Since 1998, the Ain-Karim Vineyard has been growing different grape varieties for the production of high-altitude tropical wines in the municipality of Sutamarchan, located in the Alto Ricaurte region of Boyaca (Colombia). Pruning is used to limit the number and length of branches, generating a suitable balance between plant vigor and production; thereby, regulating fruit quantity and quality and ensuring reserves for the subsequent production. This study aimed to evaluate the effect of three pruning types (short = two buds on two spurs; long = five buds on three spurs and mixed = combination of short and long pruning types) on the fruit quality of V. vinifera, Cabernet Sauvignon and Sauvignon Blanc varieties. To accomplish this, a completely randomized two-factor design was used. Physicochemical variables of fruit quality (fresh cluster weight, water content, total soluble solids (TSS), total titratable acidity (TTA), technical maturity index (TMI), and pH) were determined at harvest. The long pruning type presented the highest values for the fresh cluster weight and TSS of the fruits from both varieties and a higher TMI in the Cabernet Sauvignon variety. These results indicate that, under the conditions of the vineyard, long pruning is the most suitable.