Hibiscus sabdariffa L. is a medicinal plant that has various pharmacological activities. One of the activities of Hibiscus sabdariffa L. is the immunomodulatory activity closely connected with the anticancer activities. This study aimed to test the cytotoxic activity of Hibiscus sabdariffa L. Calyx Fractions against breast cancer cells (MDA-MB-231), determine the active fractions, and analyze the compounds of the active fraction of Hibiscus sabdariffa L. with LCMS. The Hibiscus sabdariffa

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... x was fractionated using vacuum liquid chromatography. Meanwhile, the cytotoxic activity was examined using the MTT method. The test results discovered that the Fraction A of Hibiscus sabdariffa L. Calyx is the active fraction with a score of IC50 187.89 µg/ml. The identification of Fraction A's chemical compounds using LC-MS discovered three identified compounds:tert-butyl-4-methoxy phenol Nigakilactone H and Stigmastan-3,6-dione. Since these compounds potentially function as active cytotoxic compounds,it is suggested that further studies isolate the active compounds from Fraction A of Hibiscus sabdariffa L.