Agricultura – Ştiinţă şi practică no.

3- 4(87-88)/2013 Agriculture - Science and Practice

CHANGES OF HYDROXYMETHYLFURFURAL
IN HEATED HONEY
Stan Laura, Sevastiaa Muste, Maria Tofana, Ramona Suharoschi
University of Agricultural Sciences and Veterinary Medicine Cluj-Napoca,
Faculty of Food Science and Technology Calea Floreşti 64, 400509 Cluj-Napoca, Romania,
laurastan@usamvcluj.ro

Abstract. Beekeepers use to heat crystallized honey to make it more attractive for
consumers. During this process fructose from honey is thermally degraded to hydroxymethylfurfural
(HMF). This paper is focused on evaluation of changes in HMF content of multifloral and acacia
honey due to thermal treatment performed for 2 hours at 40°C, 60°C, 80°C and 100°C. Other basic
quality parameters of heated and non-heated honey were evaluated by methods developed by
International Honey Commission: humidity, pH, acidity and diastase. Highest changes in HMF were
recorded after heating acacia honey at 100°C, about 9 times higher than non-heated honey and this
may be due to high level of fructose in this particular honey. Multifloral honey treated at 100°C
presented an increase in HMF about 5,5 times higher compared to non-heated honey.

Keywords: hydroxymethylfurfural, honey, thermal treatment.

INTRODUCTION

Natural honey crystallizes in time due to inevitable physico-chemical processes

which take place between honey components (Mărghitaş, 2005). Physical factors of
crystallization include storage temperature and the presence of pollen grains and dust in
honey. The chemical factors of crystallization relate to honey sugars and there are more
arguments to consider. First, glucose crystallizes while fructose doesn’t and fructose is
unstable in acid aqueous solution. When fructose and glucose are together in acid aqueous
solution, like in the case of honey, fructose is transformed into glucose by isomerization.
Secondly, when ratio of fructose:glucose reach 1:1 (w/w) crystallization is triggered.
Thirdly, when glucose is 2,1 times the water content, crystallization is accelerated.
Physical and chemical factors contribute and generate the so called “fast” and “slow”
crystallizing honeys. Honeys with low amount of pollen and high amount of fructose are
known as slow crystallizing honeys (e.g. acacia honey), while honeys with high amount of
pollen (e.g. sun-flower and rape) and fructose:glucose ratio about 1:1 are known as fast
crystallizing honeys.
Once honey crystallization started, there is nothing to stop it. If 10% of the whole
honey mass is crystallysed, then very soon all quantity will be a chunk of crystallized
honey. This knowledge is used in food industry to create the so called “cream honey”
designed for easier spreading on breakfast slice of bread.
However, not many consumers know that honey crystallization is a natural
process. In fact, many Romanian consumers refuse to buy crystallized honey, because they
believe it is adulterated. Therefore, beekeepers use to heat crystallized honey in order to
accelerate the bottling and successfully sell it to consumers. Unfortunately, heating causes
degradation of important components found in honey: coagulation of proteins (enzymes
and proline from honey and all proteins of the pollen within honey), sugar caramelization
due to Maillard reactions and fructose degradation to hydroxymethylfurfural (HMF).
Studies regarding the degradation of fructose into HMF in citrus, chestnut and multifloral

- 101 -
Agricultura – Ştiinţă şi practică no. 3- 4(87-88)/2013 Agriculture - Science and Practice

honey were previously reported by other researchers (Fallico, 2004, 2008, Zappala, 2005,
Chiş, 2011).
This study is focused on studying the effect of multifloral and acacia honey heating
on thermo-labile compounds from honey (HMF and diastase). Due to particular
composition of acacia honey the authors were interested to research the variation of
diastase and HMF due to thermal treatment.

MATERIAL AND METHOD

The research has been carried out in Laboratory for Food Quality Control and
Safety in USAMV Cluj-Napoca. Honey samples (Acacia - A and Multifloral - M) were
collected from Cluj county in 2013, from a stationary beekeeper. Samples of 200g were
packed in closed air-tight jars and treated at 5 different temperatures for 2 hours in a water
bath as seen in Table 1. After treatment and between determinations samples were kept at
room temperature (20°C).

Table 1
Analyzed honey samples
Heating temperature Codification of Multifloral Codification of acacia
(°C) honey samples honey samples
20 M1 A1
40 M2 A2
60 M3 A3
80 M4 A4
100 M5 A5

All samples were analyzed applying methods developed by International Honey

Commission in order to determine humidity, pH, free acidity, hydroxymethylfurfural
(White method) and diastase (Phadebas method). All chemicals (p.a. purity) were
purchased from Chemomed (Romania), diastase tablets (Pharmacia Diagnostics, UK) and
all determinations were performed in triplicate. Apparatus used in this study are: digital
refractometer Abbe; pH-meter Schott Duran, water bath Memmet, spectrophotometer UV-
VIS 1700 Schimadzu, Japan.

