The aim of present study is to discuss the taxonomic and biological history and significance of Phlebotominae sand flies giving emphasis on their collection, preservation and identification of new species as well as differentiation of male and female. Several species of Phlebotominae sand flies are known throughout the world including India. These Phlebotominae sand flies are economically important natural enemies of human beings and cause Kala-azar and visceral leishmaniasis. This causes a lot of deaths. Taxonomy plays an important role in identifying these tiny insects accurately. The accurate identification is important to apply relevant control measures. A wrong identification may lead to misleading results and may upset the entire control strategy. From the start of sand fly research various methods for collection and identification were used. In the early twentieth century, the use of internal structures particularly, cibarium, pharynx and spermathecae was introduced as taxonomic characters. The identification of female sand flies was always found to be difficult and hence scanty work on females are available. Recently female external as well as internal genital structures are used for their identification. Another important character is the genital filament/spermathecal duct ratio which could be a good point for taxonomic differentiation of male and female Phlebotominae sand flies. The detailed study of some biologically important structures of sand flies e.g., mouthparts and reproductive organs both internal as well as external have led to the introduction of few new taxonomic characters.