Plant material (Ocimum americanum, Ocimum basilicum) was purchased from local nursery of district Amravati from Maharashtra. Plant material (Ocimum kilimandscharicum, Ocimum gratissimum, Ocimum sanctum) was purchased from Nagarjun Garden Dr. P.D.K.V. Akola. Identification and authentication was made from H.O.D, Botany department, Government Vidarbha Institute of science & Humanities, Amravati (Maharashtra) Phamacognostic evaluations Macroscopic evaluation Macroscopy of whole plant was studied by observing the organoleptic characters such as color, odor, size, shape, taste and special features including touch and texture etc. Organoleptic evaluations can be done by means of organs of special sense which includes the above parameters and thereby define some specific characteristics of the material which can be considered as a first step towards establishment of identity. 25-26 Microscopic evaluation Microscopic evaluation is very helpful in the initial identification of herbs as well as in detection of adulteration by characteristics tissue features. Every plant possesses a characteristics tissue structure which can be demonstrated through study of tissue arrangement, cell walls and configurations, when properly mountain in stains, reagents and media. Thus it determines the size, shape, and relative structure of different cell Microscopical parameters ABSTRACT There are about 150 species of Ocimum in the world and out of that 68 species are found in India. Comparative pharmacognostic study of these Ocimum species is unspecified. Growing demand of Ocimum plants demands quality standards for correct identification of desired Ocimum species. Objectives: Hence aim of present study is to establish comparative pharmacognostical, phytochemical and biological standards for most commonly found and morphologically confusing five species of Ocimum. Methods: Macroscopic, microscopic, preliminary phytochemical evaluations, extraction of essential oils, TLC analysis, in vitro antioxidant and antimicrobial potency of selected five species carried out and compared. Results: This comparative study reports that O. sanctum should be preferred in medicine use among selected five species based on phytochemical composition, antioxidant and antimicrobial potency.