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Not canon, but I would suggest:
1. to prove you can do it to the guild;
2. to prove you are willing to do it to the guild.
The prestige class assumes assassins' guilds are the primary training ground. As it is an evil-only PrC (as written) I guess killing someone just to get in said PrC is also a definitively evil act, and so should make it difficult for non-evil character to qualify without alignment change.

KaeYoss |

"So, we might be the Assassin's guild, and we might be willing to let you join. Not saying we are, you see, but it's a big multiverse. If we were, though we would have to be sure you're not a guard."
"I'm not a guard. See? Must be true. Guards aren't allowed to say they're not a guard."
"Of course not, but we need a better proof. Here, kill this person. A guard is definetly not allowed to do something like that. If you did that, I'm sure any assassins' guilds around would see that you mean business."

Nero24200 |

My guess is as a test of some sorts, to prove that you're capable of assassin styled killing (i.e., reaching the target, dispatching the target, not leaving a trace and not getting caught.)
Also it enforces that anyone entering the class is evil (since killing someone for no reason other than to join a group of killers is pretty evil).

KaeYoss |

(since killing someone for no reason other than to join a group of killers is pretty evil).
What does that remind me of? Ah, yes:
BM: "Did you know that my hadoken spell is powered by love?"
WM: "Really?"
BM: "Oh yeah! Every time I use this spell, a little love dies and is forever gone from the world."
WM: "What? But that's horrible!"
BM: "Of course. Why do you think did I sacrifice a whole orphanage to the dark forces of hell? Just out of fun? That would be sick!"
WM: O_O