RESULTS AND DISCUSSION

Honey has always been considered as a healthy and natural food product. The most
important parameters to evaluate honey quality are: safety (absence of contaminants like
antibiotics, pesticides and heavy metals), indicators of authenticity (sucrose, pollen, other
minor sugars), indicators of maturity (humidity, reducing sugars), indicators of degradation
(acidity, hydroxymethylfurfural and diastase).
The indexes most used to evaluate manipulation errors by beekeeper (e.g.
overheating) are hydroxymethylfurfural and diastase. HMF is the most important
intermediate product of 2 reactions: the acid catalyzed degradation of hexose and the
decomposition of 3-deoxyosone in Maillard reaction (Fallico, 2008). Diastase is an enzyme
naturally present in fresh honeys (Persano Oddo & all, 1999) whose levels decrease during
- 102 -
Agricultura – Ştiinţă şi practică no. 3- 4(87-88)/2013 Agriculture - Science and Practice

storage and/or heating (Tosi, 2002). Literature data shows that HMF levels in honey
depend on the chemical properties and on the floral origin of honey, temperature and
heating duration (Fallico, 2008).
Different approaches have been used by international bodies to establish the
limits of HMF in honey: from no limits at all, like USA (Docket 2006P-0101), Australian
and New Zealand (ANFS, Standard 2.8.2.), to very restrictive ones – European Union
(Council Directive 2001/110 EC).
The experimental results obtained in this study are presented in Table 2 and they
meet European Council (EC110/2001) criteria for tested quality parameters only for honey
samples treated below 80°C. The samples treated at 100°C gave no measurable amounts of
diastase.
Table 2
Quality parameters of heated honey samples taken in the study

Sample Humidity Free acidity HMF Diastase

code (%) (mEq/kg) (mg/kg) (DN)
M1 17,2 5,75 ± 0,35 15,67 ± 1,85 18,43 ± 0,95
M2 17,2 6,25 ± 0,30 15,77 ± 0,11 18,37 ± 0,14
M3 17,2 6,39 ± 0,71 15,80 ± 2,30 17,2 ± 0,33
M4 18,2 6,50 ± 0,54 21,61 ± 2,20 5,1 ± 0,45
M5 18,4 7,50 ± 0,66 85,49 ± 0,09 0
A1 17,0 7,50 ± 0,71 9,26 ± 0,27 12,47 ± 0,52
A2 17,6 7,75 ± 0,35 9,86 ± 0,42 11,36 ± 0,29
A3 17,6 7,75 ± 0,42 12,26 ± 2,34 9,52 ± 0,86
A4 17,8 8,00 ± 0,73 14,57 ± 0,55 2,53 ± 0,31
A5 17,8 8,25 ± 0,55 80,13 ± 0,74 0

The increase of HMF in multifloral honeys treated at 100°C was 5,5 times higher
then the sample kept at 20°C (R2 = 0,563) and almost 9 times higher for acacia honey (R2 =
0,566). Diastase decreases during thermal treatment R2 = 0,818 for multifloral honey and
R2 = 0,918 for acacia honey.
Chiş, 2011 determined HMF after thermal treatment of chestnut honey and
concluded that this honey type can be conditioned until 60°C without crossing the admitted
limit for HMF in Europe.

CONCLUSIONS

This article is focused on evaluation of thermal degradation of diastase and HMF

from multifloral and acacia honey. There is a direct correlation between thermal treatment
and degradation of honey. Overheating honey to 100°C is destroying the diastase and
highly increasing the HMF level above the limits admitted by European legislation.
Although the study was performed for thermal treatment for only 2 hours, it well known
that heat has cumulative degrading effect. Therefore, beekeepers who wish to liquefy their
product should not exceed in duration even if they use the recommended heating
temperature.

- 103 -
 Agricultura – Ştiinţă şi practică no. 3- 4(87-88)/2013 Agriculture - Science and Practice

REFERENCES

1. ANFS – Australian and New Sealand Food Standard Code, Standard 2.8.2. Honey
issue, 53:1-2
2. Canada Agricultural Product Act. Honey Regulations. C.R.C. 287
3. Chiş, A., Purcărea, C., 2011, The effect of thermal treatment on fresh monofloral honey
types from Bihor county, Analele Univ. Oradea, Fascicula: Ecotoxicologie, Zootehnie şi Tehnologii
de Ind. Alimentară, 71-76
4. Council Directive 2001/110/EC of 20 December 2001 realting to honey, Official
Journal of the European Communities, L10/47-L10/52
5. Fallico, B., Arena, E., Zappala, M, 2008, Degradation of 5-Hydroxymethylfurfural in
honey, J. Food Sci., 73/9, 625-631
6. Fallico, B., Zappala, M., Arena E., Verzera, A. 2004, Effects of heating process on
chemical composition and HMF levels in Sicilian monofloral honeys. Food Chemistry, 85, 305–
313.
7. Mărghitaș, L.A., Albinele și produsele lor, 2005, Editura Ceres
8. Persano Oddo, L., Piazza M.G., Pulcini P., 1999, Invertase activity in honey,
Apidologie, 30, 57-66
9. Swiss Bee Research Centre, 2002, Harmonised methods of the International Honey
Commission, 1-54
10. Tosi E., M. Ciappini, E. Ré and H. Lucero, 2002, Honey thermal treatment effects on
hydroxymethylfurfural content, Food Chemistry, 77, 71–74.
11. Turhan I., N. Tetik, M. Karhan, F. Gurel and H. Reyhan Tavukcuoglu, 2008, Quality of
honeys influenced by thermal treatment LWT - Food Science and Technology, 41 (8) 2008, 1396-
1399
12. Zappala, M, Fallico, B., Arena, E., Verzera, A., 2008, Degradation of 5-
Hydroxymethylfurfural in honey, J. Food Sci., 73/9, 625-631
13. Zappala, M., Fallico, B., Arena, E. and Verzera, A., 2005, Method for the
determination of HMF in honey: a comparison, Food Control, 16, 273-277.

- 104 -