1. HMF in HONEY Biagio Fallico, Elena Arena, Mario Zappalà, Antonella Verzera University of Catania - University of Messina (Italy) 3 rd AOAC Europe – EURACHEM Symposium 3 March 2005 Brussels

2. HONEY REGULATIONS Codex Alimentarius Commission Codex Alimentarius Commission Joint FAO/WHO Food Standard Programme (24th session, Geneva, 2-7 July 2001) Report of the seventh session of the Codex Committee on Sugars (ALINORM 01/25, London 9-11 Feb. 2000) EC Directive 2001/110 Relating to Honey 3 rd AOAC Europe, Eurachem Symposium, Brussels 3 March 2005

3. Essential Composition and Quality Factors Chemical composition (moisture, sugars, insoluble solids, conductivity) Contaminants Hygiene Labelling Methods of sampling and analysis ALINORM 01/25 3 rd AOAC Europe, Eurachem Symposium, Brussels 3 March 2005

4. Additional Composition and Quality Factors Free acidity Diastase activity Hydroxymethylfurfural (HMF) After processing and/or blending:  40 mg/Kg 40 mg/Kg  80 mg/Kg for honey from Countries or Regions with tropical 80 mg/Kg for honey from Countries or Regions with tropical ambient temperatures ALINORM 01/25 3 rd AOAC Europe, Eurachem Symposium, Brussels 3 March 2005

5. EC Directive 2001/110 Annex II – Composition criteria for honey 1.sugars 2.moisture 3.water insoluble solids 4.electrical conductivity 5.free acids 6.Diastase activity and HMF (determined after processing) 3 rd AOAC Europe, Eurachem Symposium, Brussels 3 March 2005 6.a Diastase (Schade scale)  8 (in general, except bakers honey)  3 (honey with low natural enzyme content (e.g. citrus honey) and a HMF level not higher than 15 mg/kg. 6.b HMF  40 mg/Kg  80 mg/Kg in honeys of declared origin from Regions with tropical climate

6. Analyses: moisture, glucose, fructose, pH, free acids, lactones, total acidity, conductivity, ash, diastase and HMF Heating at: 50 °C up to 144 h (6 days); 70 °C up to 96 h (4 days) 100°C up to 60 h (2.5 days) HEATING TREATMENTS Citrus (Citrus Aurantum L.) Chestnut ( Castanea Sativa L.) Eucalyptus (Eucalyptus Camaldulensis L.) Sulla ( Hedysarium Coronarium L.) 3 rd AOAC Europe, Eurachem Symposium, Brussels 3 March 2005

7. Chemical composition of the Unifloral honeys ParameterCitrusChestnutEucalyptusSulla pH3.45.93.73.4 Free acids (meq/kg)22.59.72315.5 Total acidity (meq/kg)2511.432.518 Electrical conductivity(  S) 1931128413126 Diastase (DU)7.627.333.910.8 HMF (mg/kg)5.95--1.23 3 rd AOAC Europe, Eurachem Symposium, Brussels 3 March 2005

8. ChestnutEucaliptusCitrusSulla HMF formation during heating at 100°C hours of heating 0 4000 8000 12000 16000 0481224364860 mg/kg 3 rd AOAC Europe, Eurachem Symposium, Brussels 3 March 2005

9. HMF formation during heating at 70°C ChestnutEucaliptusCitrusSulla 3 rd AOAC Europe, Eurachem Symposium, Brussels 3 March 2005

10. HMF formation during heating at 50°C ChestnutEucaliptusCitrusSulla 3 rd AOAC Europe, Eurachem Symposium, Brussels 3 March 2005

11. Influencing factors on HMF levels at 50°C ( Results applying ANOVA and Multiple linear regression model analyses) Main factors: pH of honey temperature and time of heating Specific factors: Eucalyptus and Sulla were correlated to acidity Citrus was correlated, at the same time, to: time, pH and free acidity. 3 rd AOAC Europe, Eurachem Symposium, Brussels 3 March 2005

12. HMF is clearly an heating index, but the statement that the amount of HMF is independent of honey type and composition could be wrong. Among the studied honeys, Citrus, at the same time of heating, gives the highest amount of HMF 3 rd AOAC Europe, Eurachem Symposium, Brussels 3 March 2005

13. Citrus honey is a low natural enzyme content 9.3±2.7 (Range: 3.4-16.3) D.U. (Persano Oddo et al., 1995, 1999) The half life of Diastase in honey at 30°C is: 6/7 months This means 15 mg/kg as HMF limit 3 rd AOAC Europe, Eurachem Symposium, Brussels 3 March 2005

14. Storage of Citrus Honey (24  4°C) 0 15 30 45 60 75 056810151718 Citrus 1Citrus 2Citrus 3 HMF (mg/Kg) Months

15. This does not sound as “scientific based” because: a very low limit when there is a natural attitude to form HMF, independently from heating at the same time, honeys of different origin (chestnut) can be overheated without reaching the limit (40 mg/Kg). 3 rd AOAC Europe, Eurachem Symposium, Brussels 3 March 2005

16. Practical consequences (2004): High honeys sampling levels 10 not conform samples in 6 months 35 samples (25 citrus) EU Alert 3 rd AOAC Europe, Eurachem Symposium, Brussels 3 March 2005

17. Practical consequences (2004): Direct Effects 10 criminal proceedings 1000€ each as penalties Legal expenses Product declassification or destruction Side Effects costumers complaints No honey to sell Toxic or contaminated samples 3 rd AOAC Europe, Eurachem Symposium, Brussels 3 March 2005

18. Conclusions: More attention to: Hygiene aspects and Contaminants Eliminate the limit of 15 mg/kg for low enzyme honeys Correlate HMF limits to chemical parameters of honeys and not to the diastase activity 3 rd AOAC Europe, Eurachem Symposium, Brussels 3 March 2005