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Mycosporine-like amino acids (MAAs) are natural UV-absorbing compounds found in microalgae and macroalgae. The content of MAAs in algae varies with the seasons and environmental factors. Red alga dulse in Usujiri (Hokkaido, Japan) is an underutilized resource. Therefore, we investigated the amount of MAAs in Usujiri dulse in 2022 to clarify the suitable months for MAA extraction. In addition, we also evaluated the extraction method focusing on the extraction volume. MAAs were prepared via the 20 volumes of 25% ethanol extraction method and detected via HPLC. The results showed that the amount of MAAs on 25 March 2022 showed the highest value (40.4 μmol/g DW) among the samples from 24 January to 13 May. The tendency of suitable samples for MAA preparation corresponded to the term from mid-February to early April, which was the same as the previous three years. Although the surveys from 2019–2021 were performed by using the successive water–methanol method, it was found that the improved method also reflected the monthly variation in MAAs. The extraction of MAAs was performed via 20 or 40 volumes of 25% ethanol at 4 °C for 24 h. The amount of MAAs with 40 volumes of 25% ethanol extraction increased 1.3-fold compared to that with 20 volumes of 25% ethanol extraction. These data are useful information for valuable compound extraction from Usujiri dulse.