Pediatric chronic pain is common and responsible for significant healthcare burden. However, the mechanisms underlying the development and/or maintenance of pediatric chronic pain remain poorly understood. Allostatic load (AL), or wear and tear on the nervous system following significant or prolonged stress, has been proposed to play a role in the maintenance of chronic pain, but minimal research has examined this possibility. This gap in research is particularly notable given the high exposure to adverse childhood experiences (ACEs; abuse/neglect, etc.) and psychological stress in this population. Accordingly, the current study aimed to preliminarily examine the measurement of AL in a treatment-seeking pediatric pain population. Biomarkers were collected during an already scheduled new patient pain evaluation and included salivary cortisol, dehydroepiandrosterone (DHEA), and C-reactive protein, in addition to waist–hip ratio, body-mass index, and blood pressure. A total of 61 children and adolescents with chronic pain (Mage = 14.47 years; 88.5% female and white/Caucasian) completed study procedures and were included in analyses. Preliminary results indicated that a multifactorial AL composite is feasible to assess for in a tertiary pain treatment setting and that over 50% of youth with chronic pain were classified as high risk for AL (two or more risk factors). Further, it was found that individual AL risk factors were significantly associated with functional disability and that AL may moderate the association between psychosocial and functional outcomes. Given the pilot nature of this study, results should be used to inform future investigations with larger and more diverse pediatric pain samples.