Ultraviolet- visible absorption spectrophotometry is still increasingly finding its way in pharmaceutical analysis worldwide. With the advancement in this method, it achieves precision and accuracy in analysis of active pharmaceutical ingredients (API) as well as commercial dosage forms. This illustrative guide is divided into two sections i.e. qualification and quantification. Qualification includes comparative calibration procedures as per difference Pharmacopoeias with various reference materials, comparative validation parameters and procedures as per different guidelines, optimum conditions for spectrophotometric measurement and factors affecting absorption. On the other hand quantification includes procedures and approaches for analyzing the pharmaceutical substance via different spectrophotometric methods including various derivative spectrophotometric and chemo-metric techniques.