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The Conceptual Inventory of Natural Selection (CINS) assesses conceptual understanding in an area with well-documented alternative conceptions. Much of its use is among undergraduates, but the information learned from its responses can be a useful resource for secondary teachers by eliciting alternative conceptions and leveraging these ideas during instruction. As a result, its application must be investigated at the secondary level. The present study further investigated the CINS’s internal properties by administering it to a group of students at an urban high school. A principal components analysis demonstrated inconsistencies between the original and present validations, but it provided insight into the rationale students use in answering certain items. Results also reveal how certain items may be revised for future use.