A single-laboratory validation was completed for a method to determine total terpene lactones in Ginkgo biloba products. The method determines terpene lactones on the basis of the main terpene lactones (Bilobalide, Ginkgolide A, Ginkgolide B, Ginkgolide C, and Ginkgolide J) by high-performance liquid chromatography with evaporative light-scattering detection after extraction. Nine matrixes were chosen for study, including crude leaf material, standardized dry powder extract, single- and multiple-entity finished products, and alcohol and glycerin tinctures. The sample purification with prepacked columns allows selective extraction of the terpene lactones with no interferences from any matrix under study. A Youden ruggedness trial testing 7 instrumental and preparation factors with the potential to affect quantitative results showed that 2 factors (volume of the column elution solvent and pH of the diluent) were the most important parameters to control during sample preparation. The method performed well in terms of precision; 4 matrixes tested in triplicate over a 3-day period showed an overall repeatability relative standard deviation (RSD) of about 3%. HorRat values were within the limits for performance acceptability, ranging from 0.5 to 1.0. Analysis of variance testing at a = 0.05 showed no significant differences among the within-or between-group sources of variation, although comparison of within-day, between-day, and total precision showed that most of the RSD came from within-day determinations except those for the Ginkgo dry extract (Gb-SLV-2). Accuracy testing at 4 concentration levels of terpene lactones obtained by spiking a negative control matrix at approximately 300, 750, 1500, and 2250 microg/mL gave recoveries of about 91% for the 300 microg/mL level, about 98% for the 750 microg/mL level, about 99% for the 1500 microg/mL level, and 97% for the 2250 microg/mL level with an overall recovery of 96% and an RSD of 3.2%.