GC-MS Analysis of Beta Vulgaris Extract

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ISSN Print: 2664-6781

ISSN Online: 2664-679X

IJACR 2025; 7(1): 43-49

Received: 22-11-2024

Institute of Pharmaceutical

Sciences, Peoples University of

Medical and Health Sciences

for Women (PUMHSW)

Nawabshah, Pakistan

Institute of Pharmaceutical

Sciences, Peoples University of

Medical and Health Sciences

for Women (PUMHSW)

Nawabshah, Pakistan

The resistance to therapeutic drugs increases the demand to discover novel chemical compounds with

potential potency. Medicinal plants provide a wide window of opportunities to discover new drug leads

through a screening process with scientific validation and optimization. Hyphenated techniques in drug

discovery made it easier to isolate the structural information. It allows analysts to distinguish between

known and unknown compounds. Advancing the research, Gas Chromatography-Mass Spectroscopy

was employed in this study to identify the bioactive compound within the ethanolic extract, marking a

notable progression. The spectrum confirms the presence of various medicinal components including

heneicosane, n-decyl-N'-isopropyl, Octacosane, Eicosane, Hexatriacontane, 1,2-Benzenedicarboxylic

acid, bis(2-ethylhexyl) ester, Tetracosane, Heptacosane, 1,2-Benzenedicarboxylic acid, diundecyl ester,

Di-isooctyl phthalate, Eicosane, 2-methyl, Nonacosane, Tetracosane, 11-decyl, responsible for the

antimicrobial activity, anti-inflammatory, cytotoxicity, antimalarial, anti-inflammatory property of

beetroot thus could play vital roles in health care programs.

Keywords: Drug discovery, medicinal plants, GC-MS, bioactive compounds, beetroot extract,

antimicrobial, anti-inflammatory, cytotoxicity

Medicinal Plants have been employed as primary sources in past decades for the

development of chemical compounds, lately utilized to manufacture herbal products but also

play an important role in drug discovery to expand the pharmaceutical formulations in the

modern therapeutic system [1]. Different industries like chemical, pharmaceutical, cosmetics,

and industrial raw materials have been utilizing plant-based chemical compounds to develop

different formulations to treat human illness [2]. The US drug market recognized Different

types of new drugs like vincristine, resin amine, vinblastine, and reserpine during 1950-1970

in approximately 100 different plants [3]. Apart from the complimentary use of herbal plants,

the ethnopharmacological properties reveal diverse biological activity like Artemisinin from

Artemisia annua, used to treat multidrug-resistant malaria [4]. Silymarin is used in liver

disease, derived from the seeds of Silybum marianum [5], with the help of analytical

techniques. Similarly, the contribution of beetroot in nutritional sciences for human health

and diet is renowned along with its distinguished phytochemical constituents which can be

used for multiple health purposes [6]. The pharmaceutical industry is creating a new

revolution in modern medicine research to discover new molecules through advanced

analytical methods and techniques [7]. Drug discovery always relies on the generation and

innovation in analytical techniques and big data converter tools for molecule recognition and

screening [8]. The major invention in analytical instrumentations includes Gas

Chromatography-Mass Spectroscopy, high-performance liquid chromatography, mass

spectrometry, capillary electrophoresis, vibrational spectroscopies (infrared and Raman), X-

ray diffractometry, and hyperspectral imaging techniques [9].

Gas Chromatography-Mass Spectroscopy is an advanced sophisticated analytical technology

with a combination of two powerful analytical tools mainly operated for Compounds

identification with low detection limits based on chemical structure and potential for

quantitative measures [10, 11].

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GC-MS has been demonstrating and uncovering the

medicinal Chemical Constituents of many plants including

evolvulus alsinoides (Dwarf morning glory) [12]. Polygonum

Chinense (Creeping smartweed or Chinese knotweed) [13].

Moringa oleifera (Drumstick tree) [14]. Justicia wynaadensis

(Moddu Soppu) [15]. For decades, numerous beneficial

chemical constituents have been waiting to be discovered.

GC-MS has enormous applications not specifically in

Pharmaceutical sciences but also in environmental analysis,

explosives investigations, food and flavor analysis, drug

testing of athletes, and forensic analysis [11]. Previous studies

reported the diversity in pharmacological properties of

beetroot and its therapeutic uses for human health. Some

studies have confirmed the presence of many chemical

compounds and phyto-components for example flavonoids,

saponins, essential oils, phenols, tannins, and fatty acids.

