The HPLC method for determining vitamin D in multivitamin preparations was studied collaboratively. Samples were distributed to 43 laboratories in 10 countries. Twenty-four laboratories gave their results. The method specifies a reverse phase system for the cleanup and a straight phase system for the determination. Difficulties arise when laboratories are equipped with HPLC apparatus with a single column. Laboratories experienced in vitamin D analysis reported good results, even if they were equipped with a single column HPLC apparatus. Eleven of the 24 laboratories reported satisfactory results. For this rather complicated method, the coefficient of variation for a single assay is acceptable. The method has been adopted official first action as an alternative procedure to the chemical method for determining vitamin D in multivitamins.