Adulteration Identification of Commercial Honey with the C-4 Sugar Content of Negative Values by an Elemental Analyzer and Liquid Chromatography Coupled to Isotope Ratio Mass Spectroscopy

According to the AOAC 998.12 method, honey is considered to contain significant C-4 sugars with a C-4 sugar content of >7%, which are naturally identified as the adulteration. However, the authenticity of honey with a C-4 sugar content of <0% calculated by the above method has been rarely investigated. A new procedure to determine delta C-13 values of honey, corresponding extracted protein and individual sugars (sucrose, glucose, and fructose), delta H-2 and delta O-18 values, sucrose content, and reducing sugar content of honey using an elemental analyzer and liquid chromatography coupled to isotope ratio mass spectroscopy, was first developed to demonstrate the authenticity of honey with a C-4 sugar content of <0%. For this purpose, 800 commercial honey samples were analyzed. A quite similar pattern on the pentagonal radar plot (isotopic compositions) between honey with -7 < C-4 sugar content (%) < 0 and 0 < C-4 sugar content (%) < 7 indicated that honey with 7 < C-4 sugar content (%) < 0 could be identified to be free of C-4 sugars as well. A very strong correlation is also observed between delta C-13 honey values and delta C-13 protein values of both honey groups. For the delta O-18 value, the C-4 sugar content (%) < 7 group has lower (p < 0.05) values (16.30 parts per thousand) compared to other honey, which could be a useful parameter for adulterated honey with a C-4 sugar content (%) < 7. The use of isotopic compositions and some systematic differences permits the honey with a C-4 sugar content of <0% to be reliably detected. The developed procedure in this study first and successfully provided favorable evidence in authenticity identification of honey with a C-4 sugar content of <0%.