Analysis of Flavanoids in Aqueous Extract of Eugenia uniflora (L.) Leaves Using HPTLC

Most of the traditional medicinal plants in India are not scientifically validated. Scientific evaluation along with traditional knowledge is essential to obtain effective drugs for commercial purpose. The aim of the study was to establish the chemical fingerprint of flavonoid profile in aqueous extract of Eugenia uniflora leaves. Fingerprint analysis by HPTLC has become an effective and powerful tool for linking the chemical constituent profile of the plants with botanical identity and for the estimation of chemical and biochemical markers. HPTLC analysis was done and profiles were developed for authentication. The aqueous extract of Eugenia uniflora showed the presence of several flavonoids. The development of such fingerprint for leaves is useful in differentiating the species from the adulterant and also acts as biomarker for this plant in pharmaceutical industry. Geedhu Daniel and S.Krishnakumari Department of Biochemistry,Kongunadu Arts and Science College,Coimbatore,Tamilnadu,India-641029 Submission: 5 August 2016 Accepted: 10 August 2016 Published: 25 August 2016 www.ijsrm. humanjournals.com Citation: S.Krishnakumari et al. Ijsrm.Human, 2016; Vol. 4 (2): 54-65. 55 INTRODUCTION Medicinal plants are of great importance to the health of individuals and communities. The medicinal value of these plants lies in some chemical substances that produce a definite physiological action on the human body. The most important of these bioactive constituents of plants are alkaloids, tannins, flavonoids, and phenolic compounds [1]. Many of these indigenous medicinal plants are used as spices and food plants. Plants have great potential uses, especially as traditional medicine and pharmacopoeial drugs. A large proportion of the world population depends on traditional medicine because of the scarcity and high costs of orthodox medicine. Medicinal plants have provided the modern medicine with numerous plant derived therapeutic agents. Many plants contain a variety of phytopharmaceuticals, which have found very important applications in the fields of agriculture, human and veterinary medicine. Natural products play a dominant role in the development of novel drug leads for the treatment and prevention of diseases [2-4]. A large number of medicinal plants and their purified constituents have shown beneficial therapeutic potentials [5]. It has been shown that in-vitro screening methods could provide the needed preliminary observations to select crude plant extracts with potentially useful properties for further chemical and pharmacological investigations [6]. Modern methods describing the identification and quantification of active constituents in the plant material may be useful for proper standardization of herbals and its formulations. Also, the WHO has emphasized the need to ensure the quality of medicinal plant products using modern controlled techniques and applying suitable standards [7, 8] . Modern high-performance TLC (HPTLC) is an efficient instrumental analysis, and optimized quantitative HPTLC using a densitometric evaluation can produce results analogous to those obtained with gas chromatography (GC) and high performance liquid chromatography (HPLC) [9,10]. Thus, HPTLC „fingerprint analysis‟ may be a powerful tool for the quality control of raw plant material and may be an alternative technique, particularly in the analysis of crude plant extracts. An improvement over conventional TLC, HPTLC is an instrumental technique whereby special plates and instrumental resources for sampling are used and the quantitative evaluation of www.ijsrm. humanjournals.com Citation: S.Krishnakumari et al. Ijsrm.Human, 2016; Vol. 4 (2): 54-65. 56 separations is aided by densitometry [11]. HPTLC offers better resolution and estimation of active constituents can be done with reasonable accuracy in a shorter duration[12]. Eugenia uniflora L. (Myrtaceae) is a tropical and subtropical shrub widely distributed in American countries [13]. It is commonly referred to as Pitanga cherry or Brazilian cherry. Regarding their effects on human health, both fruit and leaves are used as folk medicine to treat similar diseases, although the leaves show the advantage of being perennial and continuously available, while the fruits are available during a short period of the year [14]. The fresh or dried leaves have been used empirically as medicine, since the 15 th century, for treating inflammatory and stomach diseases, rheumatism, fever, and hypertension [15, 16]. Some studies have confirmed that Eugenia uniflora possesses anti-inflammatory, antimicrobial, and antifungal properties [15, 17–19]. These benefits are usually attributed to the presence of many secondary metabolites present in the leaves, which includes many volatile terpenoid oils, flavonoids, and condensed and hydrolysable tannins, leucoanthocyanidins, and steroids and/or triterpenoids [20]. Flavonoids are present in many plant extracts, being constantly the focus of pharmacological studies. Despite their well-described antioxidant activity [21-23], they have shown many other properties, such as anti-inflammatory, antimicrobial, antiaging, anticancer, and antiallergic, hypocholesterolemic and vasodilatation [21-26]. Together, the anti-inflammatory and antioxidant properties of flavonoids can explain the efficacy of plant extracts against various diseases, such as osteoporosis and rheumatism [27]. Therapeutic properties attributed to the flavonoid contents of E. uniflora against inflammatory disorders were reported. With this knowledge, the present research work was aimed to produce the fingerprint analysis using HPTLC to profile flavonoids in leaf extract of Eugenia uniflora. www.ijsrm. humanjournals.com Citation: S.Krishnakumari et al. Ijsrm.Human, 2016; Vol. 4 (2): 54-65. 57 MATERIALS AND METHODS Plant Collection and Authentication Fresh leaves of Eugenia uniflora (Linn), FamilyMyrtaceae, were collected from Wayanad district, Kerala during the month of April 2014. Taxonomic authentication was done by Dr. V.S Ramachandran, Taxonomist, Department of Botany, Bharathiar University, Coimbatore, Tamilnadu, India.