¹Department of Biochemistry, Gujarat University, Ahmedabad, Gujarat, India
²Department of Life Sciences, Food and Nutrition Science, Gujarat University, Ahmedabad, Gujarat, India

*Corresponding author:Trivedi S, Department of Biochemistry, Gujarat University, Ahmedabad, Gujarat, India. Email: savitri_trivedi@yahoo.com

Copyright: ©Trivedi S, et al. 2024. This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

Article Information:Submission: 20/05/2024; Accepted: 13/06/2024; Published: 18/06/2024

Medicinal plants are a gift to humankind as they play a significant role in preserving, maintaining, and improving our health. Aerva lanata has been known for decades for its various pharmacological activities. Despite extensive research on its medicinal properties, there have been limited studies focusing on its hepatoprotective activity. This study aims to explore the bioactive components of Aerva lanata and investigate its potential hepatoprotective properties. The review utilized various scientific databases, including PubMed and ScienceDirect, to gather information on the pharmacological activities and bioactive components of Aerva lanata. The whole plant methanolic extract of Aerva lanata was subjected to preliminary phytochemical analysis, revealing the presence of alkaloids, flavonoids, phenols, glycosides, and tannins. High-Performance Thin-Layer Chromatography (HPTLC) was used to identify prominent compounds, followed by High Resolution Liquid Chromatography-Mass Spectrometry Orbitrap (HR-LCMS, Orbitrap) for detailed component analysis. The HPTLC chromatogram indicated significant spots at Rf values of 0.026, 0.075, 0.096, 0.46, and 0.82. HR-LCMS analysis revealed several medicinally important compounds, including norharman, 10-hydroxycanthin-6-one, 9-methoxycanthin-6-one, trigonelline, methylquinoline, 8-hydroxyquinoline, quercetin, kaempferol, rutin, and artemisinin. These findings support the traditional uses of Aerva lanata and suggest the potential for new drug discoveries. The study concludes that Aerva lanata contains several bioactive compounds with medicinal properties, justifying its traditional applications. Further research is needed to fully understand its hepatoprotective mechanisms and explore new therapeutic possibilities.

Keywords:Aerva lanata; HR-LCMS; Hepatoprotective; Medicinal Plants; Phytochemicals

Aerva lanata (A. lanata) is a well-known medicinal plant belonging to the family Amaranthaceae which is found all over India as a common weed [1]. It is native to Africa, tropical Asia, Madagascar and Saudi Arabia [2]. A. lanata is named differently in different regions of India like Chhaya in West Bengal, Gorakhganja in Gujarat,Pindidonda in Telugu, Buikallan in Punjab, Bhuyi in Rajasthan, Ulinai in Tamiland Paunsia in Odissa [3]. It is known to contain various alkaloids, flavonoids, proteins, amino acids, quinones and carbohydrates in different solvent extracts [4]. In recent years, it has gained prominent importance in the field of medicine because of its different pharmacological properties. Numerous studies on its extraction ingredients from different plant parts reported its antidiuretic [5], antimicrobial [6,7], hepatoprotective [8], antiinflammatory, anti-helminthic [9], anti-HIV [10] anti-cancerous [11-12], anti-diarrheal [13], anti-urolithiatic [14], hypolipidemic[15], nephroprotective [16], analgesic [17], antidiabetic [18] and antiasthmatic activity [19].

Herbal ingredients are promising choice as medicines in Ayurveda and Unani treatment since ancient times because of fewer side effects and low costs [20,21]. The potential bioactive compounds from plant were isolated and selected based on the phytochemical, taxonomic and ethnomedicinal approach [22]. A. lanata is a gift from nature because it possesses plentiful healing properties [2,23]. It

has been observed that A. lanata has been used traditionally to treat common ailments like headache, burns, jaundice, cholera, measles, gall stones, diarrhea, cough and bronchitis, snake bites, arthritis [9]. Only few studies in literature evaluated significant hepatoprotective activity of A. lanata [8,24,25]. In the present study, an attempt is made to explore the biological components of A. lanata and zooming on few components which can be the potential active components possessing hepatoprotective activity.

