RESEARCH ARTICLE Year : 2023  |  Volume : 60  |  Issue : 1  |  Page : 94-100

Characteristic features and comparative analysis of essential oil composition of selected genus of Ocimum sanctum L. through GC-MS

Prerna Sharma1, Kumud Upadhyaya2
1 Veer Madho Singh Bhandari Uttarakhand Technical University, Dehradun; Guru Gobind Singh College of Pharmacy, Yamunanagar, Haryana, India
2 Department of Pharmacognosy, Kumaun University, Nainital, Uttarakhand, India

Date of Submission10-Jan-2021Date of Acceptance15-Nov-2021Date of Web Publication5-Apr-2023

Correspondence Address:
Prerna Sharma
Uttarakhand Technical University, Dehradun, Uttarakhand, Associate Professor, Guru Gobind Singh College of Pharmacy, Yamunanagar, Haryana
India

Source of Support: None, Conflict of Interest: None

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DOI: 10.4103/0972-9062.331404

Background & objectives: The main aim of this research is to provide literature on the Ocimum plant, and to know the significance of the Ocimum species carried out by pharmacognostic study and experimental design for GC-MS.
Ocimum genus are very important for their therapeutic potential among the most important aromatic herbs.
Methods: Extreme attention has been put on literature reports in which the utilization of tulsi and their pharmacognostic study has been done by performing morphological and microscopic leaf experimental design and by using essential oil through the GC-MS instrumentation method.
Results: The utilization of these characteristics would be important for the drug discovery scientist to develop a specific formulation of the crude drug, which will be a magical therapeutic agent in the future, with many advantages. GC-MS chromatogram of Ocimum sanctum, Ocimum canum, and Ocimum gratissimum oil showed major peaks and has been identified after comparison of the mass spectra with the NIST library, indicating the presence of three phytocomponents. From the results, the GC-MS study suggested that anethole which is well reported antimicrobial compound is more in O. canum (2.66%) in comparison to O. sanctum (1,28%) but absent in O. gratissimum. The results indicated that the antimicrobial activity is more in O. canum due to the presence ofa high amount of anethole in comparison to O. gratissimum and O. sanctum.
Interpretation & conclusion: The result revealed that O. canum has a microscopic character that can be identified by the characteristic GC MS analysis of extracts to distinguish between different species of the ocimum plant.

Keywords: O. Genus; Antibacterial; Plant; Natural source; Tulsi; GC-MS; Eugenol

How to cite this article:
Sharma P, Upadhyaya K. Characteristic features and comparative analysis of essential oil composition of selected genus of Ocimum sanctum L. through GC-MS. J Vector Borne Dis 2023;60:94-100
How to cite this URL:
Sharma P, Upadhyaya K. Characteristic features and comparative analysis of essential oil composition of selected genus of Ocimum sanctum L. through GC-MS. J Vector Borne Dis [serial online] 2023 [cited 2023 Apr 6];60:94-100. Available from: http://www.jvbd.org//text.asp?2023/60/1/94/331404   Background 

Ocimum sanctum L., commonly known as holy basil, is a plant known for its medicinal values, of the genus Ocimum belonging to the family Lamiaceae Ocimum is very important for its therapeutic potential among the most importantaromatic herbs for their enormous medicinal properties, such as anticancerous, antidiabetic, spasmolytic, carminative, cardioprotective, anthelmintic, and diaphoretic actions. Ocimum sanctum possesses various biological activities such as analgesic, antipyretic, antidiabetic, hepatoprotective, hypolipidemic, immunomodulatory, and anti-inflammatory. It is observed that some of its phytoconstituents such as eugenol, linoleic acid, luteolin, and β-sitosterol prevent skin, liver, oral, and lung cancers through increasing the anti-oxidant activity, inducing apoptosis, altering the gene expression, and inhibiting metastasis[11]. It has been observed that Ocimum sanctum leaves showed inhibition of the growth of tumour cells[10]. It is assumed that Ocimum sanctum would be an effective medicine so far in inhibiting all kinds of cancer[1],[2],[3],[4].

Ocimum sanctum is used as traditional medicine and possesses various biological activities because of the active components present in the plant such as eugenol, linoleic acid, oleic acid, rosmarinic acid, ocimarin, isorientin, orientin, aesculectin, aesculin, chlorogenic acid, galuteolin, gallic acid, citronellal, camphene, sabinene, dimethylbenzene, ethylbenzene, vitamin C, and calcium[5],[6],[7].

  MATERIAL & METHODS 

Collection, identification, and authentication of selected plants

A collection of all three types of tulsi leaves named Rama Tulsi (Ocimum sanctum), Krishna Tulsi (Ocimum canum) and Vana Tulsi (Ocimum gratissimum) of the genus Ocimum were collected for one month from the local area of Haryana, India. Identification of this was confirmed by Dr. RS Jayasomu, Head, Raw material Herbarium and Museum Division (RHMD), NISCAIR, New Delhi, India where a voucher sample (Ref.No. NISCAIR/RHMD/Consult/-2016/3000-27-2) has been deposited. For further studies in a pharmacognostical manner, phytochemical analysis, and extraction, leaves were collected, shade dried, and converted into fine powdered form.

Pharmacognostical study

The pharmacognostic study has been done by performing morphological and microscopic analysis of Leaf as per WHO guidelines.

Microscopic studies

Transverse section (TS) of leaf and powder characteristics were identified with (Phloroglucinol + HCL) reagents such as chloral hydrate and glycerine to study the cells, fibres, xylem vessels, starch grains, and calcium oxalate crystals. A permanent slide of TS of Leaf was prepared to observe the presence and arrangement of cellular structures as per the procedure of Johansen[8] and the representative figures were taken with the help of a microscopic image camera.

