Natural fibers such as flax or hemp may be used in composite materials. However, their use for this purpose is hampered by the large natural variation in tensile strength and other quality parameters. The first step in managing these variations is to develop methods for fast and reliable determination of relevant parameters. One quality parameter of the fibers is the amount of structural distortions known as dislocations or kink bands. Here, a method developed for the quantification of dislocations in pulp fibers was adapted and tested successfully for hemp yarn segments. The method is based on acid hydrolysis and subsequent determination of the fiber segment length distribution. The premise of the method is that acid hydrolysis causes fibers to break in the dislocations rather than in other places. By use of polarized light microscopy and image analysis it was found that the premise was correct, and furthermore results showed that fibers broke more often in large dislocations than in small ones. However, it was also found that the hemp fiber segments did not break in all dislocations, and strict standardization of the procedure for acid hydrolysis is therefore necessary if results from different batches of fibers are to be compared.