Comparison between official methods, optimization and validation of the analytical methodology by spectrophotometric methods have been used to determine proline conten because in which the content of proline was measured by spectrophotometry from the colour developed with ninhydrin at a wavelength of 510 nm and compares official spectrophotometric methods for the determination of proline content in honey – those of the International Honey Commission (IHC) and the Association of Official Analytical Chemists (AOAC) – with the original Ough method.