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In this work, CNTs electrochemical electrode has been integrated on a flow injection based microfluidic chip for detection of cholesterol (CHO). Metal electrodes were deposited and CNTs were grown on a glass substrate by sputtering and chemical vapor deposition at low temperature. Polydimethylsiloxane microfluidic componet was then bonded on substrate. Cholesterol oxidase/potassium hexacyanoferrate were immobilized by polyvinyl alcohol using in-channel flow processing. Cholesterol detections were made by flow injection analysis and fast response time and high sensitivity have been achieved with high throughput of more than 100 samples per hour.