Some shrimp hatcheries use artificial insemination (AI) to improve the male to female ratio in their breeding populations. We describe a sperm extender solution, which allows the short-term storage of diluted sperm in Litopenaeus vannamei, and its use in an artificial insemination process. We also evaluate its fertilization capacity. An AI experiment was designed using two, one, or half spermatophore segments. We tested four treatments involving three different male:female ratios: Natural mating (1:1), Regular and Regular diluted (1:2) and Half diluted (1:4). Data analysis revealed that the number of nauplii produced per mating was affected by treatment, with Regular (158 420) performing better than Half diluted (112 864) (P < 0.05), but with no differences between the latter and Regular diluted (130 340) (P > 0.05). A binomial variable named female success (FS) was defined as successful when the number of nauplii obtained per mate was ≥25 000. Analysis showed differences for FS across treatments (P < 0.001), but not between Regular (79.2%), the hatchery conventional AI technique and Half diluted (60.4%), maybe due to sample size. Since the number of nauplii per mate is crucial to consider AI successful, it is necessary to improve this AI technique before it can be used in the shrimp industry.