In the present research, consumption of medication is increasing that leads to environmental pollution. Thus, drug pharmaceutical control has become a routine technique in many laboratories. This project focuses on the enhancement of a method to determine insignificant amounts of ciprofloxacin in aqueous and biological solutions. This study looks at solid phase extraction of insignificant amounts of ciprofloxacin using carbon nanotubes in aqueous samples, and ultraviolet and visible (UV-Vis) spectrophotometry measurement in biological samples. These systems include two phases – the aqueous donor phase and the conjugated carbon nanotubes acceptor phase. Experiments are performed in two steps – aqueous phase extraction and ciprofloxacin resorption using methanol acidic solvent; resorbed samples are provided to UV-Vis spectrophotometry for further analysis. This method is cheap, simple, fast, and compatible with most instrumental analysis methods. Extraction parameters including resorbing organic solvent effect, pH of donor and acceptor phases, duration of extraction, duration of resorption, shaking time, volume of donor phases, and effects of surfactants are optimized and analysed; and measurements are performed under the optimized situation. The mentioned techniques have many advantages including short extraction time, low consumption of organic solvents, removal of effects of prior experiments, low diagnosis threshold, and high concentration factor. For ciprofloxacin, concentration factor and diagnosis threshold are 51 and 9.51, respectively. Linear domain and relative standard are 1.12%.