A more time efficient and simpler method than methods based upon AOAC procedure 979.24 was developed for the analysis of concentrated microencapsulated vitamin D3. The improved efficiency and simplicity begins with the use of a 10% water in DMSO solution to dissolve the sample matrix. A 2.0 mL DMSO volume is combined with 200 mg of microencapsulated vitamin D3 in a flask or bottle. The mixture stands without agitation for 10 min when testing a starch matrix sample and for 30−60 min when testing a cellulose matrix sample. Five minutes of sonication or heating is required for vitamin A/D combinations. Next, 2,2,4-trimethylpentane (isooctane) is pipetted into the DMSO solution−sample mixture. The isooctane volume used was proportional to the sample concentration; specific isooctane volumes relative to sample concentrations are provided. The mixture is stirred for ca. 5 min using a magnetic stirrer. The supernatant is filtered through a 0.2 μm PTFE membrane into a suitable LC vial and chromatographed. Validation data which support equivalence to traditional AOAC 979.24-based methodology are presented. Keywords: Vitamin D; DMSO extraction; vitamin analysis