1.1 Background of the study Nature is the primary source of man’s basic needs. It has provided him with shelter, clothing, fertilizer, flavors and fragrances and above all, food and medicine throughout the ages. Natural products have played an important role in treating and preventing human diseases. Records from traditional medical systems and modern medical practice show the invaluable role of these natural products particularly of the plant kingdom. Even before the advent of modern medical technology and the development of clinical drugs, medicinal plants have gained value in restoring and maintaining the good health of human kind. Since the ancient times, plant sources have been utilized in Homeopathy, Allopathy, and Ayurvedic medical practices and systems in countries such as India, Brazil, Chile, Mexico, Pakistan, Switzerland, and United Kingdom (Vickers & Smith, 2006). People living near these medical plant sources mostly depend on traditional medicine for treating ailments (Rasul, Saleem, & Nawaz, 1989). Plant parts were taken in as decoctions, infusions, tinctures, juices, syrups or tablets. There were also ointments and pastes directly applied to an affected area. Along with this traditional application is also the modern medical systems that multiplied the plants’ role through applications such as synthetic drug and antibiotics. Over 50 % of drugs in clinical use, together with their derivatives and analogs, are natural plant products from plant sources (Saranya et al., 2013). Few drugs developed from natural sources have undoubtedly revolutionized medicine, like antibiotics, antiparasitics, antimalarials, lipid control agents, immune-suppressants for organ transplants, and anticancer drugs (Harvey, 2008; Katiyar et al., 2012). The growing need for medicine has led to the increase in interest of studying possible plant sources of pharmacologically active compounds that can be utilized as medicines. Nowadays, a center of attraction among researchers who opt to develop new scaffolds of drugs is on seeking for plant-based products that are non-toxic, exhibit no adverse-effects, and with protective and curing properties (Shukla, Sharma,et al., 2016). These properties are determined through the analysis of health- promoting and biologically-active compounds naturally present in plants known as phytochemicals (Roberts & Moreau, 2016). As sources of direct medical agents, these phytochemicals can be derived from different parts of plants such as leaves, flowers, barks, seeds, roots, pulps, etc. They can also be raw materials for the development of more complex semi-synthetic compounds for pharmaceutical advancements (Banu & Cathrine, 2015). Hence, studies conducted for the addition to the knowledge base of characterized phytochemicals present in different plant species pose significant contribution in the field of medicine. Tiwari et.al. (2011) claims that the richest bio-resource of drugs of traditional systems of medicine and modern medicines are the medicinal plants. Information about the phytochemicals in plants are beneficial since dependence on traditional medicine is more eminent in terms of usage, despite the advent of modern medicine. The use of plant extracts and their active components as traditional medicine for primary health care need accounts to an estimated 80% of the earth’s population and 6 million inhabitants, according to the World Health Organization (Sivakumar, Moideen, Varghese, & Sheik, n.d., 2011). Thus, there is a necessity in researching information about the medicinal properties and phytochemical composition of plants. Amaranthus viridis L. belongs to Amaranthaceae family. It is a multinational genus of herbs communally known as ‘Green Amaranth’. In Greek, Amaranthus is defined as a “never- ending flower”. Some species of Amaranthus are often taken as weeds (Ahmed, Hanif, & Iftkhar, 2013). A. viridis L. is an annual herb also known as “kulitis” in the Philippines, “chilaka thotakura” in Telugu, “Chauraiya” in India and “Green amaranth” in the Americas. It is an erect, branched, and smooth plant that grows around 10 – 100 cm tall. Its leaves, about 4 – 10 cm long, are usually ovate, alternate, notched and long-petioled. Its very small, green, and densely packed flowers grow up to a millimeter long and the sepals are often ovate to linear. Also, the inflorescences are much-branched simple spikes. It can bear small indehiscent fruits with black or brown seeds. In some countries it is harvested from the wild as a food and medicinal resource (Reyad-ul-Ferdous, 2015). A. viridis L. is both a wild and cultivated pot herb (Uphof, 1968) and is rich in magnesium, calcium, iron, zinc and a significant amount of two fatty acids, linoleic and linolenic, and is a good source of vitamins B and C that are