Hooker and captain Dan Rowett was the star man for Chester as he finished the game with four tries and the man of the match honours.
Rowett was joined on the score sheet by impressive debutant Damian Tierney and prop Andy Hill with back row forward Luke Edwards adding three conversions.
Chester produced some excellent attacking rugby with fullback Mark Hanson and winger Terry Toland to the fore while the forward pack, led by Dan Timperley and Chris Young, stood up well against a solid home pack.
Gladiators arguably finished the game the stronger but ultimately paid the price for slow starts at the beginning of each half.
Little Hulton took full advantage by running in two tries in the opening 10 minutes of the game and another brace in the first 10 minutes of the second period.
The two clubs have proved tough to separate over the past couple of years and this was another competitive game played in the right spirit and one from which both teams will take many positives.
Chester's next game is on Saturday, March 2 when they travel to Widnes Tigers for their final pre-season match.