Hydrogen sulfide is found in many environments including sewage systems, petroleum extraction platforms, kraft paper mills, and exhaled breath, but its determination at ppb levels remains a challenge within the analytical chemistry field. Off-line methods for analysis of gaseous reduced sulfur compounds can suffer from a variety of biases associated with high reactivity, sorptive losses, and atmospheric oxidative reactions. Here, we present a portable, online, and disposable gas sensor platform for the in situ determination of gaseous hydrogen sulfide, employing a 470 nm light emitting diode (LED) and a microfiber optic USB spectrometer. A sensing layer was created by impregnating 2.5 μL (0.285 nmol) of fluorescein mercury acetate (FMA) onto the surface of a micropaper analytical device with dimensions of 5 × 5 mm, which was then positioned in the optical detection system. The quantitative determination of HS was based on the quenching of fluorescence intensity after direct selective reaction between the gas and FMA. This approach enabled linear calibration within the range 17-67 ppb of HS, with a limit of detection of 3 ppb. The response time of the sensor was within 60 s, and the repeatability was 6.5% (RSD). The sensor was employed to monitor HS released from a mini-scale wastewater treatment tank in a research laboratory. The appropriate integration of optoelectronic and mechanical devices, including LED, photodiode, pumps, and electronic boards, can be used to produce simple, fully automated portable sensors for the in situ determination of HS in a variety of environments.