aerva lanata using GC-MS analysis. Materials and Methods Roots of Aerva lanata was extracted by Soxhlet extraction method using n-hexane. The chromatogram was recorded by injecting the extract by splitless injection mode into the GC MS 5975 C Agilent equipped with a QP- 5000 (quadrupole) Gas Chromatography–Mass Spectrometer. Results: 23 phytoconstituents were identified by close matches with standard MS spectra and compared with NIST- 11 and WILEY library data from n-hexane extract. The major compounds reported are oleic acid (21.68%), γ-sitosterol (10.67%), Stigmasterol (5.13%), 3-β,5-α-stigmast-7-en-3-ol (4.23 %) and other constituents were found to be in traceable quantities. Conclusion: GC-MS analysis of roots of Aerva lanata revealed certain interesting facts of presentation of various phytoconstituents. The presence of various phytoconstituents contributes to the medicinal activity of the plant.

 

 

 

INTRODUCTION:

Aerva lanata (Linn) Juss. ex. Schult belongs to Amaranthaceae is an erect or prostate herbaceous weed, commonly distributed throughout the hotter parts of India and ascends to 3,000 feet on the hills¹. The genus Aerva have 22 species reported². The plant A. lanata (Fig.1) is commonly known as Astmabayda in Sanskrit and Sirupoolai, Pulai in Tamil³. The roots of the plant Aerva lanata are extensively used in Indian system of medicine to cure urinary stones. Other uses reported are demulcent, diuretic, antidiabetic and in the treatment of headache^4,5.

 

The roots of the A. lanata is used in important Ayurvedic and Siddha formulations namely Shatavaryadi ghrita, Sirugan peelai chooranam and Nerunjil kudineer^6,7.

 

Figure 1: Habit with a flowering twig of Aerva lanata

 

Figure 2: Dried roots of Aerva lanata

 

The whole plants of A. lanata reported to contain major phytochemicals such as β-sitosterol, α-amyrin, betulin, hentriacontane, sitosteryl palmitate, D-glucoside, glycosides, kaempferol-3-galactoside and kaempferol-3-rhamnogalactoside⁸. The methanolic extract of roots of Aerva lanata reported to contain 23 different compounds such as pyridine (21.74%), 1,2-benzenedicarboxylic acid dioctyl ester (7.83%), hydroquinone monobenzyl ether (5.71%), 6-trifluoroacetyl-7,11-methano-1,2,3,4,5-pentahydroazuleno[2,1-g]cycloundecene (4.39%), (5S,8S,9S)-1-benzyl-10-methyl 8,9-bis(benzoyloxy)-4, 7-dioxo-5-methyl-3, 6-diazadecane dicarboxylate (4.33%) etc.,^9,10,11,12. GC-MS analysis of n-hexane extract of roots of A. lanata provides information about presence of phytoconstituents in the plant. The present investigation is carried out to determine the possible chemical components of n-hexane extract of roots of A. lanata.

 

MATERIALS AND METHOD:

Collection of Plant Material:

The roots of Aerva lanata (Fig.2) was collected from Arignar Anna Hospital campus and authenticated by Pharmacognosy department, Captain Srinivasa Murthy Regional Ayurveda Drug Development Institute, Arumbakkam, Chennai, Govt. of India and the voucher specimen was deposited (185A/rt/23) in raw drug museum for future reference.

 

Preparation of Extract:

The shade dried and coarsely powdered (10/44) roots of  A. lanata (4g) was extracted using n-hexane (AR) by soxhlet extraction method. The extract was filtered through Whatman No.1 filter paper and concentrated in vacuum to constant weight. The yield of the extract is 0.42 %. The n-hexane extract of the plant was used for GC-MS analysis.

 

Table 1. GC-MS Data of n-hexane extract of roots of Aerva lanata

 

Figure 3:  GC-MS Chromatogram of n-hexane extract of roots of Aerva lanata

 

Gas Chromatography- Mass Spectrometer Conditions:

Gas chromatography combined with mass spectroscopy is a preferable methodology for routine analysis of compounds. n-hexane extract was injected by split less injection mode into the GC MS 5975 C Agilent equipped with a QP-5000 (quadrupole) mass spectrometer, fitted with a DB-5 MS ultra inert 30m x 250µm, film thickness 0.25µm capillary GC column, coated with polydimethyl siloxane. Helium was the carrier gas at flow rate of 1.5 ml/min. The injector port temperature was 250°C and the oven temperature was maintained at 70°C for 3 mins, 10°C/min up to 300°C for 9 mins. The ionization voltage was 70 eV. It is separated into various constituents with different retention time which are detected by mass spectrophotometer and matched with those of the NIST – 11 and WILEY library and published mass spectra.

 

RESULTS AND DISCUSSION:

GC-MS analysis of the n-hexane extracts of the roots of A. lanata is given in Table 1. Twenty-three compounds were identified by comparison with the authentic spectra obtained from GC-MS library (NIST-11 and WILEY). Even though several peaks were present in the GC-MS chromatogram (Figure 3-5), the identification of 23 constituents was established by comparison with the authentic spectra obtained from GC-MS library with the SI factor. The major compounds reported are oleic acid (21.68%) γ-sitosterol (10.67%), stigmasterol (5.13%), 3-β,5-α-stigmast-7-en-3-ol (4.23%) and other constituents were found to be in traceable quantities. γ-sitosterol has been reported to reduce LDL cholesterol and thereby reduces the cardiovascular risk¹³, potential anticancer activity against breast and Lung cancer cells by in-vitro study¹⁴, hepatoprotective and anti hyperglycaemic activity by increasing insulin level¹⁵. Among the major phytoconstituent reported, γ-sitosterol (Figure 6) and stigmasterol (Figure 7) having high medicinal value.

