Since the dawn of human civilization, many medicinal plants have been used as a source of a wide range of bioactive compounds. They have been used as pure, isolated natural compounds or as raw plant materials in the treatment of various diseases due to their content of bioactive compounds and their high efficacy even at low concentrations. Their importance in medicine is increasing and with the continued growth of biodiversity, they play a significant role in meeting this need. (Fotsing et al., 2022).  
       
In Iraq, it is called Water Hyssop or Stream Hyssop because it grows on the edges of rivers and streams and in India, it is called Hyssop, Ashnan Dawood, or Brahmi (Bharti et al., 2013). It is also called wild barberry (BARBIN BARRI) and it is widespread in marshes, wetlands and warmer regions of the world in China, Sri Lanka, throughout India and in Iraq-Basra. The genus Baccopa includes about (100) species, one of which is found in Iraq. It was formerly classified under the family Scrophulariaceae, but DNA-based molecular genetic analyses have identified it under the family plantaginaceae (Olmstead et al., 2001). Aublet (1775) was the first to describe the genus Baccopa and Austin (2004) stated that the term Baccopa was derived from a latinized form of the original name given by the indigenous Caribbean people (American Indians) in French Guiana. The type species was named Baccopa aquatica and the species name monnieri was given in honor of the French naturalist Louis-Guillaume Le Momier (Nadkarni, 1988). It has been used for centuries in Ayurvedic medicine as a memory and learning enhancer, sedative and anti-epileptic. It is used as an antioxidant, antimicrobial, antipyretic and anti-inflammatory agent and in the treatment of asthma, epilepsy, splenomegaly, leprosy, hoarseness and as a cardiac tonic (Patil et al., 2014). In India and Pakistan, it has been used to improve digestion and gastrointestinal function (Bammidi et al., 2011). It has also been used as an antidepressant, anxiolytic, anticonvulsant, antiparkinsonian agent and analgesic and is used to relax smooth muscles. Its pharmacological properties are attributed to the presence of bacosides and triterpenoid saponins (Gayathri et al., 2021). B. monnieri also contains saponins, alkaloids, flavonoids, phenylprepanoids, triterpenes, amino acids, sugars and sterols (EDA, 2024).
       
It possesses a wide range of biological and medicinal properties. It is anti-inflammatory (Balakrishnan et al., 2010), antibacterial (Malathi et al., 2011), antioxidant (Ashokkumar et al., 2008), antitumor (Cheong and Teoh, 2014), antidiabetic (Balamurugan et al., 2011) and has antipyretic and antitussive properties (Forestieri et al., 1996). It is also used to treat respiratory diseases and joint pain. These medicinal activities are attributed to the presence of secondary bioactive compounds, as it contains a wide variety of chemical compounds such as sugars, triterpenoids, flavonoids, phenols, steroids, essential oils, resins, tannins and others. It is widely used as an herbal beverage in the treatment of numerous ailments. (Khdera and Saad, 2024).

Based on the method of Washeeh et al., (2024), the chemical compounds were extracted from ground plant leaves with some modifications:-
1- The leaves of the plant species under study were thoroughly washed with distilled water to remove debris and dust, dried with a towel and left for several days to dry at room temperature and away from light.
2- The leaves were ground using an electric grinder for 10 minutes in order to obtain a fine powder.
3- 2 grams of ground leaves were extracted and dissolved in 20 ml of 99% methanol. The extract was then shaken vertically for 20 minutes and left for 9 hours at a dark room temperature.
4- The extract was filtered using a fine filter with openings of 0.45 micrometers and connected to a medical syringe.
5-  The extract was concentrated and the water was expelled from it by adding (99%) hexane in a volume of 1 ml.
6- The floating portion of the filtrate was separated from the water using hexane, then collected and its chemically active compounds were identified. The mass spectra of the separated compounds were interpreted using GC-MS based on the National Institute of Standards and Technology (NIST) data library, which contains more than 62,000 known spectra.

The results of the current study of the chemical compounds of the leaf extracts of the two species under study, which were separated and analyzed using GC-MS technology, showed an abundance of chemical content both quantitatively and qualitatively. 35 chemical compounds were recorded for both species under study and their presence varied between phenols, terpenes, fatty acids, amino acids, esters, alkanes and steroids, (Table 1, 2).




