Road cycling is an endurance sport characterized by several anthropometric performance factors, such as reduced body mass and body fat percentage. As the power to weight ratio is considered one of the most important markers of performance in this sport discipline, it is speculated that anthropometric factors could relate to the physiological parameters found in road cyclists of different performance levels. The current study aimed to describe the anthropometric differences across road cyclists of different performance levels and to assess whether anthropometric values could relate to physiological markers that are commonly used to classify road cyclists, according to their performance level. We classified 46 cyclists as recreationally trained, trained, well trained and professional, according to their VO2max, and performed graded exercise tests and complete anthropometric assessments. The results showed that there were no significant anthropometric differences between trained, well trained and professional cyclists, with only recreationally trained cyclists exhibiting larger perimeters and skinfolds than professional cyclists. Further, although physiological performance, such as VO2max and respiratory compensation point, correlated negatively with several skinfolds and perimeters, these correlations remained restrained and did not distinguish between cyclists of different specialties.