The total vitamin D content in meat, i.e., vitamin D-3 and 25-hydroxyvitamin D-3, was determined by HPLC after alkaline hydrolysation, solid-phase extraction and semi-preparative HPLC. For detection, a DAD detector between 220 and 320 nm was used and quantification was performed at 265 nm. Vitamin D-2 was used as internal standard for vitamin D-3 as well as for 25-hydroxyvitamin D-3. Precision for vitamin D-3 was determined in lean meat and lard to 9.1% and 7.1%, respectively. The corresponding values for 25-hydroxyvitamin D-3 were 8.9% and 9.9%. Accuracy was determined in spiked samples, which showed a recovery of 94.7% and 99.0% for vitamin D-3 and 25-hydroxyvitamin D-3, respectively. The method is applicable for establishing data for food composition tables. (C) 2004 Elsevier Inc. All rights reserved.