The Araneidae is among the most speciose spider families, but there are few karyotype studies (1.9%) and some species-rich clades are without any chromosomal study. Understanding the evolution of chromosome number and Sex Chromosome Systems is made more difficult by many uncertain evolutionary relationships within the family. In this work, the chromosomal analysis of eight araneid species (Acacesia benigna Glueck, 1994, Actinosoma pentacanthum (Walckenaer, 1841), Alpaida bicornuta (Taczanowski, 1878), Dubiepeira Levi, 1991 sp., Gasteracantha cancriformis (Linnaeus, 1758), Parawixia bistriata (Rengger, 1836), Verrucosa meridionalis (Keyserling, 1892) and Verrucosa scapofracta Lise, Kesster & Silva, 2015), contribute to discussions of some evolutionary scenarios of chromosome evolution. The gonads were submitted to colchicine treatment, hypotonization, slide preparation, and Giemsa staining. The species analyzed showed 2n♂ = 24 (11II + X1X2), except Dubiepeira sp. with 2n♂ = 41 (19II + X1X2X3), and both Verrucosa species, which presented 2n♂ = 47 (22II + X1X2X3) in V. meridionalis and 2n♂ = 50 (23II + X1X2X3X4) in V. scapofracta. The species analyzed possess all chromosomes with acro/telocentric chromosomal morphology. The 2n♂ = 24, X1X2 found in most species studied here is the most frequent karyotype in the Araneidae. This study presents the first chromosomal data for the diverse clade “Micrathenines”, the highest diploid number among entelegynes (2n♂ = 50), and the first record of an X1X2X3X4 in the Araneidae. The chromosome data suggest a series of fission events in the origin of Verrucosa karyotypes, and a close relationship between Dubiepeira sp. and Araneus ventricosus (L. Koch, 1878). Moreover, Alpaida bicornuta can be cytotaxonomically distinguished of other Alpaida species karyotyped up to now.