       
The leaves of B. monnieri contained (15) compounds (Table 1 and Fig 1), while the leaves of P. nodiflora contained 20 compounds (Table 2 and Fig 2). The current study recorded a variation in the chemical compounds, as they differed in their concentrations, retention times and the types of chemical compounds present and their distribution between the two species under study. One phenolic compound phenol, 2,2′-methylenebis [6-(1,1-dimethylethyl)- methyl-,was identified in the species B. monnieri, at a concentration of 2.42% at a retention time of 25.165 minutes, as well as one phenolic compound, 4-Mercaptophenol, in the species P. nodiflora, at a concentration of 7.94% at a retention time of 9.620 minutes (Table 1 and 2).




       
In Table (1,2), 6 fatty acids were recorded. 3 compounds were recorded in the type B. monnieri and the highest relative abundance was for the compound Tricosanoic acid at a concentration of 4.24% at minute 20.361 and the lowest relative abundance was for the compound 9,12,15-Octadecatrienoic acid, ethyl ester, (Z, Z, Z)- at a concentration of 3.17% at minute 21.651. Likewise, 3 compounds were recorded in the type P. nodiflora and the compound pentadecanoic acid had the highest relative abundance at a concentration of 1.78% at minute 20.170 and the compound hexadecanoic acid, methyl ester had the lowest relative abundance at a concentration of 1.65% at minute 19.459. Seven ester compounds were separated and analyzed in the two species under study in Table (1, 2), distributed as follows: 4 compounds in the species B. monnieri. The highest relative abundance was found at a concentration of 12.15% for the compound Acetic acid, cyclohexyl ester at minute 4.911, while the lowest relative abundance was for the compound Phthalic acid, isobutyl 4-methylpent-2-yl ester at a concentration of 5.31% at minute 28.125. 3 compounds were recorded in the species P. nodiflora. The compound phthalic acid, di (oct-3-yl) ester showed the highest relative abundance at a concentration of 3.60% at a retention time of 26.368 minutes, while the lowest relative abundance was recorded for the compound Methy lmethylphosphonofluoridate at a concentration of 1.69% at a retention time of 13.904 minutes.
       
Regarding steroid compounds, the results in Table  (1,2) show the presence of one steroid compound in B. monnieri, namely Cholistan-22(26)-epoxy and one compound in P. nodiflora, namely Cholistan-22(26)-isoepoxy. The highest relative abundance was for the compound Cholistan-22(26)-epoxy in B. monnieri, with a concentration of 15.60% at a retention time of 17.401 minutes (Fig 3).


       
The current study, through chemical analysis using gas chromatography-mass spectrometry (GC-MS) of plant extracts from the leaves of two species, *B. monnieri* and *P. nodiflora*, collected from plants found in the Shatt al-Shamiya area of Diwaniyah Governorate, isolated and characterized them based on their chemical content. The results showed an abundance and diversity of chemical compounds, which reinforces the medicinal importance of the two species under study (Fig 4).


       
Forms of percentage values for the relative concentrations of shared active secondary compounds among the studied species.
       
Both the Plantaginaceae and Verbenaceae families, to which the two species under study belong, have a rich and diverse chemical content of chemical compounds. Zaharie et al., (2022) and reported that genera within the first family contain a wide range of chemical compounds, including flavonoids, iridoids, terpenoids, phenolic acids, sterols and alkaloids. As noted in the studies by Rahmatullah et al., (2011) that the Verbenaceae family is rich in secondary chemical compounds such as triterpenoids, flavonoids, chromomoric acid derivatives, quinones, apocarotenoids and lignans. These compounds are associated with the biological activities of species within both families, a finding confirmed by the current study on both B. monnieri (Plantaginaceae) and P. nodiflora (Verbenaceae).
       
Regarding phenolic compounds, one phenolic compound was recorded in B. monnieri that is not found in P. nodiflora, which also contains one phenolic compound not found in the first type. The current study agreed with several researchers, including Parameswaran et al., (2018), in their study identifying the bioactive compounds present in the B. monnieri plant, which is widespread in India, using GC-MS technology in the presence of one phenolic compound. It did not agree with the study by Valotto et al., (2024), which included an analysis of previous studies on the neuroprotective effects of B. monnieri, focusing on neuroprotective mechanisms associated with the presence of the main active compounds, namely Bacosides.
       
