1.       Introduction

Plants containing bioactive phytochemicals have long been recognised as medicinal plants [1]. Their therapeutic properties are attributed to the occurrence of phytochemicals in different plant parts such as leaves, stems, and roots [2]. The phytochemicals in the different parts of plants often varies significantly between different plant organs, which explains why traditional healers selectively used specific parts of a plant as a cure for ailments [3]. This practice emphasizes the importance of understanding both the type and distribution of phytochemicals within plants to fully harness their medicinal potential [4].Phytochemicals extracted from plants have demonstrated a wide range of therapeutic properties and have historically been applied in traditional medicine [5]. In the present study, the antisickling activities of Azadirachta indica A. Juss. and Helianthus annuus L. were evaluated through both qualitative and quantitative phytochemical analyses. Comparative research indicates that the phytochemical profile of medicinal plants plays a central role in their antisickling activity [6]. Accordingly, plants containing such bioactive compounds have been classified as medicinal due to their capacity to prevent or treat specific diseases [7]. Their therapeutic efficacy depends on the presence and concentration of these phytochemicals within distinct plant parts; thus, the rational selection of the appropriate plant organ has been a common practice in traditional medicine [8].Medicinal plants remain a vital source of therapeutic agents, largely due to their diverse phytochemical profiles. Among these, Azadirachta indica (neem) and Helianthus annuus (sunflower) have been the focus of recent scientific interest, particularly within Indian research communities, owing to their rich bioactive content and traditional use.

2.       Literature Review

Azadirachta indica (commonly known as neem) has been extensively investigated for its diverse array of bioactive compounds. Among these, limonoids such as nimbolide, azadirachtin, and gedunin have been shown to exhibit potent anticancer, anti-inflammatory, and immunomodulatory effects by targeting key oncogenic and inflammatory signalling pathways [9]. Recent reviews have confirmed the presence of phytochemicals such as nimbidin, nimbin, nimbolinin, salannin, quercetin, and azadirachtin across neem’s leaves, bark, and roots, consolidating its pharmacological breadth [10]. Phytochemical screening of neem leaves has identified alkaloids, cardiac glycosides, saponins, phenols, flavonoids, and terpenoids—compounds traditionally recognised for their medicinal relevance [11]. More recently, computational modelling techniques, particularly those utilising topological indices and molecular docking, have begun to prioritise neem compounds for therapeutic exploration, thereby illustrating the growing integration of computational pharmacognosy in modern drug discovery [12, 17]. Additional reports have highlighted neem’s antimicrobial, hepatoprotective, and neuroprotective activities, thereby broadening the therapeutic spectrum associated with its phytoconstituents [18].

Helianthus annuus (sunflower) also displays a complex phytochemical profile, with both nutritional and medicinal value. A comprehensive 2024 phytochemical investigation quantified the presence of flavonoids, alkaloids, saponins, tannins, triterpenoids, glycosides, phenols, coumarins, steroids, and phytosterols in sunflower petals, with extracts exhibiting alkaloid concentrations up to 650 mg/g and flavonoid concentrations up to 300 mg/g [13]. In addition, sunflower crop residues were found to contain tannins, flavonoids, terpenes, saponins, alkaloids, and steroids, all of which are associated with antioxidant, antimicrobial, antitumour, and hypoglycaemic activities [14]. Previous investigations conducted between 2016–2017 have similarly emphasised the presence of phenolic acids, flavonoids, and tocopherols in sunflower seeds and sprouts, reinforcing their antioxidant and cardioprotective properties [15]. Furthermore, recent pharmacological reports have demonstrated significant antioxidant and anti-inflammatory activities of H. annuus leaf extracts, thereby corroborating its role as a natural therapeutic agent [16-19]. Sunflower-derived phytosterols have also been associated with hypocholesterolaemia activity, adding to its harmacological relevance [20]. Although direct evidence of antisickling activity from Azadirachta indica or Helianthus annuus is not yet documented in the literature, their phytochemical profiles provide a compelling scientific basis for further exploration. The limonoids of A. indica may interact with erythrocyte membrane proteins, potentially enhancing membrane stability and reducing haemolysis. Meanwhile, the rich antioxidant composition of H. annuus—particularly flavonoids, phenolic acids, and tocopherols—could counteract oxidative stress, a key contributor to erythrocyte sickling in sickle cell disease [21]. Importantly, oxidative stress has been directly linked to increased polymerisation of sickle haemoglobin (HbS) and consequent red cell deformation, suggesting that natural antioxidants may play a critical role in mitigating this process [22]. The limonoid-rich profile of A. indica and the diverse antioxidant phytochemicals present in H. annuus strongly suggest compelling biochemical grounds for antisickling research. Future investigations should therefore focus on targeted antisickling bioassays, isolation of active principles, in vitro and in vivo mechanistic studies, and structure–activity relationship (SAR) analysis to identify lead compounds. Such efforts may ultimately illuminate novel therapeutic avenues for the management of sickle cell disease, while also contributing to the broader field of plant-derived drug discovery [23].

