An instrumental texture analysis method has been optimized for the differentiation of grape ripening stages based on the simultaneous determination of mechanical and acoustic parameters of the seeds. Two factorial central composite design was used to optimize the most influencing operative conditions (speed and deformation) on mechanical and acoustic measurements. This experimental design in combination with response surface methodology showed that the most responsive parameters to changes in seeds during ripening were Young’s modulus of elasticity, many acoustic parameters measured with instrumental gain set to 0 (acoustic energy, linear distance, and number of peaks) and others measured at 24 dB gain (linear distance, number of peaks, and average pressure level). However, the optimal operative conditions depended on the texture parameter used. A correlation study between texture parameters and phenolic compounds of the seeds revealed that acoustic parameters like the average pressure level could be proposed as phenolic maturity indices.