A novel multifunctional two-sides silicon lab-onchip (LOC) was designed and fabricated for providing a highefficient long-term continuous analysis platform of measuring the concentration of adenosine 5'-triphosphate (ATP) based on bioluminescence. This lab-on-chip mainly consists of Y shape internal ribbed micro mixers, T-junction droplet generators, different kinds of spiral time delay channels (TD-Cs) and the chambers. Enzyme solutions and ATP were separately injected into the chip at different flow rates through 3 inlet ports. Due to the advantage of Segmented Flow Analysis (SFA), it was introduced and applied with the T-junction droplet generators in this paper, and thus oxygen was injected into the central of the bottom surface through another main inlet periodically. Numerical investigations prove that combing with SFA and Tjunction droplet generator is stable to transport the sample; micro mixer can achieve an outstanding mixing efficiency in a small space; the detection of ATP standard solution at optimal conditions was successful, keeping a good linearity between the concentration with the intensity of light during the range from 10 −5 mol /ml to 10 −9 mol/ml.