The invention discloses a portable underground water fixed-depth sampling device, a sampling system and a sampling method, and belongs to the field of site pollution investigation and risk assessment.A moving blocking air bag is arranged; stagnant water at the upper end and the lower end of an underground water sampling probe in a well pipe is effectively blocked, an underground water sample at the depth where the sampling probe is located can be accurately collected, the problem that in the well washing sampling process of a traditional method, upper-layer water samples and lower-layer watersamples in the well pipe are mixed, and consequently sample dilution is caused is solved, and the concentration detection accuracy is improved. Underground water samples of different depths can be accurately collected in the same monitoring well, fixed-depth sampling is achieved, and the cost is obviously reduced. In addition, the stagnant water on the upper layer and the lower layer of the underground water sampling probe in the well pipe is effectively blocked, the well washing volume before sampling can be reduced, the sampling time is saved, the sampling efficiency is improved, the well washing wastewater generation amount is reduced, the sampling cost is further reduced, disturbance to underground water in the well pipe can be reduced, and the accuracy of detected concentration is guaranteed.