 3. Materials and Methods

3.1. Collection of Plant Samples

Fresh leaves, stems, and seeds of Azadirachta indica A. Juss. and Helianthus annuus L. were collected from both cultivated farms and open fields in the districts of Raipur and Mahasamund. The plant materials were properly identified and authenticated before carrying out the phytochemical analysis. The collected leaves, stems, and seeds were cut into small pieces and air-dried in the shade for two weeks. Once dried, they were ground to a fine powder (1 mm particle size) using a grinder, and then subjected to phytochemical screening.

3.2 Preparation of Extracts

Four solvents of increasing polarity were used for extraction: ethanol, methanol, chloroform, and petroleum ether. Thirty grams of powdered seeds, leaves and stems of Azadirachta indica A. Juss. and Helianthus annuus L. were extracted with each solvent separately using a Soxhlet apparatus (250 ml) for 48 hours. The extracts were then concentrated by slow evaporation [24]. The resulting crude extracts were stored in closed containers for preliminary qualitative phytochemical analysis.

3.3  Phytochemical Screening

The extracts of each plant part were subjected to standard phytochemical tests to identify the constituents. The procedures described by Trease and Evans (1989) and Sofowora (1993) were followed. The presence of saponins, tannins, reducing sugars, alkaloids, terpenoids, flavonoids, cardiac glycosides, and anthraquinones was assessed using established methods [25, 27]. Quantitative estimation of phytochemicals (alkaloids, saponins, flavonoids, phenols, and tannins) was performed using standard methods [28–33].

4.       Result and Discussion

A.     Qualitative Phytochemical Analysis

II Qualitative phytochemical analysis of Azadirachta indica A. Juss-Qualitative phytochemical analysis of Azadirachta indica A. JUSS., regardless of the type of extract employed, found tannins, alkaloids, phenols, terpenoids, flavonoids and reducing sugars in all the components studied viz., leaves, fruits and stems; while saponins were present in the leaves and fruits; cardiac glycosides and anthraquinones in the stems (Table 1).

Qualitative phytochemical analysis of Helianthus annus LIN- In the other plant, Helianthus annus L., the study found flavonoids, alkaloids, phenols and reducing sugars in all the components viz., leaves, stems and seeds, while saponins are present in the leaves and seeds; and anthraquinones were only present in the seeds (Table 2).

B. Quantitative Phytochemical Analysis

I.                       Quantitative phytochemical analysis of Azadirachta indica A. JUSS-

The quantitative phytochemical analysis of Azadirachta indica A. Juss. revealed notable variations in the distribution of secondary metabolites across different plant parts. In the leaves, flavonoids were found in the highest proportion (10.1%), followed by phenols (5.2%), alkaloids (3.7%), saponins (2.6%), and tannins (1.18%).

Table 1: Phytochemical constituents of different extracts of the leaves, fruits and stems of Azadirachta indica A. Juss

^# EXTRACTS; * 1-Leaf 2-Fruit 3-Stem; + (Positive); - (Negative)

Table 2: Phytochemical constituents of different extracts of the leaves, fruits and stems of Helianthus annuus L.

^# EXTRACTS; *1-Leaf 2-Fruit 3-Stem; + (Positive); - (Negative)

The fruits showed a markedly higher concentration of flavonoids (20.1%) and alkaloids (8.1%), while phenols (4.4%), saponins (4.1%), and tannins (0.45%) were present in comparatively lower amounts. Flavonoids and phenols constituted the predominant phytochemicals, with fruits showing the richest flavonoid content, while stems were particularly rich in phenols (Table 3).

In the stems, phenols were the most abundant (7.1%), followed flavonoids (4.9%), alkaloids (1.4%), saponins (1.6%), and tannins (0.53%).

Table 3: Quantitative phytochemical estimation of Azadirachta indica A. Juss results are given as percentage.

II.                 Quantitative phytochemical analysis of Helianthus annus L

The quantitative phytochemical analysis of Helianthus annuus L. demonstrated distinct differences in secondary metabolite content across the leaves, seeds, and stems. In the leaves, alkaloids were the most abundant (5.7%), followed by phenols (4.59%), flavonoids (2.92%), saponins (2.85%), and tannins (0.62%). The seeds contained the highest concentration of alkaloids (14%) and notable amounts of flavonoids (5.35%) and saponins (4.18%). Phenols (3.16%) and tannins (0.42%) were comparatively lower. In the stems, the dominant compounds were alkaloids (4.3%), flavonoids (3.96%), and phenols (3.52%), while saponins and tannins were negligible or absent. Seeds exhibited the richest phytochemical profile, particularly in alkaloids, while leaves showed a more balanced distribution of metabolites, and stems presented moderate levels of alkaloids, phenols, and flavonoids (Table 4).

Table 4: Quantitative phytochemical estimation of Helianthus annuus L results are given as percentage.

5. Conclution:

The antisickling activities of Azadirachta indica and Helianthus annuus were examined using analytical methods involving of phytochemical evaluations of these plants. The antisickling effects recorded at different concentrations of extracts from leaves, stems, fruits, and seeds were directly related to their phytochemical contents. Screening showed a relatively higher presence of phenols, flavonoids, saponins, and alkaloids in many parts, while tannins, glycosides, and reducing sugars occurred in moderate quantities. These observations agree with earlier reports which highlighted the importance of secondary metabolites in exerting several biological activities, especially antioxidant, antimicrobial, and antisickling effects.