The high air-tightness multicore fiber optic penetration connector is a core component for the optical fiber sensing and communication technologies applied in the space environment simulator under the vacuum thermal environment. High air-tightness and insertion loss are the two key indexes of the fiber optic penetration connector. The air-tightness and insertion loss on-line synchronous detection method was proposed. First, established hardware-in-the-loop testing platform by using the vacuum pumping system, the vacuum vessel, the helium mass spectrometer leak detector and optical time-domain reflectmeter, then, described the air tightness and insertion loss on-line detection principle, finally, designed a detection test scheme and air-tightness and insertion loss were tested. Experimental results indicate that the leakage rate is lower than 1.0×10-7Pa•L/S, the minimum of which is1.0×10-10Pa•L/S and the insertion loss at wave length window 1550 nm is ±0.07db, which is less than ±0.1db. It can lay the data basis for the design of opto-mechanical combination and later period fine processing.