Not as simple as using split, however:
string input = "FirstName=ABC;LastName=XZY;Username=User1;Password=1234";
string username = Regex.Match(input, "Username=(?<username>.*?)(;|$)").Groups["username"].Value;
In this case, groups can be in any order.
And, if you like to get creative:
var answers = from tuple in input.Split(';')
where tuple.StartsWith("Username=")
select tuple.Split('=')[1];
username = answers.Count() > 0 ? answers.First() : string.Empty;
One might say the last piece has better semantics.
EDIT: Update the last piece to deal with input strings that doesn't have the required tuple.