essential in humans (Morton, 1981). Furthermore, amino acid composition of this plant meets with the protein standard of WHO. In traditional system of medicine, various plant parts of A. viridis L. such as leaves, flowers, stems, fruits, seeds, barks and even the whole plant is used in the treatment of numerous diseases. Its uses include diuretic, analgesic, antipyretic, vermifuge, antiulcer, antidiabetic, anticholesterolemic, laxative, asthma and veneral diseases (Pulipati, 2014; Reyad-ul-Ferdous, Shahjahan, Tanvir, & Mukti, 2015). Countries like India and Nepal traditionally use A. viridis L. to reduce labor pains and as an antipyretic (Kirithikar and Basu, 1986; Mark Turin, 2003). According to Eduardo and Quisumbing (1951), the Negritos of the Philippines use the bruised leaves of A. viridis L. to treat skin diseases such as eczema, psoriasis and rashes. A. viridis L. (kulitis) leaves have been used in traditional medicinal such as treating diabetes, anemia, constipation, eczema, bronchitis, eye infection, skin rashes and cleansing, psoriasis, labor pains, fever, cardiovascular diseases, dysentery, acne, abscesses, venereal diseases, inflammation, rheumatism, ulcer, leprosy, gonorrhea, diuretic, purgative, orchitis and hemorrhoids (Stuart, Jr., 2016; Fern, 2017). Other traditional uses are as an anti-inflammatory agent of the urinary tract, in venereal diseases, as a vermifuge, diuretic, anti- rheumatic, antiulcer, analgesic, antiemetic; laxative, for improving appetite, as an antileprotic, for the treatment of respiratory problems, eye treatment and for asthma (Anonymous, 1988; Agra et al., 2007; 2008; Kirithikar and Basu, 1986; Hassan Sher and Khan, 2006; Quershi et al., 2008; Muhammad Ejaz Ul Islam Dar, 2003; Muhammad Arshad, 2000; Muhammad and Amusa, 2005). A. viridis L. has been extensively used as a traditional medicine implement but according to Sivagnanam and Chandra (2016), its disease-curing effects lack sufficient proof. Hence, through phytochemical analysis, the present study aims to investigate on and determine the phytochemicals present in the leaves of A. viridis L. to validate its uses in traditional medicine and further utilize its application in pharmacology. Kulitis (A. viridis L.) has been used in the treatment of insect bites, regular burns, and sunburns. In addition, the leaf extracts of this herbal plant were found to show similar antimicrobial activity against E. coli and S. aureus measuring 15 mm zone of inhibition for both strains (Malik, Nawaz, & Nisar, 2016). Researches have been found that explains the medicinal properties of A. viridis L. leaf. According to the research conducted by Girija, et. al. (2011), methanolic extract of A. viridis L. leaf has shown beneficial effects in reducing the elevated blood glucose level and body weight changes, and improves the lipid profile of Streptozotocin induced rats. The results obtained from another study indicate that the leaves of A. viridis L. is a potential source of natural antioxidants (Jadhav & Biradar, 2016). Several studies about the active constituents of A. viridis L. have also been conducted. In the study conducted by Sadia et al. (2013), qualitative phytochemical screening resulted in the presence of alkaloids, saponins, flavonoids, tannins, total phenolics and absence of terpenoids and resins in A. viridis L. leaf extract. With this, there is a need to verify its presence using the solvents aside from methanol to confirm its use in traditional medicine on dysentery and diabetes. Kulitis (Amaranthus viridis L.) Kulitis Leaves Maceration Methanol Ethanol Aqueous PHYTOCHEMICAL SCREENING Figure 1. y Another study by Ahmed Test for: et al. (2011) suggested the possibility of using the methanolic extracts in treating diseases caused by bacteria and fungi. This property can be attributed to its flavonoid content. Yet, the •extraction of different phytochemicals using other solvents must also Flavonoids – Ferric chloride Test be taken into consideration.• With this, only a limited Alkaloids – Wagner’s Testnumber of researches have been found to study the medicinal properties of A. viridis L. leaf. • Saponins – Foam Test • Terpenoids – Salkowski’s Test The multitude of uses terms of the–medicinal properties of A. viridis L. makes it sensible • inPhytosterols Salkowski’s Test for the many researches that studies it at present. However, even with many researches about “kulitis” (A. viridis L.) and its parts, the analysis of the phytochemicals present in the leaves of this plant is very minimal, and such studies do not compare the phytochemicals that can be infused from the different extracts. The researchers, therefore, conducted this study to address the need for further information on the phytochemicals present in the leaves of A. viridis L. using