GC-MS ANALYSIS OF CURCUMA ZEDOARIA TUBER EXTRACTS AND ITS ANTIBACTERIAL ACTIVITY

GC-MS ANALYSIS OF CURCUMA ZEDOARIA TUBER EXTRACTS AND ITS ANTIBACTERIAL ACTIVITY

R. Nirmala Nithya *, R. Sai Nandhini and U. Koushi Kumar

Department of Biotechnology, Vel Tech Rangarajan Dr. Sagunthala R&D Institute of Science and Technology, Chennai, Tamil Nadu, India.

ABSTRACT: This study investigates the presence of bioactive compounds present in the tubers of Curcuma zedoaria using GC-MS analysis. Different compounds, including alkaloids, tannins, flavonoids, and proteins, were identified using the preliminary phytochemical screening for aqueous, ethanol and methanol extracts. The present study reports the antimicrobial activity analysis using aqueous, ethanol and methanol extracts. The results show that the ethanol and methanol extracts showed relatively enhanced results. The extracts were subjected to GC-MS analysis, and antibacterial properties for the extracts were analyzed. Bioactive compounds such as 1,2 cyclopentanedione, 2(5H)-furanone, hexadecanoic acid, succinic acid, oleic acid, pyrrolidine etc., were identified through GC-MS. These compounds exhibited various therapeutic functions such as anticancer, anti-inflammatory, and anti-microbial activity. The antibacterial assay showed a considerable zone of inhibition for the ethanolic extracts in the case of K. pneumonia and B. subtilis. Hence, these crude extracts can be used as a better and alternative source, which can be used in herbal medicines. The study concluded that C. zedoaria tuber contains various bioactive compounds. Therefore, this tuber can be considered of phytopharmaceutical importance.

Keywords: Bioactive compounds, Anti- bacterial, GC-MS, Therapeutic, Phytochemicals

INTRODUCTION: Bioactive compounds / secondary metabolites are major sources in conventional medicine. The most expensive resource for some potential medications is thought to be plants ^1-4. Even though the utilization of plants as medication has been known for centuries, nowadays its remedial worth is picking up centrality. Curcuma zedoaria (C. zedoaria) belongs to the class curcuma and family Zingiberaceae ^5-8. It is predominantly known as kichilikizhangu locally and white turmeric ^9-10. Pharmacological evaluation reports suggest this plant has antifungal, antiamoebic, antiulcer, anticancer, analgesic, and anti-asthmatic activity ^11-16.

In ancient days, the rhizome of C. zedoaria was used to treat bad breath, skin problems, menstrual irregularities, and digestive disorders ^17-24. The gas chromatography-mass spectrometry (GC-MS) study is worth identifying and quantifying small molecular metabolites, including sugars, acids, unsaturated fats, drugs, sterol and toxins ^18-24. This technique can easily analyze the secondary metabolites of both plant and non-plant species. As a result, the current study's main objective is to assess the various bioactive substances in the aqueous, ethanol, and methanol extracts of C. zedoaria using GC-MS along with the investigation of antibacterial activity for the same ^25-30.

MATERIALS AND METHODS:

Collection of Plants: The local market in Chennai, Avadi, where the plant material was purchased. To get rid of any adhering material, it was washed in running water, dried at RT for a week in the shade. An airtight container was used to store the dried plant materials after they had been ground into a fine powder

Preparation of Plant Extract: The coarse powder of C. zedoaria was extracted using water, ethanol and methanol solvents in a ratio of 1:5 and kept in an orbital shaker for 48 h. The extracts were filtered using Whatman no.1 filter paper and were dried in distillation unit ³¹.

The prepared extracts were used to check antibacterial efficacy, screen the preliminary phytochemical compounds, and perform GC-MS analysis to determine the pharmacological compounds.

Phytochemical Screening: The C. zedoaria extracts were assessed for the existence of phytochemical compounds such as alkaloids, phenol, flavonoids, saponins, tannins, terpenoids, protein, carbohydrate and cardiac glycosides by using the standard protocol ³¹.

Antibacterial Activity of C. zedoaria: The methanol and ethanol extracts were tested for antibacterial activity in a agar well diffusion method. Three bacterial species namely, Escherichia coli, Bacillus subtilis, Klebsiella pneumonia were grown on nutrient agar plates using spread plate technique.

Four wells were bored in 0.5 cm in diameter and 0.5 µL of the plant extract was added in the bored wells. The plates were then incubated at 37°C for 24 hours. The observation was done based on different antibiotics, and the zone of inhibition was measured using the scale around the discs ³².

GC-MS Analysis: Bioactive compounds of methanol and ethanol extracts were determined by GCMS analysis (model-GCMS-QP 2010 plus). For GC-MS detection, an electron ionization system with ionizing energy of 70 eV was used. Helium gas (99.9%) was used as the carrier gas at constant flow rate 1 ml/min and an injection volume of 2 μl was employed (split ratio of 1:10); injector temperature 280°C; ion-source temperature 250°C.

The oven temperature was programmed from 50°C (isothermal for 1 min), with an increase of 10°C/min, to 200°C, ending with a 5 min isothermal at 300°C. Mass spectra were taken at 70 eV in a scan-interval of 0.30 s. The identification of each compound was compared by their relative retention time and mass spectra.

GCMS interpretation was carried out by the NIST (National Institute Standard and Technology) database with 62000 patterns. The unknown compounds were compared with the NIST library is known compounds and its name, molecular mass, molecular weight, and chemical structure have been ascertained.

Based on the GCMS analysis, the peak in ethanol and methanol extracts of the C. zedoaria plant showed the presence of bioactive secondary metabolites ³³.

