The World Health Organization recommends measurement of glucose-6-phosphate dehydrogenase (G6PD) activity before initiation of 8-aminoquinoline therapy. A new drug for malaria prophylaxis and treatment (tafenoquine) is contraindicated in patients with G6PD deficiency or unknown G6PD status given its prolonged half-life. Assessments of percentage of normal G6PD activity using laboratory-specific result distributions are not widely available, making tafenoquine-eligibility decisions potentially challenging. Using an institutional review board-exempt protocol, a data set of quantitative G6PD results was retrieved from a national reference laboratory. G6PD testing was previously performed at 37 °C using an automated enzymatic assay configured on a Roche cobas c501 chemistry analyzer. Overall, 52,216 results from patients 18 years and older and 6,397 results from patients younger than 18 years were obtained. A modified adjusted male median of 12.7 U/g Hb was derived for adult males in this assay configuration. Result distributions showed higher G6PD activity in neonates. Retrospective data analysis can be used to determine laboratory-specific normal G6PD activity values in clinical populations and thus can assist in clinical-eligibility considerations for 8-aminoquinoline treatment.