This paper described a fast, simple, and small scale method of sample preparation followed by high-performance liquid chromatography (HPLC) coupled photodiode array (PDA) detector for simultaneous quantification of astaxanthin, canthaxanthin, and β-apo-8′-carotenoic acid ethyl ester in hen’s egg yolk. The HPLC-PDA was performed on a C18 column with an isocratic mobile phase. Analytes were extracted from the sample using a handheld ultrasonic homogenizer, and purified by MonoSpin®-SI, a centrifugal monolithic silica spin mini-columns, and quantified within 30 min. The proposed method obtained average recoveries for the three analytes in the range of 70.5–101.1% with relative standard deviations ≤5.0%. The quantitation limits were 0.3 μg g−1 for AX, 0.5 μg/g for CX, and 1.0 μg g−1 for ACAEE, respectively.