Since its first discovery more than a century ago, chromatographic separation has become increasingly essential to organic chemistry, pharmaceuticals, proteomics, the environment, and industry. However, recent developments in basic science indicate that increasing numbers of small molecules and proteins are extremely hard to separate successfully by the traditional chromatography. Herein, we propose an intelligent chromatograph that can be controlled by various conditions such as temperature, pH, and ionic strength at the same time, which is fulfilled by the functionalization of silica microparticles with multiple stimuli-responsive copolymers, poly(N-isopropylacrylamide-co-2-(diethylamino)ethyl methacrylate-co-N-tert-butylacrylamide), by surface-initiated atom transfer radical polymerization. The intelligent chromatographic properties were evaluated in a controlled manner based on adenosine nucleotides in the aqueous mobile phase. The results indicate that the influences of temperature, pH, and mobile-phase ionic strength could be well regulated by the chain length and composition of the copolymers. Moreover, the present findings can be directed toward the design and application of a multiply controlled chromatograph.