Thread-based microfluidic analytical devices have received growing attention since threads have some advantages over other materials. Compared to paper, threads are also capable of spontaneously transporting fluid due to capillary action, but they have superior mechanical strength and do not require hydrophobic barriers. Therefore, thread-based microfluidic devices can be inexpensively fabricated with no need for external pumps or sophisticated microfabrication apparatus. Despite these outstanding features, achieving a controlled and continuous flow rate is still a challenging task, mainly due to fluid evaporation. Here, we overcome this challenge by inserting a cotton thread into a polyethylene tube aiming to minimize fluid evaporation. Also, a cotton piece was inserted into the outlet reservoir to improve the wicking ability of the device. This strategy enabled the fabrication of an innovative electrochemical thread in a tubing microfluidic device that was capable to hold a consistent flow rate (0.38 μL s⁻¹) for prolonged periods, allowing up to 100 injections in a single device by simply replacing the cotton piece in the outlet reservoir. The proposed device displayed satisfactory analytical performance for selected model analytes (dopamine, hydrogen peroxide, and tert-butylhydroquinone), in addition to being successfully used for quantification of nitrite in spiked artificial saliva samples. Beyond the flow rate improvement, this “thread-in-tube” strategy ensured the protection of the fluid from external contamination while making it easier to connect the electrode array to the microchannels. Thus, we envision that the thread in a tube strategy could bring interesting improvements to thread-based microfluidic analytical devices.