1. INTRODUCTION:

Complimentary medicine continues to play a very important role in treating a variety of ailments using plants that date back to the dawn of civilization. People from all over the world have benefited from complementary medicine for a long time¹. A number of historical, cultural, and other factors have contributed to the rise in popularity of folk medicine in recent years, particularly in developing countries².

It is important to note that there are many scientific studies being conducted related to medicinal plants, however, this lack may have significant consequences. In order to better understand and assess the therapeutic effects of herbs, herbal research involves isolating and elucidating plant chemical structures³.

In order to achieve greater recoveries, less solvent intake, and a more precise analysis of these active herbal constituents, the experimental design needs to be improved as a result of an increasing approach to immediately identifying active phytochemical constituents from various matrices⁴. To study plant active chemicals, many extraction and analytical methods have been developed, such as spectrophotometers, high-performance liquid chromatography, capillary electrophoresis, and gas chromatography⁵.

The efficacy of herbal medicine was unlike that of conventional medicine. Plants have been studied for biological function only 6% of the time, and phytochemicals have only been explored a few times⁶. Accordingly, many medicinal plants would benefit from further research into their activity and pharmacological properties to better understand their use and benefits⁷. A gas chromatography mass spectrometric technique was utilized for the hydro chemical analysis of the phytochemical components, which was further followed by qualitative and quantitative assessments of the individual components⁸.

1.1 Mass spectroscopy (MS)based on following principles:

An analytical method involves determining the mass-to-charge ratio of electrically charged species. Various species' mass is determined by it⁹. There are various processes involved in MS that can be used to detect the mass to charge ratio of chemical components by means of ionizing them in order to produce charged species or molecules or their fragments¹⁰.

1.2 Gas Chromatography (GC) is Based on the Following Principles:

As a result of partitioning performance between stationary and mobile phases, chromatography separates mixtures of components¹¹.The chromatographic separation of volatile substances is one of the most common chromatography processes. An absorbent solid is coated with helium, which acts as the mobile phase. The stationary phase is a liquid that has a high boiling point. Analytical chemistry relies heavily on it¹². Mobile and stationary phases separate mixtures of different substances. Mobile-phase material is attracted to and passes through the stationary phase as it is attracted to the stationary phase¹³.

1.2.1 Basic principle of Gas chromatography (GC)- Mass spectroscopy (MS):

1. The injection of sample solution into the inlet of a gas chromatograph, followed by the vaporization of the sample solution into a chromatographic column by a carrier gas¹⁴.

2. As the sample moves through the column (mobile phase), the compounds that make up the mixture of interest interact with the column coating (stationary phase).

3. At the inlet of the ion source, where the eluting chemicals are converted into ions, the final segment of the column passes through a heated transfer line¹⁵.

1.3 Analyzing phytoconstituents in herbal plants with GC-MS:

1.3.1. Alkaloids are determined as follows:

· After fifteen days at room temperature, Datura stramonium leaves were dried to a fine powder and packed in an airtight container to prevent humid effects. After this stage, the container was stored at room temperature. Analyzing Daturastramonium extract with a GC-MS instrument (Agilent 7890A series, USA) was conducted using the ethanol extract. Identifying the alkaloids in the narcissus as being alkaloids of the Amaryllidaceae family will yield galanthamine, lycoramine, narwedine, haemanthamine, and tazzetine¹⁶.

· In order to analyze Withania somnifera and Withania obtusifolia using a Perkin Elmer Clarus 500 gas chromatography system equipped with an Elite-5 capillary column (30 nm x 0.25mm) accompanied by a turbo mass gold mass detector, helium was utilized as a carrier gas¹⁷. A temperature of 200°C was used for the injector, and a temperature of 600°C was used for 15 minutes before progressively increasing to 270°C for three minutes afterward. Using the mass spectra, the components of the sample were identified using Wiley and NIST libraries¹⁸.