A. lanata was collected from natural habitats, Godhra, Gujarat, India. Plant authenticated by Dr. Bharat Maitreya (Professor, Department of Botany, Bioinformatics and Climate Change Impact Management, Gujarat University). The specimens collected was deposited for reference (No. GU/BOT/A/A21) at the herbarium, Gujarati University. The extraction of active ingredients from different parts of A. lanata was prepared using standard hot and dried methodology. Accurately Weighed 1gm of dried plant material was transferred to a 250 ml volumetric flask dissolving in 100 ml of solvent and kept on hot plate for 5 hours at 70 °C. The process was repeated until the solution become colorless. The content of the flask was filtered through Whatman No.1 paper (Merck, Mumbai, India). The residue was dried, concentrated and stored in desiccator for further analysis. The qualitative phytochemical analysis was carried out using standardized tests to identify polyphenols, tannins, flavonoids, steroids, alkaloids, saponins, terpenoids etc. [26].

An HPTLC system is equipped with a TLC scanner 3, Linomat V sample applicator and CATS V 4.06 software. All the solvents and chemicals procured were of HPTLC grade (Merck, Mumbai, India. The sample extract was prepared in 90% ethanol. The sample was applied as bands of 0.2 mm eachby CamagLinomat V sample applicator (Muttenz, Switzerland) equipped with 100 μl Hamilton syringe on 20 cm * 10 cm aluminum plates coated with silica gel 60F254 (Merck, Mumbai, India). Ascending development was performed at room temperature with Toluene: Ethyl acetate solution in ration of 9:1 (v/v) as a mobile phase in a Camag glass twin-trough chamber saturated with mobile phase vapor for 30 mins prior. The developed plates were air dried and scanned at 366 nm with a Camag TLC scanner with CATS V4.06 software, using deuterium lamp.

HR-LCMS was done to determine the bioactive compounds present in methanolic extract of A. lanata. The LC system consist of two pumps and an automated injector. Separation of components achieved in Syncronis C-18 column (Size: 5 μm, 100 mm x 4.6 mm) (Thermo Fisher Scientific, Massachusetts, USA). Before analysis, the sample was prepared by centrifuging at 12000 rpm for 10 mins. The separated components enter into QExactive Hybrid Quadrupole- Orbitrap mass spectrometer (Thermo Fisher Scientific, Massachusetts, USA) equipped with an electrospray ionization interface (ESI). This system has Xcalibur,Version 4.2.28.14 data acquisition software. Two mobile phases; A-0.1% formic acid in Milli-Q water and DAcetonitrile in Milli-Q water were used as mobile phase at a flow rate of 500 μl min−1.

The present study was designed to evaluate the phytochemical screening and HR-LCMS analysis of the crude methanolic extract fraction of A. lanata. In plant extraction process, the percentage yield of aqueous and methanolic extract of A. lanata whole plant was obtained 5.5% and 5.2%, for leaves 4.2% and 2.7%, for stem 2.5% and 3.7%, and for flower 3.4% and 2.9% w/w, respectively.(Table 2) showed the preliminary qualitative phytochemical analysis of methanolic extract of A. lanata and its plant parts, which confirm the presence of Alkaloids, carbohydrates, glycosides, phenols, protein, amino acids, and flavonoids in whole plant extract. Numerous solvent compositions were tested for HPTLC analysis, from which Toluene: Ethyl acetate (v/v) gave the desired aim results. HPTLC chromatogramof whole plant methanolic extract of A. lanata determined high resolution and reproducible peaks under 366 nm, which revealed the presence of five prominent bands with Rf value ranged from 0.026-0.82 [Figure 1].

