Amoxicillin is one of the broad-spectrum beta-lactam antibiotics widely used in the treatment of many diseases. It is inevitable that 4-hydroxyphenylglycine (4-HPG) and 6-Aminopenicylanic acid (6-APA), which are used during the production of this antibiotic, are incorporated into the molecular lattice of the product as impurities. Today, many expensive methods and chemical devices are used for the purification of Amoxicillin by determining 6-APA and 4-HPG, which are defined as impurities. In this study, it was aimed to develop a fast, simple, and specific UV-spectrophotometric method for the determination of 4-HPG and 6-APA. Another aim of this article is to cause as little harm as possible to the environment and human health by using as few chemicals as possible throughout the study. In this study, all attempts to determine 6-APA and 4-HPG, which are impurities in the production of amoxicillin, were carried out with the help of a UV/VIS spectrophotometer. Also, Four different concentrations of NaOH were used as a solvent for each impurity. UV spectra of 4-HPG and 6-APA concentrations between 210 and 400 nm were measured. In the literature, the UV spectrum of 4-HPG has been revealed for the first time in this study and examined in detail. The UV spectrum of 4-HPG was characterized in 3 regions. Again, the response of 6-APA to different NaOH concentrations was demonstrated for the first time in this study. It was determined that the peaks of 6-APA dissolved in NaOH shifted from 222 nm to 227 nm depending on the concentration amount. In addition, it is an ideal green procedure that makes a difference in the literature, as the study is carried out for the control and determination of impurities without the use of any organic solvents or chemicals harmful to the environment.