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Compared with other racial/ethnic groups in the United States (US), American Indians/Alaska Natives have one of the fastest climbing rates of drug overdose deaths involving stimulants. Validating the substances self-reported by Indigenous people who use injection drugs (IPWIDs) can present logistical and cultural challenges. While the collection of biospecimens (e.g., urine, blood, hair follicle) can be one way to cross-validate the substances self-reported by IPWIDs, the collection of biospecimens has been historically problematic when conducting substance use research with Indigenous North Americans. In our National Institutes of Health (NIH)-supported pilot research conducted with IPWIDs, we have documented low willingness to provide a biospecimen to a research team. This article demonstrates an alternative method for validating self-reported substances injected by IPWIDs that does not require the extraction of biospecimens from Indigenous bodies and spaces. The method described includes: • Collecting used, unwashed syringes from IPWIDs at the time of behavioral assessment, • Sampling the used syringe by washing the syringe needle/barrel with methanol, • Analyzing the samples with gas chromatography mass spectrometry (GC–MS) and liquid chromatography coupled to triple-quadrupole mass spectrometry (LC-QQQ-MS).This method offers a more culturally appropriate alternative to validate substances self-reported by IPWIDs during behavioral assessments.