Experimental design for GC-MS

GC-MS analysis of the Rama Tulsi (Ocimum sanctum), Krishna Tulsi (Ocimum canum), and Vana Tulsi (Ocimum gratissimum) essential oil was performed using the below-given instruments information.

Thermo Trace 1300GC coupled with Thermo TSQ 800 Triple Quadrupole MS.

For GC - THERMO TRACE 1300 GC

For MS - THERMO TSQ 8000

Software used: XCalibur 2.2SP1 with Foundation 2.0SP1Column: BP 5MS (30m X 0.25mm, 0.25μm)Column Makeup: 5% Phenyl Polysilphenylene- siloxaneInjector: S (Split)Injection volume:2μLSplit Ratio: 20:1Injector temp: 250°CMS transfer line temp:230°CIon source temp: 230°CMass Range:40-700Carrier Flow:1.0ml/minOven Program:

Initial Temp: 50°C Hold time: 1.0 min

Temp 1: 220°C Hold Time: 5.0 min

Rate: 5°C/min

Detector: MS TSQ 8000Library used: NIST 2.0

Microscopical evaluation

The powder microscopy of selected varieties of Tulsi such as Rama Tulsi (Ocimum sanctum), Krishna Tulsi (Ocimum canum), and Vana Tulsi (Ocimum gratissimum) was done and evaluated for the characterization of the drug through pictures in their respective sequence. These pictures are critically evaluated with specific microscopic features [Figure 1].

Krishna Tulsi (Ocimum canum) and Vana Tulsi (Ocimum gratissimum) were done and evaluated for the characterization of the drug through pictures in their respective sequence. These pictures are critically evaluated with specific microscopic features. For this, Freehand sections of the plant material were stained with Phloroglucinol and HCl reagents such as chloral hydrate and glycerine to study the cells, bre, xylem vessels, starch grains, and calcium oxalate crystals. A permanent slide of TS of Leaf was prepared to observe the presence and arrangement of cellular structures, the representative figures were taken with the help of a microscopic image camera [Figure 2].

Krishna Tulsi (Ocimum canum) and Vana Tulsi (Ocimum gratissimum) were done and evaluated for the characterization of the drug through the below-given pictures in their respective sequence. These pictures are critically evaluated with specific microscopic features. For this, Freehand sections of the plant material were stained with Phloroglucinol and HCl reagents such as chloral hydrate and glycerine to study the cells, bre, xylem vessels, starch grains, and calcium oxalate crystals. A permanent slide of TS of Leaf was prepared to observe the presence and arrangement of cellular structures, the representative figures were taken with the help of a microscopic image camera [Figure 3].

Characterization of Ocimum

Gas Chromatography-Mass Spectrometry analysis

A Perkin-Elmer gas chromatograph (model 8700), with a flame ionization detector (FID), was used for the chemical analysis of the Rama Tulsi (O. sanctum), Krishna Tulsi (O. canum) and Vana Tulsi (O. gratissimum) essential oil. The temperatures of the injector and detector were set at 220°C and 290°C, respectively. The column thermostat temperature was started from 80°C and raised to 220°C at the rate of 4°C min-1, whereas initial and final temperatures were held for 3 and 10 min, respectively. The carrier gas was helium with a flow of 1.5 mL min-1. A sample of 1.0 μL was injected (split ratio 100:1). For quantification purposes a built-in data-handling program of the equipment (Perkin-Elmer) was used. The essential oil composition was reported as a relative percentage of the total peak area[12],[13],[14][Figure 4], [Figure 5] & [Figure 6] [Table 1], [Table 2] & [Table 3].

Table 1: Chemical Composition of volatile oil extract from O. gratissimum analyzed by GC-MS

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Table 2: Chemical Composition of volatile oil extract from O. sanctum analyzed by GC-MS

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Table 3: Chemical Composition of volatile oil extract from O. canum analyzed by GC-MS

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Ethical statement: Not applicable

  Results 

The hydro-distillation of essential oil from leaf extract of Ocimum sanctum L. yielded pale yellow aromatic oil. GC-MS chromatogram of the Ocimum sanctum, Ocimum canum, and Ocimum gratissimum oil showed major peaks and has been identified after comparison of the mass spectra with the NIST library, indicating the presence of three phytocomponents. The GC-MS study suggested that anethole which is well reported antimicrobial compound is more in O. canum (2.66%) in comparison to O. sanctum (1,28%) but absent in O. gratissimum. The results indicated that the antimicrobial activity is more in O. canum due to the presence high amount of anethole in comparison to O. gratissimum, and O. sanctum [Table 4].

  Conclusion 

The summarized information has been focused on the microscopic character of O. canum, with the characteristic GC-MS analysis of the extracts[9], to identify different species of the Ocimum plant. The retention time of each chemical constituent is reported for future identification of the plant and its variant. In the future, it will be a source of literature to identify variant plant species for the quality production of the plant. Ocimum sanctum L. and further experiments are required for better exploitation of essential oil for its commercial purposes.

  Abbreviations 

OS = Ocimum sanctum

O. = Ocimum genus

COX-1 = Cyclooxygenase-1

COX-2 = Cyclooxygenase-2

VEGF = Vascular endothelium growth factor

WHO = World Health Organization

COPD = Chronic obstructive pulmonary disease),

IBD = Inflammatory bowel syndrome

DMBA= 7, 12- dimethylbenz(a)anthracene

VEGFR-3 = Vascular Endothelial Growth Factor Receptor 3

VLA-4 = Very Late Antigen-4

Conflict of interest: None

 

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  [Figure 1], [Figure 2], [Figure 3], [Figure 4], [Figure 5], [Figure 6]
 
 
  [Table 1], [Table 2], [Table 3], [Table 4]