2000 Italian Open – Women's singles

Monica Seles defeated Amélie Mauresmo in the final, 6–2, 7–6(7–4) to win the women's singles tennis title at the 2000 Italian Open. It was her third title of the year, and the 47th of her career.

Venus Williams was the defending champion, but lost in the third round to Jelena Dokic.

Seeds
The first eight seeds received a bye into the second round. • # 🇺🇸 Lindsay Davenport (third round, withdrew)

• # 🇫🇷 Nathalie Tauziat (quarterfinals)

• # 🇺🇸 Venus Williams (third round)

• # 🇫🇷 Mary Pierce (third round)

• # 🇺🇸 Monica Seles (champion)

• # 🇪🇸 Arantxa Sánchez Vicario (quarterfinals)

• # 🇫🇷 Julie Halard-Decugis (second round)

• # 🇫🇷 Sandrine Testud (second round)

• # 🇩🇪 Anke Huber (second round)

• # 🇺🇸 Jennifer Capriati (first round)

• # 🇷🇺 Elena Likhovtseva (first round)

• # 🇫🇷 Amélie Mauresmo (final)

• # 🇯🇵 Ai Sugiyama (second round)

• # 🇫🇷 Nathalie Dechy (first round)

• # 🇧🇪 Dominique van Roost (second round)

• # Silvija Talaja (first round)