The Serbian was downed by German Michael Berrer, 7-6 (7-5), 6-3, on the hard court floors of Dom Sportova.
Meanwhile, Sixth-seeded German Benjamin Becker was able to escape Tipsarevic's fate, hanging on to overcome Swedish qualifier Andreas Vinciguerra, 1-6, 6-1, 7-6 (7-4).
The highlight for the home crowd on the second day of Zagreb was a win by Croatian wild card Petar Jelenic, who topped Serbian qualifier Ilia Bozoljac 7-6 (8-6), 7-5.
Also notching first-round wins were German Philipp Petzschner, Slovakian Lukas Lacko, Austria's Daniel Koellerer and French qualifier Alexandre Sidorenko.
Campus cop fatally shoots Texas student during traffic stop
Puzzle-maker slips 'Murdoch Is Evil' into Rupert Murdoch's Sunday Telegraph