The 2009 Challenger Ciudad de Guayaquil was a professional tennis tournament played on outdoor red clay courts. It was the fifth edition of the tournament which was part of the 2009 ATP Challenger Tour. It took place in Guayaquil, Ecuador between 9 and 15 November 2009.
| Country | Player | Rank1 | Seed | |
|---|---|---|---|---|
 Colombia | Santiago Giraldo | 111 | 1 | |
 Ecuador | Nicolás Lapentti | 114 | 2 | |
| Rui Machado | 118 | 3 | ||
| Nicolás Massú | 133 | 4 | ||
 Argentina | Sebastián Decoud | 145 | 5 | |
| Argentina | Sergio Roitman | 146 | 6 | |
 Brazil | Júlio Silva | 149 | 7 | |
| Argentina | Brian Dabul | 153 | 8 |
The following players received wildcards into the singles main draw:
Emilio Gómez Roberto Quiroz
Daniil Sirota Mariano ZabaletaThe following players received entry from the qualifying draw:
See main article: 2009 Challenger Ciudad de Guayaquil – Singles. Nicolás Lapentti def. Santiago Giraldo, 6–2, 2–6, 7–6(4)
See main article: 2009 Challenger Ciudad de Guayaquil – Doubles. Júlio César Campozano / Emilio Gómez def. 
Andreas Haider-Maurer / Lars Pörschke, 6–7(2), 6–3, [10–8]