Regarding terpene compounds, the current study also showed an important feature in the isolation and identification of the two species, through which the B. monnieri species can be isolated by the presence of one compound not found in the P. nodiflora species, which can be distinguished by the presence of 5 compounds not found in the first species. It was also observed that the two species share two compounds, namely squalene and Vitamin E (Fig 5, 6), with different concentrations and retention times. The B. monnieri species was distinguished by the highest relative abundance of the Squalene compound, as its concentration reached 7.44% at a retention time of 28.973 minutes. The highest relative abundance of the Vitamin E compound was also in the B. monnieri species, at a concentration of 3.57% at a retention time of 33.344 minutes. The presence of common compounds among plant species belonging to different families may be due to the fact that secondary metabolites are produced by plants and accumulate within them in response to abiotic and biotic stresses. This enhances the plant’s defense mechanisms and its ability to adapt. This may lead to the appearance of similar compounds in different plant species as a result of the similarity of the metabolic defense responses against stresses, since they are found in similar environmental conditions in the study area. This was confirmed by Khan et al., (2025) and Vyas et al., (2020).




       
The current study also agrees with that of Altaf et al., (2025); Mohamed et al., (2024) in their investigation of the antioxidant and anticancer effects of ethanol extract and n-hexane extract of B. monnieri. GC-MS analysis revealed the presence of several terpene compounds, including squalene and vitamin E, which were also identified in the current study for the aforementioned species. Furthermore, the study by Jain et al., (2017) concurred with their findings regarding the presence of vitamin E.
       
The current study also agrees with the study by Attanayake et al., (2025) in India, which identified squalene in their investigation of the potential bioactive compounds for therapeutic applications in P. nodiflora using GC-MS. The current study agreed with many studies on the appearance of fatty acids, including the Subashri and Pillai (2014) study in India on the type B. monnieri, despite their differences in the numbers and types of compounds recorded, as (23) compounds were recorded, including tannins, saponins, quinones, terpenes, steroids, flavotoids, alkanes, glycosides, coumarins, anthocyanins and betacyanins, but it agreed with them on the appearance of the fatty acid Hexadecanoic acid, methyl ester. The study by Attanayake et al., (2025) also agreed with it in the appearance of the above compound, which is one of the important antibacterial compounds. It also agreed with the study by Seenivasan et al., (2024); Sinkar et al., (2020) in the appearance of the above compound in the P. nodiflora species. This was confirmed by Truong et al., (2019), who stated that variations in the types and proportions of aqueous-alcoholic solvents used lead to differences in the concentrations and types of chemical compounds recorded.
       
As for the ester compounds, the B. monnieri species was isolated with four ester compounds that were not observed in the P. nodiflora species, which was distinguished by recording two compounds that were not recorded in the first species. Both species shared the recording of the Phthalic acid, di (oct-3-yl) ester compound (Fig 7) and the B. monnieri species was distinguished by the highest relative abundance of it at a concentration of 8.32% at a retention time of 26.350 minutes. The species B. monnieri contained two alkane compounds not observed in P. nodiflora, which contained four compounds absent in the former. This finding aligns with the findings of Subashri and Pillai (2014), who also identified terpene and alkane compounds in their investigation of *B. monnieri*’s antioxidant activity using extracts in different solvents, analyzed by GC-MS. Likewise, the B. monnieri species was distinguished by a single steroid compound not found in the P. nodiflora species, which in turn also contained a single steroid compound not found in the first species.


       
Mohammed and Al-Obaidi (1989) confirmed that the stronger and more widespread the root system in the soil, the greater the absorption of water and nutrients, thus increasing photosynthesis and the production of secondary metabolic products. They also noted that the type of column used in the GC-MS instrument has a significant impact on the types of compounds separated. The choice of column depends primarily on the type of solvent used and the polarity of the extract. A non-polar column is used to separate non-polar compounds such as esters, alkanes, fatty acids and terpenes, while a polar column is used to separate polar compounds such as alcohols and phenols. For both types of compounds, a general column consisting of 5% polar and 95% non-polar fractions is used. The general column used in the current study was used.

The study concludes that the richness of the two species in the presence of various chemical compounds, including phenols, terpenes, fatty acids, amino acids, esters, alkanes and steroids. The leaves of the species B. monnieri contained 15 compounds, while the leaves of the species P. nodiflora contained 20 compounds. Some of these compounds appeared in the species B. monnieri, while they were lost from the species P. nodiflora and vice versa, as they appeared in the second species and were lost from the first,

The authors declare that there is no conflict of interest.