The main objective of this study is to isolate the bioactive

compound in beetroot.

The study's ethics approval was sought from the Ethical

Committee of the Institute of Pharmaceutical Science of

Peoples University of Medical and Health Sciences for

Women (PUMHSW) (Reference #: PUMHS-032-2024

The fresh beetroot (Beta vulgaris) was purchased from the

local market in March 2024, Shaheed Benazirabad, Sindh,

Pakistan, identified by Dr Tehseen for research at Peoples

University of Medical and Health Sciences for Women

(PUMHSW), Shaheed Benazirabad, Sindh, Pakistan.

The Fresh beetroot was cleaned, dried, decorticated, and

ground into coarser powder using a mechanical grinder. The

powder was weighed about 5 gm, dissolved in 100 ml of

ethanol, and then macerated for 72 hours at room

temperature. The extract was filtered using cellulose filter

paper. A 10 µg/ml sample was submitted for GC-MS

analysis at the Institute of Pharmaceutical Science Liaquat

University of Medical and Health Sciences Jamshoro Sindh

Pakistan.

The GC-MS analysis of ethanol extract of Beta Vulgaris

was performed by using Shimadzu GC 2010 comprising

AOC-20i+s autosampler, a Shimadzu single quadrupole

GCMS-QP2020 NX gas chromatograph interfaced with a

mass spectrometer (GC-MS) equipped with an Elite 5-MS

capillary column 30 m x 0.25 mm I.D. x 0.25 µm thick film

with a composition of (5% 1,4-bis (Dimethylsiloxy)

phenylene, 95% dimethyl polysiloxane). The injection mode

was normal with 0.3 sec injection port dwell time and 3

times pumping. Column Temperature was 80 ºC for 2 min.,

rising to 300 °C then held for 4mins. Linear Flow Control

Mode was maintained throughout the process, 95.8 KPa

Column Pressure, GCMS-QP 2020 with TCD detector,

Injector and detector temperature 200 ºC and 300 ºC

respectively. Helium utilized as a carrier gas with a constant

Flow rate at 1.40 ml/min and Injection Volume 1µl was

employed with Injection split mode (Ratio of 10:0). The

total flow was 18.4 ml/min. Mass spectra were taken at 1.10

kv+0.20 kv, with a scan time of 0.30 sec, and fragments

from 42 to 550 in ACQ mode with a scan speed of 2000 cm.

The solvent cut time was 3min and the total GC-MS running

time was 27 min.

GC-MS Chromatogram of ethanolic extract of Beta vulgaris

showed different peaks (figure1) which indicated the

presence of phytochemicals constituents. The mass spectra

are presented in (Figure 2). The Molecular formula,

retention time, molecular weight, and bioactivities of beta

vulgaris are shown in Table 1.

Fig 1: Shows the Chromatogram of Beta vulgaris

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Target >>

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Fig 2: shows the mass spectra of different compounds by GC-MS analysis

Table 1: Shows the name, Retention time, Molecular formula, Molecular weight, bioactivity, and Molecular structure of compounds

identified in Beta vulgaris by GCMS analysis.

1 Heneicosane 14.665 C21H44 296 Antibacterial

2 n-decyl-N'-isopropyl 12.005 C14H30N2O 242

Unknown

3 Octacosane 15.625 C28H58 394 Antimalarial

Anti-inflammatory

4 Eicosane 16.505 C20H42 282 Anti-inflammatory

5 Hexatriacontane 17.325 C36H74 506 Unknown

6 1,2-Benzenedicarboxylic acid, bis(2-

ethylhexyl) ester 17.580 C24H38O4 390

Unknown

7 Tetracosane 18.810 C24H50 338 Antibacterial

antioxidant

8

Heptacosane 19.490 C27H56 380

Cytotoxic, and

antibacterial activity

9 1,2-Benzenedicarboxylic acid,

diundecyl ester 19.785 C30H50O4 474

Unknown

10 Di-isooctyl phthalate 19.970 C24H38O4 390

Unknown

11 Eicosane, 2-methyl

21.585 C21H44 296 Carrier compound

12 Nonacosane

22.460 C29H60 408 Antibacterial

13 Tetracosane, 11-decyl 23.495

C34H70 478

Antibacterial

Bioactivity source: PubChem, Molecular Structure source: Chemdraw

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Interpretation on mass-spectrum GC-MS was conducted

using the database of the National Institute of Standard and

Technology (NIST) having more than 60,000 patterns of

different components. GC-MS chromatogram of ethanolic

extract of beetroot demonstrated mass peaks at different

retention times The unknown components were analyzed by

comparing their spectra with the known components stored

in the NIST library. After collecting, assembling, and

screening the results, 13 mass peaks were found and

resembled the compound present in the NIST library. The

identified compounds and chromatogram details are

mentioned above in Table 1.

