There is provided a microneedle bio-sampling apparatus including: an bio-sampling body enabling a microneedle to be inserted into human skin; and a blocking unit installed in the bio-sampling body and preventing the microneedle from moving to the skin again to prevent repeated bio-sampling by the microneedle. Since the microneedle bio-sampling apparatus is configured to prevent repeated bio-sampling of a single-use microneedle, secondary infection to other part of a human body or a different person due to blood, human anatomy, or the like, that may remain in a microneedle after the microneedle is used once otherwise in case of repeated bio-sampling of a microneedle, can be prevented.