Honey is the most produced and consumed bee product and is a gift from the Creator to humanity. However, regular chemical analysis is necessary to determine the quality of honey and whether it is fake. Proline and diastase analyzes are also included in the basis of honey-related analyzes. Proline is an amino acid found in honey, while diastase is an enzyme. The quality of honey is determined by these two analyzes. The methods of these analyzes are generally applied spectrophotometrically. Therefore, in this chapter, spectrophotometric applications of proline and diastase analysis, which are important quality parameters in honey, are included.