HR-LCMS analysis of methanolic extract of A. lanata lead to the identification of 47 bioactive compounds at different retention time that could attribute towards the pharmacological properties (Table 3). The big molecules fragments into small compounds giving rise to appearance of peaks at different m/z ratio. We have identified these components by comparing with the authentic m/z spectra available in HR-LCMS data library. These identified components correspond to 80 % of the chromatogram [Figure 2]. The first predominant peak observed with less retention time 0.97 was identified as Adenine whereas Hexadecanamide was the last compound identified which took longest retention time 26.37. HR-LCMS data of A. lanata confirms that it comprises of active alkaloids such as Norharman, aervine (10-hydroxycanthin-6-one), 9-methoxycanthin-6-one and Trigonelline. The heterocyclic compounds such as 6-Methylquinoline and 8-Hydroxyquinoline also been observed. A. lanata comprises of flavanols like Quercetin and Kaempferol [Table 3]. This plant also has flavonoid glucoside named rutin and artemisinin which is a terpene. Additionally, we have identified carbohydrates, few phenolic compounds, and amino acids like phenyl alanine and tyrosine.

Phytochemicals play vital role in treating various diseases and are still used in modern medicine. The identification of new constituents is not only important for therapeutic agents but also helps in finding

new economically important components like oil, saponins, tannins, flavonoids, gums etc. [27-29]. Nowadays it becomes easier with different methods to isolate and identify the bioactive compounds present in plant. Hot and dried method of plant extraction is very old method. Among 28 species of Aerva genus, only three species A. lanata, A. javanica, A. persica are known to have the medicinal properties [30].

The results of the present preliminary study on A.lanata’s whole plant methanolic extract evidenced the presence of alkaloids, flavonoids, phenols, terpenes and glycosides. Thus, this shows that whole plant extract prepared using methanol as a solvent had the maximum bioactive compounds. These phytoconstituents could be responsible for the medicinal properties of A.lanata [31-33]. Hence, for further confirmation of these different-phytoconstituents, we conducted HR-LCMS analysis for the methanolic whole plant extract. The obtained biological molecules in methanolic extract have therapeutic values. The hexadecanoic acid, which is methyl ester of palmitic acid, is already known for its anti-oxidant, hypocholesterolemic, antis-androgenic, hemolytic, and 5-Alpha-reductase inhibitor and pesticide activity [34]. Alkaloids are known for their role in plant defense system against pathogenic organisms. Similarly, A. lanata also contains many alkaloids like 10-hydroxycanthin-6- one (Aervine), 13-methoxy-1,6-diazatetracyclo hexadeca-heptaen- 2-one, and Betaine. Presence of aervine, 9-methoxycanthin-6-one and β-carboline-like alkaloid has also been reported previously [35,36]. The compound 9-Oxo-10(E),12(E)-octadecadienoic acidfound in A. lanata as a significant role in prostaglandin biosynthesis and possesses other biological activities such as anti-inflammatory, antiarthritic and hepatoprotective [37,38]. A previous study showed that A. lanata hydroalcoholic extract was effective in protecting liver damage in rats caused by paracetamol [8,24]. Rutin, a flavanol, is known for its anti-inflammatory, anti-cancerous, neuro-protective, cardio protective and anti-oxidative activity [39,40]. One of the studies confirmed the hepatoprotective activity of alkaloid present in petroleum ether extract of A. lanata [12]. Considering the presence of different biologically active compounds in the methanolic extract of A. lanata, few compounds can be chosen for bioinformatic studies to select the best molecule which could be studied further in-vitro and in-vivo for hepatoprotective activity.

The developed HPTLC profile revealed fingerprint of phytochemicals present in A. lanata and confirming the purity and identity of the species. HR-LCMS revealed the presence of many diverse bioactive components which justifies the use of whole plant extract for treating different diseases in older times. However, a new area of research will open if isolation and identification of novel pharmacologically active compounds is conducted done. Further, Invitro and in-vivo study is needed to prove the hepatoprotective activity of those phytoconstituents present in A. lanata.