Eye glass beads are the most typical type of glass artifacts from the Spring and Autumn Period and the Warring States Period to the Han Dynasties in ancient China. They played a crucial role in cultural exchanges and technological dissemination between China and foreign countries, as well as in the origination and evolution of ancient Chinese glass technology. A large number of glass artifacts dated from the Warring States Period have been unearthed in Hunan Province, with more than 200 eye glass beads alone. Many scholars have paid attention to and conducted various studies on the glass products unearthed in Hunan Province over the past decades; however, there is still a lack of specialized scientific characterization and research on eye glass beads in general. The Chu State was one of the key regions for the origination and development of ancient glass technology in ancient China, as well as for technological and cultural exchanges. Hunan Province, located in the core area of the Chu State, is of great significance to the study of the status of Hunan in the origination and development of glass handicraft technology during the Warring States Period. In order to scientifically reveal the chemical composition systems, manufacturing techniques, provenances and cultural exchanges of eye glass beads dated to the Warring States Period unearthed in Hunan Province, eleven typical eye glass beads from Changsha, Changde and Hengyang of this period were selected for scientific analysis. A portable energy dispersive X-ray fluorescence spectrometer and a 3D digital microscopic system were used for non-destructive analysis of the chemical composition, surface and cross-sectional micro-structures. Based on a combination of the obtained scientific information with previous relevant research results, this study discusses and determines the flux system, manufacturing techniques and provenance of this batch of eye glass beads.  Based on these results, it was found that the laminated inlay manufacturing technique was used for all the eleven eye glass beads. The matrix was first produced, and then inlaid with different glass strips or slices during the incomplete solidification state. In terms of the chemical composition system, the eleven eye glass beads belong to three systems:soda-lime silicate glass, lead-barium silicate glass and potash-lime silicate glass. Among them, three eye glass beads are natron sub-type soda-lime glass, and the one flat eye glass bead is plant-ash sub-type soda-lime glass. Both the two types of glass beads were probably produced in Egypt or eastern Mediterranean area, and were then transported into Hunan via the land Silk Road. Another three eye glass beads are lead-barium glass, and the remaining four beads are potash-lime glass. Both of the types are typical native Chinese glasses during the Warring States Period. These seven eye glass beads were produced by local craftsmen in the Chu State using two characteristic native Chinese formulas. However, they also drew on and imitated the shape and laminated inlay manufacturing technique of eye glass beads imported from abroad. At the same time, the potash-lime glass beads were also influenced by the technology of traditional Chinese potash-rich faience beads, demonstrating the exquisite craftsmanship and boundless creativity of ancient Chinese artisans. The research in this article, to a certain extent, scientifically reveals the main flux system and manufacturing techniques of typical eye glass beads unearthed in Hunan Province dated to the Warring States Period, and explores the possible provenances of these glass beads. The results could provide a valuable perspective on the development level of glass handicraft technology in the Chu State and surrounding areas during the Warring States Period, as well as on regional and Sino-foreign exchanges.