RESULTS AND DISCUSSION:

Phytochemical Screening: Preliminary phytochemical analysis of C. zedoaria revealed the presence of secondary metabolites such as alkaloids, flavonoids, glycosides, terpenoids, proteins, and reducing sugars, phenolic compounds, and saponins were absent. The phytochemical screening of methanol extract, ethanol extract, and aqueous extract has been listed in Table 1, with their corresponding test method.

TABLE 1: PHYTOCHEMICAL SCREENING OF C. ZEDOARIA USING DIFFERENT EXTRACTS

++ denotes highly present, + denotes mildly present and - denotes absence.

Gas Chromatography and Mass Spectroscopy: Gas chromatography and mass spectroscopy of ethanol and methanol extracts of C. zedoaria were carried out to detect the constituents of esters, volatile matter, long-chain, and alcohol acids in Fig. 1 & 2.

FIG. 1: GC-MS CHROMATOGRAM ANALYSIS OF C. ZEDOARIA ETHANOLIC EXTRACT

FIG. 2: GC-MS CHROMATOGRAM ANALYSIS OF C. ZEDOARIA METHANOLIC EXTRACT

GC-MS chromatogram analysis of C. zedoaria ethanolic extract is shown in Fig. 1 and methanolic extract in Fig. 2. Fig. 1 and 2 illustrate the GC-MS spectrum and confirm the presence of various bioactive components with different retention times. The mass spectrometer analyzed the nature and structure of these compounds were the compounds eluted at different times ⁴¹.

The GC-MS study of C. zedoaria had shown the presence of many phytochemicals which added their contribution to the medicinal importance of that plant. The results showed ten major peaks in ethanolic extract and eight in methanolic extract.

The properties of the phytochemical compounds are listed in Table 2 and Table 3 for ethanolic and methanolic extract, respectively. The various phytochemical identified here contributes to medicinal activities. The compound name, molecular formula, retention time, and biological activity have been. Many of the compounds proved to have antimicrobial, antioxidant, anti-inflammatory, and antitumor activities ^42-44.

Among the compounds identified, the most predominant compounds acknowledged in ethanolic compounds are 2(3H)-furanone, allyl (ethoxy) dimethylsilane, hexadecanoic acid ethyl ester, ethyl 9-hexadecenoate, docosanoic acid, cyclopentane carboxylic acid, ethyl p-methoxycinnamate, propanoic acid, succinic acid and 3-bromocholest-5-ene.

Moreover, the compounds including cyclopent-4-ene, cylopentanone, pyrrolidine, 2(3H)-furanone, 5 methyl 2-hydroxycyclopent-2-en-1-one, 1-butene, 4-isothiocyanato, phenol, formic acid 2-propenyl ester were identified in methanol extract of C. zedoaria.

TABLE 2: PHYTOCOMPONENTS IDENTIFIED IN THE ETHANOLIC EXTRACT OF C. ZEDOARIA BY GC-MS

TABLE 3: PHYTOCOMPONENTS IDENTIFIED IN THE METHANOLIC EXTRACT OF C. ZEDOARIA BY GC-MS

Since, the aqueous extract did not respond to much of the compounds, the antibacterial study was carried out for ethanol and methanol extracts. The zone of inhibition was observed around the well in the plates containing microorganisms Escherichia coli, Bacillus subtilis, and Klebsiella pneumonia. The results were compared with standard antibiotics such as ampicillin and streptomycin. Compared to ethanol and methanol extract, ethanol extract exhibited the highest zone of inhibition against Bacillus subtilis (30 mm) and Klebsiella pneumonia (35 mm). From the tested bacteria, E. coli shows resistance towards the plant extract even at higher concentrations. In the case of the plant extracts, depending on the bacterial zone of inhibition, they can either act alone or in combination to inhibit bacterial growth as it is due to the presence of active metabolites ⁴⁵. Plant-derived medicines are vital to human health and are effective against microbial infections.

TABLE 4: ZONE OF INHIBITION BY THE WELL DIFFUSION METHOD

CONCLUSION: Traditional & home remedial medicine use a wide variety of natural materials. Among these, medicinal plants are widely used to treat various diseases caused by microorganisms. Based on this, the current study has tried to reveal the importance of C. zedoaria tubers extracts. GC-MS studies have shown various bioactive compounds in C. zedoaria tubers extracts. Moreover, these bioactive compounds importance have been conformed via antibacterial assay. The antibacterial activities against the growth of the selected organisms have been confirmed will the formation of clear zones. The study confirms the effectiveness of C. zedoaria ethanolic extract on K. Pneumonia and B. subtilis compared to E. coli. Moreover, the C. zedoaria tubers extract's antibacterial activity might be linked with alkaloids, terpenoids, flavonoids, and other bioactive compounds present in the C. zedoaria tuber. From the results, it is concluded that C. zedoaria tuber contains various bioactive compounds. Therefore, this tuber can be considered as phytopharmaceutical importance.

ACKNOWLEDGMENT: We take this opportunity to express our profound gratitude to Department of Biotechnology, Vel Tech Rangarajan Dr. Sagunthala R&D Institute of Science and Technology, for providing all the facilities required for the completion of this work.

CONFLICTS OF INTEREST: There are no conflicts of interest. The authors alone are responsible for the content and writing of this article.

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    How to cite this article:

    Nithya RN, Nandhini RS and Kumar UK: GC-MS analysis of Curcuma zedoaria tuber extracts and its antibacterial activity. Int J Pharm Sci & Res 2023; 14(10): 4854-59. doi: 10.13040/IJPSR.0975-8232.14(10).4854-59.

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