Mistrz Świata i aktualny lider klasyfikacji generalnej, Francesco Bagnaia wykręcił najszybszy czas jednego okrążenia podczas kwalifikacji do Grand Prix Włoch na torze Mugello! Pierwszą linię uzupełnią bracia Marquez, Marc i Alex.
| POS | RIDER | NAT | TEAM | TIME/DIFF | LAP | MAX |
| 1 | Francesco Bagnaia | ITA | Ducati Lenovo (GP23) | 1’44.855s | 6/8 | 356k |
| 2 | Marc Marquez | SPA | Repsol Honda (RC213V) | +0.078s | 5/7 | 355k |
| 3 | Alex Marquez | SPA | Gresini Ducati (GP22) | +0.152s | 6/6 | 354k |
| 4 | Jack Miller | AUS | Red Bull KTM (RC16) | +0.331s | 7/7 | 358k |
| 5 | Jorge Martin | SPA | Pramac Ducati (GP23) | +0.413s | 3/8 | 358k |
| 6 | Marco Bezzecchi | ITA | Mooney VR46 Ducati (GP22) | +0.435s | 5/7 | 358k |
| 7 | Aleix Espargaro | SPA | Aprilia Racing (RS-GP23) | +0.525s | 5/8 | 354k |
| 8 | Johann Zarco | FRA | Pramac Ducati (GP22) | +0.772s | 6/6 | 354k |
| 9 | Alex Rins | SPA | LCR Honda (RC213V) | +0.847s | 7/8 | 352k |
| 10 | Brad Binder | RSA | Red Bull KTM (RC16) | +0.876s | 3/4 | 362k |
| 11 | Luca Marini | ITA | Mooney VR46 Ducati (GP22) | +1.264s | 3/7 | 356k |
| 12 | Enea Bastianini | ITA | Ducati Lenovo (GP23) | +2.029s | 5/6 | 360k |
| Qualifying 1: | ||||||
| 13 | Maverick Viñales | SPA | Aprilia Racing (RS-GP23) | 1’45.591s | 6/7 | 354k |
| 14 | Franco Morbidelli | ITA | Monster Yamaha (YZR-M1) | 1’45.754s | 3/7 | 355k |
| 15 | Fabio Quartararo | FRA | Monster Yamaha (YZR-M1) | 1’45.755s | 7/7 | 354k |
| 16 | Takaaki Nakagami | JPN | LCR Honda (RC213V) | 1’45.86s | 7/7 | 353k |
| 17 | Michele Pirro | ITA | Aruba.it Ducati (GP23) | 1’46.002s | 2/7 | 358k |
| 18 | Miguel Oliveira | POR | RNF Aprilia (RS-GP22) | 1’46.003s | 7/7 | 354k |
| 19 | Fabio Di Giannantonio | ITA | Gresini Ducati (GP22) | 1’46.17s | 7/8 | 354k |
| 20 | Raul Fernandez | SPA | RNF Aprilia (RS-GP22) | 1’46.347s | 2/3 | 354k |
| 21 | Augusto Fernandez | SPA | Tech3 GASGAS (RC16)* | 1’46.359s | 7/7 | 353k |
| 22 | Lorenzo Savadori | ITA | Aprilia Factory (RS-GP23) | 1’47.244s | 6/6 | 353k |
| 23 | Jonas Folger | GER | Tech3 GASGAS (RC16) | 1’47.806s | 7/7 | 348k |