Rice straw derived biochar was fabricated and applied as a purification agent. The adsorption kinetics, isotherms, and thermodynamics for adsorbates were determined using the biochar. Adsorption kinetics and isotherms were best fitted by the pseudo-second order and Langmuir models. Biochar could effectively remove chlorophyll in 9 different solutions. Biochar was employed as a clean-up reagent for 149 pesticides detection, which revealed that biochar had a higher phytochrome removal capacity than graphitized carbon black and 123 pesticides had satisfactory recovery values. The biochar was prepared into a sample pad by electrospinning and was then used for online sample clean-up in a test strip, and it showed high ability of removing phytochrome and improving detection sensitivity. Thus, biochar could be applied as a purification agent to remove pigmentation, making it a promising candidate not only for sample pretreatment but also in the fields of food, agriculture and environment.