This study tests whether native speakers of American English exhibit a glide-vowel distinction ([j]-[i]) in a speech elicitation experiment. When reading sentences out loud, participants’ pronunciations of 4 near-minimal pairs of pre-existing lexical items (e.g., Eston[iə] vs. pneumon[jə]) exhibit significant differences when acoustically measured, confirming the presence of a [j]-[i] distinction. This distinction is also found to be productively extended to the production of 20 near-minimal pairs of nonce words (e.g., Súmia → [sumiə] vs. Fímya → [fimjə]), diversified and balanced along different phonologically relevant factors of the surrounding environment. Multiple acoustic measurements are compared to test what aspects most consistently convey the distinction: F2 (frontness), F1 (height), intensity, vocalic sequence duration, transition earliness, and transition speed. This serves the purpose of documenting the distinction’s acoustic phonetic realization. It also serves in the comparison of phonological representations. Multiple types of previously proposed phonological representations are considered along with the competing predictions they generate regarding the acoustic measurements performed. Results suggest that the primary and most consistent characteristic of the distinction is earliness of transition into the following vowel, with results also suggesting that the [j] glide has a greater degree of constriction. The [j] glide is found to have a significantly less anterior articulation, challenging the application of a representation based on place or articulator differences that would predict [j] to be more anterior.