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Listen to this siteWednesday 2 August 2006
This research project aims to develop a method for the determination of HMF in a range of honey types.
Study Duration : September 2002 to August 2003.
Contractor : Central Science Laboratory.
The European Honey Directive and the Revised Codex Standard for Honey specify standards for honey, which include criteria for quality factors including moisture, acidity and hydroxymethylfurfural (HMF) concentration.
HMF is a sugar breakdown product. Its presence was once considered to be a marker for the adulteration of honey with commercial invert syrup, however it has since been found that HMF can occur naturally in honey through improper heat processing or extended storage. As such, HMF is used as indicator of honey quality since it increases with temperature and storage time.
The EU Honey Directive 2001/110 and the Honey (England) Regulations 2003 (SI 2003/2243) state maximum levels of HMF in different honey types. The measurement of HMF is therefore of importance in determining honey quality and compliance with regulations.
Both spectroscopic and chromatographic methods can be used for the determination of the HMF content of honey, however, problems have been reported for spectroscopic methods. These include the use of carcinogenic reagents, and the fact the methods are non-specific and therefore measure components in honey that are not HMF leading to overestimates of HMF content.
Interference problems can occur and steps to reduce this such as filtration or clarification have been used. A method that separates the HMF from other components in the honey would allow more accurate measurement of HMF to be made.
This project aims to compare the White spectroscopic method and chromatographic (HPLC) methods for HMF determination.
This work compared the White spectroscopic method and chromatographic (HPLC) methods for HMF determination. The spectroscopic method was found to consistently produce higher HMF results than the two HPLC methods investigated, which closely agreed with each other.
In order to improve the selectivity of the HPLC procedure appropriate clean-up of samples was evaluated. Three strategies were tested:
The SPE method was found to provide the best clean-up for a variety of honey types and removed impurities that could potentially co-elute or interfere with HMF during HPLC analysis.
The SPE-HPLC method was subject to single laboratory validation. Six types of honey were used containing a range of HMF concentrations. The method was successfully validated and shown to have with acceptable chromatographic performance, trueness, linearity and precision.
The limit of detection was calculated as 1.4mg/kg and the measurement uncertainty associated with the method at the legal limit of 40mg/kg was calculated as ± 9.0 mg/kg at 95% confidence.
This work has shown that the SPE-HPLC method performs better than current spectroscopic methods, which overestimate HMF content. The HPLC method has been validated, and comparisons of the measurement uncertainty with HORRAT values (statistically predicted method performance) demonstrate the method performance to be acceptable.
The HPLC HMF procedure has subsequently been subjected to full validation through collaborative trial according to the Harmonised IUPAC Guidelines. It is hoped that the collaborative trial results will be published in a peer-reviewed journal.
Driffield M, Chan D, MacArthur R, MacDonald S, Brereton P, Wood R, Single laboratory validation of a method for the determination of hydroxymethylfurfural in honey by using solid-phase extraction cleanup and liquid chromatography, J. Assoc. Off. Anal. Chem., 88, (1), 121-27, 2005
The final report is available from the FSA Library and Information centre.
To obtain a copy, please contact the Enquiry Desk, Information Services, Food Standards Agency (020 7276 8181/8182 or at
library&info@foodstandards.gsi.gov.uk
).
Contact
: For any enquiries concerning this research project, please contact the relevant Programme contact or email
science@foodstandards.gsi.gov.uk
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