· Isoquinoline alkaloids are found in Fumariaagraria. According to gas chromatography and mass spectrometry, the iso-quinoline alkaloid in Fumariaagraria consists of protopine, cryptopine, sinactine, stylopine, bicuculline, adlumine, perfume, andfumariline¹⁹.

· In an 7890 A GC-MSD mass spectrometry system, Pyrrolizidine alkaloid in honey was determined using a 5975 C mass spectrometry detector. It was heated to 250°C at the injection port, 300°C at the interface, 230°C at the source, and 150°C at the quadrupole, respectively²⁰. It was 70 eV with electron collision ionization. Injection of two microliters of material at a split ratio of 10:1 was used to separate compounds on the DB-5 MS column. As a carrier gas, helium was used²¹.

· As a result of operating in EI mode on a Shimadzu GC-MS-2010 instrument, alkaloids isolated from epipremnumaureum were found at a potential of 70 eV. This experiment was performed with a Restek-5MS column (30m x 0.25mm x 0.25m) from Restek. I set the oven temperature to 25°C at 5°C min-1 for a period of 5 minutes and held it at 250°C for another 5 minutes²². I then extended it to 280°C at 10°C min-1 and held it for 10 minutes. The injector's temperature was 2500C when the injection mode was in a typical injection mode²³. By comparing mass spectral data from real compounds with literature and real substance data, helium has been discovered as a carrier gas for alkaloids ²⁴.

1.3.2. Flavonoids are determined as follows:

· Isoliquiritegenin, liquiritegenin, formononetin, vestitol, neovestitol and medicarpin were determined using GC in seven samples of Red Cuban Propolis.

· Using PerkinElmer GC CLARUS 500 equipment, the flavonoids in amaranthuscaudatus were analysed with an elite fused silica capillary column that was operated in electron impact mode with 70 eV electrons. Helium was used as a carrier gas for the experiments, and 0.5 EI of injection were injected into the column (a split ratio of 10:1). The injector temperature was 250°C and the ion source temperature was 280°C. It was preset for a temperature rise of 110°C to 200°C for 2 minutes, then 5°C/min to 280°C for 9 minutes, followed by another 9-minute isothermal at 280°C. Several pieces were scanned at 0.5 s with a scan distance of 70 eV and sizes ranging from 40 to 550 Da.

· After consuming cranberry juice for two weeks, GC-MS analysis shows that there are anthocyanins detected in the plasma after consumption of flavonoids²⁵.

1. 3. 3. Terpenoids are determined as follows:

· A GC-MS analysis revealed that cannabis sativa contains monoterpenes, myrcene, sesquiterpenes, β-caryophyllene and α-humulene.

· An indole terpenoid alkaloid of Rhazya Stricta was found by using GC-MS for determining the alkaloid, which was derived from terpenoid glycosides²⁶.

1.3.4. Glycosides are determined as follows:

To quantify the saponin content in Cassytha fiiformis leaves, we used a GC-MSD apparatus and split injection system (50:1). Gas chromatography was performed using an Agilent 19091S-433HP-5MS capillary column with a 0.25mm inner diameter and 0.25-micrometer phase thickness²⁷.

2. CONCLUSION:

In recent years, herbal plants, the basis of traditional herbal medicine, have been used for very useful pharmacological studies which have served as a guide to the acceptance value of herbal plants as effective sources of new chemical compounds with therapeutic properties; these studies have been very useful in determining their acceptance value as a source of new chemicals. An analysis of GC-MS was used to identify the biologically active compounds in herbal plants, including alkaloids, flavonoids, terpenoids, and glucosidic compounds. Due to the presence of these phytoconstituents and biologically active compounds identified by gas chromatography in this study, herbal plants may be a new effective source of chemicals.

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Received on 30.08.2022 Revised on 12.10.2024

Accepted on 17.05.2025 Published on 08.11.2025

Available online from November 13, 2025

Research J. Pharmacy and Technology. 2025;18(11):5601-5604.

DOI: 10.52711/0974-360X.2025.00808

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