The eye spear, or an absorbent sponge-like material, has been proposed as a useful method of obtaining repeated saliva samples from infants and young children for cortisol determination. This brief report examines possible interference effects of different types of eye spears under conditions of relatively high and low cortisol levels, with or without the use of oral stimulant, and using two common assays. In Study 1, one type of eye spear was compared to passively collected drool using two different assays (EIA, DELFIA), across high and low concentrations of cortisol. No differences were found between methods for either assay or cortisol level, indicating that the spears are potentially a viable method of collecting saliva. Study 2 compared three other types of absorbent eye spears to passive drooling under the presence or absence of oral stimulant use. This study revealed that the degree of interference varied as a function of the specific type of eye spear that was employed; stimulant use had no effect. Taken together, the results raise important considerations to take into account when selecting collection materials and procedures in the measurement of salivary cortisol.