different Identification of Phytochemicals Present in extracts. Kulitis (Amaranthus viridis L.) Leaves The maceration extraction technique and the phytochemical screening methods will be utilized by the researchers to meet the goals of the study. Maceration is a conventional extraction technique that has become a popular and inexpensive way to get essential oils and bioactive compounds from plants (Azmir et al., 2013). The different phytochemical screening methods using the extracts – methanol, ethanol, and aqueous – will follow after the maceration process. The study of Kanerkar et. al., 2016 states that the roots of A. viridis L. are known for its antifertility activity in Ayurveda. The term antifertility many times is used loosely in ethnic literature denoting abortifacient, anti-implantation and anti-ovulatory activity. Aqueous root extract was administered orally at the dose of 50 mg/kg, 100 mg/kg and 150 mg/kg body weight respectively for 5 days from 11-15 days of pregnancy to female albino rats. The results show that the antifertility effect is expressed as abortifacient activity which is dose dependent increasing with higher dose. Methanol extracts of the dried leaves and seeds of A. viridis L. suggest the possibility of using the methanolic extracts in treating the diseases caused by the test organisms. Phytochemical investigation of the said extract determined that the leaves contain glycosides, proteins, alkaloids, saponins, and tannins (Ahmed et. al., 2013). The ethanol extract of A. viridis L. leaves contain high amount of flavonoids and phenolic contents and exhibited high antioxidant activity (Sadia et. al., 2016). Despite various studies about the different parts of A. viridis L. using a single solvent, there are few available researches regarding its active components. There are no studies that compares the presence of phytochemicals in methanol, ethanol and aqueous solvents. Knowledge of the chemical constituents of plants is desirable, not only for the discovery of therapeutic agents, but also because such information will be of value in disclosing new resources of such chemical substances. It is of great interest to carry out a screening of these plants in order to validate their use in folk medicine and to reveal the active principle by isolation and characterization of their constituent (Sivagnanam, 2016). This means that there is a gap between the type of active compounds present in the A. viridis L. and the different extracts to be used for testing the presence of these chemically active compounds. The present study finds its relevance in the context of this pertinent research gap. Scope and Limitation The study employed qualitative analysis of the phytochemicals present in A. viridis leaf extracts. This method posed limits to the study to the identification of the phytochemicals present wherein a quantitative analysis on the extracts could further present data conclusive for intensive phytochemical profiling for A. viridis leaves. The quantitative analysis of the phytochemicals in the A. viridis leaves was not pursued due to the unavailability of instruments required. While this study investigates on the phytochemicals present in the leaves of A. viridis, examination of the other parts of the plant such as the roots , stem, and flowers could also provide further data useful for comparison with the phytochemicals present in the leaves. The study was also limited to three solvents used for the extraction process (Methanol, Ethanol, and Water) whereas the results from other solvents could also be investigated. These variables were not incorporated into the research design due to constraints on the allotted research time and available resources. Scope and Limitation The study employed qualitative analysis of the phytochemicals present in A. viridis leaf extracts. This method posed limits to the study to the identification of the phytochemicals present wherein a quantitative analysis on the extracts could further present data conclusive for intensive phytochemical profiling for A. viridis leaves. The quantitative analysis of the phytochemicals in the A. viridis leaves was not pursued due to the unavailability of instruments required. While this study investigates on the phytochemicals present in the leaves of A. viridis, examination of the other parts of the plant such as the roots , stem, and flowers could also provide further data useful for comparison with the phytochemicals present in the leaves. The study was also limited to three solvents used for the extraction process (Methanol, Ethanol, and Water) whereas the results from other solvents could also be investigated. These variables were not incorporated into the research design due to constraints on the allotted research time and available resources.