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Currently, microalgae have attracted as potential feedstock for biofuel production. Hydrothermal liquefaction was proposed as technology to convert microalgae into bio-crude oil. Microalgae used in this study was Indonesia-cultivated Chlorella sp., This work investigated the effect of temperature (200°C, 225°C, 250°C), biomass weight-water ratio (1:20, 2:20, 3:20), and residence time (10, 20, 30 minutes) on bio-crude oil yield of non-catalytic hydrothermal liquefaction. The highest bio-crude oil yield was 2.25%, obtained at temperature of 250°C biomass weight-water ratio of 1:20, and residence time of 10 minutes. The highest component of bio-crude oil was alcohols. The low bio-crude oil yield was caused by the longer residence time of cooling step (driving gas conversion), low amount of carbon-hydrogen content and high amount of oxygen-ash content in biomass. Furthermore, the highest component of bio-crude oil was alcohols, stimulated by low carbon content coupled with high oxygen content in Chlorella sp.