~ 130 ~ Journal of Pharmacognosy and Phytochemistry 2015; 3(6): 130-139 E-ISSN: 2278-4136 P-ISSN: 2349-8234 JPP 2015; 3(6): 130-139 Received: 15-01-2015 Accepted: 02-02-2015 Aveen Nozad Adham Department Of Pharmacognosy, College Of Pharmacy, Hawler Medical University, Erbil, Kurdisatn Region, (IRAQ). Correspondence: Aveen Nozad Adham Department Of Pharmacognosy, College Of Pharmacy, Hawler Medical University, Erbil, Kurdisatn Region, (IRAQ). Department Of Pharmacognosy, College Of Pharmacy, Hawler Medical University, Erbil, Kurdisatn Region, (IRAQ). Comparative extraction methods, phytochemical constituents, fluorescence analysis and HPLC validation of rosmarinic acid content in Mentha piperita, Mentha longifolia and Osimum basilicum Aveen Nozad Adham Abstract The present investigation is aimed to compare the percentage value of Mentha Piperita, Mentha longifolia and Osimum basilicum extracts by using different solvent and extraction methods, phytochemical screening of constituents present in dry leaves, also to study fluorescent characteristics of leaves powder which are responsible for the medicinal properties of the plant using various standard tests and HPLC analysis of rosmarinic acid content in three labiatae species. The highest percentage extract was yielded from Mentha longifolia (18.5%) by using maceration methods and ethanol 80% solvent. Phytochemical analysis revealed the presence of alkaloids, saponins, phenols, flavonoids, terpenoids, cardioactive glycosides, tannins, carbohydrates in dry leaf extracts. Presence of alkaloids and phenolic acid was determined in high quantity in Mentha longifolia, while saponins present in high quantity in Mentha Piperita and flavonoids in Osimum basilicum. The fluorescent characteristics of leaves powder with various chemical reagents were noted under visible and UV light. HPLC analysis of dry leaf extracts revealed presence of rosmarinic acid among the investigated plants and in higher percentage in Osimum basilicum (0.306%). Keywords: Labiatae species; phytochemical constituents; Fluorescence analysis; HPLC validation; Rosmarinic acid. 1. Introduction The Labiatae family is one of the most employed medicinal plants as a worldwide source of spices and also as a consolidated source of extracts with strong antibacterial and antioxidant properties. Within this family, the genus Ocimum and Mentha provides various species, and Ocimum basilicum is one of the more than 60 Ocimum species [1, 2] . The Ocimum basilicum is an annual herb which grows in several regions around the world. The plant probably originated in India, Afghanistan, Pakistan, Northern India and Iran, and now is cultivated worldwide. Traditionally has been extensively utilized in food as a flavoring agent, and in perfumery and medical industries [3] . The leaves and flowering tops of the plant are perceived as carminative, galactogogue, stomachic and antispasmodic in folk medicine [4] . However, recently the potential uses of O. basilicum essential oil, particularly as antimicrobial and antioxidant agents have also been investigated [5, 6] . Externally, they have been applied for the treatment of acne, loss of smell, insect stings, snake bites, and skin infections [7] . The activity is chiefly attributed to a variety of phenolic compounds and composition of essential oil. The main compounds responsible for typical aroma are chavicol methyl ether (estragol), linalool, eugenol, 1, 8- cineole and methyl cinnamate [8] . The non-volatile compounds were found to be rich in phenolic acids with major part of caffeic and rosmarinic acid [9] . The genus Mentha belonging to the family Labiatae, and consists of about 25-30 species, are herbaceous, perennial plants, widespread distribution in Iraq [10] Europe, Australia and South Africa [11, 12] . Mentha longifolia which can reach up to 1.5 m high in favourable conditions [10] . Mentha piperita which can reach 100 cm in height (40 inches) has four-sided stem [13] . Various biological activities have been reported for species of Mentha, such as antibacterial [14, 15] , antifungal [16] , and insecticidal properties [17, 18] . Mentha spp possesses antioxidant properties due to the presence of active constituents like menthone, menthol, flavonoids, rosmarinic acid and carvone [19] . The aim of this research is the evaluate the influence of using different extraction methods on the yield of extracted substances, also phytochemical study, fluorescence analysis of active constituents and HPLC for rosmarinic acid content in Mentha Piperita, Mentha longifolia and Osimum basilicum and compare the results among these three labiatae species.