The article's studies focus on utilizing Gas Chromatography-

Mass Spectroscopy (GC-MS) to investigate the bioactive

compounds observed in the ethanolic extract of Beta

vulgaris (Beetroot). This examination addresses the

escalating demand for novel chemical entities with potent

pharmacological properties due to the increasing resistance

to conventional therapeutic drugs. The observation confirms

the existence of many bioactive components and also sheds

light on its role in the major activity associated with beetroot

i.e. antibacterial activity. The employment of hyphenated

techniques for example GC-MS enables the accurate

identification and isolation of compounds and presents a

significant advancement inside the realm of drug discovery

within medicinal plants.

The compounds heneicosane [16] tetracosane [17],

heptacosane [18], nonacosane [19], Tetracosane, 11-decyl [20]

with retention time 14.665, 18.810, 19.490, 22.460, 23.495

found to possess antibacterial activity because of the

presence of long chain alkane compound. Most compounds

have microbial activity, this plant is mainly beneficial as

herbal medicine against antibacterial resistance.

Additionally, tetracosane [21] and heptacosane also showed

the ability of anti-oxidant and cytotoxic [22]. The compound

octacosane with a retention time of 15.625 was found to

possess antimalarial [23] and anti-inflammatory activity [24],

while eicosane at 16.505 exhibits only anti-inflammatory

activity [24]. therefore, this plant could be beneficial in the

management of inflammation. The eicosane-2methyl at

21.585 retention time is not a bioactive compound but could

work as a carrier compound [25] and serve as a reservoir in a

well-controlled environment for bioactive compounds in

drug formulation for industrial purposes.

The biological activity of Compound n-decyl-N'-isopropyl,

Hexatriacontane, 1,2-Benzenedicarboxylic acid, bis(2-

ethylhexyl) ester, 1,2-Benzenedicarboxylic acid, diundecyl

ester, Di-isooctyl phthalate is unknown.

The Importance of this research extends beyond the mere

identification of many bioactive compounds in beetroot. It

underscores the extensive capacity of natural sources in drug

development and the pivotal role of advanced analytical

methodologies in uncovering novel bioactive compounds.

The GC-MS evaluation was carried out to exemplify how

cutting-edge scientific tools can unveil hidden therapeutic

agents inside natural sources. Employing elucidating the

presence of many compounds and linking them to the

antimicrobial properties of beetroot, this examination not

only contributes to the understanding of the

pharmacological and therapeutic properties of this plant but,

additionally, exemplifies the strength of analytical technique

in unlocking the pharmaceutical potential of natural

substances.

In the last decades’ medicinal plants have been the subject

of drug discovery. It caught the interest of researchers

worldwide to discover new lead compounds with the help of

analytical techniques. GC-MS performs a vital role in

assisting the investigations and Preliminary information

which provides a reliable source for the researcher. This

research article highlights the presence of bioactive alkanes

and different compounds in Beta vulgaris. Figuring out the

compounds contributes to understanding beetroot's

medicinal chemistry diversity and promotes its potential use

in antimicrobial, anti-inflammatory, antioxidant, and

cytotoxicity applications. Furthermore, this work provides a

foundation for future investigations aimed at isolating and

characterizing other extensive compounds present in

beetroot.

The Author like to express sincere gratitude to the

supervisor Sir Tehseen Channa for his guidance advice, and

continuous assistance and support throughout the research

process. The Author also appreciates the Head of Pharmacy

Department of Peoples University of Medical and Health

Sciences for Women for Providing the Lab facilities and

encouragement and the Head of Pharmacy Department of

Liaquat University of Medical & Health Sciences for GC-

MS analysis.

None declared.

The author received no financial support for this article's

research, authorship, and publication.

 GC-MS: Gas Chromatography-Mass Spectroscopy

 AOC-20i+s: Automatic operation controller

(autosampler)

 GCMS-QP2020 NX: Equipment number

 Elite 5-MS: Column

 TCD: Thermal conductivity detector

 ACQ mode: Acquisition mode

 NIST: National Institute of Standard and Technology

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