Sesquiterpene lactones (SLs) are a group of over 500 compounds, characteristic of the Asteraceae. They are interesting from the chemical and chemotaxonomic point of view, and show antitumour, anti-leukaemic, anti-cardiovascular disease, reduction of inflammation, and anti-microbial activities. The SLs, free lactucin and lactucopicrin, content in 572 accessions of lettuce (Lactuca sp.) germplasm introduced from 30 countries were quantified using high-performance liquid chromatography. Variation range of free lactucin content exhibited trace ~ 235.3 μg from 1g of dried leaves and average was 41.2 ± 1.2 μg (Avr ± SE), variation range 55.0 ~ 3,041.0 μg and average 526.9 ± 17.5 μg in free lactucopicrin content, and variation range 66.3 ~ 3,188.5 μg and average 568.1 ± 18.1 μg in total free SLs content. Lactucopicrin occupied 92.7% of the total SLs content. Among the varietal types, crisphead type exhibited the highest average total free SLs content, next is leaf lettuce, and butterhead type lettuce exhibited the lowest that of content. German accessions exhibited the lowest average total free SLs content, Korean accessions exhibited the highest, and European origin accessions exhibited lower that of content. Red leaf color accessions having higher SLs content than that of green color. Seven accessions having more than 2,000 μg·g-1 dwt of total free SLs content and five accessions having less than 100 μg·g- 1dwt that of content. These accessions can be used as low SLs content cultivar breeding or high SLs content cultivar breeding sources as well as research materials for medical treatment such as, anti-tumour, anti-leukaemic, and antimicrobial activities, etc.