An internal validation study was conducted using the PowerPlex® 16 HS system to ensure proper performance on the Applied Biosystems 3130 Genetic Analyzer in the University of Central Oklahoma laboratory. Manual extraction with the DNA IQ^TM system was performed. The Quantifiler^TM Human Quantification kit was used to quantify the samples. Promega Corporation’s PowerPlex® 16 HS system was used to amplify DNA samples on a GeneAmp® PCR System 9700 thermal cycler. Separation occurred through capillary electrophoresis on an Applied Biosystems 3130 Genetic Analyzer. Following parameters established through the validation, an environmental study was conducted to simulate casework samples. The environmental study included ultraviolet treatment, tannic acid, humic acid, and hematin. The results support the multiplexing system is capable of handling DNA samples.