 

 

Figure 4: GC-MS Spectra of γ-sitosterol (Retention time-28.287)

 

 

 

Figure 5: GC-MS Spectra of Stigmasterol (Retention time-27.704)

 

                

Figure 6: γ-sitosterol                                                                                    Figure 7: Stigmasterol

 

CONCLUSION:

In the present study twenty-three chemical constituents have been identified from n-hexane extract of the roots of A. lanata by Gas Chromatogram Mass Spectrometry (GC-MS) analysis. The presence of various phytochemicals contributes to the medicinal activity of the plant.

 

ACKNOWLEDGEMENT:

The authors are very grateful to the Director General, Central Council for Research in Ayurvedic Sciences, New Delhi for providing opportunity to conduct this study and Dr. D. Ramasamy, Assistant Professor, Department of Chemistry, Govt. Thirumagal Mill’s College, Gudiyattam, Vellore District, Tamil Nadu for his encouragement and guidance to carry out the study.

 

CONFLICT OF INTEREST:

Nil.

 

ABBREVIATIONS USED:

AR: Analytical Reagent; Fig: Figure; GC-MS: Gas Chromatography and Mass specrtroscopy; SI: Super imposability; t_R : Retention time.

 

REFERENCES:

1.      Chadha YR, editor. The Wealth of India, Raw materials, Volume 1,  A and B, National Institute of Science Communication and Information Resources, CSIR, New Delhi. 1948; p.34.

2.      The Plant List (2013). Version 1.1. Published on the Internet 2018 [cited 2018 January 3]. Available from: http://www.theplantlist.org/.

3.      Narayana AK and Kolammal M. Pharmacognosy of Ayurvedic Drugs Kerala. Volume 6. Department of Pharmacognosy, University of Kerala, Kerala. 1st ed: 1963; pp.25-6.

4.      Chopra RN, Nayar SL, and Chopra IC. Glossary of Indian Medicinal Plants, National Institute of Science Communication (CSIR), New Delhi. 5^th ed: 1999; p.8.

5.      Ayurvedic Formulary of India, Minstry of Health and Family Welfare, Dept of ISM and H., Govt. of India, New Delhi. 1st eng. Ed: 2000; p.2.

6.      Sharma PC, Yelne MB, Dennis TJ and Aruna Joshi editors. Database on Medicinal Plants Used in Ayurveda and Siddha, Volume 8. CCRAS, Min. of Health and Family Welfare, Dept of AYUSH; New Delhi. 2007; pp.85-95.

7.      Gupta A K, Neeraj Tandon and Madhu Sharma editors. Quality standards of Indian Medicinal Plants. Volume 3. Indian Council of Medical Research, New Delhi. 2005; pp.9-19.

8.      Yamunadevi M, Wesely EG and Johnson M. Chromatographic finger print analysis of steroids in Aerva lanata L. by HPTLC technique. Asian Pacific Journal of Tropical Biomedicine. 2011; 1(6): 428-433.

9.      Yamunadevi M, Wesely EG and Johnson M. Chromatographic finger print studies on Saponins of Aerva lanata (L.) juss. Ex schultes by using HPTLC. International Journal of Current Pharmaceutical Research. 2012; 4(2): 52-57.

10.   Yamuna DM, Wesely EG and Johnson M. Chromatographic Studies on the Tannins of Aerva lanata (L.) Juss. Ex Schultes. IOSR Journal of Pharmacy. 2012; 2(1): 41-51.

11.   Chauhan VS and Swapna M. Isolation characterization and cytotoxic activity of diterpenoid and flavonoids of Aerva lanata. International Journal of Pharmaceutical and Chemical Sciences. 2014; 3(3): 766-769.

12.   U.S. National Library of Medicine, clinicaltrials.gov. 2017 [cited 2018 July 11]. Available from:https://clinicaltrials.gov/ct2/show/NCT00153738.

13.   Shenbagamoorthy S, Ramar T, Vellingiri S, Krishnasamy K, Palani G, Shanmugam A, Soundarapandian K. Sitosterol from Acacia nilotica L. induces G2/M cell cycle arrest and apoptosis through c-Myc suppression in MCF-7 and A549 cells. Journal of Ethnopharmacology. 2012; 803– 809.

14.   Rangachari B, Veeramuthu D and Savarimuthu I. Antidiabetic activity of γ-sitosterol isolated from Lippia nodiflora L. in streptozotocin induced diabetic rats. European Journal of Pharmacology. 2011; 410–418.

15.   Navpreet K, Jasmine C, Akash J and Lalit K. Stigmasterol: A Comprehensive Review, IJPSR, 2011; 2(9): 2259-2265.

 

 

 

Received on 06.05.2020           Modified on 25.06.2020

Accepted on 23.07.2020         © RJPT All right reserved